Electrospun polylactic acid and polyvinyl alcohol fibers as efficient and stable nanomaterials for immobilization of lipases.

PubMed

Sóti, Péter Lajos; Weiser, Diana; Vigh, Tamás; Nagy, Zsombor Kristóf; Poppe, László; Marosi, György

2016-03-01

Electrospinning was applied to create easy-to-handle and high-surface-area membranes from continuous nanofibers of polyvinyl alcohol (PVA) or polylactic acid (PLA). Lipase PS from Burkholderia cepacia and Lipase B from Candida antarctica (CaLB) could be immobilized effectively by adsorption onto the fibrous material as well as by entrapment within the electrospun nanofibers. The biocatalytic performance of the resulting membrane biocatalysts was evaluated in the kinetic resolution of racemic 1-phenylethanol (rac-1) and 1-phenylethyl acetate (rac-2). Fine dispersion of the enzymes in the polymer matrix and large surface area of the nanofibers resulted in an enormous increase in the activity of the membrane biocatalyst compared to the non-immobilized crude powder forms of the lipases. PLA as fiber-forming polymer for lipase immobilization performed better than PVA in all aspects. Recycling studies with the various forms of electrospun membrane biocatalysts in ten cycles of the acylation and hydrolysis reactions indicated excellent stability of this forms of immobilized lipases. PLA-entrapped lipases could preserve lipase activity and enantiomer selectivity much better than the PVA-entrapped forms. The electrospun membrane forms of CaLB showed high mechanical stability in the repeated acylations and hydrolyses than commercial forms of CaLB immobilized on polyacrylamide beads (Novozyme 435 and IMMCALB-T2-150).

I-NERI-2007-004-K, DEVELOPMENT AND CHARACTERIZATION OF NEW HIGH-LEVEL WASTE FORMS FOR ACHIEVING WASTE MINIMIZATION FROM PYROPROCESSING

DOE Office of Scientific and Technical Information (OSTI.GOV)

S.M. Frank

Work describe in this report represents the final year activities for the 3-year International Nuclear Energy Research Initiative (I-NERI) project: Development and Characterization of New High-Level Waste Forms for Achieving Waste Minimization from Pyroprocessing. Used electrorefiner salt that contained actinide chlorides and was highly loaded with surrogate fission products was processed into three candidate waste forms. The first waste form, a high-loaded ceramic waste form is a variant to the CWF produced during the treatment of Experimental Breeder Reactor-II used fuel at the Idaho National Laboratory (INL). The two other waste forms were developed by researchers at the Korean Atomicmore » Energy Research Institute (KAERI). These materials are based on a silica-alumina-phosphate matrix and a zinc/titanium oxide matrix. The proposed waste forms, and the processes to fabricate them, were designed to immobilize spent electrorefiner chloride salts containing alkali, alkaline earth, lanthanide, and halide fission products that accumulate in the salt during the processing of used nuclear fuel. This aspect of the I-NERI project was to demonstrate 'hot cell' fabrication and characterization of the proposed waste forms. The outline of the report includes the processing of the spent electrorefiner salt and the fabrication of each of the three waste forms. Also described is the characterization of the waste forms, and chemical durability testing of the material. While waste form fabrication and sample preparation for characterization must be accomplished in a radiological hot cell facility due to hazardous radioactivity levels, smaller quantities of each waste form were removed from the hot cell to perform various analyses. Characterization included density measurement, elemental analysis, x-ray diffraction, scanning electron microscopy and the Product Consistency Test, which is a leaching method to measure chemical durability. Favorable results from this demonstration project will provide additional options for fission product immobilization and waste management associated the electrochemical/pyrometallurgical processing of used nuclear fuel.« less

The immobilization of lipase on PVDF-co-HFP membrane

NASA Astrophysics Data System (ADS)

Kayhan, Naciye; Eyüpoǧlu, Volkan; Adem, Şevki

2016-04-01

Lipase is an enzyme having a lot of different industrial applications such as biodiesel production, biopolymer synthesis, enantiopure pharmaceutical productions, agrochemicals, etc. Its immobilized form on different substances is more conventional and useful than its free form. Supporting material was prepared using PVDF-co-HFP in laboratory conditions and attached 1,4-diaminobutane (DA) and epichlorohydrin (EPI) ligands to the membrane to immobilize lipase enzyme. The immobilization conditions such as enzyme amount, pH, the concentration of salt, thermal stability and activity were stabilized for our experimental setup. Then, biochemical characterizations were performed on immobilized lipase PVDF-co-HFP regarding optimal pH activity, temperature and thermal stability. Also, the desorption ratios of immobilized enzyme in two different pathway were investigated to confirm immobilization stability for 24 hours.

Selective cesium removal from radioactive liquid waste by crown ether immobilized new class conjugate adsorbent.

PubMed

Awual, Md Rabiul; Yaita, Tsuyoshi; Taguchi, Tomitsugu; Shiwaku, Hideaki; Suzuki, Shinichi; Okamoto, Yoshihiro

2014-08-15

Conjugate materials can provide chemical functionality, enabling an assembly of the ligand complexation ability to metal ions that are important for applications, such as separation and removal devices. In this study, we developed ligand immobilized conjugate adsorbent for selective cesium (Cs) removal from wastewater. The adsorbent was synthesized by direct immobilization of dibenzo-24-crown-8 ether onto inorganic mesoporous silica. The effective parameters such as solution pH, contact time, initial Cs concentration and ionic strength of Na and K ion concentrations were evaluated and optimized systematically. This adsorbent was exhibited the high surface area-to-volume ratios and uniformly shaped pores in case cavities, and its active sites kept open functionality to taking up Cs. The obtained results revealed that adsorbent had higher selectivity toward Cs even in the presence of a high concentration of Na and K and this is probably due to the Cs-π interaction of the benzene ring. The proposed adsorbent was successfully applied for radioactive Cs removal to be used as the potential candidate in Fukushima nuclear wastewater treatment. The adsorbed Cs was eluted with suitable eluent and simultaneously regenerated into the initial form for the next removal operation after rinsing with water. The adsorbent retained functionality despite several cycles during sorption-elution-regeneration operations. Copyright © 2014 Elsevier B.V. All rights reserved.

Supplemental Immobilization of Hanford Low-Activity Waste: Cast Stone Screening Tests

DOE Office of Scientific and Technical Information (OSTI.GOV)

Westsik, Joseph H.; Piepel, Gregory F.; Lindberg, Michael J.

2013-09-30

More than 56 million gallons of radioactive and hazardous waste are stored in 177 underground storage tanks at the U.S. Department of Energy’s (DOE’s) Hanford Site in southeastern Washington State. The Hanford Tank Waste Treatment and Immobilization Plant (WTP) is being constructed to treat the wastes and immobilize them in a glass waste form. The WTP includes a pretreatment facility to separate the wastes into a small volume of high-level waste (HLW) containing most of the radioactivity and a larger volume of low-activity waste (LAW) containing most of the nonradioactive chemicals. The HLW will be converted to glass in themore » HLW vitrification facility for ultimate disposal at an offsite federal repository. At least a portion (~35%) of the LAW will be converted to glass in the LAW vitrification facility and will be disposed of onsite at the Integrated Disposal Facility (IDF). The pretreatment and HLW vitrification facilities will have the capacity to treat and immobilize the wastes destined for each facility. However, a second LAW immobilization facility will be needed for the expected volume of LAW requiring immobilization. A cementitious waste form known as Cast Stone is being considered to provide the required additional LAW immobilization capacity. The Cast Stone waste form must be acceptable for disposal in the IDF. The Cast Stone waste form and immobilization process must be tested to demonstrate that the final Cast Stone waste form can comply with the waste acceptance criteria for the disposal facility and that the immobilization processes can be controlled to consistently provide an acceptable waste form product. Further, the waste form must be tested to provide the technical basis for understanding the long-term performance of the waste form in the disposal environment. These waste form performance data are needed to support risk assessment and performance assessment (PA) analyses of the long-term environmental impact of the waste disposal in the IDF. The PA is needed to satisfy both Washington State IDF Permit and DOE Order requirements. Cast Stone has been selected for solidification of radioactive wastes including WTP aqueous secondary wastes treated at the Effluent Treatment Facility (ETF) at Hanford. A similar waste form called Saltstone is used at the Savannah River Site (SRS) to solidify its LAW tank wastes.« less

Enhancement of L(+)-Lactic Acid Production of Immobilized Rhizopus Oryzae Implanted by Ion Beams

NASA Astrophysics Data System (ADS)

Fan, Yonghong; Yang, Yingge; Zheng, Zhiming; Li, Wen; Wang, Peng; Yao, Liming; Yu, Zengliang

2008-02-01

Immobilized Rhizopus oryzae culturing may be a solution to the inhibited production of L(+)-lactic acid in submerged fermentation, which is caused by aggregated mycelia floc. In the present study, a R. oryzae mutant (RL6041) with a 90% conversion rate of glucose into L-lactic acid was obtained by N+ implantation under the optimized conditions of a beam energy of 15 keV and a dose of 2.6 × 1015 ions/cm2. Using polyurethane foam as the immobilization matrix, the optimal L-lactic acid production conditions were determined as 4 mm polyurethane foam, 150 r/min, 50 g/L ~ 80 g/L of initial glucose, 38°C and pH 6.0. 15-cycle repeated productions of L-lactic acid by immobilized RL6041 were performed under the optimized culturing conditions and over 80% of the glucose was converted into L-lactic acid in 30 hours on average. The results show that immobilized RL6041 is a promising candidate for continuous L-lactic acid production.

A glucose biosensor based on direct electrochemistry of glucose oxidase immobilized on nitrogen-doped carbon nanotubes.

PubMed

Deng, Shengyuan; Jian, Guoqiang; Lei, Jianping; Hu, Zheng; Ju, Huangxian

2009-10-15

A novel biosensor for glucose was prepared by immobilizing glucose oxidase (GOx) on nitrogen-doped carbon nanotubes (CNx-MWNTs) modified electrode. The CNx-MWNTs membrane showed an excellent electrocatalytic activity toward the reduction of O(2) due to its diatomic side-on adsorption on CNx-MWNTs. The nitrogen doping accelerated the electron transfer from electrode surface to the immobilized GOx, leading to the direct electrochemistry of GOx. The biofunctional surface showed good biocompatibility, excellent electron-conductive network and large surface-to-volume ratio, which were characterized by scanning electron microscopy, contact angle and electrochemical impedance technique. The direct electron transfer of immobilized GOx led to stable amperometric biosensing for glucose with a linear range from 0.02 to 1.02 mM and a detection limit of 0.01 mM (S/N=3). These results indicated that CNx-MWNTs are good candidate material for construction of the third-generation enzyme biosensors based on the direct electrochemistry of immobilized enzymes.

Pineapple stem bromelain immobilized on different supports: catalytic properties in model wine.

PubMed

Ilaria, Benucci; Marco, Esti; Katia, Liburdi; Maria Vittoria, Garzillo Anna

2012-01-01

Bromelain from pineapple stem has been covalently immobilized on different supports to select the more efficient biocatalyst that should be applied toward unstable proteins in real white wine. In this preliminary study, catalytic properties of different immobilized bromelain forms were compared under wine-like conditions, against a synthetic substrate (Bz-Phe-Val-Arg-pNA).Covalent immobilization affected protease kinetic properties, even if all immobilized forms presented both a better substrate affinity and higher half-life (with the exception of a few procedures) with respect to the free enzyme. Stem bromelain was successfully immobilized on chitosan beads without glutaraldehyde thus yielding a food-safe and promising biocatalyst for unstable real wine future application. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

Comparative study of thermostability and ester synthesis ability of free and immobilized lipases on cross linked silica gel.

PubMed

Kumari, Annapurna; Mahapatra, Paramita; Kumar, Garlapati Vijay; Banerjee, Rintu

2008-06-01

A novel support has been utilized for immobilization of lipase, which was prepared by amination of silica with ethanolamine followed by cross linking with glutaraldehyde. Lipases from Rhizopus oryzae 3562 and Enterobacter aerogenes were immobilized on activated silica gel, where they retained 60 and 50% of respective original activity. The thermal stability of the immobilized lipases was significantly improved in comparison to the free forms while the pH stability remained unchanged. E. aerogenes and R. oryzae 3562 lipases retained 75 and 97% of respective initial activity on incubation at 90 degrees C, whereas both the free forms became inactive at this temperature. The conversion yield of isoamyl acetate was found to be higher with the immobilized fungal (90 vs. 21%) and bacterial lipases (64 vs. 18%) than the respective free forms. Immobilized R. oryzae 3562 lipases retained 50% activity for isoamyl acetate synthesis up to ten cycles whereas it was eight cycles for E. aerogenes.

Status of plutonium ceramic immobilization processes and immobilization forms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ebbinghaus, B.B.; Van Konynenburg, R.A.; Vance, E.R.

1996-05-01

Immobilization in a ceramic followed by permanent emplacement in a repository or borehole is one of the alternatives currently being considered by the Fissile Materials Disposition Program for the ultimate disposal of excess weapons-grade plutonium. To make Pu recovery more difficult, radioactive cesium may also be incorporated into the immobilization form. Valuable data are already available for ceramics form R&D efforts to immobilize high-level and mixed wastes. Ceramics have a high capacity for actinides, cesium, and some neutron absorbers. A unique characteristic of ceramics is the existence of mineral analogues found in nature that have demonstrated actinide immobilization over geologicmore » time periods. The ceramic form currently being considered for plutonium disposition is a synthetic rock (SYNROC) material composed primarily of zirconolite (CaZrTi{sub 2}O{sub 7}), the desired actinide host phase, with lesser amounts of hollandite (BaAl{sub 2}Ti{sub 6}O{sub 16}) and rutile (TiO{sub 2}). Alternative actinide host phases are also being considered. These include pyrochlore (Gd{sub 2}Ti{sub 2}O{sub 7}), zircon (ZrSiO{sub 4}), and monazite (CePO{sub 4}), to name a few of the most promising. R&D activities to address important technical issues are discussed. Primarily these include moderate scale hot press fabrications with plutonium, direct loading of PuO{sub 2} powder, cold press and sinter fabrication methods, and immobilization form formulation issues.« less

Cell-Free Expression and In Situ Immobilization of Parasite Proteins from Clonorchis sinensis for Rapid Identification of Antigenic Candidates

PubMed Central

Ju, Jung Won; Kim, Ho-Cheol; Shin, Hyun-Il; Kim, Yu Jung; Kim, Dong-Myung

2015-01-01

Progress towards genetic sequencing of human parasites has provided the groundwork for a post-genomic approach to develop novel antigens for the diagnosis and treatment of parasite infections. To fully utilize the genomic data, however, high-throughput methodologies are required for functional analysis of the proteins encoded in the genomic sequences. In this study, we investigated cell-free expression and in situ immobilization of parasite proteins as a novel platform for the discovery of antigenic proteins. PCR-amplified parasite DNA was immobilized on microbeads that were also functionalized to capture synthesized proteins. When the microbeads were incubated in a reaction mixture for cell-free synthesis, proteins expressed from the microbead-immobilized DNA were instantly immobilized on the same microbeads, providing a physical linkage between the genetic information and encoded proteins. This approach of in situ expression and isolation enables streamlined recovery and analysis of cell-free synthesized proteins and also allows facile identification of the genes coding antigenic proteins through direct PCR of the microbead-bound DNA. PMID:26599101

Immobilization of Pseudomonas sp. DG17 onto sodium alginate–attapulgite–calcium carbonate

PubMed Central

Wang, Hong Qi; Hua, Fei; Zhao, Yi Cun; Li, Yi; Wang, Xuan

2014-01-01

A strain of Pseudomonas sp. DG17, capable of degrading crude oil, was immobilized in sodium alginate–attapulgite–calcium carbonate for biodegradation of crude oil contaminated soil. In this work, proportion of independent variables, the laboratory immobilization parameters, the micromorphology and internal structure of the immobilized granule, as well as the crude oil biodegradation by sodium alginate–attapulgite–calcium carbonate immobilized cells and sodium alginate–attapulgite immobilized cells were studied to build the optimal immobilization carrier and granule-forming method. The results showed that the optimal concentrations of sodium alginate–attapulgite–calcium carbonate and calcium chloride were 2.5%–3.5%, 0.5%–1%, 3%–7% and 2%–4%, respectively. Meanwhile, the optimal bath temperature, embedding cell amount, reaction time and multiplication time were 50–60 °C, 2%, 18 h and 48 h, respectively. Moreover, biodegradation was enhanced by immobilized cells with a total petroleum hydrocarbon removal ranging from 33.56% ± 3.84% to 56.82% ± 3.26% after 20 days. The SEM results indicated that adding calcium carbonate was helpful to form internal honeycomb-like pores in the immobilized granules. PMID:26019567

In situ assembly in confined spaces of coated particle scaffolds as thermal underfills with extraordinary thermal conductivity.

PubMed

Hong, Guo; Schutzius, Thomas M; Zimmermann, Severin; Burg, Brian R; Zürcher, Jonas; Brunschwiler, Thomas; Tagliabue, Giulia; Michel, Bruno; Poulikakos, Dimos

2015-01-14

In situ assembly of high thermal conductivity materials in severely confined spaces is an important problem bringing with it scientific challenges but also significant application relevance. Here we present a simple, affordable, and reproducible methodology for synthesizing such materials, composed of hierarchical diamond micro/nanoparticle scaffolds and an ethylenediamine coating. An important feature of the assembly process is the utilization of ethylenediamine as an immobilizing agent to secure the integrity of the microparticle scaffolds during and after each processing step. After other liquid components employed in the scaffolds assembly dry out, the immobilization agent solidifies forming a stable coated particle scaffold structure. Nanoparticles tend to concentrate in the shell and neck regions between adjacent microparticles. The interface between core and shell, along with the concentrated neck regions of nanoparticles, significantly enhance the thermal conductivity, making such materials an excellent candidate as thermal underfills in the electronics industry, where efficient heat removal is a major stumbling block toward increasing packing density. We show that the presented structures exhibit nearly 1 order of magnitude improvement in thermal conductivity, enhanced temperature uniformity, and reduced processing time compared to commercially available products for electronics cooling, which underpins their potential utility.

Melter Technologies Assessment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Perez, J.M. Jr.; Schumacher, R.F.; Forsberg, C.W.

1996-05-01

The problem of controlling and disposing of surplus fissile material, in particular plutonium, is being addressed by the US Department of Energy (DOE). Immobilization of plutonium by vitrification has been identified as a promising solution. The Melter Evaluation Activity of DOE`s Plutonium Immobilization Task is responsible for evaluating and selecting the preferred melter technologies for vitrification for each of three immobilization options: Greenfield Facility, Adjunct Melter Facility, and Can-In-Canister. A significant number of melter technologies are available for evaluation as a result of vitrification research and development throughout the international communities for over 20 years. This paper describes an evaluationmore » process which will establish the specific requirements of performance against which candidate melter technologies can be carefully evaluated. Melter technologies that have been identified are also described.« less

The effects of below-elbow immobilization on driving performance.

PubMed

Jones, Evan M; Barrow, Aaron E; Skordas, Nic J; Green, David P; Cho, Mickey S

2017-02-01

There is limited research to guide physicians and patients in deciding whether it is safe to drive while wearing various forms of upper extremity immobilization. The purpose of this study is to evaluate the effect of below-elbow removable splints and fiberglass casts on automobile driving performance. 20 healthy subjects completed 10 runs through a closed, cone-marked driving course while wearing a randomized sequence of four different types of immobilization on each extremity (short arm thumb spica fiberglass cast, short arm fiberglass cast, short arm thumb spica splint, and short arm wrist splint). The first and last driving runs were without immobilization and served as controls. Performance was measured based on evaluation by a certified driving instructor (pass/fail scoring), cones hit, run time, and subject-perceived driving difficulty (1-10 analogue scoring). The greatest number of instructor-scored failures occurred while immobilized in right arm spica casts (n=6; p=0.02) and left arm spica casts (n=5; p=0.049). The right arm spica cast had the highest subject-perceived difficulty (5.2±1.9; p<0.001). All forms of immobilization had significantly increased perceived difficulty compared to control, except for the left short arm splint (2.5±1.6; p>0.05). There was no significant difference in number of cones hit or driving time between control runs and runs with any type of immobilization. Drivers should use caution when wearing any of the forms of upper extremity immobilization tested in this study. All forms of immobilization, with exception of the left short arm splint significantly increased perceived driving difficulty. However, only the fiberglass spica casts (both left and right arm), significantly increased drive run failures due to loss of vehicle control. We recommend against driving when wearing a below-elbow fiberglass spica cast on either extremity. Copyright © 2016. Published by Elsevier Ltd.

A facile strategy for enzyme immobilization with highly stable hierarchically porous metal-organic frameworks.

PubMed

Liu, Xiao; Qi, Wei; Wang, Yuefei; Su, Rongxin; He, Zhimin

2017-11-16

Metal-organic frameworks (MOFs) have drawn extensive research interest as candidates for enzyme immobilization owing to their tunable porosity, high surface area, and excellent chemical/thermal stability. Herein, we report a facile and universal strategy for enzyme immobilization using highly stable hierarchically porous metal-organic frameworks (HP-MOFs). The HP-MOFs were stable over a wide pH range (pH = 2-11 for HP-DUT-5) and met the catalysis conditions of most enzymes. The as-prepared hierarchical micro/mesoporous MOFs with mesoporous defects showed a superior adsorption capacity towards enzymes. The maximum adsorption capacity of HP-DUT-5 for glucose oxidase (GOx) and uricase was 208 mg g -1 and 225 mg g -1 , respectively. Furthermore, we constructed two multi-enzyme biosensors for glucose and uric acid (UA) by immobilizing GOx and uricase with horseradish peroxidase (HRP) on HP-DUT-5, respectively. These sensors were efficiently applied in the colorimetric detection of glucose and UA and showed good sensitivity, selectivity, and recyclability.

Horseradish peroxidase-immobilized magnetic mesoporous silica nanoparticles as a potential candidate to eliminate intracellular reactive oxygen species.

PubMed

Shen, Yajing; Zhang, Ye; Zhang, Xiang; Zhou, Xiuhong; Teng, Xiyao; Yan, Manqing; Bi, Hong

2015-02-21

Horseradish peroxidase-immobilized magnetic mesoporous silica nanoparticles (MMSNs-HRP) have been synthesized by a NHS/EDC coupling between the amino groups of horseradish peroxidase (HRP) and the carboxyl groups on the MMSNs surface. It is found that the immobilized HRP on MMSNs still retain high activity and the MMSNs-HRP can eliminate the reactive oxygen species (ROS) in Chinese hamster ovary (CHO) cells induced by the addition of H2O2 aqueous solution. Further, the fluorescent MMSN-HRP-CD nanoparticles have been prepared by attaching biocompatible, fluorescent carbon dots (CDs) to MMSNs-HRP. We have also investigated the effect of an applied magnetic field on cellular uptake of MMSNs-HRP-CDs and found that the internalization of MMSNs-HRP-CDs by CHO cells could be enhanced within 2 hours under the magnetic field. This work provides us with a novel and efficient method to eliminate ROS in living cells by using HRP-immobilized nanoparticles.

Vitrified chemically bonded phosphate ceramics for immobilization of radioisotopes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wagh, Arun S.

2016-04-05

A method of immobilizing a radioisotope and vitrified chemically bonded phosphate ceramic (CBPC) articles formed by the method are described. The method comprises combining a radioisotope-containing material, MgO, a source of phosphate, and optionally, a reducing agent, in water at a temperature of less than 100.degree. C. to form a slurry; curing the slurry to form a solid intermediate CBPC article comprising the radioisotope therefrom; comminuting the intermediate CBPC article, mixing the comminuted material with glass frits, and heating the mixture at a temperature in the range of about 900 to about 1500.degree. C. to form a vitrified CBPC articlemore » comprising the radioisotope immobilized therein.« less

Immobilization of pectinase onto chitosan magnetic nanoparticles by macromolecular cross-linker.

PubMed

Sojitra, Uttam V; Nadar, Shamraja S; Rathod, Virendra K

2017-02-10

Pectinase was immobilized onto chitosan magnetic nanoparticles (CMNPs) by dextran polyaldehyde as a macromolecular cross-linking agent. The parameters like cross-linking concentration, time and CMNPs to enzyme ratio were optimized. Further, prepared magnetic pectinase nanobiocatalyst was characterized by FT-IR and XRD. The thermal kinetic studies for immobilized pectinase showed two folds improved thermal stability in the range of 55-75°C as compared to free form. The V max and K m values of immobilized pectinase were found to be nearly equal to native form which indicated that conformational flexibility of pectinase was retained even after immobilization. The residual activity of immobilized pectinase was 85% after seven successive cycles of reuse, while it retained upto 89% residual activity on storage of fifteen days which exhibited excellent stability and durability. The conformational changes in pectinase after immobilization were evaluated by FT-IR spectroscopy data analysis tools. Finally, magnetic pectinase nanobiocatalyst was employed for apple juice clarification which showed turbidity reduction upto 74% after 150min treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

A SPR strategy for high-throughput ligand screenings based on synthetic peptides mimicking a selected subdomain of the target protein: a proof of concept on HER2 receptor.

PubMed

Monfregola, Luca; Vitale, Rosa Maria; Amodeo, Pietro; De Luca, Stefania

2009-10-01

The discovery of pharmaceutical agents is a complex, lengthy and costly process, critically depending on the availability of rapid and efficient screening methods. In particular, when targets are large, multidomain proteins, their complexity may affect unfavorably technical feasibility, costs and unambiguity of binding test interpretation. A possible strategy to overcome these problems relies on molecular design of receptor fragments that are: sensible targets for ligand screenings, conformationally stable also as standalone domains, easily synthesized and immobilized on chip for Biacore experiments. An additional desirable feature for new ligands is the ability of selectively targeting alternative conformational states typical of many proteins. To test the feasibility of such approach on a case with potential applicative interest, we developed a surface plasmon resonance (SPR)-based screening method for drug candidates toward HER2, a Tyr-kinase receptor targeted in anticancer therapies. HER2 was mimicked by HER2-DIVMP, a modified fragment of it immobilized onto the sensor surface specifically modeling HER2 domain IV in its bounded form, designed by structural comparison of HER2 alone and in complex with Herceptin, a monoclonal therapeutic anti-HER2 antibody. This design and its implementation in SPR devices was validated by investigating Herceptin- HER2-DIVMP affinity, measuring its dissociation constant (K(D)=19.2 nM). An efficient synthetic procedure to prepare the HER2-DIVMP peptide was also developed. The HER2-DIVMP conformational stability suggested by experimental and computational results, makes it also a valuable candidate as a mold to design new molecules selectively targeting domain IV of HER2.

In vitro and in vivo evaluation of bone morphogenetic protein-2 (BMP-2) immobilized collagen-coated polyetheretherketone (PEEK)

NASA Astrophysics Data System (ADS)

Du, Ya-Wei; Zhang, Li-Nan; Ye, Xin; Nie, He-Min; Hou, Zeng-Tao; Zeng, Teng-Hui; Yan, Guo-Ping; Shang, Peng

2015-03-01

Polyetheretherketone (PEEK) is regarded as one of the most potential candidates of biomaterials in spinal implant applications. However, as a bioinert material, PEEK plays a limited role in osteoconduction and osseointegration. In this study, recombinant human bone morphogenetic protein-2 (rhBMP-2) was immobilized onto the surface of collagen-coated PEEK in order to prepare a multi-functional material. After adsorbed onto the PEEK surface by hydrophobic interaction, collagen was cross-linked with N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). EDC/NHS system also contributed to the immobilization of rhBMP-2. Water contact angle tests, XPS and SEM clearly demonstrated the surface changes. ELISA tests quantified the amount of rhBMP-2 immobilized and the release over a period of 30 d. In vitro evaluation proved that the osteogenesis differentiation rate was higher when cells were cultured on modified PEEK discs than on regular ones. In vivo tests were conducted and positive changes of major parameters were presented. This report demonstrates that the rhBMP-2 immobilized method for PEEK modification increase bioactivity in vitro and in vivo, suggesting its practicability in orthopedic and spinal clinical applications.

Potential Applications of Immobilized β-Galactosidase in Food Processing Industries

PubMed Central

Panesar, Parmjit S.; Kumari, Shweta; Panesar, Reeba

2010-01-01

The enzyme β-galactosidase can be obtained from a wide variety of sources such as microorganisms, plants, and animals. The use of β-galactosidase for the hydrolysis of lactose in milk and whey is one of the promising enzymatic applications in food and dairy processing industries. The enzyme can be used in either soluble or immobilized forms but the soluble enzyme can be used only for batch processes and the immobilized form has the advantage of being used in batch wise as well as in continuous operation. Immobilization has been found to be convenient method to make enzyme thermostable and to prevent the loss of enzyme activity. This review has been focused on the different types of techniques used for the immobilization of β-galactosidase and its potential applications in food industry. PMID:21234407

Protein Immobilization Capabilities of Sucrose and Trehalose Glasses: The Effect of Protein/Sugar Concentration Unraveled by High-Field EPR.

PubMed

Malferrari, Marco; Savitsky, Anton; Lubitz, Wolfgang; Möbius, Klaus; Venturoli, Giovanni

2016-12-01

Disaccharide glasses are increasingly used to immobilize proteins at room temperature for structural/functional studies and long-term preservation. To unravel the molecular basis of protein immobilization, we studied the effect of sugar/protein concentration ratios in trehalose or sucrose matrixes, in which the bacterial photosynthetic reaction center (RC) was embedded as a model protein. The structural, dynamical, and H-bonding characteristics of the sugar-protein systems were probed by high-field W-band EPR of a matrix-dissolved nitroxide radical. We discovered that RC immobilization and thermal stabilization, being independent of the protein concentration in trehalose, occur in sucrose only at sufficiently low sugar/protein ratios. EPR reveals that only under such conditions does sucrose form a microscopically homogeneous matrix that immobilizes, via H-bonds, the nitroxide probe. We conclude that the protein immobilization capability depends critically on the propensity of the glass-forming sugar to create intermolecular H-bond networks, thus establishing long-range, homogeneous connectivity within the matrix.

Immobilization of Superoxide Dismutase on Polyelectrolyte-Functionalized Titania Nanosheets.

PubMed

Rouster, Paul; Pavlovic, Marko; Szilagyi, Istvan

2018-02-16

The superoxide dismutase (SOD) enzyme was successfully immobilized on titania nanosheets (TNS) functionalized with the poly(diallyldimethylammonium chloride) (PDADMAC) polyelectrolyte. The TNS-PDADMAC solid support was prepared by hydrothermal synthesis followed by self-assembled polyelectrolyte layer formation. It was found that SOD strongly adsorbed onto oppositely charged TNS-PDADMAC through electrostatic and hydrophobic interactions. The TNS-PDADMAC-SOD material was characterized by light scattering and microscopy techniques. Colloidal stability studies revealed that the obtained nanocomposites possessed good resistance against salt-induced aggregation in aqueous suspensions. The enzyme kept its functional integrity upon immobilization; therefore, TNS-PDADMAC-SOD showed excellent superoxide radical anion scavenging activity. The developed system is a promising candidate for applications in which suspensions of antioxidant activity are required in the manufacturing processes. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

I-NERI Annual Technical Progress Report 2007-004-K Development and Characterization of New High-Level Waste Forms for Achieving Waste Minimization from Pyroprocessing

DOE Office of Scientific and Technical Information (OSTI.GOV)

S. Frank

The current method for the immobilization of fission products that accumulate in electrorefiner salt during the electrochemical processing of used metallic nuclear fuel is to encapsulate the electrorefiner salt in a glass-bonded sodalite ceramic waste form. This process was developed by Argonne National Laboratory in the USA and is currently performed at the Idaho National Laboratory for the treatment of Experimental Breeder Reactor-II (EBR-II) used fuel. This process utilizes a “once-through” option for the disposal of spent electrorefiner salt; where, after the treatment of the EBR-II fuel, the electrorefiner salt containing the active fission products will be disposed of inmore » the ceramic waste form (CWF). The CWF produced will have low fission product loading of approximately 2 to 5 weight percent due to the limited fuel inventory currently being processed. However; the design and implementation of advanced electrochemical processing facilities to treat used fuel would process much greater quantities fuel. With an advanced processing facility, it would be necessary to selectively remove fission products from the electrorefiner salt for salt recycle and to concentrate the fission products to reduce the volume of high-level waste from the treatment facility. The Korean Atomic Energy Research Institute and the Idaho National Laboratory have been collaborating on I-NERI research projects for a number of years to investigate both aspects of selective fission product separation from electrorefiner salt, and to develop advanced waste forms for the immobilization of the collected fission products. The first joint KAERI/INL I-NERI project titled: 2006-002-K, Separation of Fission Products from Molten LiCl-KCl Salt Used for Electrorefining of Metal Fuels, was successfully completed in 2009 by concentrating and isolating fission products from actual electrorefiner salt used for the treated used EBR-II fuel. Two separation methods were tested and from these tests were produced concentrated salt products that acted as the feed material for development of advanced waste forms investigated in this proposal. Accomplishments from the first year activities associated with this I-NERI project included the down selection of candidate waste forms to immobilize fission products separated from electrorefiner salt, and the design of equipment to fabricate actual waste forms in the Hot Fuels Examination Facility (HFEF) at the INL. Reported in this document are accomplishments from the second year (FY10) work performed at the INL, and includes the testing of waste form fabrication equipment, repeating the fission product precipitation experiment, and initial waste form fabrication efforts.« less

Bioreactors with immobilized lipases: state of the art.

PubMed

Balcão, V M; Paiva, A L; Malcata, F X

1996-05-01

This review attempts to provide an updated compilation of studies reported in the literature pertaining to reactors containing lipases in immobilized forms, in a way that helps the reader direct a bibliographic search and develop an integrated perspective of the subject. Highlights are given to industrial applications of lipases (including control and economic considerations), as well as to methods of immobilization and configurations of reactors in which lipases are used. Features associated with immobilized lipase kinetics such as enzyme activities, adsorption properties, optimum operating conditions, and estimates of the lumped parameters in classical kinetic formulations (Michaelis-Menten model for enzyme action and first-order model for enzyme decay) are presented in the text in a systematic tabular form.

Label-Free Detection of Cardiac Troponin-I Using Carbon Nanofiber Based Nanoelectrode Arrays

NASA Technical Reports Server (NTRS)

Periyakaruppan, Adaikkappan; Koehne, Jessica Erin; Gandhiraman, Ram P.; Meyyappan, M.

2013-01-01

A sensor platform based on vertically aligned carbon nanofibers (CNFs) has been developed. Their inherent nanometer scale, high conductivity, wide potential window, good biocompatibility and well-defined surface chemistry make them ideal candidates as biosensor electrodes. A carbon nanofiber (CNF) multiplexed array has been fabricated with 9 sensing pads, each containing 40,000 carbon nanofibers as nanoelectrodes. Here, we report the use of vertically aligned CNF nanoelectrodes for the detection of cardiac Troponin-I for the early diagnosis of myocardial infarction. Antibody, antitroponin, probe immobilization and subsequent binding to human cardiac troponin-I were characterized using electrochemical impedance spectroscopy and cyclic voltammetry techniques. Each step of the modification process resulted in changes in electrical capacitance or resistance to charge transfer due to the changes at the electrode surface upon antibody immobilization and binding to the specific antigen. This sensor demonstrates high sensitivity, down to 0.2 ng/mL, and good selectivity making this platform a good candidate for early stage diagnosis of myocardial infarction.

Lysozyme-immobilized electrospun PAMA/PVA and PSSA-MA/PVA ion-exchange nanofiber for wound healing.

PubMed

Tonglairoum, Prasopchai; Ngawhirunpat, Tanasait; Rojanarata, Theerasak; Opanasopit, Praneet

2014-08-27

Abstract This research was aimed to develop the lysozyme immobilized ion-exchange nanofiber mats for wound healing. To promote the healing process, the PSSA-MA/PVA and PAMA ion-exchange nanofiber mats were fabricated to mimic the extracellular matrix structure using electrospinning process followed by thermally crosslinked. Lysozyme was immobilized on the ion-exchane nanofibers by an adsorption method. The ion-exchange nanofibers were investigated using SEM, FTIR and XRPD. Moreover, the lysozyme-immobilized ion-exchange nanofibers were further investigated for lysozyme content and activity, lysozyme release and wound healing activity. The fiber diameters of the mats were in the nanometer range. Lysozyme was gradually absorbed into the PSSA-MA/PVA nanofiber with higher extend than that is absorbed on the PAMA/PVA nanofiber and exhibited higher activity than lysozyme-immobilized PAMA/PVA nanofiber. The total contents of lysozyme on the PSSA-MA/PVA and PAMA/PVA nanofiber were 648 and 166 µg/g, respectively. FTIR and lysozyme activity results confirmed the presence of lysozyme on the nanofiber mats. The lysozyme was released from the PSSA-MA/PVA and PAMA/PVA nanofiber in the same manner. The lysozyme-immobilized PSSA-MA/PVA nanofiber mats and lysozyme-immobilized PAMA/PVA nanofiber mats exhibited significantly faster healing rate than gauze and similar to the commercial antibacterial gauze dressing. These results suggest that these nanofiber mats could provide the promising candidate for wound healing application.

Development of Alternative Technetium Waste Forms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Czerwinski, Kenneth

2013-09-13

The UREX+1 process is under consideration for the separation of transuranic elements from spent nuclear fuel. The first steps of this process extract the fission product technicium-99 ({sup 99}Tc) into an organic phase containing tributylphosphate together with uranium. Treatment of this stream requires the separation of Tc from U and placement into a suitable waste storage form. A potential candidate waste form involves immobilizing the Tc as an alloy with either excess metallic zirconium or stainless steel. Although Tc-Zr alloys seem to be promising waste forms, alternative materials must be investigated. Innovative studies related to the synthesis and behavior ofmore » a different class of Tc materials will increase the scientific knowledge related to development of Tc waste forms. These studies will also provide a better understanding of the behavior of {sup 99}Tc in repository conditions. A literature survey has selected promising alternative waste forms for further study: technetium metallic alloys, nitrides, oxides, sulfides, and pertechnetate salts. The goals of this project are to 1) synthesize and structurally characterize relevant technetium materials that may be considered as waste forms, 2) investigate material behavior in solution under different conditions of temperature, electrochemical potential, and radiation, and 3) predict the long-term behavior of these materials.« less

Effects of immobilization on spermiogenesis

NASA Technical Reports Server (NTRS)

Meitner, E. R.

1980-01-01

The influence of immobilization stress on spermiogenesis in rats was investigated. After 96 hour immobilization, histological changes began to manifest themselves in the form of practically complete disappearance of cell population of the wall of seminiferous tubule as well as a markedly increased number of cells with pathologic mitoses. Enzymological investigations showed various changes of activity (of acid and alkaline phosphatase and nonspecific esterase) in the 24, 48, and 96 hour immobilization groups.

Sulfur cycling, retention, and mobility in soils: A review

Treesearch

Pamela J. Edwards

1998-01-01

Sulfur inputs to forests originate from mineral weathering, atmospheric deposition, and organic matter decomposition. In the soil, sulfur occurs in organic and inorganic forms and is cycled within and between those forms via mobilization, immobilization, mineralization, oxidation, and reduction processes. Organic sulfur compounds are largely immobile. Inorganic sulfur...

Nanoporous Gold for Enzyme Immobilization.

PubMed

Stine, Keith J; Jefferson, Kenise; Shulga, Olga V

2017-01-01

Nanoporous gold (NPG) is a material of emerging interest for immobilization of biomolecules, especially enzymes. The material provides a high surface area form of gold that is suitable for physisorption or for covalent modification by self-assembled monolayers. The material can be used as a high surface area electrode and with immobilized enzymes can be used for amperometric detection schemes. NPG can be prepared in a variety of formats from alloys containing between 20 and 50 % atomic composition of gold and less noble element(s) by dealloying procedures. Materials resembling NPG can be prepared by hydrothermal and electrodeposition methods. Related high surface area gold structures have been prepared using templating approaches. Covalent enzyme immobilization can be achieved by first forming a self-assembled monolayer on NPG bearing a terminal reactive functional group followed by conjugation to the enzyme through amide linkages to lysine residues. Enzymes can also be entrapped by physisorption or immobilized by electrostatic interactions.

Protein Surface Structural Recognition in Inactive Areas: A New Immobilization Strategy for Acetylcholinesterase.

PubMed

Diao, Jianxiong; Yu, Xiaolu; Ma, Lin; Li, Yuanqing; Sun, Ying

2018-05-16

This work reported a new method of design for the immobilization of acetylcholinesterase (AChE) based on its molecular structure to improve its sensitivity and stability. The immobilization binding site on the surface of AChE was determined using MOLCAD's multi-channel functionality. Then, 11 molecules ((+)-catechin, (-)-epicatechin, (-)-gallocatechin, hesperetin, naringenin, quercetin, taxifolin, (-)-epicatechin gallate, flupirtine, atropine, and hyoscyamine) were selected from the ZINC database (about 50 000 molecules) as candidate affinity ligands for AChE. The fluorescence results showed that the binding constant K b between AChE and the ligands ranged from 0.01344 × 10 4 to 4.689 × 10 4 M -1 and there was one independent class of binding site for the ligands on AChE. The AChE-ligand binding free energy ranged from -12.14 to -26.65 kJ mol -1 . Naringenin, hesperetin, and quercetin were the three most potent immobilized affinity ligands. In addition, it was confirmed that the binding between the immobilized ligands only occurred at a single site, located in an inactive area on the surface of AChE, and did not affect the enzymatic activity as shown through a competition experiment and enzyme assay. This method based on protein surface structural recognition with high sensitivity and stability can be used as a generic approach for design of the enzyme immobilization and biosensor development.

Enzyme immobilization techniques on poly(glycidyl methacrylate-co-ethylene dimethacrylate) carrier with penicillin amidase as model.

PubMed

Drobník, J; Saudek, V; Svec, F; Kálal, J; Vojtísek, V; Bárta, M

1979-08-01

Two types of bead-form macroporous carriers based on glycidyl methacrylate with ethylene dimethacrylate copolymers were used for the immobilization of penicillin amidase either directly or after chemical modification. Direct binding through oxirane groups, which is equally efficient at pH 4.2 and 7, is relatively slow and brings about an activity loss at low enzyme concentrations. The most efficient immobilization was achieved on glutaraldehyde-activated amino carrier, irrespective of whether the amino groups were formed by ammonia or 1,6-diaminohexane treatment of the original oxirane carrier. Hydrazine treatment gave lower immobilization yields. The same is true of the azide method independent of the length of the spacer. Most enzyme activity was preserved by coupling the carbodiimide-activated enzyme to the carrier with alkyl or arylamino groups at the end of a longer substituent. Immobilization on diazo-modified carrier gave average results. Rapid immobilization by a lysine-modified phosgene-treated carrier resulted in an activity loss. It is suggested that multipoint and very tight attachment of the enzyme molecule to the matrix decreased the activity. The immobilized activity is quite stable in solution and very stable upon lyophilization with sucrose.

DuraLith geopolymer waste form for Hanford secondary waste: correlating setting behavior to hydration heat evolution.

PubMed

Xu, Hui; Gong, Weiliang; Syltebo, Larry; Lutze, Werner; Pegg, Ian L

2014-08-15

The binary furnace slag-metakaolin DuraLith geopolymer waste form, which has been considered as one of the candidate waste forms for immobilization of certain Hanford secondary wastes (HSW) from the vitrification of nuclear wastes at the Hanford Site, Washington, was extended to a ternary fly ash-furnace slag-metakaolin system to improve workability, reduce hydration heat, and evaluate high HSW waste loading. A concentrated HSW simulant, consisting of more than 20 chemicals with a sodium concentration of 5 mol/L, was employed to prepare the alkaline activating solution. Fly ash was incorporated at up to 60 wt% into the binder materials, whereas metakaolin was kept constant at 26 wt%. The fresh waste form pastes were subjected to isothermal calorimetry and setting time measurement, and the cured samples were further characterized by compressive strength and TCLP leach tests. This study has firstly established quantitative linear relationships between both initial and final setting times and hydration heat, which were never discovered in scientific literature for any cementitious waste form or geopolymeric material. The successful establishment of the correlations between setting times and hydration heat may make it possible to efficiently design and optimize cementitious waste forms and industrial wastes based geopolymers using limited testing results. Copyright © 2014 Elsevier B.V. All rights reserved.

DNA hybridization sensor based on pentacene thin film transistor.

PubMed

Kim, Jung-Min; Jha, Sandeep Kumar; Chand, Rohit; Lee, Dong-Hoon; Kim, Yong-Sang

2011-01-15

A DNA hybridization sensor using pentacene thin film transistors (TFTs) is an excellent candidate for disposable sensor applications due to their low-cost fabrication process and fast detection. We fabricated pentacene TFTs on glass substrate for the sensing of DNA hybridization. The ss-DNA (polyA/polyT) or ds-DNA (polyA/polyT hybrid) were immobilized directly on the surface of the pentacene, producing a dramatic change in the electrical properties of the devices. The electrical characteristics of devices were studied as a function of DNA immobilization, single-stranded vs. double-stranded DNA, DNA length and concentration. The TFT device was further tested for detection of λ-phage genomic DNA using probe hybridization. Based on these results, we propose that a "label-free" detection technique for DNA hybridization is possible through direct measurement of electrical properties of DNA-immobilized pentacene TFTs. Copyright © 2010 Elsevier B.V. All rights reserved.

pH-dependent immobilization of urease on glutathione-capped gold nanoparticles.

PubMed

Garg, Seema; De, Arnab; Mozumdar, Subho

2015-05-01

Urease is a nickel-dependent metalloenzyme that catalyzes the hydrolysis of urea to form ammonia and carbon dioxide. Although the enzyme serves a significant role in several detoxification and analytical processes, its usability is restricted due to high cost, availability in small amounts, instability, and a limited possibility of economic recovery from a reaction mixture. Hence, there is a need to develop an efficient, simple, and reliable immobilization strategy for the enzyme. In this study, the carboxyl terminated surface of glutathione-capped gold nanoparticles have been utilized as a solid support for the covalent attachment of urease. The immobilization has been carried out at different pH conditions so as to elucidate its effect on the immobilization efficiency and enzyme bioactivity. The binding of the enzyme has been quantitatively and qualitatively analyzed through techniques like ultraviolet-visible spectroscopy, intrinsic steady state fluorescence, and circular dichorism. The bioactivity of the immobilized enzyme was investigated with respect to the native enzyme under different thermal conditions. Recyclability and shelf life studies of the immobilized enzyme have also been carried out. Results reveal that the immobilization is most effective at pH of 7.4 followed by that in an acidic medium and is least in alkaline environment. The immobilized enzyme also exhibits enhance activity in comparison to the native form at physiological temperature. The immobilized urease (on gold glutathione nanoconjugates surface) can be effectively employed for biosensor fabrication, immunoassays and as an in vivo diagnostic tool in the future. © 2014 Wiley Periodicals, Inc.

Simultaneous immobilization of cadmium and lead in contaminated soils by hybrid bio-nanocomposites of fungal hyphae and nano-hydroxyapatites.

PubMed

Yang, Zhihui; Liang, Lifen; Yang, Weichun; Shi, Wei; Tong, Yunping; Chai, Liyuan; Gao, Shikang; Liao, Qi

2018-04-01

Self-aggregation of bulk nano-hydroxyapatites (n-HAPs) undermines their immobilization efficiencies of heavy metals in the contaminated soils. Here, the low-cost, easily obtained, and environment-friendly filamentous fungi have been introduced for the bio-matrices of the hybrid bio-nanocomposites to potentially solve such problem of n-HAPs. According to SEM, TEM, XRD, and FT-IR analyses, n-HAPs were successfully coated onto the fungal hyphae and their self-aggregation was improved. The immobilization efficiencies of diethylene-triamine-pentaacetic acid (DTPA)-extractable Cd and Pb in the contaminated soils by the bio-nanocomposites were individually one to four times of that by n-HAPs or the fungal hyphae. Moreover, the Aspergillus niger-based bio-nanocomposite (ANHP) was superior to the Penicillium Chrysogenum F1-based bio-nanocomposite (PCHP) in immobilization of Cd and Pb in the contaminated soils. In addition, the results of XRD showed that one of the potential mechanisms of metal immobilization by the hybrid bio-nanocomposites was dissolution of n-HAPs followed by precipitation of new metal phosphate minerals. Our results suggest that the hybrid bio-nanocomposite (ANHP) can be recognized as a promising soil amendment candidate for effective remediation on the soils simultaneously contaminated by Cd and Pb.

Improved catalytic properties of Penicillium notatum lipase immobilized in nanoscale silicone polymeric films.

PubMed

Rehman, Saima; Wang, Ping; Bhatti, Haq Nawaz; Bilal, Muhammad; Asgher, Muhammad

2017-04-01

Lipases are one of the most proficient biocatalysts having enormous biotechnological prospective. Immobilization offers a potential solution to improve the stability and recycling characteristics of lipases. An extracellular lipase from Penicillium notatum (PNL) was immobilized in silicon polymers (SiP) through entrapment, and subsequently coated this matrix on the network of fibers in the sponges. The silicone polymers-immobilized lipase (SiP-lipase) displayed highest apparent activity and entrapment efficiency of 1.19Ug -1 polymers and 92.3%, respectively. It also exhibited greater catalytic activity in broad-working pHs and higher temperature than equivalent free-state of enzyme. Immobilization caused an improvement in thermo-stability of the lipase with an increase in energy of activation. The recycling potential of SiP-lipase was investigated. After reusing the sponge pieces for ten reaction cycles, the SiP preserved its structure without leakage of enzyme, and retained around 90% of its original activity. The SiP surface analysis was envisaged by scanning electron microscopy that further confirmed the recycling efficiency of SiP-lipase. Overall, SiP-lipase displayed a number of useful properties that make it a promising candidate for future applications in different chemical processes. Copyright © 2017 Elsevier B.V. All rights reserved.

Immobilization of Cr (VI) in stainless steel slag and Cd, As, and Pb in wastewater using blast furnace slag via a hydrothermal treatment

NASA Astrophysics Data System (ADS)

Tae, Soon-Jae; Morita, Kazuki

2017-05-01

The immobilization of hexavalent chromium in stainless steel slag using blast furnace slag as the immobilizing agent and by performing a hydrothermal treatment was investigated. The results showed that there was no immobilization in the absence of the blast furnace slag. On the other hand, the hexavalent chromium in stainless steel slag could be immobilized through the hydrothermal reaction when blast furnace slag was used at 250 °C for 24 h. A leaching test was performed to evaluate the degree of immobilization of hexavalent chromium in the products formed by the hydrothermal reaction. It was found that the degree of immobilization was very high. Based on the results obtained, the immobilization mechanism of hexavalent chromium in stainless steel slag, resulting from the hydrothermal treatment of blast furnace slag, could be elucidated. Finally, the immobilization of cadmium, lead, and arsenic using blast furnace slag as the immobilization agent was also studied while focusing on the effects of the hydrothermal treatment.

Novel functionalized fluorescent polymeric nanoparticles for immobilization of biomolecules

NASA Astrophysics Data System (ADS)

Jain, Swati; Chattopadhyay, Sruti; Jackeray, Richa; Abid, C. K. V. Zainul; Singh, Harpal

2013-07-01

Novel, size controlled fluorescent polymeric nanoparticles (FPNP) were synthesized having acetoacetoxy functionality on the surface for immobilization of biomolecules which can be utilized as biomarkers and labels in fluoroimmunoassays. Core-shell nanoparticles of poly(styrene, St-methyl methacrylate, MMA-acetoacetoxy ethyl methacrylate, AAEM), stabilized by various concentrations of surfactant, sodium lauryl sulphate (SLS), were obtained by facile miniemulsion co-polymerization encapsulated with pyrene molecules in their hydrophobic core. Analytical, spectroscopic and imaging characterization techniques revealed the formation of stable, monodisperse, spherical nano sized particles exhibiting high luminescence properties. Particles with 1% SLS (S1) showed good dispersion stability and fluorescence intensity and were chosen as ideal candidates for further immobilization studies. Steady state fluorescence studies showed 10 times higher fluorescence intensity of S1 nanoparticles than that of pyrene solution in solvent-toluene at the same concentration. Environmental factors such as pH, ionic strength and time were found to have no effect on fluorescence intensity of FPNPs. Surface β-di-ketone groups were utilized for the covalent immobilization of enzyme conjugated antibodies without any activation or pre-treatment of nanoparticles.Novel, size controlled fluorescent polymeric nanoparticles (FPNP) were synthesized having acetoacetoxy functionality on the surface for immobilization of biomolecules which can be utilized as biomarkers and labels in fluoroimmunoassays. Core-shell nanoparticles of poly(styrene, St-methyl methacrylate, MMA-acetoacetoxy ethyl methacrylate, AAEM), stabilized by various concentrations of surfactant, sodium lauryl sulphate (SLS), were obtained by facile miniemulsion co-polymerization encapsulated with pyrene molecules in their hydrophobic core. Analytical, spectroscopic and imaging characterization techniques revealed the formation of stable, monodisperse, spherical nano sized particles exhibiting high luminescence properties. Particles with 1% SLS (S1) showed good dispersion stability and fluorescence intensity and were chosen as ideal candidates for further immobilization studies. Steady state fluorescence studies showed 10 times higher fluorescence intensity of S1 nanoparticles than that of pyrene solution in solvent-toluene at the same concentration. Environmental factors such as pH, ionic strength and time were found to have no effect on fluorescence intensity of FPNPs. Surface β-di-ketone groups were utilized for the covalent immobilization of enzyme conjugated antibodies without any activation or pre-treatment of nanoparticles. Electronic supplementary information (ESI) available: Resulting ATR-FTIR spectrum and procedure to study fluorescence of nanoparticles, effect of particle size, concentration, pH, ionic strength and time on Fl intensity of FPNP. See DOI: 10.1039/c3nr34100c

Secondary Waste Cast Stone Waste Form Qualification Testing Plan

DOE Office of Scientific and Technical Information (OSTI.GOV)

Westsik, Joseph H.; Serne, R. Jeffrey

2012-09-26

The Hanford Tank Waste Treatment and Immobilization Plant (WTP) is being constructed to treat the 56 million gallons of radioactive waste stored in 177 underground tanks at the Hanford Site. The WTP includes a pretreatment facility to separate the wastes into high-level waste (HLW) and low-activity waste (LAW) fractions for vitrification and disposal. The LAW will be converted to glass for final disposal at the Integrated Disposal Facility (IDF). Cast Stone – a cementitious waste form, has been selected for solidification of this secondary waste stream after treatment in the ETF. The secondary-waste Cast Stone waste form must be acceptablemore » for disposal in the IDF. This secondary waste Cast Stone waste form qualification testing plan outlines the testing of the waste form and immobilization process to demonstrate that the Cast Stone waste form can comply with the disposal requirements. Specifications for the secondary-waste Cast Stone waste form have not been established. For this testing plan, Cast Stone specifications are derived from specifications for the immobilized LAW glass in the WTP contract, the waste acceptance criteria for the IDF, and the waste acceptance criteria in the IDF Permit issued by the State of Washington. This testing plan outlines the testing needed to demonstrate that the waste form can comply with these waste form specifications and acceptance criteria. The testing program must also demonstrate that the immobilization process can be controlled to consistently provide an acceptable waste form product. This testing plan also outlines the testing needed to provide the technical basis for understanding the long-term performance of the waste form in the disposal environment. These waste form performance data are needed to support performance assessment analyses of the long-term environmental impact of the secondary-waste Cast Stone waste form in the IDF« less

Engineered Materials for Cesium and Strontium Storage Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sean M. McDeavitt

2010-04-14

Closing the nuclear fuel cycle requires reprocessing spent fuel to recover the long-lived components that still have useful energy content while immobilizing the remnant waste fission products in stable forms. At the genesis of this project, next generation spent fuel reprocessing methods were being developed as part of the U.S. Department of Energy's Advanced Fuel Cycle Initiative. One of these processes was focused on solvent extraction schemes to isolate cesium (Cs) and strontium (Sr) from spent nuclear fuel. Isolating these isotopes for short-term decay storage eases the design requirements for long-term repository disposal; a significant amount of the radiation andmore » decay heat in fission product waste comes from Cs-137 and Sr-90. For the purposes of this project, the Fission Product Extraction (FPEX) process is being considered to be the baseline extraction method. The objective of this project was to evaluate the nature and behavior of candidate materials for cesium and strontium immobilization; this will include assessments with minor additions of yttrium, barium, and rubidium in these materials. More specifically, the proposed research achieved the following objectives (as stated in the original proposal): (1) Synthesize simulated storage ceramics for Cs and Sr using an existing labscale steam reformer at Purdue University. The simulated storage materials will include aluminosilicates, zirconates and other stable ceramics with the potential for high Cs and Sr loading. (2) Characterize the immobilization performance, phase structure, thermal properties and stability of the simulated storage ceramics. The ceramic products will be stable oxide powders and will be characterized to quantify their leach resistance, phase structure, and thermophysical properties. The research progressed in two stages. First, a steam reforming process was used to generate candidate Cs/Sr storage materials for characterization. This portion of the research was carried out at Purdue University and is detailed in Appendix A. Steam reforming proved to be too rigorous for efficient The second stage of this project was carried out at Texas A&M University and is Detailed in Appendix B. In this stage, a gentler ceramic synthesis process using Cs and Sr loaded kaolinite and bentonite clays was developed in collaboration with Dr. M. Kaminski at Argonne National Laboratory.« less

Immobilization of bacterial proteases on water-solved polymer by means of electron beam

NASA Astrophysics Data System (ADS)

Gonchar, A. M.; Auslender, V. L.

1996-12-01

Possibility of electron beam usage for proteases' immobilization on 1,4-polyalkylene oxide (1,4-PAO) was studied to obtain biologically active complex for multi-purpose usage. It is shown that immobilization of Bacillus Subtilis protease takes place due to free-radical linking of enzyme and carrier with formation of mycellium-like structures. Immobilization improves heat resistance of enzyme up to 60°C without substrate and up to 80°C in presence of substrate, widens range of pH activity in comparison with non-immobilized forms. Immobilized proteases do not contain peroxides or long-live radicals. Our results permitted to create technologies for production of medical and veterinary preparations, active components for wool washing agents and leather fabrication technology.

Combination of a recombinant bacterium with organonitrile-degrading and biofilm-forming capability and a positively charged carrier for organonitriles removal.

PubMed

Li, Chunyan; Sun, Yueling; Yue, Zhenlei; Huang, Mingyan; Wang, Jinming; Chen, Xi; An, Xuejiao; Zang, Hailian; Li, Dapeng; Hou, Ning

2018-04-10

The immobilization of organonitrile-degrading bacteria via the addition of biofilm-forming bacteria represents a promising technology for the treatment of organonitrile-containing wastewater, but biofilm-forming bacteria simply mixed with degrading bacteria may reduce the biodegradation efficiency. Nitrile hydratase and amidase genes, which play critical roles in organonitriles degradation, were cloned and transformed into the biofilm-forming bacterium Bacillus subtilis N4 to construct a recombinant bacterium B. subtilis N4/pHTnha-ami. Modified polyethylene carriers with positive charge was applied to promote bacterial adherence and biofilm formation. The immobilized B. subtilis N4/pHTnha-ami was resistant to organonitriles loading shocks and could remove organic cyanide ion with a initial concentration of 392.6 mg/L for 24 h in a moving bed biofilm reactor. The imputed quorum-sensing signal and the high-throughput sequencing analysis of the biofilm indicated that B. subtilis N4/pHTnha-ami was successfully immobilized and became dominant. The successful application of the immobilized recombinant bacterium offers a novel strategy for the biodegradation of recalcitrant compounds. Copyright © 2018 Elsevier B.V. All rights reserved.

Radiation-induced polymerization for the immobilization of penicillin acylase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Boccu, E.; Carenza, M.; Lora, S.

The immobilization of Escherichia coli penicillin acylase was investigated by radiation-induced polymerization of 2-hydroxyethyl methacrylate at low temperature. A leak-proof composite that does not swell in water was obtained by adding the cross-linking agent trimethylolpropane trimethacrylate to the monomer-aqueous enzyme mixture. Penicillin acylase, which was immobilized with greater than 70% yield, possessed a higher Km value toward the substrate 6-nitro-3-phenylacetamidobenzoic acid than the free enzyme form (Km = 1.7 X 10(-5) and 1 X 10(-5) M, respectively). The structural stability of immobilized penicillin acylase, as assessed by heat, guanidinium chloride, and pH denaturation profiles, was very similar to that ofmore » the free-enzyme form, thus suggesting that penicillin acylase was entrapped in its native state into aqueous free spaces of the polymer matrix.« less

Preparation, characterization and catalytic behavior of pectinase covalently immobilized onto sodium alginate/graphene oxide composite beads.

PubMed

Dai, Xiao-Yan; Kong, Li-Min; Wang, Xiao-Ling; Zhu, Qing; Chen, Kai; Zhou, Tao

2018-07-01

Pectinase was immobilized onto sodium alginate/graphene oxide beads via amide bonds by using N,N'-dicyclohexylcarbodiimide/N-hydroxysuccinimide as the activating agent. The immobilized pectinase was characterized by Fourier transform infrared spectra and scanning electron microscopy analyses. Immobilization conditions were optimized by Box-Behnken design and the response surface method. The activity of the immobilized pectinase prepared under optimal conditions reached 1236.86 ± 40.21 U/g, with an enzyme activity recovery of 83.5%. The optimal pH of free pectinase was 4.5, while that of immobilized pectinase was shifted to 4.0. The optimal temperature of immobilized pectinase was increased to 60 °C, which was 10 °C higher than that of free form. Furthermore, the immobilized pectinase possessed a superior thermal stability and storage stability to those of free pectinase. Reusability studies indicated that the immobilized pectinase retained 73% of initial activity after six times cycles. Due to these good properties, such immobilized pectinase may find application in food industry. Copyright © 2018 Elsevier Ltd. All rights reserved.

Spectral, molecular, in vivo cytotoxicity and immobilization of β-galactosidase on poly(o-toluidine)-titanium dioxide nanocomposite

NASA Astrophysics Data System (ADS)

Shakir, M.; Khan, Mohd Shoeb; Alam, Md Fazle; Younus, H.; Alam, Mahboob; Lee, Dong-Ung

2017-06-01

The nanocomposites of poly(o-toluidine)-titanium dioxide (POT/TiO2) have been synthesized by in-situ oxidative polymerization of o-toluidine incorporating TiO2 nanoparticles. Subsequently, β-Galactosidase (BGAL) has been immobilized on POT/TiO2. The comparative spectral studies like FTIR, SEM, TEM and XRD and TGA revealed the synergistic interaction between POT and TiO2 in POT/TiO2 nanocomposite. The molecular docking simulation predicts the modes of interactions of POT and POT/TiO2 with BGAL. BGAL is successfully immobilized on POT/TiO2 with loading efficiency of 84.51%. The immobilized POT/TiO2 has enhanced its stability, recycling efficiency and residual activity making it an ideal candidate for industrial applications. Furthermore, density functional theory (6-311G (d,p) basic set was used to investigate the structures, theoretical vibrational frequencies, the HOMO-LUMO and other properties like ionization potential and electron affinity of the monomer to pentamer of o-toluidine. The in vivo cytotoxicity studies of POT and POT/TiO2 have also been examined by brine shrimp.

A new insight into the immobilization mechanism of Zn on biochar: the role of anions dissolved from ash

PubMed Central

Qian, Tingting; Wang, Yujun; Fan, Tingting; Fang, Guodong; Zhou, Dongmei

2016-01-01

Biochar is considered to be a promising material for heavy metal immobilization in soil. However, the immobilization mechanisms of Zn2+ on biochars derived from many common waste biomasses are not completely understood. Herein, biochars (denoted as PN350, PN550, WS350, and WS550) derived from pine needle (PN) and wheat straw (WS) were prepared at two pyrolysis temperatures (350 °C and 550 °C). The immobilization behaviors and mechanisms of Zn2+ on these biochars were systematically investigated. The results show that compared with biochars produced at low temperature, biochars produced at high temperature contained higher amounts of ash and exhibited much higher sorption capacities of Zn2+. By using Zn K-edge EXAFS spectroscopy, we find that the formation of various Zn precipitates/minerals, which was caused by the release of OH−, CO32−, and Si species from biochar, was the immobilization mechanism of Zn2+ on PN and WS biochars. Hydrozincite and Zn(OH)2 were the main species formed on PN350, PN550, and WS350; while on WS550, besides hydrozincite, a large fraction of hemimorphite was formed. The occurrence of hydrozincite and hemimorphite on biochar during Zn2+ immobilization is firstly reported in our study, which provides a new insight into the immobilization mechanism of Zn2+ on biochar. PMID:27641899

A new insight into the immobilization mechanism of Zn on biochar: the role of anions dissolved from ash

NASA Astrophysics Data System (ADS)

Qian, Tingting; Wang, Yujun; Fan, Tingting; Fang, Guodong; Zhou, Dongmei

2016-09-01

Biochar is considered to be a promising material for heavy metal immobilization in soil. However, the immobilization mechanisms of Zn2+ on biochars derived from many common waste biomasses are not completely understood. Herein, biochars (denoted as PN350, PN550, WS350, and WS550) derived from pine needle (PN) and wheat straw (WS) were prepared at two pyrolysis temperatures (350 °C and 550 °C). The immobilization behaviors and mechanisms of Zn2+ on these biochars were systematically investigated. The results show that compared with biochars produced at low temperature, biochars produced at high temperature contained higher amounts of ash and exhibited much higher sorption capacities of Zn2+. By using Zn K-edge EXAFS spectroscopy, we find that the formation of various Zn precipitates/minerals, which was caused by the release of OH-, CO32-, and Si species from biochar, was the immobilization mechanism of Zn2+ on PN and WS biochars. Hydrozincite and Zn(OH)2 were the main species formed on PN350, PN550, and WS350; while on WS550, besides hydrozincite, a large fraction of hemimorphite was formed. The occurrence of hydrozincite and hemimorphite on biochar during Zn2+ immobilization is firstly reported in our study, which provides a new insight into the immobilization mechanism of Zn2+ on biochar.

A new insight into the immobilization mechanism of Zn on biochar: the role of anions dissolved from ash.

PubMed

Qian, Tingting; Wang, Yujun; Fan, Tingting; Fang, Guodong; Zhou, Dongmei

2016-09-19

Biochar is considered to be a promising material for heavy metal immobilization in soil. However, the immobilization mechanisms of Zn(2+) on biochars derived from many common waste biomasses are not completely understood. Herein, biochars (denoted as PN350, PN550, WS350, and WS550) derived from pine needle (PN) and wheat straw (WS) were prepared at two pyrolysis temperatures (350 °C and 550 °C). The immobilization behaviors and mechanisms of Zn(2+) on these biochars were systematically investigated. The results show that compared with biochars produced at low temperature, biochars produced at high temperature contained higher amounts of ash and exhibited much higher sorption capacities of Zn(2+). By using Zn K-edge EXAFS spectroscopy, we find that the formation of various Zn precipitates/minerals, which was caused by the release of OH(-), CO3(2-), and Si species from biochar, was the immobilization mechanism of Zn(2+) on PN and WS biochars. Hydrozincite and Zn(OH)2 were the main species formed on PN350, PN550, and WS350; while on WS550, besides hydrozincite, a large fraction of hemimorphite was formed. The occurrence of hydrozincite and hemimorphite on biochar during Zn(2+) immobilization is firstly reported in our study, which provides a new insight into the immobilization mechanism of Zn(2+) on biochar.

Osteoblast Differentiation on Collagen Scaffold with Immobilized Alkaline Phosphatase.

PubMed

Jafary, F; Hanachi, P; Gorjipour, K

2017-01-01

In tissue engineering, scaffold characteristics play an important role in the biological interactions between cells and the scaffold. Cell adhesion, proliferation, and activation depend on material properties used for the fabrication of scaffolds. In the present investigation, we used collagen with proper characteristics including mechanically stability, biodegradability and low antigenicity. Optimization of the scaffold was done by immobilization of alkaline phosphatase on the collagen surface via cross-linking method, because this enzyme is one of the most important markers of osteoblast, which increases inorganic phosphate concentration and promote mineralization of bone formation. Alkaline phosphatase was immobilized on a collagen surface by 1-ethyl-3-(dimethylaminopropyl) carbodiimide hydrochloride, as a reagent. Then, rat mesenchymal stem cells were cultured in osteogenic medium in control and treated groups. The osteogenesis-related genes were compared between treatments (differentiated cells with immobilized alkaline phosphatase/collagen scaffold) and control groups (differentiated cells on collagen surface without alkaline phosphatase) on days 3 and 7 by quantitative real-time PCR (QIAGEN software). Several genes, including alkaline phosphatase, collagen type I and osteocalcine associated with calcium binding and mineralization, showed upregulation in expression during the first 3 days, whereas tumor necrosis factor-α, acting as an inhibitor of differentiation, was down-regulated during osteogenesis. Collagen scaffold with immobilized alkaline phosphatase can be utilized as a good candidate for enhancing the differentiation of osteoblasts from mesenchymal stem cells.

Immobilization of l-Asparaginase on Carrier Materials: A Comprehensive Review.

PubMed

Ulu, Ahmet; Ates, Burhan

2017-06-21

There are two major applications of l-asparaginase (L-ASNase): leukemia therapy and the food industry. Especially, its chemotherapeutic effect has attracted interest from the scientific community and individual scientists. Therefore, to protect the intrinsic activity and half-time of L-ASNase, several carriers and immobilization techniques for immobilization of L-ASNase have been described in articles. Unfortunately, a comprehensive review about immobilization of L-ASNase has not been written until now. In this review, we have thoroughly discussed the carriers for L-ASNase by illustrating immobilization findings including both past and present applications. In addition, we have revealed advantages and disadvantages of immobilized enzyme and related it to free form. We believe that this review will not only provide background information, but also guide future developments.

Evaluation of free and immobilized Aspergillus niger NRC1ami pectinase applicable in industrial processes.

PubMed

Esawy, Mona A; Gamal, Amira A; Kamel, Zeinat; Ismail, Abdel-Mohsen S; Abdel-Fattah, Ahmed F

2013-02-15

The Aspergillus niger NRC1ami pectinase was evaluated according to its hydrolysis efficiency of dry untreated orange peels (UOP), HCl-treated orange peels and NaOH-treated orange peels (HOP and NOP). Pectinase was entrapped in polyvinyl alcohol (PVA) sponge and the optimum pH and temperature of the free and immobilized enzymes were shifted from 4, 40 °C to 6, 50 °C respectively. The study of pH stability of free and immobilized pectinase showed that the immobilization process protected the enzyme strongly from severe alkaline pHs. The immobilization process improved the enzyme thermal stability to great instant. The unique feature of the immobilization process is its ability to solve the orange juice haze problem completely. Immobilized enzyme was reused 12 times in orange juice clarification with 9% activity loss from the original activity. Maximum reaction rate (V(max)) and Michaelis-Menten constant (K(m)) of the partially purified form were significantly changed after immobilization. Copyright © 2012 Elsevier Ltd. All rights reserved.

Nitrogen-Doped Carbon Dots as A New Substrate for Sensitive Glucose Determination.

PubMed

Ji, Hanxu; Zhou, Feng; Gu, Jiangjiang; Shu, Chen; Xi, Kai; Jia, Xudong

2016-05-04

Nitrogen-doped carbon dots are introduced as a novel substrate suitable for enzyme immobilization in electrochemical detection metods. Nitrogen-doped carbon dots are easily synthesised from polyacrylamide in just one step. With the help of the amino group on chitosan, glucose oxidase is immobilized on nitrogen-doped carbon dots-modified carbon glassy electrodes by amino-carboxyl reactions. The nitrogen-induced charge delocalization at nitrogen-doped carbon dots can enhance the electrocatalytic activity toward the reduction of O₂. The specific amino-carboxyl reaction provides strong and stable immobilization of GOx on electrodes. The developed biosensor responds efficiently to the presence of glucose in serum samples over the concentration range from 1 to 12 mM with a detection limit of 0.25 mM. This novel biosensor has good reproducibility and stability, and is highly selective for glucose determination under physiological conditions. These results indicate that N-doped quantum dots represent a novel candidate material for the construction of electrochemical biosensors.

Sugarcane bagasse as support for immobilization of Bacillus pumilus HZ-2 and its use in bioremediation of mesotrione-contaminated soils.

PubMed

Liu, Jie; Chen, Shaohua; Ding, Jie; Xiao, Ying; Han, Haitao; Zhong, Guohua

2015-12-01

The degrading microorganisms isolated from environment usually fail to degrade pollutants when used for bioremediation of contaminated soils; thus, additional treatments are needed to enhance biodegradation. In the present study, the potential of sugarcane bagasse as bacteria-immobilizing support was investigated in mesotrione biodegradation. A novel isolate Bacillus pumilus HZ-2 was applied in bacterial immobilization, which was capable of degrading over 95 % of mesotrione at initial concentrations ranging from 25 to 200 mg L(-1) within 4 days in flask-shaking tests. Scanning electron microscope (SEM) images showed that the bacterial cells were strongly absorbed and fully dispersed on bagasse surface after immobilization. Specially, 86.5 and 82.9 % of mesotrione was eliminated by bacteria immobilized on bagasse of 100 and 60 mesh, respectively, which indicated that this immobilization was able to maintain a high degrading activity of the bacteria. Analysis of the degradation products determined 2-amino-4-methylsulfonylbenzoic acid (AMBA) and 4-methylsulfonyl-2-nitrobenzoic acid (MNBA) as the main metabolites in the biodegradation pathway of mesotrione. In the sterile soil, approximately 90 % of mesotrione was degraded after supplementing 5.0 % of molasses in bacteria-bagasse composite, which greatly enhanced microbial adaptability and growth in the soil environment. In the field tests, over 75 % of mesotrione in soil was degraded within 14 days. The immobilized preparation demonstrated that mesotrione could be degraded at a wide range of pH values (5.0-8.0) and temperatures (25-35 °C), especially at low concentrations of mesotrione (5 to 20 mg kg(-1)). These results showed that sugarcane bagasse might be a good candidate as bacteria-immobilizing support to enhance mesotrione degradation by Bacillus p. HZ-2 in contaminated soils.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Asmussen, R. Matthew; Pearce, Carolyn I.; Miller, Brian W.

A cementitious waste form, Cast Stone, is a possible candidate technology for the immobilization of low activity nuclear waste (LAW) at the Hanford site. This paper focuses on the addition of getter materials to Cast Stone that can sequester Tc from the LAW, and in turn, lower Tc release from the Cast Stone. Two getters which produce different products upon sequestering Tc from LAW were tested: Sn(II) apatite (Sn-A) that removes Tc as a Tc(IV)-oxide and potassium metal sulfide (KMS-2) that removes Tc as a Tc(IV)-sulfide species, allowing for a comparison of stability of the form of Tc upon enteringmore » the waste form. The Cast Stone with KMS-2 getter had the best performance with addition equivalent to ~0.08 wt% of the total waste form mass. The observed diffusion (D obs) of Tc decreased from 4.6 ± 0.2 × 10 -12 cm 2/s for Cast Stone that did not contain a getter to 5.4 ± 0.4 × 10 -13 cm 2/s for KMS-2 containing Cast Stone. Finally, it was found that Tc-sulfide species are more stable against re-oxidation within getter containing Cast Stone compared with Tc-oxide and is the origin of the decrease in Tc D obs when using the KMS-2.« less

Immobilized sialyltransferase fused to a fungal biotin-binding protein: Production, properties, and applications.

PubMed

Kajiwara, Hitomi; Tsunashima, Masako; Mine, Toshiki; Takakura, Yoshimitsu; Yamamoto, Takeshi

2016-04-01

A β-galactoside α2,6-sialyltransferase (ST) from the marine bacterium Photobacterium sp. JT-ISH-224 with a broad acceptor substrate specificity was fused to a fungal biotin-binding protein tamavidin 2 (TM2) to produce immobilized enzyme. Specifically, a gene for the fusion protein, in which ST from Photobacterium sp. JT-ISH-224 and TM2 were connected via a peptide linker (ST-L-TM2) was constructed and expressed in Escherichia coli. The ST-L-TM2 was produced in the soluble form with a yield of approximately 15,000 unit/300 ml of the E. coli culture. The ST-L-TM2 was partially purified and part of it was immobilized onto biotin-bearing magnetic microbeads. The immobilized ST-L-TM2 onto microbeads could be used at least seven consecutive reaction cycles with no observed decrease in enzymatic activity. In addition, the optimum pH and temperature of the immobilized enzyme were changed compared to those of a free form of the ST. Considering these results, it was strongly expected that the immobilized ST-L-TM2 was a promising tool for the production of various kind of sialoligosaccharides. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Protein immobilization onto various surfaces using a polymer-bound isocyanate

NASA Astrophysics Data System (ADS)

Kang, Hyun-Jin; Cha, Eun Ji; Park, Hee-Deung

2015-01-01

Silane coupling agents have been widely used for immobilizing proteins onto inorganic surfaces. However, the immobilization method using silane coupling agents requires several treatment steps, and its application is limited to only surfaces containing hydroxyl groups. The aim of this study was to develop a novel method to overcome the limitations of the silane-based immobilization method using a polymer-bound isocyanate. Initially, polymer-bound isocyanate was dissolved in organic solvent and then was used to dip-coat inorganic surfaces. Proteins were then immobilized onto the dip-coated surfaces by the formation of urea bonds between the isocyanate groups of the polymer and the amine groups of the protein. The reaction was verified by FT-IR in which NCO stretching peaks disappeared, and CO and NH stretching peaks appeared after immobilization. The immobilization efficiency of the newly developed method was insensitive to reaction temperatures (4-50 °C), but the efficiency increased with reaction time and reached a maximum after 4 h. Furthermore, the method showed comparable immobilization efficiency to the silane-based immobilization method and was applicable to surfaces that cannot form hydroxyl groups. Taken together, the newly developed method provides a simple and efficient platform for immobilizing proteins onto surfaces.

Method of immobilizing weapons plutonium to provide a durable, disposable waste product

DOEpatents

Ewing, Rodney C.; Lutze, Werner; Weber, William J.

1996-01-01

A method of atomic scale fixation and immobilization of plutonium to provide a durable waste product. Plutonium is provided in the form of either PuO.sub.2 or Pu(NO.sub.3).sub.4 and is mixed with and SiO.sub.2. The resulting mixture is cold pressed and then heated under pressure to form (Zr,Pu)SiO.sub.4 as the waste product.

Estimation of affinities of ligands in mixtures via magnetic recovery of target-ligand complexes and chromatographic analyses: chemometrics and an experimental model

PubMed Central

2011-01-01

Abstract Background The combinatorial library strategy of using multiple candidate ligands in mixtures as library members is ideal in terms of cost and efficiency, but needs special screening methods to estimate the affinities of candidate ligands in such mixtures. Herein, a new method to screen candidate ligands present in unknown molar quantities in mixtures was investigated. Results The proposed method involves preparing a processed-mixture-for-screening (PMFS) with each mixture sample and an exogenous reference ligand, initiating competitive binding among ligands from the PMFS to a target immobilized on magnetic particles, recovering target-ligand complexes in equilibrium by magnetic force, extracting and concentrating bound ligands, and analyzing ligands in the PMFS and the concentrated extract by chromatography. The relative affinity of each candidate ligand to its reference ligand is estimated via an approximation equation assuming (a) the candidate ligand and its reference ligand bind to the same site(s) on the target, (b) their chromatographic peak areas are over five times their intercepts of linear response but within their linear ranges, (c) their binding ratios are below 10%. These prerequisites are met by optimizing primarily the quantity of the target used and the PMFS composition ratio. The new method was tested using the competitive binding of biotin derivatives from mixtures to streptavidin immobilized on magnetic particles as a model. Each mixture sample containing a limited number of candidate biotin derivatives with moderate differences in their molar quantities were prepared via parallel-combinatorial-synthesis (PCS) without purification, or via the pooling of individual compounds. Some purified biotin derivatives were used as reference ligands. This method showed resistance to variations in chromatographic quantification sensitivity and concentration ratios; optimized conditions to validate the approximation equation could be applied to different mixture samples. Relative affinities of candidate biotin derivatives with unknown molar quantities in each mixture sample were consistent with those estimated by a homogenous method using their purified counterparts as samples. Conclusions This new method is robust and effective for each mixture possessing a limited number of candidate ligands whose molar quantities have moderate differences, and its integration with PCS has promise to routinely practice the mixture-based library strategy. PMID:21545719

Process for immobilizing radioactive boric acid liquid wastes

DOEpatents

Greenhalgh, Wilbur O.

1986-01-01

A method of immobilizing boric acid liquid wastes containing radionuclides by neutralizing the solution and evaporating the resulting precipitate to near dryness. The dry residue is then fused into a reduced volume, insoluble, inert, solid form containing substantially all the radionuclides.

Efficient magnetic recycling of covalently attached enzymes on carbon-coated metallic nanomagnets.

PubMed

Zlateski, Vladimir; Fuhrer, Roland; Koehler, Fabian M; Wharry, Scott; Zeltner, Martin; Stark, Wendelin J; Moody, Thomas S; Grass, Robert N

2014-04-16

In the pursuit of robust and reusable biocatalysts for industrial synthetic chemistry, nanobiotechnology is currently taking a significant part. Recently, enzymes have been immobilized on different nanoscaffold supports. Carbon coated metallic nanoparticles were found to be a practically useful support for enzyme immobilization due to their large surface area, high magnetic saturation, and manipulatable surface chemistry. In this study carbon coated cobalt nanoparticles were chemically functionalized (diazonium chemistry), activated for bioconjugation (N,N-disuccinimidyl carbonate), and subsequently used in enzyme immobilization. Three enzymes, β-glucosidase, α-chymotrypsin, and lipase B were successfully covalently immobilized on the magnetic nonsupport. The enzyme-particle conjugates formed retained their activity and stability after immobilization and were efficiently recycled from milliliter to liter scales in short recycle times.

Method of identity analyte-binding peptides

DOEpatents

Kauvar, Lawrence M.

1990-01-01

A method for affinity chromatography or adsorption of a designated analyte utilizes a paralog as the affinity partner. The immobilized paralog can be used in purification or analysis of the analyte; the paralog can also be used as a substitute for antibody in an immunoassay. The paralog is identified by screening candidate peptide sequences of 4-20 amino acids for specific affinity to the analyte.

Getters for improved technetium containment in cementitious waste forms

DOE PAGES

Asmussen, R. Matthew; Pearce, Carolyn I.; Miller, Brian W.; ...

2017-07-26

A cementitious waste form, Cast Stone, is a possible candidate technology for the immobilization of low activity nuclear waste (LAW) at the Hanford site. This paper focuses on the addition of getter materials to Cast Stone that can sequester Tc from the LAW, and in turn, lower Tc release from the Cast Stone. Two getters which produce different products upon sequestering Tc from LAW were tested: Sn(II) apatite (Sn-A) that removes Tc as a Tc(IV)-oxide and potassium metal sulfide (KMS-2) that removes Tc as a Tc(IV)-sulfide species, allowing for a comparison of stability of the form of Tc upon enteringmore » the waste form. The Cast Stone with KMS-2 getter had the best performance with addition equivalent to ~0.08 wt% of the total waste form mass. The observed diffusion (D obs) of Tc decreased from 4.6 ± 0.2 × 10 -12 cm 2/s for Cast Stone that did not contain a getter to 5.4 ± 0.4 × 10 -13 cm 2/s for KMS-2 containing Cast Stone. Finally, it was found that Tc-sulfide species are more stable against re-oxidation within getter containing Cast Stone compared with Tc-oxide and is the origin of the decrease in Tc D obs when using the KMS-2.« less

Getters for improved technetium containment in cementitious waste forms.

PubMed

Asmussen, R Matthew; Pearce, Carolyn I; Miller, Brian W; Lawter, Amanda R; Neeway, James J; Lukens, Wayne W; Bowden, Mark E; Miller, Micah A; Buck, Edgar C; Serne, R Jeffery; Qafoku, Nikolla P

2018-01-05

A cementitious waste form, Cast Stone, is a possible candidate technology for the immobilization of low activity nuclear waste (LAW) at the Hanford site. This work focuses on the addition of getter materials to Cast Stone that can sequester Tc from the LAW, and in turn, lower Tc release from the Cast Stone. Two getters which produce different products upon sequestering Tc from LAW were tested: Sn(II) apatite (Sn-A) that removes Tc as a Tc(IV)-oxide and potassium metal sulfide (KMS-2) that removes Tc as a Tc(IV)-sulfide species, allowing for a comparison of stability of the form of Tc upon entering the waste form. The Cast Stone with KMS-2 getter had the best performance with addition equivalent to ∼0.08wt% of the total waste form mass. The observed diffusion (D obs ) of Tc decreased from 4.6±0.2×10 -12 cm 2 /s for Cast Stone that did not contain a getter to 5.4±0.4×10 -13 cm 2 /s for KMS-2 containing Cast Stone. It was found that Tc-sulfide species are more stable against re-oxidation within getter containing Cast Stone compared with Tc-oxide and is the origin of the decrease in Tc D obs when using the KMS-2. Copyright © 2017 Elsevier B.V. All rights reserved.

Process for immobilizing radioactive boric acid liquid wastes

DOEpatents

Greenhalgh, W.O.

1984-05-10

Disclosed is a method of immobilizing boric acid liquid wastes containing radionuclides by neutralizing the solution and evaporating the resulting precipitate to near dryness. The dry residue is then fused into a reduced volume, insoluble, inert, solid form containing substantially all the radionuclides.

Metal Immobilization Influence On Bioavailability And Remediation For Urban Environments

EPA Science Inventory

Immobilization of soil contaminants, such as lead, via phosphate amendments to alter the chemical environment of metals into highly insoluble forms is a well established process. The literature has documented numerous examples of highly contaminated Pb sites at shooting ranges, b...

Immobilization of microbial cell and yeast cell and its application to biomass conversion using radiation techniques

NASA Astrophysics Data System (ADS)

Kaetsu, Isao; Kumakura, Minoru; Fujimura, Takashi; Kasai, Noboru; Tamada, Masao

The recent results of immobilization of cellulase-producing cells and ethanol-fermentation yeast by radiation were reported. The enzyme of cellulase produced by immobilized cells was used for saccharification of lignocellulosic wastes and immobilized yeast cells were used for fermentation reaction from glucose to ethanol. The wastes such as chaff and bagasse were treated by γ-ray or electron-beam irradiation in the presence of alkali and subsequent mechanical crushing, to form a fine powder less than 50 μm in diameter. On the other hand, Trichoderma reesei as a cellulase-producing microbial cell was immobilized on a fibrous carrier having a specific porous structure and cultured to produce cellulase. The enzymatic saccharification of the pretreated waste was carried out using the produced cellulase. The enhanced fermentation process to produce ethanol from glucose with the immobilized yeast by radiation was also studied. The ethanol productivity of immobilized growing yeast cells thus obtained was thirteen times that of free yeast cells in a 1:1 volume of liquid medium to immobilized yeast cells.

Petroleum oil removal by immobilized bacterial cells on polyurethane foam under different temperature conditions.

PubMed

Alessandrello, Mauricio J; Juárez Tomás, María S; Raimondo, Enzo E; Vullo, Diana L; Ferrero, Marcela A

2017-09-15

In this work, a mixed biofilm composed by Pseudomonas monteilii P26 and Gordonia sp. H19 was formed using polyurethane foam (PUF) as immobilization support, for crude oil removal from artificial sea water. Fresh immobilized cells and immobilized cells that were stored at 4°C for two months before use were assessed. The oil removal assays were carried out at microcosm scale at 4, 15 and 30°C. A viability loss of P. monteilii P26 was observed after the storage. The highest removal value (75%) was obtained at 30°C after 7days using fresh immobilized cells on PUF. Enhanced oil bioremoval was obtained at 4°C and 15°C with the previously stored immobilized cells compared to the fresh immobilized cells. Crude oil sorption on the different systems was responsible for the removal of 22-33% oil at the different temperatures. In conclusion, an economic tool for petroleum bioremediation is proposed. Copyright © 2017 Elsevier Ltd. All rights reserved.

Immobilization of R-ω-transaminase on MnO2 nanorods for catalyzing the conversion of (R)-1-phenylethylamine.

PubMed

Sun, Jian; Cui, Wen-Hui; Du, Kun; Gao, Qian; Du, Mengmeng; Ji, Peijun; Feng, Wei

2017-03-10

R-ɷ-transaminases transfer an amino group from an amino donor (e.g. (R)-1-phenylethylamine) onto an amino acceptor (e.g. pyruvate), resulting a co-product (e.g. d-alanine). This work intends to immobilize R-ɷ-Transaminase on MnO 2 nanorods to achieve multienzyme catalysis. R-ɷ-Transaminase (RTA) and d-amino acid oxidase (DAAO) have been fused to an elastin-like polypeptide (ELP) separately through genetic engineering of the enzymes. ELP-RTA and ELP-DAAO have been separately immobilized on polydopamine-coated MnO 2 nanorods. When the two immobilized enzymes were used together in one pot, the transformation of (R)-1-phenylethylamine was catalyzed by the immobilized ELP-RTA, and the co-product d-alanine was converted back to pyruvate under the catalysis of the immobilized ELP-DAAO, achieving the recycling of pyruvate in situ. Thus pyruvate was maintained at a low concentration in order to reduce its negative effect. On the other hand, the generated H 2 O 2 of ELP-DAAO was decomposed by the MnO 2 nanorods, and the evolved oxygen oxidized the reduced cofactors of ELP-DAAO. Forming the circles of hydrogen peroxide→oxygen→hydrogen peroxide accelerated the deamination reaction. The highly efficient conversion of the co-product d-alanine back to pyruvate accelerated the forming of the pyruvate→d-alanine→pyruvate cycle between the two immobilized enzymes. The coordination of the pyruvate→d-alanine→pyruvate and hydrogen peroxide→oxygen→hydrogen peroxide cycles accelerated the transformation of (R)-1-phenylethylamine. As a result, As a result, the immobilized enzymes achieved a conversion of 98±1.8% in comparison to 69.6±1.2% by free enzymes. Copyright © 2017 Elsevier B.V. All rights reserved.

Improvement of Leaching Resistance of Low-level Waste Form in Korea

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, J.Y.; Lee, B.C.; Kim, C.L.

2006-07-01

Low-level liquid concentrate wastes including boric acid have been immobilized with paraffin wax using concentrate waste drying system in Korean nuclear power plants since 1995. Small amount of low density polyethylene (LDPE) was added to increase the leaching resistance of the existing paraffin waste form and the influence of LDPE on the leaching behavior of waste form was investigated. It was observed that the leaching of nuclides immobilized within paraffin waste form remarkably reduced as the content of LDPE increased. The acceptance criteria of paraffin waste form associated with leachability index and compressive strength after the leaching test were successfullymore » satisfied with the help of LDPE. (authors)« less

Stabilization of α-amylase by using anionic surfactant during the immobilization process

NASA Astrophysics Data System (ADS)

El-Batal, A. I.; Atia, K. S.; Eid, M.

2005-10-01

This work describes the entrapment of α-amylase into butylacrylate-acrylic acid copolymer (BuA/AAc) using γ irradiation. The effect of an anionic surfactant (AOT), the reuse efficiency, and kinetic behavior of immobilized α-amylase were studied. Covering of α-amylase with bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT) made the enzyme more stable than the uncovered form. The hydrolytic activity of the pre-coated immobilized α-amylase was increased below the critical micelle concentration (cmc) (10 mmol/L). The results showed an increase in the relative activity with increase in the degree of hydration. The pre-coated immobilized α-amylase showed a higher k/K and lower activation energy compared to the free and uncoated-immobilized preparation, respectively. The results suggest that the immobilization of α-amylase is a potentially useful approach for commercial starch hydrolysis in two-phase systems.

Glucose uptake and glycogen synthesis in muscles from immobilized limbs

NASA Technical Reports Server (NTRS)

Nicholson, W. F.; Watson, P. A.; Booth, F. W.

1984-01-01

Defects in glucose metabolism in muscles of immobilized limbs of mice were related to alterations in insulin binding, insulin responsiveness, glucose supply, and insulin activation of glycogen synthase. These were tested by in vitro methodology. A significant lessening in the insulin-induced maximal response of 2-deoxyglucose uptake into the mouse soleus muscle occurred between the 3rd and 8th h of limb immobilization, suggesting a decreased insulin responsiveness. Lack of change in the specific binding of insulin to muscles of 24-h immobilized limbs indicates that a change in insulin receptor number did not play a role in the failure of insulin to stimulate glucose metabolism. Its inability to stimulate glycogen synthesis in muscle from immobilized limbs is due, in part, to a lack of glucose supply to glycogen synthesis and also to the ineffectiveness of insulin to increase the percentage of glycogen synthase in its active form in muscles from 24-h immobilized limbs.

Selective detection and recovery of gold at tannin-immobilized non-conducting electrode.

PubMed

Banu, Khaleda; Shimura, Takayoshi; Sadeghi, Saman

2015-01-01

A tannin-immobilized glassy carbon electrode (TIGC) was prepared via electrochemical oxidation of the naturally occurring polyphenolic mimosa tannin, which generated a non-conducting polymeric film (NCPF) on the electrode surface. The fouling of the electrode surface by the electropolymerized film was evaluated by monitoring the electrode response of ferricyanide ions as a redox marker. The NCPF was permselective to HAuCl4, and the electrochemical reduction of HAuCl4 to metallic gold at the TIGC electrode was evaluated by recording the reduction current during cyclic voltammetry measurement. In the mixed electrolyte containing HAuCl4 along with FeCl3 and/or CuCl2, the NCPF remained selective toward the electrochemical reduction of HAuCl4 into the metallic state. The chemical reduction of HAuCl4 into metallic gold was also observed when the NCPF was inserted into an acidic gold solution overnight. The adsorption capacity of Au(III) on tannin-immobilized carbon fiber was 29±1.45 mg g(-1) at 60°C. In the presence of excess Cu(II) and Fe(III), tannin-immobilized NCPF proved to be an excellent candidate for the selective detection and recovery of gold through both electrochemical and chemical processes. Copyright © 2014 Elsevier B.V. All rights reserved.

Performance of the Fluidized Bed Steam Reforming Product Under Hydraulically Unsaturated Conditions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Neeway, James J.; Qafoku, Nikolla; Williams, Benjamin D.

2014-05-01

Currently, several candidates for secondary waste immobilization at the Hanford site in the State of Washington, USA are being considered. To demonstrate the durability of the product in the unsaturated Integrated Disposal Facility (IDF) at the site, a series of tests have been performed one of the candidate materials using the Pressurized Unsaturated Flow (PUF) system. The material that was tested was the Fluidized Bed Steam Reformer (FBSR) granular product and the granular product encapsulated in a geopolymer matrix. The FBSR product is composed primarily of an insoluble sodium aluminosilicate matrix with the dominant phases being feldspathoid minerals mostly nepheline,more » sodalite, and nosean. The PUF test method allows for the accelerated weathering of materials, including radioactive waste forms, under hydraulically unsaturated conditions, thus mimicking the open-flow and transport properties that most likely will be present at the IDF. The experiments show a trend of decreasing tracer release as a function of time for several of the elements released from the material including Na, Si, Al, and Cs. However, some of the elements, notably I and Re, show a steady release throughout the yearlong test. This result suggests that the release of these minerals from the sodalite cage occurs at a different rate compared with the dissolution of the predominant nepheline phase.« less

Release characteristics and bioactivity of gelatin-tricalcium phosphate membranes covalently immobilized with nerve growth factors.

PubMed

Chen, Pei-Ru; Chen, Ming-Hong; Lin, Feng-Huei; Su, Wen-Yu

2005-11-01

The gelatin-tricalcium phosphate membranes were cross-linking with low concentration glutaraldehyde solution (GTG). This material has good mechanical property, biocompatibility, and is feasible for surgical manipulation. For axonal regeneration, nerve growth factors (NGF) were immobilized onto the composite (GTG) with carbodiimide. The purpose of this study was to evaluate the release characteristics and bioactivity of NGF after covalent immobilization onto the GTG membranes (GEN). NGF immobilized onto and released from the composite was quantified using ELISA method. PC 12 cells were cultured on the GTG and GEN composites. Cell survival, cytotoxicity, and cellular activity were evaluated by total protein content, LDH activity, and MTT assay respectively. Neurite outgrowth assay was used to evaluate the biological activity of NGF released from GEN composite. From ELISA measurement, the releasing curve for NGF showing two distinctive parts with different slopes indicated that NGF were released from the composite in diffusion-controlled mechanism and degradation-controlled mechanism respectively. While culturing with PC 12 cells, LDH leakage results implied that whether GTG composite cross-linked with NGF or not showed little cytotoxicity. The total protein content and cellular activity of PC 12 cells were lower on GTG and GEN membranes than control group. However, 56%+/-3.98 of PC 12 cells showed significant neurite outgrowth on GEN membranes which was statistically higher than GTG without NGF immobilization. In addition, sustained release of bioactive NGF for two months had been demonstrated by neurite outgrowth assay. From these experiments, it can be concluded that the technique used in the present study is capable of immobilizing NGF onto GTG membranes covalently and remaining the bioactivity of NGF. Therefore, GEN composite can be materials for sustained release of bioactive NGF and a candidate for future therapeutic application in nerve repair.

Materials and processes for the effective capture and immobilization of radioiodine: A review

DOE PAGES

Riley, Brian J.; Vienna, John D.; Strachan, Denis M.; ...

2015-12-02

In this study, the immobilization of radioiodine produced from reprocessing used nuclear fuel is a growing priority for research and development of nuclear waste forms. This review provides a comprehensive summary of the current issues surrounding processing and containment of 129I, the isotope of greatest concern due to its long half-life of 1.6 × 10 7 y and potential incorporation into the human body. Strategies for disposal of radioiodine, captured by both wet scrubbing and solid sorbents, are discussed, as well as potential iodine waste streams for insertion into an immobilization process. Next, consideration of direct disposal of salts, incorporationmore » into glasses, ceramics, cements, and other phases is discussed. The bulk of the review is devoted to an assessment of various sorbents for iodine and of waste forms described in the literature, particularly inorganic minerals, ceramics, and glasses. This review also contains recommendations for future research needed to address radioiodine immobilization materials and processes.« less

Comparison of Zirconium Phosphonate-Modified Surfaces for Immobilizing Phosphopeptides and Phosphate-Tagged Proteins.

PubMed

Forato, Florian; Liu, Hao; Benoit, Roland; Fayon, Franck; Charlier, Cathy; Fateh, Amina; Defontaine, Alain; Tellier, Charles; Talham, Daniel R; Queffélec, Clémence; Bujoli, Bruno

2016-06-07

Different routes for preparing zirconium phosphonate-modified surfaces for immobilizing biomolecular probes are compared. Two chemical-modification approaches were explored to form self-assembled monolayers on commercially available primary amine-functionalized slides, and the resulting surfaces were compared to well-characterized zirconium phosphonate monolayer-modified supports prepared using Langmuir-Blodgett methods. When using POCl3 as the amine phosphorylating agent followed by treatment with zirconyl chloride, the result was not a zirconium-phosphonate monolayer, as commonly assumed in the literature, but rather the process gives adsorbed zirconium oxide/hydroxide species and to a lower extent adsorbed zirconium phosphate and/or phosphonate. Reactions giving rise to these products were modeled in homogeneous-phase studies. Nevertheless, each of the three modified surfaces effectively immobilized phosphopeptides and phosphopeptide tags fused to an affinity protein. Unexpectedly, the zirconium oxide/hydroxide modified surface, formed by treating the amine-coated slides with POCl3/Zr(4+), afforded better immobilization of the peptides and proteins and efficient capture of their targets.

Production of α-keto acids Part I. Immobilized cells ofTrigonopsis variabilis containing D-amino acid oxidase.

PubMed

Brodelius, P; Nilsson, K; Mosbach, K

1981-12-01

Whole cells ofTrigonopsis variabilis were immobilized by entrapment in Ca(2+)-alginate and used for the production of α-keto acids from the corresponding D-amino acids. The D-amino acid oxidase within the immobilized cells has a broad substrate specificity. Hydrogen peroxide formed in the enzymatic reaction was efficiently hydrolyzed by manganese oxide co-immobilized with the cells. The amino acid oxidase activity was assayed with a new method based on reversed-phase HPLC. Oxygen requirements, bead size, concentration of cells in the beads, flow rate, and other factors were investigated in a " trickle-bed " reactor.

Noncovalent immobilization of electrocatalysts on carbon electrodes for fuel production.

PubMed

Blakemore, James D; Gupta, Ayush; Warren, Jeffrey J; Brunschwig, Bruce S; Gray, Harry B

2013-12-11

We show that molecular catalysts for fuel-forming reactions can be immobilized on graphitic carbon electrode surfaces via noncovalent interactions. A pyrene-appended bipyridine ligand (P) serves as the linker between each complex and the surface. Immobilization of a rhodium proton-reduction catalyst, [Cp*Rh(P)Cl]Cl (1), and a rhenium CO2-reduction catalyst, Re(P)(CO)3Cl (2), afford electrocatalytically active assemblies. X-ray photoelectron spectroscopy and electrochemistry confirm catalyst immobilization. Reduction of 1 in the presence of p-toluenesulfonic acid results in catalytic H2 production, while reduction of 2 in the presence of CO2 results in catalytic CO production.

Characterization of Proteus vulgaris K80 lipase immobilized on amine-terminated magnetic microparticles.

PubMed

Natalia, Agnes; Kristiani, Lidya; Kim, Hyung Kwoun

2014-10-01

Proteus vulgaris K80 lipase was expressed in Escherichia coli BL21 (DE3) cells and immobilized on amine-terminated magnetic microparticles (Mag-MPs). The immobilization yield and activity retention were 84.15% and 7.87%, respectively. A homology model of lipase K80 was constructed using P. mirabilis lipase as the template. Many lysine residues were located on the protein surface, remote from active sites. The biochemical characteristics of immobilized lipase K80 were compared with the soluble free form of lipase K80. The optimum temperature of K80-Mag-MPs was 60°C, which was 20°C higher than that of the soluble form. K80-Mag-MPs also tended to be more stable than the soluble form at elevated temperatures and a broad range of pH. K80-Mag-MP maintained its stable form at up to 40°C and in a pH range of 5.0- 10.0, whereas soluble K80 maintained its activity up to 35°C and pH 6.0-10.0. K80-Mag-MPs had broader substrate specificity compared with that of soluble K80. K80-Mag-MPs showed about 80% residual relative activity after five recovery trials. These results indicate the potential benefit of K80-Mag-MPs as a biocatalyst in various industries.

Data Packages for the Hanford Immobilized Low Activity Tank Waste Performance Assessment 2001 Version [SEC 1 THRU 5

DOE Office of Scientific and Technical Information (OSTI.GOV)

MANN, F.M.

Data package supporting the 2001 Immobilized Low-Activity Waste Performance Analysis. Geology, hydrology, geochemistry, facility, waste form, and dosimetry data based on recent investigation are provided. Verification and benchmarking packages for selected software codes are provided.

Immobilization of radioactive and hazardous wastes in a developed sulfur polymer cement (SPC) matrix

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wagdy, M.; Azim, Abdel; El-Gammal, Belal

Available in abstract form only. Full text of publication follows: A process has been developed for the immobilization Cs, Sr, Ce, Pb, and Cr in forms that is non-dispersible and could be safely immobilized. The simulated radioactive wastes of Cs, Sr, and Ce, and the hazardous wastes of Cr, and Pb were immobilized in the stable form of sulfur polymer cement (SPC). In this process, the contaminants (in a single form) were added to the sulfur mixture of sulfur and aromatic /or aliphatic hydrocarbons that used as polymerizing agents for sulfur (95% S, and 5% organic polymer by weight). Durabilitymore » of the fabricated SPC matrices was assessed in terms of their water of immersion, porosity, and compressive strength. The water immersion, and open porosity were found to be less than 2.5% for all the prepared matrices, whereas the compressive strength was in the range between 62.4 and 142.3 Kg.cm{sup -2}, depending on the composition of the prepared matrix. The prepared SPC matrices that characterized by X-ray diffraction (XRD) showed that the different added contaminants were stabilized during the solidification process during their reaction with sulfur and the organic polymer to form the corresponding metal sulfides. Toxicity Characteristic Leaching Procedure (TCLP), and the IAEA standard method have assessed the leachability of the prepared waste matrices. The TCLP results showed that most the concentration of the contaminants released were under their detection limit. The leach index for the investigated metals from the prepared SPC matrices was in the range of 9-11. The order of release of the investigated metals was Sr>Cs>Pb>Cr>Ce for the aliphatic polymer, and Sr>Cr>Pb>Cs>Ce for the aromatic one. The results obtained revealed a high performance for the prepared SPC matrices, as they are of low cost effect, highly available materials, and possessed good mechanical and leaching properties. Key Words: SPC/ Matrices/ Immobilization/ Wastes/ Leachability. (authors)« less

Method of identity analyte-binding peptides

DOEpatents

Kauvar, L.M.

1990-10-16

A method for affinity chromatography or adsorption of a designated analyte utilizes a paralog as the affinity partner. The immobilized paralog can be used in purification or analysis of the analyte; the paralog can also be used as a substitute for antibody in an immunoassay. The paralog is identified by screening candidate peptide sequences of 4--20 amino acids for specific affinity to the analyte. 5 figs.

Graphene oxide-mediated electrochemistry of glucose oxidase on glassy carbon electrodes.

PubMed

Castrignanò, Silvia; Valetti, Francesca; Gilardi, Gianfranco; Sadeghi, Sheila J

2016-01-01

Glucose oxidase (GOD) was immobilized on glassy carbon electrodes in the presence of graphene oxide (GO) as a model system for the interaction between GO and biological molecules. Lyotropic properties of didodecyldimethylammonium bromide (DDAB) were used to stabilize the enzymatic layer on the electrode surface resulting in a markedly improved electrochemical response of the immobilized GOD. Transmission electron microscopy images of the GO with DDAB confirmed the distribution of the GO in a two-dimensional manner as a foil-like material. Although it is known that glassy carbon surfaces are not ideal for hydrogen peroxide detection, successful chronoamperometric titrations of the GOD in the presence of GO with β-d-glucose were performed on glassy carbon electrodes, whereas no current response was detected upon β-d-glucose addition in the absence of GO. The GOD-DDAB-GO system displayed a high turnover efficiency and substrate affinity as a glucose biosensor. The simplicity and ease of the electrode preparation procedure of this GO/DDAB system make it a good candidate for immobilizing other biomolecules for fabrication of amperometric biosensors. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

Immobilization technique for enhanced production of the immunosuppressant mycophenolic acid by ultraviolet and gamma-irradiated Penicillium roqueforti.

PubMed

Ismaiel, A A; Ahmed, A S; El-Sayed, E R

2015-07-01

Different entrapment matrices were screened to immobilize two strains of Penicillium roqueforti (AG101 and LG109) for more effective production of mycophenolic acid (MPA). Further improvement in the MPA productivity from immobilization of spores and mycelia was adopted by UV and gamma irradiation. Penicillium roqueforti strains were immobilized in different entrapping carriers and used for MPA production in shake flask cultures. Maximum MPA production was achieved on using an alginate concentration of 3·0% (w/v) and a mycelial fresh weight of 10% (w/v). MPA produced by alginate-immobilized spores and mycelia was almost double in comparison to the free system. The MPA-producing ability of immobilized AG101 and LG109 strain was significantly enhanced by mutagenesis through irradiation by UV (254 nm) for 120 and 90 min, respectively and gamma rays at 0·75 KGy. The feasibility of MPA production in a semi-continuous form by immobilized cells as affected by irradiation was adopted. MPA production by immobilized spores and mycelia was more intensified by UV and gamma irradiation. Moreover, the immobilized cell culture was superior to free-cell culture. These findings indicate the future possibility to reduce the cost of producing fermentation-based drugs. © 2015 The Society for Applied Microbiology.

Utilization of radiation technique on the saccharification and fermentation of biomass

NASA Astrophysics Data System (ADS)

Kaetsu, I.; Kumakura, M.; Fujimura, T.; Yoshii, F.; Kojima, T.; Tamada, M.

The application of irradiation technique to the process of saccharification and subsequent fermentation of cellulosic wastes such as chaff and rice straw to obtain ethanol, was investigated. It was found that when waste raw materials were irradiated by ?-ray or electron beam, they became accessible to the subsequent enzymatic saccharification reaction. Irradiation of 10 7-10 8 Rad was enough for this effect. Some kind of additives reduced necessary dosage for this pretreatment. Cellulase, Trichoderma reesei which produce cellulase, and yeast were immobilized as biocatalysts for biomass conversion by radiation-induced polymerization of glass-forming monomer at low temperature. The immobilized cellulase showed almost same activity of glucose production as the native cellulase. Continuous saccharification reaction was carried out by using the immobilized cellulase. The immobilized Trichoderma reesei and the immobilized yeast showed almost same activity as the intact biocatalysts. It was concluded that the continuous saccharification and subsequent fermentation could be carried out effectively by using the immobilized biocatalysts. Spinach chloroplasts were immobilized by the same method as the first step for the conversion of water into hydrogen gas using solar energy. The immobilized chloroplasts kept the O 2 evolution activity in storage more than 30 days at 4°C. Thermostatility of chloroplasts was also improved greatly by the immobilization.

Nuclear fuel cycle waste stream immobilization with cermets for improved thermal properties and waste consolidation

NASA Astrophysics Data System (ADS)

Ortega, Luis H.; Kaminski, Michael D.; Zeng, Zuotao; Cunnane, James

2013-07-01

In the pursuit of methods to improve nuclear waste form thermal properties and combine potential nuclear fuel cycle wastes, a bronze alloy was combined with an alkali, alkaline earth metal bearing ceramic to form a cermet. The alloy was prepared from copper and tin (10 mass%) powders. Pre-sintered ceramic consisting of cesium, strontium, barium and rubidium alumino-silicates was mixed with unalloyed bronze precursor powders and cold pressed to 300 × 103 kPa, then sintered at 600 °C and 800 °C under hydrogen. Cermets were also prepared that incorporated molybdenum, which has a limited solubility in glass, under similar conditions. The cermet thermal conductivities were seven times that of the ceramic alone. These improved thermal properties can reduce thermal gradients within the waste forms thus lowering internal temperature gradients and thermal stresses, allowing for larger waste forms and higher waste loadings. These benefits can reduce the total number of waste packages necessary to immobilize a given amount of high level waste and immobilize troublesome elements.

Optimization of Enzyme Co-Immobilization with Sodium Alginate and Glutaraldehyde-Activated Chitosan Beads.

PubMed

Gür, Sinem Diken; İdil, Neslihan; Aksöz, Nilüfer

2018-02-01

In this study, two different materials-alginate and glutaraldehyde-activated chitosan beads-were used for the co-immobilization of α-amylase, protease, and pectinase. Firstly, optimization of multienzyme immobilization with Na alginate beads was carried out. Optimum Na alginate and CaCl 2 concentration were found to be 2.5% and 0.1 M, respectively, and optimal enzyme loading ratio was determined as 2:1:0.02 for pectinase, protease, and α-amylase, respectively. Next, the immobilization of multiple enzymes on glutaraldehyde-activated chitosan beads was optimized (3% chitosan concentration, 0.25% glutaraldehyde with 3 h of activation and 3 h of coupling time). While co-immobilization was successfully performed with both materials, the specific activities of enzymes were found to be higher for the enzymes co-immobilized with glutaraldehyde-activated chitosan beads. In this process, glutaraldehyde was acting as a spacer arm. SEM and FTIR were used for the characterization of activated chitosan beads. Moreover, pectinase and α-amylase enzymes immobilized with chitosan beads were also found to have higher activity than their free forms. Three different enzymes were co-immobilized with these two materials for the first time in this study.

Characterization and immobilization on nickel-chelated Sepharose of a glutamate decarboxylase A from Lactobacillus brevis BH2 and its application for production of GABA.

PubMed

Lee, Ji-Yeon; Jeon, Sung-Jong

2014-01-01

A gene encoding glutamate decarboxylase A (GadA) from Lactobacillus brevis BH2 was expressed in a His-tagged form in Escherichia coli cells, and recombinant protein exists as a homodimer consisting of identical subunits of 53 kDa. GadA was absolutely dependent on the ammonium sulfate concentration for catalytic activity and secondary structure formation. GadA was immobilized on the metal affinity resin with an immobilization yield of 95.8%. The pH optima of the immobilized enzyme were identical with those of the free enzyme. However, the optimum temperature for immobilized enzyme was 5 °C higher than that for the free enzyme. The immobilized GadA retained its relative activity of 41% after 30 reuses of reaction within 30 days and exhibited a half-life of 19 cycles within 19 days. A packed-bed bioreactor with immobilized GadA showed a maximum yield of 97.8% GABA from 50 mM l-glutamate in a flow-through system under conditions of pH 4.0 and 55 °C.

Co-immobilization of cellulase and lysozyme on amino-functionalized magnetic nanoparticles: An activity-tunable biocatalyst for extraction of lipids from microalgae.

PubMed

Chen, Qingtai; Liu, Dong; Wu, Chongchong; Yao, Kaisheng; Li, Zhiheng; Shi, Nan; Wen, Fushan; Gates, Ian D

2018-05-03

An activity-tunable biocatalyst for Nannochloropsis sp. cell-walls degradation was prepared by co-immobilization of cellulase and lysozyme on the surface of amino-functionalized magnetic nanoparticles (MNPs) employing glutaraldehyde. The competition between cellulase and lysozyme during immobilization was caused by the limited active sites of the MNPs. The maximum recovery of activities (cellulase: 78.9% and lysozyme: 69.6%) were achieved due to synergistic effects during dual-enzyme co-immobilization. The thermal stability in terms of half-life of the co-immobilized enzymes was three times higher than that in free form and had higher catalytic efficiency for hydrolysis of cell walls. Moreover, the co-immobilized enzymes showed greater thermal stability and wider pH tolerance than free enzymes under harsh conditions. Furthermore, the co-immobilized enzymes retained up to 60% of the residual activity after being recycled 6 times. This study provides a feasible approach for the industrialization of enzyme during cell-walls disruption and lipids extraction from Nannochloropsis sp. Copyright © 2018. Published by Elsevier Ltd.

Immobilization of Cell-Adhesive Laminin Peptides in Degradable PEGDA Hydrogels Influences Endothelial Cell Tubulogenesis

PubMed Central

Ali, Saniya; Saik, Jennifer E.; Gould, Dan J.; Dickinson, Mary E.

2013-01-01

Abstract Attachment, spreading, and organization of endothelial cells into tubule networks are mediated by interactions between cells in the extracellular microenvironment. Laminins are key extracellular matrix components and regulators of cell adhesion, migration, and proliferation. In this study, laminin-derived peptides were conjugated to poly(ethylene glycol) (PEG) monoacrylate and covalently incorporated into degradable PEG diacrylate (PEGDA) hydrogels to investigate the influence of these peptides on endothelial cellular adhesion and function in organizing into tubule networks. Degradable PEGDA hydrogels were synthesized by incorporating a matrix metalloproteinase (MMP)–sensitive peptide, GGGPQGIWGQGK (abbreviated PQ), into the polymer backbone. The secretion of MMP-2 and MMP-9 by endothelial cells promotes polymer degradation and consequently cell migration. We demonstrate the formation of extensive networks of tubule-like structures by encapsulated human umbilical vein endothelial cells in hydrogels with immobilized synthetic peptides. The resulting structures were stabilized by pericyte precursor cells (10T1/2s) in vitro. During tubule formation and stabilization, extracellular matrix proteins such as collagen IV and laminin were deposited. Tubules formed in the matrix of metalloproteinase sensitive hydrogels were visualized from 7 days to 4 weeks in response to different combination of peptides. Moreover, hydrogels functionalized with laminin peptides and transplanted in a mouse cornea supported the ingrowth and attachment of endothelial cells to the hydrogel during angiogenesis. Results of this study illustrate the use of laminin-derived peptides as potential candidates for modification of biomaterials to support angiogenesis. PMID:23914330

Reusable glucose sensing using carbon nanotube-based self-assembly

NASA Astrophysics Data System (ADS)

Bhattacharyya, Tamoghna; Samaddar, Sarbani; Dasgupta, Anjan Kr.

2013-09-01

Lipid functionalized single walled carbon nanotube-based self assembly forms a super-micellar structure. This assemblage has been exploited to trap glucose oxidase in a molecular cargo for glucose sensing. The advantage of such a molecular trap is that all components of this unique structure (both the trapping shell and the entrapped enzyme) are reusable and rechargeable. The unique feature of this sensing method lies in the solid state functionalization of single walled carbon nanotubes that facilitates liquid state immobilization of the enzyme. The method can be used for soft-immobilization (a new paradigm in enzyme immobilization) of enzymes with better thermostability that is imparted by the strong hydrophobic environment provided through encapsulation by the nanotubes.Lipid functionalized single walled carbon nanotube-based self assembly forms a super-micellar structure. This assemblage has been exploited to trap glucose oxidase in a molecular cargo for glucose sensing. The advantage of such a molecular trap is that all components of this unique structure (both the trapping shell and the entrapped enzyme) are reusable and rechargeable. The unique feature of this sensing method lies in the solid state functionalization of single walled carbon nanotubes that facilitates liquid state immobilization of the enzyme. The method can be used for soft-immobilization (a new paradigm in enzyme immobilization) of enzymes with better thermostability that is imparted by the strong hydrophobic environment provided through encapsulation by the nanotubes. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr02609d

Immobilization of organic radioactive and non-radioactive liquid waste in a composite matrix

DOE Office of Scientific and Technical Information (OSTI.GOV)

Galkin, Anatoliy; Gelis, Artem V.; Castiglioni, Andrew J.

A method for immobilizing liquid radioactive waste is provided, the method having the steps of mixing waste with polymer to form a non-liquid waste; contacting the non-liquid waste with a solidifying agent to create a mixture, heating the mixture to cause the polymer, waste, and filler to irreversibly bind in a solid phase, and compressing the solid phase into a monolith. The invention also provides a method for immobilizing liquid radioactive waste containing tritium, the method having the steps of mixing liquid waste with polymer to convert the liquid waste to a non-liquid waste, contacting the non-liquid waste with amore » solidifying agent to create a mixture, heating the mixture to form homogeneous, chemically stable solid phase, and compressing the chemically stable solid phase into a final waste form, wherein the polymer comprises approximately a 9:1 weight ratio mixture of styrene block co-polymers and cross linked co-polymers of acrylamides.« less

Immobilization of Rhodococcus rhodochrous BX2 (an acetonitrile-degrading bacterium) with biofilm-forming bacteria for wastewater treatment.

PubMed

Li, Chunyan; Li, Yue; Cheng, Xiaosong; Feng, Liping; Xi, Chuanwu; Zhang, Ying

2013-03-01

In this study, a unique biofilm consisting of three bacterial strains with high biofilm-forming capability (Bacillus subtilis E2, E3, and N4) and an acetonitrile-degrading bacterium (Rhodococcus rhodochrous BX2) was established for acetonitrile-containing wastewater treatment. The results indicated that this biofilm exhibited strong resistance to acetonitrile loading shock and displayed a typical spatial and structural heterogeneity and completely depleted the initial concentration of acetonitrile (800mgL(-1)) within 24h in a moving-bed-biofilm reactor (MBBR) after operation for 30days. The immobilization of BX2 cells in the biofilm was confirmed by PCR-DGGE. It has been demonstrated that biofilm-forming bacteria can promote the immobilization of contaminant-degrading bacteria in the biofilms and can subsequently improve the degradation of contaminants in wastewater. This approach offers a novel strategy for enhancing biological oxidation of toxic pollutants in wastewater. Copyright © 2012 Elsevier Ltd. All rights reserved.

Immobilization patterns and dynamics of acetate-utilizing methanogens immobilized in sterile granular sludge in upflow anaerobic sludge blanket reactors.

PubMed

Schmidt, J E; Ahring, B K

1999-03-01

Sterile granular sludge was inoculated with either Methanosarcina mazeii S-6, Methanosaeta concilii GP-6, or both species in acetate-fed upflow anaerobic sludge blanket (UASB) reactors to investigate the immobilization patterns and dynamics of aceticlastic methanogens in granular sludge. After several months of reactor operation, the methanogens were immobilized, either separately or together. The fastest immobilization was observed in the reactor containing M. mazeii S-6. The highest effluent concentration of acetate was observed in the reactor with only M. mazeii S-6 immobilized, while the lowest effluent concentration of acetate was observed in the reactor where both types of methanogens were immobilized together. No changes were observed in the kinetic parameters (Ks and mumax) of immobilized M. concilii GP-6 or M. mazeii S-6 compared with suspended cultures, indicating that immobilization does not affect the growth kinetics of these methanogens. An enzyme-linked immunosorbent assay using polyclonal antibodies against either M. concilii GP-6 or M. mazeii S-6 showed significant variations in the two methanogenic populations in the different reactors. Polyclonal antibodies were further used to study the spatial distribution of the two methanogens. M. concilii GP-6 was immobilized only on existing support material without any specific pattern. M. mazeii S-6, however, showed a different immobilization pattern: large clumps were formed when the concentration of acetate was high, but where the acetate concentration was low this strain was immobilized on support material as single cells or small clumps. The data clearly show that the two aceticlastic methanogens immobilize differently in UASB systems, depending on the conditions found throughout the UASB reactor.

Immobilization Patterns and Dynamics of Acetate-Utilizing Methanogens Immobilized in Sterile Granular Sludge in Upflow Anaerobic Sludge Blanket Reactors

PubMed Central

Schmidt, Jens Ejbye; Ahring, Birgitte Kjær

1999-01-01

Sterile granular sludge was inoculated with either Methanosarcina mazeii S-6, Methanosaeta concilii GP-6, or both species in acetate-fed upflow anaerobic sludge blanket (UASB) reactors to investigate the immobilization patterns and dynamics of aceticlastic methanogens in granular sludge. After several months of reactor operation, the methanogens were immobilized, either separately or together. The fastest immobilization was observed in the reactor containing M. mazeii S-6. The highest effluent concentration of acetate was observed in the reactor with only M. mazeii S-6 immobilized, while the lowest effluent concentration of acetate was observed in the reactor where both types of methanogens were immobilized together. No changes were observed in the kinetic parameters (Ks and μmax) of immobilized M. concilii GP-6 or M. mazeii S-6 compared with suspended cultures, indicating that immobilization does not affect the growth kinetics of these methanogens. An enzyme-linked immunosorbent assay using polyclonal antibodies against either M. concilii GP-6 or M. mazeii S-6 showed significant variations in the two methanogenic populations in the different reactors. Polyclonal antibodies were further used to study the spatial distribution of the two methanogens. M. concilii GP-6 was immobilized only on existing support material without any specific pattern. M. mazeii S-6, however, showed a different immobilization pattern: large clumps were formed when the concentration of acetate was high, but where the acetate concentration was low this strain was immobilized on support material as single cells or small clumps. The data clearly show that the two aceticlastic methanogens immobilize differently in UASB systems, depending on the conditions found throughout the UASB reactor. PMID:10049862

Combined Spectroscopic and Calorimetric Studies to Reveal Absorption Mechanisms and Conformational Changes of Protein on Nanoporous Biomaterials

PubMed Central

Ahmadi, Saharnaz; Farokhi, Maryam; Padidar, Parisa; Falahati, Mojtaba

2015-01-01

In this study the effect of surface modification of mesoporous silica nanoparticles (MSNs) on its adsorption capacities and protein stability after immobilization of beta-lactoglobulin B (BLG-B) was investigated. For this purpose, non-functionalized (KIT-6) and aminopropyl-functionalized cubic Ia3d mesoporous silica ([n-PrNH2-KIT-6]) nanoparticles were used as nanoporous supports. Aminopropyl-functionalized mesoporous nanoparticles exhibited more potential candidates for BLG-B adsorption and minimum BLG leaching than non-functionalized nanoparticles. It was observed that the amount of adsorbed BLG is dependent on the initial BLG concentration for both KIT-6 and [n-PrNH2-KIT-6] mesoporous nanoparticles. Also larger amounts of BLG-B on KIT-6 was immobilized upon raising the temperature of the medium from 4 to 55 °C while such increase was undetectable in the case of immobilization of BLG-B on the [n-PrNH2-KIT-6]. At temperatures above 55 °C the amounts of adsorbed BLG on both studied nanomaterials decreased significantly. By Differential scanning calorimetry or DSC analysis the heterogeneity of the protein solution and increase in Tm may indicate that immobilization of BLG-B onto the modified KIT-6 results in higher thermal stability compared to unmodified one. The obtained results provide several crucial factors in determining the mechanism(s) of protein adsorption and stability on the nanostructured solid supports and the development of engineered nano-biomaterials for controlled drug-delivery systems and biomimetic interfaces for the immobilization of living cells. PMID:26230687

Activity of select dehydrogenases with sepharose-immobilized N(6)-carboxymethyl-NAD.

PubMed

Beauchamp, Justin; Vieille, Claire

2015-01-01

N(6)-carboxymethyl-NAD (N(6)-CM-NAD) can be used to immobilize NAD onto a substrate containing terminal primary amines. We previously immobilized N(6)-CM-NAD onto sepharose beads and showed that Thermotoga maritima glycerol dehydrogenase could use the immobilized cofactor with cofactor recycling. We now show that Saccharomyces cerevisiae alcohol dehydrogenase, rabbit muscle L-lactate dehydrogenase (type XI), bovine liver L-glutamic dehydrogenase (type III), Leuconostoc mesenteroides glucose-6-phosphate dehydro-genase, and Thermotoga maritima mannitol dehydrogenase are active with soluble N(6)-CM-NAD. The products of all enzymes but 6-phospho-D-glucono-1,5-lactone were formed when sepharose-immobilized N(6)-CM-NAD was recycled by T. maritima glycerol dehydrogenase, indicating that N(6)-immobilized NAD is suitable for use by a variety of different dehydrogenases. Observations of the enzyme active sites suggest that steric hindrance plays a greater role in limiting or allowing activity with the modified cofactor than do polarity and charge of the residues surrounding the N(6)-amine group on NAD.

Hooking horseradish peroxidase by using the affinity Langmuir-Blodgett technique for an oriented immobilization

NASA Astrophysics Data System (ADS)

Peng, Ye; Ling-Ling, Hu; Yu-Zhi, Du; Yong-Juan, Xu; Hua-Gang, Ni; Cong, Chen; Xiao-Lin, Lu; Xiao-Jun, Huang

2017-05-01

A novel method of oriented immobilization was presented: affinity Langmuir-Blodgett (LB) technique. Firstly, a long carbon chain was bond to a ligand of Horseradish Peroxidase (HRP). The ligand derivative appears surface activity with the hydrophobic carbon chain oriented to air and the hydrophilic ligand faced to water. Then, this derivative was put onto the water/air surface to assemble a LB film and formed the affinity interaction with the active site of HRP. After that, the affinity LB film with the enzyme was transferred onto the support to obtain the oriented immobilized HRP. The specific activity of HRP immobilized by affinity LB (182.1 ± 14 U/mg) was higher than that by adsorption (40.5 ± 5 U/mg). HRP immobilized by affinity LB could maintain a more native conformation, compared to that by adsorption. This method could be effectively used to immobilize protein with orientation and show widely promising applications in many fields including biosensor and bioreactor.

In situ immobilization on the silica gel surface and adsorption capacity of polymer-based azobenzene on toxic metal ions

NASA Astrophysics Data System (ADS)

Savchenko, Irina; Yanovska, Elina; Sternik, Dariusz; Kychkyruk, Olga; Ol'khovik, Lidiya; Polonska, Yana

2018-03-01

In situ immobilization of poly[(4-methacryloyloxy-(4'-carboxy)azobenzene] on silica gel surface has been performed by radical polymerization of monomer. The fact of polymer immobilization is confirmed by IR spectroscopy. TG and DSC-MS analysis showed that the mass of the immobilized polymer was 10.61%. The SEM-microphotograph-synthesized composite analysis showed that the immobilized polymer on the silica gel surface is placed in the form of fibers. It has been found that the synthesized composite exhibits the sorption ability in terms of microquantities of Cu(II), Cd(II), Pb(II), Mn(II) and Fe(III) ions in a neutral aqueous medium. The quantitative sorption of microquantities of Pb(II) and Fe(III) ions has been recorded. It has been found that immobilization of the silica gel surface leads to an increase in its sorption capacitance for Fe(III), Cu(II) and Pb(II) ions by half.

Efficacy and reusability of alginate-immobilized live and heat-inactivated Trichoderma asperellum cells for Cu (II) removal from aqueous solution.

PubMed

Tan, Wei Shang; Ting, Adeline Su Yien

2012-11-01

Cu(II) removal efficacies of alginate-immobilized Trichoderma asperellum using viable and non-viable forms were investigated with respect to time, pH, and initial Cu(II) concentrations. The reusability potential of the biomass was determined based on sorption/desorption tests. Cu(II) biosorption by immobilized heat-inactivated T. asperellum cells was the most efficient, with 134.22mg Cu(II) removed g(-1) adsorbent, compared to immobilized viable cells and plain alginate beads (control) with 105.96 and 94.04mg Cu(II) adsorbed g(-1) adsorbent, respectively. Immobilized non-viable cells achieved equilibrium more rapidly within 4h. For all biosorbents, optimum pH for Cu(II) removal was between pH 4 and 5. Reusability of all biosorbents were similar, with more than 90% Cu(II) desorbed with HCl. These alginate-immobilized cells can be applied to reduce clogging and post-separation process incurred from use of suspended biomass. Copyright © 2012 Elsevier Ltd. All rights reserved.

Cast Stone Formulation for Nuclear Waste Immobilization at Higher Sodium Concentrations

DOE PAGES

Fox, Kevin; Cozzi, Alex; Roberts, Kimberly; ...

2014-11-01

Low activity radioactive waste at U.S. Department of Energy sites can be immobilized for permanent disposal using cementitious waste forms. This study evaluated waste forms produced with simulated wastes at concentrations up to twice that of currently operating processes. The simulated materials were evaluated for their fresh properties, which determine processability, and cured properties, which determine waste form performance. The results show potential for greatly reducing the volume of material. Fresh properties were sufficient to allow for processing via current practices. Cured properties such as compressive strength meet disposal requirements. Leachability indices provide an indication of expected long-term performance.

TOXIC METALS IN THE ENVIRONMENT: THERMODYNAMIC CONSIDERATIONS FOR POSSIBLE IMMOBILIZATION STRATEGIES FOR PB, CD, AS, AND HG

EPA Science Inventory

The contamination of soils by toxic metals is a widespread, serious problem that demands immediate action either by removal or immobilization, which is defined as a process which puts the metal into a chemical form, probably as a mineral, which will be inert and highly insoluble ...

Natural wax oleogels-A method to prevent oxidation of fish oil

USDA-ARS?s Scientific Manuscript database

Oleogels (or organogels) are formed by immobilization of oil by an oleogelator. Therefore, it was hypothesized that immobilization of oil could also prevent the oxidation of oil. This study was focused on preventing oxidation of fish oil, an omega-3 oil, due to its beneficial health effects such as ...

Uranium Immobilization in an Iron-Rich Rhizosphere of a Native Wetland Plant from the Savannah River Site under Reducing Conditions

EPA Science Inventory

The hypothesis of this study was that iron plaque formed on the roots of wetland plants and their rhizospheres create environmental conditions favorable for iron reducing bacteria that promote the in situ immobilization of uranium. Greenhouse microcosm studies were conducted usin...

Immobilization of Ag nanoparticles/FGF-2 on a modified titanium implant surface and improved human gingival fibroblasts behavior.

PubMed

Ma, Qianli; Mei, Shenglin; Ji, Kun; Zhang, Yumei; Chu, Paul K

2011-08-01

The objective of this study was to form a rapid and firm soft tissue sealing around dental implants that resists bacterial invasion. We present a novel approach to modify Ti surface by immobilizing Ag nanoparticles/FGF-2 compound bioactive factors onto a titania nanotubular surface. The titanium samples were anodized to form vertically organized TiO(2) nanotube arrays and Ag nanoparticles were electrodeposited onto the nanotubular surface, on which FGF-2 was immobilized with repeated lyophilization. A uniform distribution of Ag nanoparticles/FGF-2 was observed on the TiO(2) nanotubular surface. The L929 cell line was used for cytotoxicity assessment. Human gingival fibroblasts (HGFs) were cultured on the modified surface for cytocompatibility determination. The Ag/FGF-2 immobilized samples displayed excellent cytocompatibility, negligible cytotoxicity, and enhanced HGF functions such as cell attachment, proliferation, and ECM-related gene expression. The Ag nanoparticles also exhibit some bioactivity. In conclusion, this modified TiO(2) nanotubular surface has a large potential for use in dental implant abutment. Copyright © 2011 Wiley Periodicals, Inc.

Letter Report: LAW Simulant Development for Cast Stone Screening Test

DOE Office of Scientific and Technical Information (OSTI.GOV)

Russell, Renee L.; Westsik, Joseph H.; Swanberg, David J.

2013-03-27

More than 56 million gallons of radioactive and hazardous waste are stored in 177 underground storage tanks at the U.S. Department of Energy’s (DOE’s) Hanford Site in southeastern Washington State. The Hanford Tank Waste Treatment and Immobilization Plant (WTP) is being constructed to treat the wastes and immobilize them in a glass waste form. The WTP includes a pretreatment facility to separate the wastes into a small volume of high-level waste (HLW) containing most of the radioactivity and a larger volume of low-activity waste (LAW) containing most of the nonradioactive chemicals. The HLW will be converted to glass in themore » HLW vitrification facility for ultimate disposal at an offsite federal repository. At least a portion (~35%) of the LAW will be converted to glass in the LAW vitrification facility and will be disposed of onsite at the Integrated Disposal Facility (IDF). The pretreatment and HLW vitrification facilities will have the capacity to treat and immobilize the wastes destined for each facility. However, a second facility will be needed for the expected volume of additional LAW requiring immobilization. A cementitious waste form known as Cast Stone is being considered to provide the required additional LAW immobilization capacity. The Cast Stone waste form must be acceptable for disposal in the IDF. The Cast Stone waste form and immobilization process must be tested to demonstrate that the final Cast Stone waste form can comply with waste acceptance criteria for the IDF disposal facility and that the immobilization processes can be controlled to consistently provide an acceptable waste form product. Further, the waste form must be tested to provide the technical basis for understanding the long term performance of the waste form in the IDF disposal environment. These waste form performance data are needed to support risk assessment and performance assessment (PA) analyses of the long-term environmental impact of the waste disposal in the IDF. A testing program was developed in fiscal year (FY) 2012 describing in some detail the work needed to develop and qualify Cast Stone as a waste form for the solidification of Hanford LAW (Westsik et al. 2012). Included within Westsik et al. (2012) is a section on the near-term needs to address Tri-Party Agreement Milestone M-062-40ZZ. The objectives of the testing program to be conducted in FY 2013 and FY 2014 are to: • Determine an acceptable formulation for the LAW Cast Stone waste form. • Evaluate sources of dry materials for preparing the LAW Cast Stone. • Demonstrate the robustness of the Cast Stone waste form for a range of LAW compositions. • Demonstrate the robustness of the formulation for variability in the Cast Stone process. • Provide Cast Stone contaminant release data for PA and risk assessment evaluations. The first step in determining an acceptable formulation for the LAW Cast Stone waste form is to conduct screening tests to examine expected ranges in pretreated LAW composition, waste stream concentrations, dry-materials sources, and mix ratios of waste feed to dry blend. A statistically designed test matrix will be used to evaluate the effects of these key parameters on the properties of the Cast Stone as it is initially prepared and after curing. The second phase of testing will focus on selection of a baseline Cast Stone formulation for LAW and demonstrating that Cast Stone can meet expected waste form requirements for disposal in the IDF. It is expected that this testing will use the results of the screening tests to define a smaller suite of tests to refine the composition of the baseline Cast Stone formulation (e.g. waste concentration, water to dry mix ratio, waste loading).« less

Rapid purification of circular DNA by triplex-mediated affinity capture

DOEpatents

Ji, Huamin; Smith, Lloyd M.

1997-01-01

A single-step capture of a target supercoiled double-stranded DNA molecule is accomplished by forming a local triple-helix among two strands of the supercoiled circular DNA and an oligonucleotide probe. The oligonucleotide is bound to an immobilizing support which facilitates the immobilization and purification of target DNA molecules. Non-target DNA molecules and other contaminating cellular material are easily removed by washing. The triple-helical structure is destabilized by raising the pH, leaving purified target DNA in the supernatant and reusable affinity capture oligonucleotide secured to the immobilizing support.

Porphyrin-based Photocatalytic Nanolithography

PubMed Central

Bearinger, Jane P.; Stone, Gary; Dugan, Lawrence C.; El Dasher, Bassem; Stockton, Cheryl; Conway, James W.; Kuenzler, Tobias; Hubbell, Jeffrey A.

2009-01-01

Nanoarray fabrication is a multidisciplinary endeavor encompassing materials science, chemical engineering, and biology. We formed nanoarrays via a new technique, porphyrin-based photocatalytic nanolithography. The nanoarrays, with controlled features as small as 200 nm, exhibited regularly ordered patterns and may be appropriate for (a) rapid and parallel proteomics screening of immobilized biomolecules, (b) protein-protein interactions, and/or (c) biophysical and molecular biology studies involving spatially dictated ligand placement. We demonstrated protein immobilization utilizing nanoarrays fabricated via photocatalytic nanolithography on silicon substrates where the immobilized proteins are surrounded by a non-fouling polymer background. PMID:19406753

Yeast Immobilization Systems for Alcoholic Wine Fermentations: Actual Trends and Future Perspectives

PubMed Central

Moreno-García, Jaime; García-Martínez, Teresa; Mauricio, Juan C.; Moreno, Juan

2018-01-01

Yeast immobilization is defined as the physical confinement of intact cells to a region of space with conservation of biological activity. The use of these methodologies for alcoholic fermentation (AF) offers many advantages over the use of the conventional free yeast cell method and different immobilization systems have been proposed so far for different applications, like winemaking. The most studied methods for yeast immobilization include the use of natural supports (e.g., fruit pieces), organic supports (e.g., alginate), inorganic (e.g., porous ceramics), membrane systems, and multi-functional agents. Some advantages of the yeast-immobilization systems include: high cell densities, product yield improvement, lowered risk of microbial contamination, better control and reproducibility of the processes, as well as reuse of the immobilization system for batch fermentations and continuous fermentation technologies. However, these methods have some consequences on the behavior of the yeasts, affecting the final products of the fermentative metabolism. This review compiles current information about cell immobilizer requirements for winemaking purposes, the immobilization methods applied to the production of fermented beverages to date, and yeast physiological consequences of immobilization strategies. Finally, a recent inter-species immobilization methodology has been revised, where yeast cells are attached to the hyphae of a Generally Recognized As Safe fungus and remain adhered following loss of viability of the fungus. The bio-capsules formed with this method open new and promising strategies for alcoholic beverage production (wine and low ethanol content beverages). PMID:29497415

Immobilization of α-amylase onto a calix[4]arene derivative: Evaluation of its enzymatic activity.

PubMed

Veesar, Irshad Ali; Solangi, Imam Bakhsh; Memon, Shahabuddin

2015-06-01

In order to enhance the cost-effectiveness practicability of enzymes in many industries such as pharmaceutical, food, medical and some other technological processes, there is great need to immobilize them onto a solid supports. In this study, a new and efficient immobilization of α-amylase from Saccharomyces cerevisiae has been developed by using the surface functionalization of calix[4]arene as support. A glutaraldehyde-containing amino group functionalized calix[4]arene was used to immobilize α-amylase covalently. In this procedure, imide bonds are formed between amino groups on the protein and aldehyde groups on the calix[4]arene surface. The surface modified support was characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM). The effect of various preparation conditions on the immobilized α-amylase process such as immobilization time, enzyme concentration, temperature and pH were investigated. The influence of pH and temperature on the activity of free and immobilized α-amylase was also studied using starch as substrate. The optimum reaction temperature and pH value for the enzymatic conversion catalyzed by the immobilized α-amylase were 25°C and 7, respectively. Compared to the free enzyme, the immobilized α-amylase retained 85% of its original activity and exhibited significant thermal stability than the free one and excellent durability. Copyright © 2015 Elsevier Inc. All rights reserved.

Immobilized lysozyme for the continuous lysis of lactic bacteria in wine: Bench-scale fluidized-bed reactor study.

PubMed

Cappannella, Elena; Benucci, Ilaria; Lombardelli, Claudio; Liburdi, Katia; Bavaro, Teodora; Esti, Marco

2016-11-01

Lysozyme from hen egg white (HEWL) was covalently immobilized on spherical supports based on microbial chitosan in order to develop a system for the continuous, efficient and food-grade enzymatic lysis of lactic bacteria (Oenococcus oeni) in white and red wine. The objective is to limit the sulfur dioxide dosage required to control malolactic fermentation, via a cell concentration typical during this process. The immobilization procedure was optimized in batch mode, evaluating the enzyme loading, the specific activity, and the kinetic parameters in model wine. Subsequently, a bench-scale fluidized-bed reactor was developed, applying the optimized process conditions. HEWL appeared more effective in the immobilized form than in the free one, when the reactor was applied in real white and red wine. This preliminary study suggests that covalent immobilization renders the enzyme less sensitive to the inhibitory effect of wine flavans. Copyright © 2016 Elsevier Ltd. All rights reserved.

Supplemental Immobilization Cast Stone Technology Development and Waste Form Qualification Testing Plan

DOE Office of Scientific and Technical Information (OSTI.GOV)

Westsik, Joseph H.; Serne, R. Jeffrey; Pierce, Eric M.

2013-05-31

The Hanford Tank Waste Treatment and Immobilization Plant (WTP) is being constructed to treat the 56 million gallons of radioactive waste stored in 177 underground tanks at the Hanford Site. The WTP includes a pretreatment facility to separate the wastes into high-level waste (HLW) and low-activity waste (LAW) fractions for vitrification and disposal. The LAW will be converted to glass for final disposal at the Integrated Disposal Facility (IDF). The pretreatment facility will have the capacity to separate all of the tank wastes into the HLW and LAW fractions, and the HLW Vitrification Facility will have the capacity to vitrifymore » all of the HLW. However, a second immobilization facility will be needed for the expected volume of LAW requiring immobilization. A number of alternatives, including Cast Stone—a cementitious waste form—are being considered to provide the additional LAW immobilization capacity.« less

Cellulase immobilization on magnetic nanoparticles encapsulated in polymer nanospheres.

PubMed

Lima, Janaina S; Araújo, Pedro H H; Sayer, Claudia; Souza, Antonio A U; Viegas, Alexandre C; de Oliveira, Débora

2017-04-01

Immobilization of cellulases on magnetic nanoparticles, especially magnetite nanoparticles, has been the main approach studied to make this enzyme, economically and industrially, more attractive. However, magnetite nanoparticles tend to agglomerate, are very reactive and easily oxidized in air, which has strong impact on their useful life. Thus, it is very important to provide proper surface coating to avoid the mentioned problems. This study aimed to investigate the immobilization of cellulase on magnetic nanoparticles encapsulated in polymeric nanospheres. The support was characterized in terms of morphology, average diameter, magnetic behavior and thermal decomposition analyses. The polymer nanospheres containing encapsulated magnetic nanoparticles showed superparamagnetic behavior and intensity average diameter about 150 nm. Immobilized cellulase exhibited broader temperature stability than in the free form and great reusability capacity, 69% of the initial enzyme activity was maintained after eight cycles of use. The magnetic support showed potential for cellulase immobilization and allowed fast and easy biocatalyst recovery through a single magnet.

Enzyme nanoparticle fabrication: magnetic nanoparticle synthesis and enzyme immobilization.

PubMed

Johnson, Patrick A; Park, Hee Joon; Driscoll, Ashley J

2011-01-01

Immobilized enzymes are drawing significant attention for potential commercial applications as biocatalysts by reducing operational expenses and by increasing process utilization of the enzymes. Typically, immobilized enzymes have greater thermal and operational stability at various pH values, ionic strengths and are more resistant to denaturation that the soluble native form of the enzyme. Also, immobilized enzymes can be recycled by utilizing the physical or chemical properties of the supporting material. Magnetic nanoparticles provide advantages as the supporting material for immobilized enzymes over competing materials such as: higher surface area that allows for greater enzyme loading, lower mass transfer resistance, less fouling effect, and selective, nonchemical separation from the reaction mixture by an applied a magnetic field. Various surface modifications of magnetic nanoparticles, such as silanization, carbodiimide activation, and PEG or PVA spacing, aid in the binding of single or multienzyme systems to the particles, while cross-linking using glutaraldehyde can also stabilize the attached enzymes.

Improvement of nuclide leaching resistance of paraffin waste form with low density polyethylene.

PubMed

Kim, Chang Lak; Park, Joo Wan; Kim, Ju Youl; Chung, Chang Hyun

2002-01-01

Low-level liquid borate wastes have been immobilized with paraffin wax using a concentrate waste drying system (CWDS) in Korean nuclear power plants. The possibility for improving chemical durability of paraffin waste form was suggested in this study. A small amount of low density polyethylene (LDPE) was added to increase the leaching resistance of the existing paraffin waste form. The influence of LDPE on the leaching behavior of waste form was investigated by performing leaching test according to ANSI/ANS-16.1 procedure during 325 days. It was observed that the leaching of nuclides immobilized within paraffin waste form made a marked reduction although little content of LDPE was added to waste form. The acceptance criteria of paraffin waste form associated with leachability index (LI) and compressive strength after the leaching test were fully satisfied with the help of LDPE.

Options for the Separation and Immobilization of Technetium

DOE Office of Scientific and Technical Information (OSTI.GOV)

Serne, R Jeffrey; Crum, Jarrod V.; Riley, Brian J.

Among radioactive constituents present in the Hanford tank waste, technetium-99 (Tc) presents a unique challenge in that it is significantly radiotoxic, exists predominantly in the liquid low-activity waste (LAW), and has proven difficult to effectively stabilize in a waste form for ultimate disposal. Within the Hanford Tank Waste Treatment and Immobilization Plant, the LAW fraction will be converted to a glass waste form in the LAW vitrification facility, but a significant fraction of Tc volatilizes at the high glass-melting temperatures and is captured in the off-gas treatment system. This necessitates recycle of the off-gas condensate solution to the LAW glassmore » melter feed. The recycle process is effective in increasing the loading of Tc in the immobilized LAW (ILAW), but it also disproportionately increases the sulfur and halides in the LAW melter feed, which have limited solubility in the LAW glass and thus significantly reduce the amount of LAW (glass waste loading) that can be vitrified and still maintain good waste form properties. This increases both the amount of LAW glass and either the duration of the LAW vitrification mission or requires the need for supplemental LAW treatment capacity. Several options are being considered to address this issue. Two approaches attempt to minimize the off-gas recycle by removing Tc at one of several possible points within the tank waste processing flowsheet. The separated Tc from these two approaches must then be dispositioned in a manner such that the Tc can be safely disposed. Alternative waste forms that do not have the Tc volatility issues associated with the vitrification process are being sought for immobilization of Tc for subsequent storage and disposal. The first objective of this report is to provide insights into the compositions and volumes of the Tc-bearing waste streams including the ion exchange eluate from processing LAW and the off-gas condensate from the melter. The first step to be assessed will be the processing of ion exchange eluate. The second objective of this report is to assess the compatibility of the available waste forms with the anticipated waste streams. Two major categories of Tc-specific waste forms are considered in this report including mineral and metal waste forms. Overall, it is concluded that a metal alloy waste form is the most promising and mature Tc-specific waste form and offers several benefits. One obvious advantage of the disposition of Tc in the metal alloy waste form is the significant reduction of the generated waste form volume, which leads to a reduction of the required storage facility footprint. Among mineral waste forms, glass-bonded sodalite and possibly goethite should also be considered for the immobilization of Tc.« less

Immobilization of single argon atoms in nano-cages of two-dimensional zeolite model systems.

PubMed

Zhong, Jian-Qiang; Wang, Mengen; Akter, Nusnin; Kestell, John D; Boscoboinik, Alejandro M; Kim, Taejin; Stacchiola, Dario J; Lu, Deyu; Boscoboinik, J Anibal

2017-07-17

The confinement of noble gases on nanostructured surfaces, in contrast to bulk materials, at non-cryogenic temperatures represents a formidable challenge. In this work, individual Ar atoms are trapped at 300 K in nano-cages consisting of (alumino)silicate hexagonal prisms forming a two-dimensional array on a planar surface. The trapping of Ar atoms is detected in situ using synchrotron-based ambient pressure X-ray photoelectron spectroscopy. The atoms remain in the cages upon heating to 400 K. The trapping and release of Ar is studied combining surface science methods and density functional theory calculations. While the frameworks stay intact with the inclusion of Ar atoms, the permeability of gasses (for example, CO) through them is significantly affected, making these structures also interesting candidates for tunable atomic and molecular sieves. These findings enable the study of individually confined noble gas atoms using surface science methods, opening up new opportunities for fundamental research.

Graphene/SiO2 nanocomposites: The enhancement of photocatalytic and biomedical activity of SiO2 nanoparticles by graphene

NASA Astrophysics Data System (ADS)

Arshad, Aqsa; Iqbal, Javed; Mansoor, Qaisar; Ahmed, Ishaq

2017-06-01

The exceptional conducting nature of graphene makes it a viable candidate for enhancing the effectiveness of photocatalytic and biomedical nanomaterials. Herein, the immobilization of monodispersed silicon dioxide (SiO2) nanoparticles on multiple graphene layers is demonstrated for intercalation of graphene nanoplatelets. Interestingly, the addition of graphene nanoplatelets with SiO2 nanoparticles enhances the photocatalytic efficiency from 46% to 99%. For biomedical applications, it is found that 75% of Gram positive and 50% of Gram negative bacteria have been killed; hence, bacterial proliferation is significantly restricted. Further, the cytotoxicity study reveals that the synthesised nanocomposites are non-toxic for both normal (human corneal epithelial cells) and cancerous (MCF-7, HEp-2) cell lines which signify their potential as carriers for drug delivery. The prepared nanocomposites with a controlled amount of carbon in the form of graphene can be employed for photocatalysis based waste water remediation, biomedicine, and nanodrug delivery.

Immobilization of single argon atoms in nano-cages of two-dimensional zeolite model systems

DOE PAGES

Zhong, Jian-Qiang; Wang, Mengen; Akter, Nusnin; ...

2017-07-17

The confinement of noble gases on nanostructured surfaces, in contrast to bulk materials, at non-cryogenic temperatures represents a formidable challenge. Here, individual Ar atoms are trapped at 300 K in nano-cages consisting of (alumino)silicate hexagonal prisms forming a two-dimensional array on a planar surface. The trapping of Ar atoms is detected in situ using synchrotron-based ambient pressure X-ray photoelectron spectroscopy. The atoms remain in the cages upon heating to 400 K. The trapping and release of Ar is studied combining surface science methods and density functional theory calculations. While the frameworks stay intact with the inclusion of Ar atoms, themore » permeability of gasses (for example, CO) through them is significantly affected, making these structures also interesting candidates for tunable atomic and molecular sieves. Our findings enable the study of individually confined noble gas atoms using surface science methods, opening up new opportunities for fundamental research.« less

Testing the Feasibility of a Low-Cost Network Performance Measurement Infrastructure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chevalier, Scott; Schopf, Jennifer M.; Miller, Kenneth

2016-07-01

Todays science collaborations depend on reliable, high performance networks, but monitoring the end-to-end performance of a network can be costly and difficult. The most accurate approaches involve using measurement equipment in many locations, which can be both expensive and difficult to manage due to immobile or complicated assets. The perfSONAR framework facilitates network measurement making management of the tests more reasonable. Traditional deployments have used over-provisioned servers, which can be expensive to deploy and maintain. As scientific network uses proliferate, there is a desire to instrument more facets of a network to better understand trends. This work explores low costmore » alternatives to assist with network measurement. Benefits include the ability to deploy more resources quickly, and reduced capital and operating expenditures. Finally, we present candidate platforms and a testing scenario that evaluated the relative merits of four types of small form factor equipment to deliver accurate performance measurements.« less

Immobilization of single argon atoms in nano-cages of two-dimensional zeolite model systems

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhong, Jian-Qiang; Wang, Mengen; Akter, Nusnin

The confinement of noble gases on nanostructured surfaces, in contrast to bulk materials, at non-cryogenic temperatures represents a formidable challenge. Here, individual Ar atoms are trapped at 300 K in nano-cages consisting of (alumino)silicate hexagonal prisms forming a two-dimensional array on a planar surface. The trapping of Ar atoms is detected in situ using synchrotron-based ambient pressure X-ray photoelectron spectroscopy. The atoms remain in the cages upon heating to 400 K. The trapping and release of Ar is studied combining surface science methods and density functional theory calculations. While the frameworks stay intact with the inclusion of Ar atoms, themore » permeability of gasses (for example, CO) through them is significantly affected, making these structures also interesting candidates for tunable atomic and molecular sieves. Our findings enable the study of individually confined noble gas atoms using surface science methods, opening up new opportunities for fundamental research.« less

Immobilized laccase mediated dye decolorization and transformation pathway of azo dye acid red 27.

PubMed

Chhabra, Meenu; Mishra, Saroj; Sreekrishnan, Trichur Ramaswamy

2015-01-01

Laccases have good potential as bioremediating agents and can be used continuously in the immobilized form like many other enzymes. In the present study, laccase from Cyathus bulleri was immobilized by entrapment in Poly Vinyl Alcohol (PVA) beads cross-linked with either nitrate or boric acid. Immobilized laccase was used for dye decolorization in both batch and continuous mode employing a packed bed column. The products of degradation of dye Acid Red 27 were identified by LC MS/MS analysis. The method led to very effective (90%) laccase immobilization and also imparted significant stability to the enzyme (more than 70% after 5 months of storage at 4°C). In batch decolorization, 90-95% decolorization was achieved of the simulated dye effluent for up to 10-20 cycles. Continuous decolorization in a packed bed bioreactor led to nearly 90% decolorization for up to 5 days. The immobilized laccase was also effective in decolorization and degradation of Acid Red 27 in the presence of a mediator. Four products of degradation were identified by LC-MS/MS analysis. The immobilized laccase in PVA-nitrate was concluded to be an effective agent in treatment of textile dye effluents.

Mitigation of Hydrogen Gas Generation from the Reaction of Uranium Metal with Water in K Basin Sludge and Sludge Waste Forms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sinkov, Sergey I.; Delegard, Calvin H.; Schmidt, Andrew J.

2011-06-08

Prior laboratory testing identified sodium nitrate and nitrite to be the most promising agents to minimize hydrogen generation from uranium metal aqueous corrosion in Hanford Site K Basin sludge. Of the two, nitrate was determined to be better because of higher chemical capacity, lower toxicity, more reliable efficacy, and fewer side reactions than nitrite. The present lab tests were run to determine if nitrate’s beneficial effects to lower H2 generation in simulated and genuine sludge continued for simulated sludge mixed with agents to immobilize water to help meet the Waste Isolation Pilot Plant (WIPP) waste acceptance drainable liquid criterion. Testsmore » were run at ~60°C, 80°C, and 95°C using near spherical high-purity uranium metal beads and simulated sludge to emulate uranium-rich KW containerized sludge currently residing in engineered containers KW-210 and KW-220. Immobilization agents tested were Portland cement (PC), a commercial blend of PC with sepiolite clay (Aquaset II H), granulated sepiolite clay (Aquaset II G), and sepiolite clay powder (Aquaset II). In all cases except tests with Aquaset II G, the simulated sludge was mixed intimately with the immobilization agent before testing commenced. For the granulated Aquaset II G clay was added to the top of the settled sludge/solution mixture according to manufacturer application directions. The gas volumes and compositions, uranium metal corrosion mass losses, and nitrite, ammonia, and hydroxide concentrations in the interstitial solutions were measured. Uranium metal corrosion rates were compared with rates forecast from the known uranium metal anoxic water corrosion rate law. The ratios of the forecast to the observed rates were calculated to find the corrosion rate attenuation factors. Hydrogen quantities also were measured and compared with quantities expected based on non-attenuated H2 generation at the full forecast anoxic corrosion rate to arrive at H2 attenuation factors. The uranium metal corrosion rates in water alone and in simulated sludge were near or slightly below the metal-in-water rate while nitrate-free sludge/Aquaset II decreased rates by about a factor of 3. Addition of 1 M nitrate to simulated sludge decreased the corrosion rate by a factor of ~5 while 1 M nitrate in sludge/Aquaset II mixtures decreased the corrosion rate by ~2.5 compared with the nitrate-free analogues. Mixtures of simulated sludge with Aquaset II treated with 1 M nitrate had uranium corrosion rates about a factor of 8 to 10 lower than the water-only rate law. Nitrate was found to provide substantial hydrogen mitigation for immobilized simulant sludge waste forms containing Aquaset II or Aquaset II G clay. Hydrogen attenuation factors of 1000 or greater were determined at 60°C for sludge-clay mixtures at 1 M nitrate. Hydrogen mitigation for tests with PC and Aquaset II H (which contains PC) were inconclusive because of suspected failure to overcome induction times and fully enter into anoxic corrosion. Lessening of hydrogen attenuation at ~80°C and ~95°C for simulated sludge and Aquaset II was observed with attenuation factors around 100 to 200 at 1 M nitrate. Valuable additional information has been obtained on the ability of nitrate to attenuate hydrogen gas generation from solution, simulant K Basin sludge, and simulant sludge with immobilization agents. Details on characteristics of the associated reactions were also obtained. The present testing confirms prior work which indicates that nitrate is an effective agent to attenuate hydrogen from uranium metal corrosion in water and simulated K Basin sludge to show that it is also effective in potential candidate solidified K Basin waste forms for WIPP disposal. The hydrogen mitigation afforded by nitrate appears to be sufficient to meet the hydrogen generation limits for shipping various sludge waste streams based on uranium metal concentrations and assumed waste form loadings.« less

One-step production of immobilized alpha-amylase in recombinant Escherichia coli.

PubMed

Rasiah, Indira A; Rehm, Bernd H A

2009-04-01

Industrial enzymes are often immobilized via chemical cross-linking onto solid supports to enhance stability and facilitate repeated use in bioreactors. For starch-degrading enzymes, immobilization usually places constraints on enzymatic conversion due to the limited diffusion of the macromolecular substrate through available supports. This study describes the one-step immobilization of a highly thermostable alpha-amylase (BLA) from Bacillus licheniformis and its functional display on the surface of polyester beads inside engineered Escherichia coli. An optimized BLA variant (Termamyl) was N-terminally fused to the polyester granule-forming enzyme PhaC of Cupriavidus necator. The fusion protein lacking the signal sequence mediated formation of stable polyester beads exhibiting alpha-amylase activity. The alpha-amylase beads were assessed with respect to alpha-amylase activity, which was demonstrated qualitatively and quantitatively. The immobilized alpha-amylase showed Michaelis-Menten enzyme kinetics exerting a V(max) of about 506 mU/mg of bead protein with a K(m) of about 5 microM, consistent with that of free alpha-amylase. The stability of the enzyme at 85 degrees C and the capacity for repeated usage in a starch liquefaction process were also demonstrated. In addition, structural integrity and functionality of the beads at extremes of pH and temperature, demonstrating their suitability for industrial use, were confirmed by electron microscopy and protein/enzyme analysis. This study proposes a novel, cost-effective method for the production of immobilized alpha-amylase in a single step by using the polyester granules forming protein PhaC as a fusion partner in engineered E. coli.

One-Step Production of Immobilized α-Amylase in Recombinant Escherichia coli▿ †

PubMed Central

Rasiah, Indira A.; Rehm, Bernd H. A.

2009-01-01

Industrial enzymes are often immobilized via chemical cross-linking onto solid supports to enhance stability and facilitate repeated use in bioreactors. For starch-degrading enzymes, immobilization usually places constraints on enzymatic conversion due to the limited diffusion of the macromolecular substrate through available supports. This study describes the one-step immobilization of a highly thermostable α-amylase (BLA) from Bacillus licheniformis and its functional display on the surface of polyester beads inside engineered Escherichia coli. An optimized BLA variant (Termamyl) was N-terminally fused to the polyester granule-forming enzyme PhaC of Cupriavidus necator. The fusion protein lacking the signal sequence mediated formation of stable polyester beads exhibiting α-amylase activity. The α-amylase beads were assessed with respect to α-amylase activity, which was demonstrated qualitatively and quantitatively. The immobilized α-amylase showed Michaelis-Menten enzyme kinetics exerting a Vmax of about 506 mU/mg of bead protein with a Km of about 5 μM, consistent with that of free α-amylase. The stability of the enzyme at 85°C and the capacity for repeated usage in a starch liquefaction process were also demonstrated. In addition, structural integrity and functionality of the beads at extremes of pH and temperature, demonstrating their suitability for industrial use, were confirmed by electron microscopy and protein/enzyme analysis. This study proposes a novel, cost-effective method for the production of immobilized α-amylase in a single step by using the polyester granules forming protein PhaC as a fusion partner in engineered E. coli. PMID:19201981

Immobilized unfolded cytochrome c acts as a catalyst for dioxygen reduction.

PubMed

Tavagnacco, Claudio; Monari, Stefano; Ranieri, Antonio; Bortolotti, Carlo Augusto; Peressini, Silvia; Borsari, Marco

2011-10-21

Unfolding turns immobilized cytochrome c into a His-His ligated form endowed with catalytic activity towards O(2), which is absent in the native protein. Dioxygen could be used by naturally occurring unfolded cytochrome c as a substrate for the production of partially reduced oxygen species (PROS) contributing to the cell oxidative stress.

Arsenate immobilization associated with microbial oxidation of ferrous ion in complex acid sulfate water.

PubMed

Ma, Yingqun; Lin, Chuxia

2012-05-30

Chemical, XRD, SEM, RS, FTIR and XPS techniques were used to investigate arsenate immobilization associated with microbial Fe(2+) oxidation in a complex acid sulfate water system consisting of a modified 9 K solution (pH 2.0) plus As, Cu, Cd, Pb, Zn and Mn. At a 1:12.5:70 molar ratio of As:Fe:S, schweretmannite formation was impeded. This was in contrast with the predominant presence of schwertmannite when the heavy metals were absent, suggesting that a schwertmannite binding model is not valid for explaining arsenate immobilization in the complex system. In this study, arsenate was initially immobilized through co-precipitation with non-Fe metals and phosphate. Subsequently when sufficient Fe(3+) was produced from Fe(2+) oxidation, formation of a mixed iron, arsenate and phosphate phase predominated. The last stage involved surface complexation of arsenate species. Pb appeared to play an insignificant role in arsenate immobilization due to its strong affinity for sulfate to form anglesite. Phosphate strongly competed with arsenate for the available binding sites. However, As exhibited an increased capacity to compete with P and S for available binding sites from the co-precipitation to surface complexation stage. Adsorbed As tended to be in HAsO(4)(2-) form. The scavenged arsenate species was relatively stable after 2464-h aging. Copyright © 2012 Elsevier B.V. All rights reserved.

Zinc tetraaminophthalocyanine-Fe3O4 nanoparticle composite for laccase immobilization

PubMed Central

Huang, Jun; Liu, Cheng; Xiao, Haiyan; Wang, Juntao; Jiang, Desheng; GU, Erdan

2007-01-01

Zinc tetraaminophthalocyanine-Fe3O4 nanoparticle composites were prepared by organic-inorganic complex technology and characterized. It has been proved that the ZnTAPc dispersed randomly onto the surface of Fe3O4 nanoparticles to form molecular dispersion layer and there was a relatively strong bond between central zinc cation and oxygen. The nanoparticle composite took the shape of roundish spheres with the mean diameter of about 15 nm. Active amino groups of magnetic carriers could be used to bind laccase via glutaraldehyde. The optimal pH for the activity of the immobilized laccases and free laccase were the same at pH 3.0 and the optimal temperature for laccase immobilization on ZnTAPc-Fe3O4 nanoparticle composite was 45°. The immobilization yields and Km value of the laccase immobilized on ZnTAPc-Fe3O4 nanoparticle composite were 25% and 20.1 μM, respectively. This kind of immobilized laccase has good thermal, storage and operation stability, and could be used as the sensing biocomponent for the fiber optic biosensor based on enzyme catalysis. PMID:18203444

Layer uniformity in glucose oxidase immobilization on SiO 2 surfaces

NASA Astrophysics Data System (ADS)

Libertino, Sebania; Scandurra, Antonino; Aiello, Venera; Giannazzo, Filippo; Sinatra, Fulvia; Renis, Marcella; Fichera, Manuela

2007-09-01

The goal of this work was the characterization, step by step, of the enzyme glucose oxidase (GOx) immobilization on silicon oxide surfaces, mainly by means of X-Ray photoelectron spectroscopy (XPS). The immobilization protocol consists of four steps: oxide activation, silanization, linker molecule deposition and GOx immobilization. The linker molecule, glutaraldehyde (GA) in this study, must be able to form a uniform layer on the sample surface in order to maximize the sites available for enzyme bonding and achieve the best enzyme deposition. Using a thin SiO 2 layer grown on Si wafers and following the XPS Si2p signal of the Si substrate during the immobilization steps, we demonstrated both the glutaraldehyde layer uniformity and the possibility to use XPS to monitor thin layer uniformity. In fact, the XPS substrate signal, not shielded by the oxide, is suppressed only when a uniform layer is deposited. The enzyme correct immobilization was monitored using the XPS C1s and N1s signals. Atomic force microscopy (AFM) measurements carried out on the same samples confirmed the results.

Nickel-impregnated silica nanoparticle synthesis and their evaluation for biocatalyst immobilization.

PubMed

Prakasham, Reddy Shetty; Devi, G Sarala; Rao, Chaganti Subba; Sivakumar, V S S; Sathish, T; Sarma, P N

2010-04-01

In the present investigation, impact of nickel-impregnated silica paramagnetic particles (NSP) as biocatalyst immobilization matrices was investigated. These nanoparticles were synthesized by sol-gel route using a nonionic surfactant block co polymer [poly (ethylene glycol)-block-poly-(propylene glycol)-block-poly (ethylene glycol)]. Diastase enzyme was immobilized on these particles (enzyme-impregnated NSP) as model enzyme and characterized using Fourier-transform infrared spectroscopy and X-ray crystallography. Analysis of enzyme-binding nature with these nanoparticles at different physiological conditions revealed that binding pattern and activity profile varied with the pH of the reaction mixture. The immobilized enzyme was further characterized for its biocatalytic activity with respect to kinetic properties such as Km and Vmax and compared with free enzyme. Paramagnetic nanoparticle-immobilized enzyme showed more affinity for substrate compared to free one. The nature of silica and nickel varied from amorphous to crystalline nature and vice versa upon immobilization of enzyme. To the best of our knowledge, this is the first report of its kind for change of nature from one form to other under normal temperatures upon diastase interaction with NSP.

Bioprocessing of Biodiesel Industry Effluent by Immobilized Bacteria to Produce Value-Added Products.

PubMed

Prakash, Jyotsana; Gupta, Rahul Kumar; Xx, Priyanka; Kalia, Vipin Chandra

2018-05-01

Biodiesel industrial effluent rich in crude glycerol (CG) was processed to produce value-added product. Under continuous culture system, Bacillus amyloliquefaciens strain CD16 immobilized within its biofilm, produced 3.2 L H 2 /day/L feed, over a period of 60 days at a hydraulic retention time of 2 days. The effective H 2 yield by B. amyloliquefaciens strain CD16 was 165 L/L CG. This H 2 yield was 1.18-fold higher than that observed with non-biofilm forming Bacillus thuringiensis strain EGU45. Bioprocessing of the effluent released after this stage, by recycling it up to 25% did not have any adverse effect on H 2 production by strain EGU45; however, a 25% reduction in yield was recorded with strain CD16. Biofilm forming H 2 producers thus proved effective as self-immobilizing system leading to enhanced process efficiency.

A comprehensive review of techniques for biofunctionalization of titanium

PubMed Central

2011-01-01

A number of surface modification techniques using immobilization of biofunctional molecules of Titanium (Ti) for dental implants as well as surface properties of Ti and Ti alloys have been developed. The method using passive surface oxide film on titanium takes advantage of the fact that the surface film on Ti consists mainly of amorphous or low-crystalline and non-stoichiometric TiO2. In another method, the reconstruction of passive films, calcium phosphate naturally forms on Ti and its alloys, which is characteristic of Ti. A third method uses the surface active hydroxyl group. The oxide surface immediately reacts with water molecules and hydroxyl groups are formed. The hydroxyl groups dissociate in aqueous solutions and show acidic and basic properties. Several additional methods are also possible, including surface modification techniques, immobilization of poly(ethylene glycol), and immobilization of biomolecules such as bone morphogenetic protein, peptide, collagen, hydrogel, and gelatin. PMID:22324003

Study of Horseradish Peroxidase Fixed on Mesoporous Materials as a Chemical Reaction Catalyst

NASA Astrophysics Data System (ADS)

Gao, Mengdan; Dai, Rongji

2017-12-01

Nanostructured mesoporous materials is a new type of porous materials, which has been widely used. It has excellent capability in enzymes immobilization, but modification on the chemical bonds of the enzyme reduce the enzymatic activity and rarely used in chemical reactions. The horseradish peroxidase was immobilized on the mesoporous materials with appropriate aperture and its activity and stability was evaluated when catalyzing the nitration reaction of amines and oxidation reaction of thiourea. The optimum mesoporous material to fix the horseradish peroxidase can be obtained by mixing polyoxyethylene - polyoxypropylene-pol, yoxyethylene(P123), 1,3,5-trimethylbenzene(TMB), and tetramethoxysilane (TMOS) at a ratio of 10:1:1, whose surface area and pore volume and pore diameter calculated by BET and BJH model were 402.903m2/g, 1.084cm2/g, 1.084cm2/g respectively. The horseradish peroxidase, immobilized on the mesoporous materials, was applied for catalyzing the nitration reaction of anilines and oxidation reaction of thiourea, produced a high product yield and can be recycled. Thus, it is a strong candidate as a catalysts for oxidation reactions, to be produced at industral scale, due to its high efficiency and low cost.

Microbial Desulfurization of Gasoline in a Mycobacterium goodii X7B Immobilized-Cell System

PubMed Central

Li, Fuli; Xu, Ping; Feng, Jinhui; Meng, Ling; Zheng, Yuan; Luo, Lailong; Ma, Cuiqing

2005-01-01

Mycobacterium goodii X7B, which had been primarily isolated as a bacterial strain capable of desulfurizing dibenzothiophene to produce 2-hydroxybiphenyl via the 4S pathway, was also found to desulfurize benzothiophene. The desulfurization product was identified as o-hydroxystyrene by gas chromatography (GC)-mass spectrometry analysis. This strain appeared to have the ability to remove organic sulfur from a broad range of sulfur species in gasoline. When Dushanzi straight-run gasoline (DSRG227) containing various organic sulfur compounds was treated with immobilized cells of strain X7B for 24 h, the total sulfur content significantly decreased, from 227 to 71 ppm at 40°C. GC flame ionization detection and GC atomic emission detection analysis were used to qualitatively evaluate the effects of M. goodii X7B treatment on the contents of gasoline. In addition, when immobilized cells were incubated at 40°C with DSRG275, the sulfur content decreased from 275 to 54 ppm in two consecutive reactions. With this excellent efficiency, strain X7B is considered a good potential candidate for industrial applications for the biodesulfurization of gasoline. PMID:15640198

Degradation of diesel oil by immobilized Candida tropicalis and biofilm formed on gravels.

PubMed

Chandran, Preethy; Das, Nilanjana

2011-11-01

The performance of diesel oil degradation by Candida tropicalis immobilized on various conventional matrices (sodium alginate, carboxyl methyl cellulose, chitosan) and biowaste materials (wheat bran, sawdust, peanut hull powder) was investigated using the method of entrapment and physical adsorption. The yeast species immobilized in wheat bran showed enhanced efficiency in degrading diesel oil (98%) compared to free cells culture (80%) over a period of 7 days. Copious amount of exopolysaccharides were also produced in the presence of diesel oil. The biofilm forming ability of C. tropicalis on PVC strips was evaluated using XTT (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide) reduction assay and monitored by scanning electron microscopy and atomic force microscopy. Yeast biofilm formed on gravels showed 97% degradation of diesel oil over a period of 10 days. The potential use of the biofilms for preparing trickling filters (gravel particles), for attenuating hydrocarbons in oily liquid wastes before their disposal in the open environment is suggested and discussed. This is the first successful attempt for 'artificially' establishing hydrocarbon degrading yeast biofilm on solid substrates.

Carrier free immobilization and characterization of trypsin.

PubMed

Menfaatli, Esra; Zihnioglu, Figen

2015-04-01

Pancreatic trypsin was immobilized by cross-linked enzyme aggregates (CLEA) which is a carrier free immobilization method. Ammonium sulfate was chosen for enzyme precipitation which was followed by cross linking of formed aggregates via glutaraldehyde. Concentrations of precipitant and cross linker were respectively optimized as 60% ammonium sulfate and 1% glutaraldehyde. Optimum pH and temperature for CLEA was increased compared to free enzyme. Furthermore, pH, thermal and storage stability were improved. Presence of additives had no effects on enzyme activity. Prepared cross-linked trypsin aggregates are convenient for in situ protein fragmentation and can be used for protein identification.

Rapid purification of circular DNA by triplex-mediated affinity capture

DOEpatents

Ji, H.; Smith, L.M.

1997-01-07

A single-step capture of a target supercoiled double-stranded DNA molecule is accomplished by forming a local triple-helix among two strands of the supercoiled circular DNA and an oligonucleotide probe. The oligonucleotide is bound to an immobilizing support which facilitates the immobilization and purification of target DNA molecules. Non-target DNA molecules and other contaminating cellular material are easily removed by washing. The triple-helical structure is destabilized by raising the pH, leaving purified target DNA in the supernatant and reusable affinity capture oligonucleotide secured to the immobilizing support. 3 figs.

Actinide Waste Forms and Radiation Effects

NASA Astrophysics Data System (ADS)

Ewing, R. C.; Weber, W. J.

Over the past few decades, many studies of actinides in glasses and ceramics have been conducted that have contributed substantially to the increased understanding of actinide incorporation in solids and radiation effects due to actinide decay. These studies have included fundamental research on actinides in solids and applied research and development related to the immobilization of the high level wastes (HLW) from commercial nuclear power plants and processing of nuclear weapons materials, environmental restoration in the nuclear weapons complex, and the immobilization of weapons-grade plutonium as a result of disarmament activities. Thus, the immobilization of actinides has become a pressing issue for the twenty-first century (Ewing, 1999), and plutonium immobilization, in particular, has received considerable attention in the USA (Muller et al., 2002; Muller and Weber, 2001). The investigation of actinides and

Evaluation of Technetium Getters to Improve the Performance of Cast Stone

DOE Office of Scientific and Technical Information (OSTI.GOV)

Neeway, James J.; Qafoku, Nikolla P.; Serne, R. Jeffrey

2015-11-01

Cast Stone has been selected as the preferred waste form for solidification of aqueous secondary liquid effluents from the Hanford Tank Waste Treatment and Immobilization Plant (WTP) process condensates and low-activity waste (LAW) melter off-gas caustic scrubber effluents. Cast Stone is also being evaluated as a supplemental immobilization technology to provide the necessary LAW treatment capacity to complete the Hanford tank waste cleanup mission in a timely and cost effective manner. One of the major radionuclides that Cast Stone has the potential to immobilize is technetium (Tc). The mechanism for immobilization is through the reduction of the highly mobile Tc(VII)more » species to the less mobile Tc(IV) species by the blast furnace slag (BFS) used in the Cast Stone formulation. Technetium immobilization through this method would be beneficial because Tc is one of the most difficult contaminants to address at the U.S. Department of Energy (DOE) Hanford Site due to its complex chemical behavior in tank waste, limited incorporation in mid- to high-temperature immobilization processes (vitrification, steam reformation, etc.), and high mobility in subsurface environments. In fact, the Tank Closure and Waste Management Environmental Impact Statement for the Hanford Site, Richland, Washington (TC&WM EIS) identifies technetium-99 ( 99Tc) as one of the radioactive tank waste components contributing the most to the environmental impact associated with the cleanup of the Hanford Site. The TC&WM EIS, along with an earlier supplemental waste-form risk assessment, used a diffusion-limited release model to estimate the release of different contaminants from the WTP process waste forms. In both of these predictive modeling exercises, where effective diffusivities based on grout performance data available at the time, groundwater at the 100-m down-gradient well exceeded the allowable maximum permissible concentrations for 99Tc. (900 pCi/L). Recent relatively short-term (63 day) leach tests conducted on both LAW and secondary waste Cast Stone monoliths indicated that 99Tc diffusivities were at or near diffusivities where the groundwater at the 100-m down-gradient well would exceed the allowable maximum permissible 99Tc concentrations. There is, therefore, a need and an opportunity to improve the retention of Tc in the Cast Stone waste form. One method to improve the performance of the Cast Stone waste form is through the addition of “getters” that selectively sequester Tc inside Cast Stone.« less

Comparison of Free and Immobilized L-asparaginase Synthesized by Gamma-Irradiated Penicillium cyclopium.

PubMed

El-Refai, Heba A; Shafei, Mona S; Mostafa, Hanan; El-Refai, Abdel-Monem H; Araby, Eman M; El-Beih, Fawkia M; Easa, Saadia M; Gomaa, Sanaa K

2016-01-01

Gamma irradiation is used on Penicillium cyclopium in order to obtain mutant cells of high L-asparaginase productivity. Using gamma irradiation dose of 4 KGy, P. cyclopium cells yielded L-asparaginase with extracellular enzyme activity of 210.8 ± 3 U/ml, and specific activity of 752.5 ± 1.5 U/mg protein, which are 1.75 and 1.53 times, respectively, the activity of the wild strain. The enzyme was partially purified by 40-60% acetone precipitation. L-asparaginase was immobilized onto Amberlite IR-120 by ionic binding. Both free and immobilized enzymes exhibited maximum activity at pH 8 and 40 degrees C. The immobilization process improved the enzyme thermal stability significantly. The immobilized enzyme remained 100% active at temperatures up to 60 degrees C, while the free asparaginase was less tolerant to high temperatures. The immobilized enzyme was more stable at pH 9.0 for 50 min, retaining 70% of its relative activity. The maximum reaction rate (V(max)) and Michaelis-Menten constant (K(m)) of the free form were significantly changed after immobilization. The K(m) value for immobilized L-asparaginase was about 1.3 times higher than that of free enzyme. The ions K+, Ba2+ and Na+ showed stimulatory effect on enzyme activity with percentages of 110%, 109% and 106% respectively.

Enhanced stability of catalase covalently immobilized on functionalized titania submicrospheres.

PubMed

Wu, Hong; Liang, Yanpeng; Shi, Jiafu; Wang, Xiaoli; Yang, Dong; Jiang, Zhongyi

2013-04-01

In this study, a novel approach combing the chelation and covalent binding was explored for facile and efficient enzyme immobilization. The unique capability of titania to chelate with catecholic derivatives at ambient conditions was utilized for titania surface functionalization. The functionalized titania was then used for enzyme immobilization. Titania submicrospheres (500-600 nm) were synthesized by a modified sol-gel method and functionalized with carboxylic acid groups through a facile chelation method by using 3-(3,4-dihydroxyphenyl) propionic acid as the chelating agent. Then, catalase (CAT) was covalently immobilized on these functionalized titania submicrospheres through 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) coupling reaction. The immobilized CAT retained 65% of its free form activity with a loading capacity of 100-150 mg/g titania. The pH stability, thermostability, recycling stability and storage stability of the immobilized CAT were evaluated. A remarkable enhancement in enzyme stability was achieved. The immobilized CAT retained 90% and 76% of its initial activity after 10 and 16 successive cycles of decomposition of hydrogen peroxide, respectively. Both the Km and the Vmax values of the immobilized CAT (27.4 mM, 13.36 mM/min) were close to those of the free CAT (25.7 mM, 13.46 mM/min). Copyright © 2012 Elsevier B.V. All rights reserved.

Solid polymer electrolyte lithium batteries

DOEpatents

Alamgir, M.; Abraham, K.M.

1993-10-12

This invention pertains to Lithium batteries using Li ion (Li[sup +]) conductive solid polymer electrolytes composed of solvates of Li salts immobilized in a solid organic polymer matrix. In particular, this invention relates to Li batteries using solid polymer electrolytes derived by immobilizing solvates formed between a Li salt and an aprotic organic solvent (or mixture of such solvents) in poly(vinyl chloride). 3 figures.

Solid polymer electrolyte lithium batteries

DOEpatents

Alamgir, Mohamed; Abraham, Kuzhikalail M.

1993-01-01

This invention pertains to Lithium batteries using Li ion (Li.sup.+) conductive solid polymer electrolytes composed of solvates of Li salts immobilized in a solid organic polymer matrix. In particular, this invention relates to Li batteries using solid polymer electrolytes derived by immobilizing solvates formed between a Li salt and an aprotic organic solvent (or mixture of such solvents) in poly(vinyl chloride).

Degradation of TCE using sequential anaerobic biofilm and aerobic immobilized bed reactor

NASA Technical Reports Server (NTRS)

Chapatwala, Kirit D.; Babu, G. R. V.; Baresi, Larry; Trunzo, Richard M.

1995-01-01

Bacteria capable of degrading trichloroethylene (TCE) were isolated from contaminated wastewaters and soil sites. The aerobic cultures were identified as Pseudomonas aeruginosa (four species) and Pseudomonas fluorescens. The optimal conditions for the growth of aerobic cultures were determined. The minimal inhibitory concentration values of TCE for Pseudomonas sps. were also determined. The aerobic cells were immobilized in calcium alginate in the form of beads. Degradation of TCE by the anaerobic and dichloroethylene (DCE) by aerobic cultures was studied using dual reactors - anaerobic biofilm and aerobic immobilized bed reactor. The minimal mineral salt (MMS) medium saturated with TCE was pumped at the rate of 1 ml per hour into the anaerobic reactor. The MMS medium saturated with DCE and supplemented with xylenes and toluene (3 ppm each) was pumped at the rate of 1 ml per hour into the fluidized air-uplift-type reactor containing the immobilized aerobic cells. The concentrations of TCE and DCE and the metabolites formed during their degradation by the anaerobic and aerobic cultures were monitored by GC. The preliminary study suggests that the anaerobic and aerobic cultures of our isolates can degrade TCE and DCE.

Immobilized lipid-bilayer materials

DOEpatents

Sasaki, Darryl Y.; Loy, Douglas A.; Yamanaka, Stacey A.

2000-01-01

A method for preparing encapsulated lipid-bilayer materials in a silica matrix comprising preparing a silica sol, mixing a lipid-bilayer material in the silica sol and allowing the mixture to gel to form the encapsulated lipid-bilayer material. The mild processing conditions allow quantitative entrapment of pre-formed lipid-bilayer materials without modification to the material's spectral characteristics. The method allows for the immobilization of lipid membranes to surfaces. The encapsulated lipid-bilayer materials perform as sensitive optical sensors for the detection of analytes such as heavy metal ions and can be used as drug delivery systems and as separation devices.

Horseradish peroxidase-nanoclay hybrid particles of high functional and colloidal stability.

PubMed

Pavlovic, Marko; Rouster, Paul; Somosi, Zoltan; Szilagyi, Istvan

2018-08-15

Highly stable dispersions of enzyme-clay nanohybrids of excellent horseradish peroxidase activity were developed. Layered double hydroxide nanoclay was synthesized and functionalized with heparin polyelectrolyte to immobilize the horseradish peroxidase enzyme. The formation of a saturated heparin layer on the platelets led to charge inversion of the positively charged bare nanoclay and to highly stable aqueous dispersions. Great affinity of the enzyme to the surface modified platelets resulted in strong horseradish peroxidase adsorption through electrostatic and hydrophobic interactions as well as hydrogen bonding network and prevented enzyme leakage from the obtained material. The enzyme kept its functional integrity upon immobilization and showed excellent activity in decomposition of hydrogen peroxide and oxidation of an aromatic compound in the test reactions. In addition, remarkable long term functional stability of the enzyme-nanoclay hybrid was observed making the developed colloidal system a promising antioxidant candidate in biomedical treatments and industrial processes. Copyright © 2018 Elsevier Inc. All rights reserved.

"Clickable" Polymeric Nanofibers through Hydrophilic-Hydrophobic Balance: Fabrication of Robust Biomolecular Immobilization Platforms.

PubMed

Kalaoglu-Altan, Ozlem I; Sanyal, Rana; Sanyal, Amitav

2015-05-11

Fabrication of hydrophilic polymeric nanofibers that undergo facile and selective functionalization through metal catalyst-free Diels-Alder "click" reaction in aqueous environment is outlined. Electrospinning of copolymers containing an electron-rich furan moiety, hydrophobic methyl methacrylate units and hydrophilic poly(ethylene glycol)s as side chains provide specifically functionalizable yet antibiofouling fibers that remain stable in aqueous media due to appropriate hydrophobic hydrophilic balance. Efficient functionalization of these nanofibers is accomplished through the Diels-Alder reaction by exposing them to maleimide-containing molecules and ligands. Diels-Alder conjugation based functionalization is demonstrated through attachment of fluorescein-maleimide and a maleimide tethered biotin ligand. Biotinylated nanofibers were utilized to mediate immobilization of the protein streptavidin, as well as streptavidin coated quantum dots. Facile fabrication from readily available polymers and their effective functionalization under mild and reagent-free conditions in aqueous media make these "clickable" nanofibers attractive candidates as functionalizable scaffolds for various biomedical applications.

Enzyme-MOF (metal-organic framework) composites.

PubMed

Lian, Xizhen; Fang, Yu; Joseph, Elizabeth; Wang, Qi; Li, Jialuo; Banerjee, Sayan; Lollar, Christina; Wang, Xuan; Zhou, Hong-Cai

2017-06-06

The ex vivo application of enzymes in various processes, especially via enzyme immobilization techniques, has been extensively studied in recent years in order to enhance the recyclability of enzymes, to minimize enzyme contamination in the product, and to explore novel horizons for enzymes in biomedical applications. Possessing remarkable amenability in structural design of the frameworks as well as almost unparalelled surface tunability, Metal-Organic Frameworks (MOFs) have been gaining popularity as candidates for enzyme immobilization platforms. Many MOF-enzyme composites have achieved unprecedented results, far outperforming free enzymes in many aspects. This review summarizes recent developments of MOF-enzyme composites with special emphasis on preparative techniques and the synergistic effects of enzymes and MOFs. The applications of MOF-enzyme composites, primarily in transferation, catalysis and sensing, are presented as well. The enhancement of enzymatic activity of the composites over free enzymes in biologically incompatible conditions is emphasized in many cases.

Facile fabrication of a stable and recyclable lipase@amine-functionalized ZIF-8 nanoparticles for esters hydrolysis and transesterification

NASA Astrophysics Data System (ADS)

Cheong, Ling-Zhi; Wei, Yayu; Wang, Hongbin; Wang, Zhiying; Su, Xiurong; Shen, Cai

2017-08-01

Zeolitic imidazolate frameworks (ZIF) represent one of the metal organic frameworks (MOF) with high potential for enzyme immobilization due to their exceptional chemical and thermal stability, negligible cytotoxicity, and easy synthesis under mild biocompatible conditions. Amine-functionalized ZIF-8 (An-ZIF-8) are capable of forming multipoint attachment via hydrogen bonding with lipase which will immobilize and further enhance stabilization of lipase. In addition, increased hydrophilicity of An-ZIF-8 will increase partitioning of An-ZIF-8 immobilized lipase at the aqueous/organic interface which enable lipase to expose its active site and retain its catalytic activity at its highest. Present study reports the use of ZIF-8 and An-ZIF-8 nanoparticles as carrier for Burkholderia cepacia lipase (BCL), compares the ester hydrolysis and transesterification activities of immobilized lipase with those of free lipase, and evaluates the reusability and recovery rate of the immobilized lipase. An-ZIF-8 nanoparticles (average 130.42 ± 0.55 nm) were facilely synthesized via mixing ZIF-8 nanoparticles with ammonia hydroxide solution. Despite having similar characteristics of high crystallinity and forming cuboid-like particles, An-ZIF-8 demonstrated significantly ( P < 0.05) lower Brunauer-Emmett-Teller (BET) surface area and higher thermal stability than ZIF-8. BCL were successfully immobilized on ZIF-8 (BCL@ZIF-8) and An-ZIF-8 (BCL@An-ZIF-8) nanoparticles with an average lipase loading rate of 8 mg/g MOF. The immobilized BCL demonstrated no significant differences in terms of esters hydrolysis and transesterification activities with those of free BCL. BCL@An-ZIF-8 demonstrated superior catalytic stability in comparison to BCL@ZIF-8 with retainment of more than 80% of its initial hydrolysis and transesterification activity for at least 10 repeated runs. In addition, more than 80% of the BCL@An-ZIF-8 can be easily recovered during each cycle of the reusability test through simple centrifugation.

The role of surfaces, chemical interfaces, and disorder on plutonium incorporation in pyrochlores

DOE PAGES

Perriot, Romain; Dholabhai, Pratik P.; Uberuaga, Blas P.

2016-07-27

Pyrochlores, a class of complex oxides with formula A 2B 2O 7, are one of the candidates for nuclear waste encapsulation, due to the natural occurrence of actinide-bearing pyrochlore minerals and laboratory observations of high radiation tolerance. In this work, we use atomistic simulations to determine the role of surfaces, chemical interfaces, and cation disorder on the plutonium immobilization properties of pyrochlores as a function of pyrochlore chemistry. We find that both Pu 3+ and Pu 4+ segregate to the surface for the four low-index pyrochlore surfaces considered, and that the segregation energy varies with the chemistry of the compound.more » We also find that pyrochlore/pyrochlore bicrystals A 2B 2O 7/A 2'B 2'O 7 can be used to immobilize Pu 3+ and Pu 4+ either in the same or separate phases of the compound, depending on the chemistry of the material. Finally, we find that Pu 4+ segregates to the disordered phase of an order/disorder bicrystal, driven by the occurrence of local oxygen-rich environments. However, Pu 3+ is weakly sensitive to the oxygen environment, and therefore only slightly favors the disordered phase. This behavior suggests that, at some concentration, Pu incorporation can destabilize the pyrochlore structure. Together, these results provide new insight into the ability of pyrochlore compounds to encapsulate Pu and suggest new considerations in the development of waste forms based on pyrochlores. Particularly, the phase structure of a multi-phase pyrochlore composite can be used to independently getter decay products based on their valence and size.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Perriot, Romain; Dholabhai, Pratik P.; Uberuaga, Blas P.

Pyrochlores, a class of complex oxides with formula A 2B 2O 7, are one of the candidates for nuclear waste encapsulation, due to the natural occurrence of actinide-bearing pyrochlore minerals and laboratory observations of high radiation tolerance. In this work, we use atomistic simulations to determine the role of surfaces, chemical interfaces, and cation disorder on the plutonium immobilization properties of pyrochlores as a function of pyrochlore chemistry. We find that both Pu 3+ and Pu 4+ segregate to the surface for the four low-index pyrochlore surfaces considered, and that the segregation energy varies with the chemistry of the compound.more » We also find that pyrochlore/pyrochlore bicrystals A 2B 2O 7/A 2'B 2'O 7 can be used to immobilize Pu 3+ and Pu 4+ either in the same or separate phases of the compound, depending on the chemistry of the material. Finally, we find that Pu 4+ segregates to the disordered phase of an order/disorder bicrystal, driven by the occurrence of local oxygen-rich environments. However, Pu 3+ is weakly sensitive to the oxygen environment, and therefore only slightly favors the disordered phase. This behavior suggests that, at some concentration, Pu incorporation can destabilize the pyrochlore structure. Together, these results provide new insight into the ability of pyrochlore compounds to encapsulate Pu and suggest new considerations in the development of waste forms based on pyrochlores. Particularly, the phase structure of a multi-phase pyrochlore composite can be used to independently getter decay products based on their valence and size.« less

Enzyme Engineering for In Situ Immobilization.

PubMed

Rehm, Fabian B H; Chen, Shuxiong; Rehm, Bernd H A

2016-10-14

Enzymes are used as biocatalysts in a vast range of industrial applications. Immobilization of enzymes to solid supports or their self-assembly into insoluble particles enhances their applicability by strongly improving properties such as stability in changing environments, re-usability and applicability in continuous biocatalytic processes. The possibility of co-immobilizing various functionally related enzymes involved in multistep synthesis, conversion or degradation reactions enables the design of multifunctional biocatalyst with enhanced performance compared to their soluble counterparts. This review provides a brief overview of up-to-date in vitro immobilization strategies while focusing on recent advances in enzyme engineering towards in situ self-assembly into insoluble particles. In situ self-assembly approaches include the bioengineering of bacteria to abundantly form enzymatically active inclusion bodies such as enzyme inclusions or enzyme-coated polyhydroxyalkanoate granules. These one-step production strategies for immobilized enzymes avoid prefabrication of the carrier as well as chemical cross-linking or attachment to a support material while the controlled oriented display strongly enhances the fraction of accessible catalytic sites and hence functional enzymes.

Immobilization of Paecilomyces variotii tannase and properties of the immobilized enzyme.

PubMed

Schons, Patrícia Fernanda; Lopes, Fernanda Cristina Rezende; Battestin, Vania; Macedo, Gabriela Alves

2011-01-01

Tannase produced by Paecilomyces variotii was encapsulated in sodium alginate beads and used for the effective hydrolysis of tannic acid; the efficiency of hydrolysis was comparable to that of the free enzyme. The alginate beads retained 100% of their efficiency in the first three rounds of successive use and 60% in rounds 4 and 5. The response surface methodology showed that the best conditions to hydrolysis of tannic acid by immobilized tannase were: sodium alginate 5.2%, CaCl₂ 0.55 M and 9 h to curing time. The optimized process resulted in 2.4 times more hydrolysed tannic acid than that obtained before optimization. The optimum pH for the actions of both the encapsulated and the free enzymes was 5.5. The optimum temperature of the reaction was determined to be 40 °C for the free enzyme and 60 °C for the immobilized form. The immobilization process improved the stability at low pH.

Synthesis of carbohydrates in a continuous flow reactor by immobilized phosphatase and aldolase.

PubMed

Babich, Lara; Hartog, Aloysius F; van Hemert, Lieke J C; Rutjes, Floris P J T; Wever, Ron

2012-12-01

Herein, we report a new flow process with immobilized enzymes to synthesize complex chiral carbohydrate analogues from achiral inexpensive building blocks in a three-step cascade reaction. The first reactor contained immobilized acid phosphatase, which phosphorylated dihydroxyacetone to dihydroxyacetone phosphate using pyrophosphate as the phosphate donor. The second flow reactor contained fructose-1,6-diphosphate aldolase (RAMA, rabbit muscle aldolase) or rhamnulose-1-phosphate aldolase (RhuA from Thermotoga maritima) and acid phosphatase. The immobilized aldolases coupled the formed dihydroxyacetone phosphate to aldehydes, resulting in phosphorylated carbohydrates. A final reactor containing acid phosphatase that dephosphorylated the phosphorylated product yielded the final product. Different aldehydes were used to synthesize carbohydrates on a gram scale. To demonstrate the feasibility of the flow systems, we synthesized 0.6 g of the D-fagomine precursor. By using immobilized aldolase RhuA we were also able to obtain other stereoisomers of the D-fagomine precursor. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Halloysite Clay Nanotubes for Enzyme Immobilization.

PubMed

Tully, Joshua; Yendluri, Raghuvara; Lvov, Yuri

2016-02-08

Halloysite clay is an aluminosilicate nanotube formed by rolling flat sheets of kaolinite clay. They have a 15 nm lumen, 50-70 nm external diameter, length of 0.5-1 μm, and different inside/outside chemistry. Due to these nanoscale properties, they are used for loading, storage, and controlled release of active chemical agents, including anticorrosions, biocides, and drugs. We studied the immobilization in halloysite of laccase, glucose oxidase, and lipase. Overall, negatively charged proteins taken above their isoelectric points were mostly loaded into the positively charged tube's lumen. Typical tube loading with proteins was 6-7 wt % from which one-third was released in 5-10 h and the other two-thirds remained, providing enhanced biocatalysis in nanoconfined conditions. Immobilized lipase showed enhanced stability at acidic pH, and the optimum pH shifted to more alkaline pH. Immobilized laccase was more stable with respect to time, and immobilized glucose oxidase showed retention of enzymatic activity up to 70 °C, whereas the native sample was inactive.

Immobilization of metals in contaminated soils using natural polymer-based stabilizers.

PubMed

Tao, Xue; Li, Aimin; Yang, Hu

2017-03-01

Three low-cost natural polymer materials, namely, lignin (Ln), carboxymethyl cellulose, and sodium alginate, were used for soil amendment to immobilize lead and cadmium in two contaminated soil samples collected from a mining area in Nanjing, China. The remediation effects of the aforementioned natural polymers were evaluated by toxicity characteristic leaching procedure (TCLP) and sequential extractions. The stabilizers could lower the bioavailability of Pb and Cd in the contaminated soils, and the amount of the exchangeable forms of the aforementioned two metals were reduced evidently. TCLP results showed that the leaching concentrations of Pb and Cd were decreased by 5.46%-71.1% and 4.25%-49.6%, respectively, in the treated soils. The contents of the organic forms of the two metals both increased with the increase in stabilizer dose on the basis of the redistribution of metal forms by sequential extractions. These findings were due to the fact that the abundant oxygen-containing groups on the polymeric amendments were effective in chelating and immobilizing Pb and Cd, which have been further confirmed from the metal adsorptions in aqueous solutions. Moreover, Ln achieved the greatest effect among the three polymers under study because of the former's distinct three-dimensional molecular structure, showing the preferential immobilization of Pb over Cd in soils also. Thus, the above-mentioned natural polymers hold great application potentials for reducing metal ion entry into the food chain at a field scale. Copyright © 2016 Elsevier Ltd. All rights reserved.

Development of a dual-analyte fluorescent sensor for the determination of bioactive nitrite and selenite in water samples.

PubMed

Martínez-Tomé, M J; Esquembre, R; Mallavia, R; Mateo, C R

2010-01-20

Nitrite and selenium are two bioactive compounds found in the environment which show beneficial effects for health at low levels but have toxic effects at higher doses. Consequently, quantification of both analytes in water samples results of great interest in areas such as biomedicine, food technology and environmental analysis. In a recent paper, we immobilized the inclusion complex formed between 2,3-diaminonaphthalene (DAN) and 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) in a sol-gel matrix, in order to prepare a highly sensitive reagentless fluorescence-based sensor for the specific measurement of nitrite. Here we have explored the possibility of using the sol-gel immobilized complex to quantify selenite (Se (IV)), the more toxic form of selenium, as well as to act as a dual-analyte chemical sensor for simultaneous quantification of both nitrite and selenite in aqueous samples. Results show that (a) inclusion of DAN in HP-beta-CD and its subsequent immobilization in a sol-gel matrix do not modify the reactivity of DAN against selenite, (b) the reaction product formed (4,5-benzopiazselenol) remains into the cyclodextrin increasing considerably its fluorescence quantum yield and avoiding, therefore, its extraction into organic solvents, (c) the developed sensor can detect selenite concentrations at submicromolar level with a minimum detection limit of 13 nM, (d) the immobilized system is able to simultaneously quantify nitrite and selenite at submicromolar concentrations in natural water samples with no further sample pre-treatment.

Simultaneous immobilization of borate, arsenate, and silicate from geothermal water derived from mining activity by co-precipitation with hydroxyapatite.

PubMed

Sasaki, Keiko; Hayashi, Yoshikazu; Toshiyuki, Kenta; Guo, Binglin

2018-09-01

The treatment of the geothermal water discharged through mining activity is a critical issue because the rate of discharge is 12,000 m 3 per day and the discharge contains high concentrations of borate (>20 mg/L) and arsenate (ca. 0.4 mg/L) as well as silicate and carbonate. The simultaneous reduction of borate and arsenate concentrations to acceptable levels was successfully performed by co-precipitation with hydroxyapatite (HAp). Although the coexisting high concentrations of carbonate act as a disturbing element, the co-precipitation equilibrium of borate was shifted to lower values by adjusting the P/Ca molar ratio, and the removal rate of borate was accelerated by using Al 3+ additives, resulting in the efficient reduction of borate within 1 h. The initially immobilized boron in HAp is in the tetragonal form, which probably occupies the hydroxyl sites in HAp, gradually transforming into the trigonal form in the solid state, as interpreted by 1 H NMR and 11 B-NMR. The coexisting silicate was also immobilized in an ellestadite form, as confirmed by 29 Si-NMR measurements. Arsenate and silicate were immobilized before borate in geothermal water. A dissolution assay of borate in the solid residues after co-precipitation with HAp verified the acceptable stability of borate, which is independent of the amount of added Al 3+ . Copyright © 2018 Elsevier Ltd. All rights reserved.

Magnetic nanoparticles coated with polyaniline to stabilize immobilized trypsin

NASA Astrophysics Data System (ADS)

Maciel, J. C.; D. Mercês, A. A.; Cabrera, M.; Shigeyosi, W. T.; de Souza, S. D.; Olzon-Dionysio, M.; Fabris, J. D.; Cardoso, C. A.; Neri, D. F. M.; C. Silva, M. P.; Carvalho, L. B.

2016-12-01

It is reported the synthesis of magnetic nanoparticles via the chemical co-precipitation of Fe 3+ ions and their preparation by coating them with polyaniline. The electronic micrograph analysis showed that the mean diameter for the nanoparticles is ˜15 nm. FTIR, powder X-ray diffraction and Mössbauer spectroscopy were used to understand the chemical, crystallographic and 57Fe hyperfine structures for the two samples. The nanoparticles, which exhibited magnetic behavior with relatively high spontaneous magnetization at room temperature, were identified as being mainly formed by maghemite ( γFe2O3). The coated magnetic nanoparticles (sample labeled "mPANI") presented a real ability to bind biological molecules such as trypsin, forming the magnetic enzyme derivative (sample "mPANIG-Trypsin"). The amount of protein and specific activity of the immobilized trypsin were found to be 13±5 μg of protein/mg of mPANI (49.3 % of immobilized protein) and 24.1±0.7 U/mg of immobilized protein, respectively. After 48 days of storage at 4 ∘C, the activity of the immobilized trypsin was found to be 89 % of its initial activity. This simple, fast and low-cost procedure was revealed to be a promising way to prepare mPANI nanoparticles if technological applications addressed to covalently link biomolecules are envisaged. This route yields chemically stable derivatives, which can be easily recovered from the reaction mixture with a magnetic field and recyclable reused.

A motor-driven syringe-type gradient maker for forming immobilized pH gradient gels.

PubMed

Fawcett, J S; Sullivan, J V; Chidakel, B E; Chrambach, A

1988-05-01

A motor driven gradient maker based on the commercial model (Jule Inc., Trumbull, CT) was designed for immobilized pH gradient gels to provide small volumes, rapid stirring and delivery, strict volume and temperature control and air exclusion. The device was constructed and by a convenient procedure yields highly reproducible gradients either in solution or on polyacrylamide gels.

Sweat collection capsule

NASA Technical Reports Server (NTRS)

Greenleaf, J. E.; Delaplaine, R. W. (Inventor)

1980-01-01

A sweat collection capsule permitting quantitative collection of sweat is described. The device consists of a frame held immobile on the skin, a closure secured to the frame and absorbent material located next to the skin in a cavity formed by the frame and the closure. The absorbent material may be removed from the device by removing the closure from the frame while the frame is held immobile on the skin.

The fate of nitrogen mineralized from leaf litter — Initial evidence from 15N-labeled litter

Treesearch

Kathryn B. Piatek

2011-01-01

Decomposition of leaf litter includes microbial immobilization of nitrogen (N), followed by N mineralization. The fate of N mineralized from leaf litter is unknown. I hypothesized that N mineralized from leaf litter will be re-immobilized into other forms of organic matter, including downed wood. This mechanism may retain N in some forests. To test this hypothesis, oak...

Liquid secondary waste. Waste form formulation and qualification

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cozzi, A. D.; Dixon, K. L.; Hill, K. A.

The Hanford Site Effluent Treatment Facility (ETF) currently treats aqueous waste streams generated during Site cleanup activities. When the Hanford Tank Waste Treatment and Immobilization Plant (WTP) begins operations, a liquid secondary waste (LSW) stream from the WTP will need to be treated. The volume of effluent for treatment at the ETF will increase significantly. Washington River Protection Solutions is implementing a Secondary Liquid Waste Immobilization Technology Development Plan to address the technology needs for a waste form and solidification process to treat the increased volume of waste planned for disposal at the Integrated Disposal Facility IDF). Waste form testingmore » to support this plan is composed of work in the near term to demonstrate the waste form will provide data as input to a performance assessment (PA) for Hanford’s IDF.« less

Immobilization of simulated radioactive soil waste containing cerium by self-propagating high-temperature synthesis

NASA Astrophysics Data System (ADS)

Mao, Xianhe; Qin, Zhigui; Yuan, Xiaoning; Wang, Chunming; Cai, Xinan; Zhao, Weixia; Zhao, Kang; Yang, Ping; Fan, Xiaoling

2013-11-01

A simulated radioactive soil waste containing cerium as an imitator element has been immobilized by a thermite self-propagating high-temperature synthesis (SHS) process. The compositions, structures, and element leaching rates of products with different cerium contents have been characterized. To investigate the influence of iron on the chemical stability of the immobilized products, leaching tests of samples with different iron contents with different leaching solutions were carried out. The results showed that the imitator element cerium mainly forms the crystalline phases CeAl11O18 and Ce2SiO5. The leaching rate of cerium over a period of 28 days was 10-5-10-6 g/(m2 day). Iron in the reactants, the reaction products, and the environment has no significant effect on the chemical stability of the immobilized SHS products.

Immobilization of CdS nanoparticles formed in reverse micelles onto aluminosilicate supports and their photocatalytic properties.

PubMed

Hirai, Takayuki; Bando, Yoko

2005-08-15

CdS nanoparticles, prepared in reverse micellar system, were immobilized onto thiol-modified aluminosilicate particles (ASSH) by a simple operation: addition of ASSH in the micellar solution and mild stirring. The resulting CdS nanoparticles-aluminosilicate composites (ASCdS) were used as photocatalysts for H2 generation from 2-propanol aqueous solution. The chemical properties of the aluminosilicate, such as affinity for water and other reactants, were found to affect the photocatalytic property of the CdS nanoparticles immobilized. Zeolite particles, having affinity for water and 2-propanol, gave a good ASCdS photocatalyst with respect to H2 generation.

Gamma-Aminobutyric Acid Production Using Immobilized Glutamate Decarboxylase Followed by Downstream Processing with Cation Exchange Chromatography

PubMed Central

Lee, Seungwoon; Ahn, Jungoh; Kim, Yeon-Gu; Jung, Joon-Ki; Lee, Hongweon; Lee, Eun Gyo

2013-01-01

We have developed a gamma-aminobutyric acid (GABA) production technique using his-tag mediated immobilization of Escherichia coli-derived glutamate decarboxylase (GAD), an enzyme that catalyzes the conversion of glutamate to GABA. The GAD was obtained at 1.43 g/L from GAD-overexpressed E. coli fermentation and consisted of 59.7% monomer, 29.2% dimer and 2.3% tetramer with a 97.6% soluble form of the total GAD. The harvested GAD was immobilized to metal affinity gel with an immobilization yield of 92%. Based on an investigation of specific enzyme activity and reaction characteristics, glutamic acid (GA) was chosen over monosodium glutamate (MSG) as a substrate for immobilized GAD, resulting in conversion of 2.17 M GABA in a 1 L reactor within 100 min. The immobilized enzymes retained 58.1% of their initial activities after ten consecutive uses. By using cation exchange chromatography followed by enzymatic conversion, GABA was separated from the residual substrate and leached GAD. As a consequence, the glutamic acid was mostly removed with no detectable GAD, while 91.2% of GABA was yielded in the purification step. PMID:23322022

Preformulation considerations for controlled release dosage forms. Part III. Candidate form selection using numerical weighting and scoring.

PubMed

Chrzanowski, Frank

2008-01-01

Two numerical methods, Decision Analysis (DA) and Potential Problem Analysis (PPA) are presented as alternative selection methods to the logical method presented in Part I. In DA properties are weighted and outcomes are scored. The weighted scores for each candidate are totaled and final selection is based on the totals. Higher scores indicate better candidates. In PPA potential problems are assigned a seriousness factor and test outcomes are used to define the probability of occurrence. The seriousness-probability products are totaled and forms with minimal scores are preferred. DA and PPA have never been compared to the logical-elimination method. Additional data were available for two forms of McN-5707 to provide complete preformulation data for five candidate forms. Weight and seriousness factors (independent variables) were obtained from a survey of experienced formulators. Scores and probabilities (dependent variables) were provided independently by Preformulation. The rankings of the five candidate forms, best to worst, were similar for all three methods. These results validate the applicability of DA and PPA for candidate form selection. DA and PPA are particularly applicable in cases where there are many candidate forms and where each form has some degree of unfavorable properties.

Site-Specific, Covalent Immobilization of Dehalogenase ST2570 Catalyzed by Formylglycine-Generating Enzymes and Its Application in Batch and Semi-Continuous Flow Reactors.

PubMed

Jian, Hui; Wang, Yingwu; Bai, Yan; Li, Rong; Gao, Renjun

2016-07-11

Formylglycine-generating enzymes can selectively recognize and oxidize cysteine residues within the sulfatase sub motif at the terminus of proteins to form aldehyde-bearing formylglycine (FGly) residues, and are normally used in protein labeling. In this study, an aldehyde tag was introduced to proteins using formylglycine-generating enzymes encoded by a reconstructed set of the pET28a plasmid system for enzyme immobilization. The haloacid dehalogenase ST2570 from Sulfolobus tokodaii was used as a model enzyme. The C-terminal aldehyde-tagged ST2570 (ST2570CQ) exhibited significant enzymological properties, such as new free aldehyde groups, a high level of protein expression and improved enzyme activity. SBA-15 has widely been used as an immobilization support for its large surface and excellent thermal and chemical stability. It was functionalized with amino groups by aminopropyltriethoxysilane. The C-terminal aldehyde-tagged ST2570 was immobilized to SBA-15 by covalent binding. The site-specific immobilization of ST2570 avoided the chemical denaturation that occurs in general covalent immobilization and resulted in better fastening compared to physical adsorption. The site-specific immobilized ST2570 showed 3-fold higher thermal stability, 1.2-fold higher catalytic ability and improved operational stability than free ST2570. The site-specific immobilized ST2570 retained 60% of its original activity after seven cycles of batch operation, and it was superior to the ST2570 immobilized to SBA-15 by physical adsorption, which loses 40% of its original activity when used for the second time. It is remarkable that the site-specific immobilized ST2570 still retained 100% of its original activity after 10 cycles of reuse in the semi-continuous flow reactor. Overall, these results provide support for the industrial-scale production and application of site-specific, covalently immobilized ST2570.

Immobilization of plutonium from solutions on porous matrices by the method of high temperature sorption

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nardova, A.K.; Filippov, E.A.; Glagolenko, Y.B.

1996-05-01

This report presents the results of investigations of plutonium immobilization from solutions on inorganic matrices with the purpose of producing a solid waste form. High-temperature sorption is described which entails the adsorption of radionuclides from solutions on porous, inorganic matrices, as for example silica gel. The solution is brought to a boil with additional thermal process (calcination) of the saturated granules.

[The action of bemitil on the self-regulation systems during short-term immobilization].

PubMed

Kirichek, L T; Bobkov, Iu G

1991-01-01

In the immobilized rats bemithyl (50 mg/kg in a single dose) was shown to normalize the state of the musculomotor and cardiovascular systems exerting the positive influence on the key links of the process of autoregulation in the form of the sedative (the central nervous system), stress-protective (hormonal regulation) and antihypoxic (metabolism) effects that characterizes it as the drug with the distinct antistress activity.

Enhanced heavy metal immobilization in soil by grinding with addition of nanometallic Ca/CaO dispersion mixture.

PubMed

Mallampati, Srinivasa Reddy; Mitoma, Yoshiharu; Okuda, Tetsuji; Sakita, Shogo; Kakeda, Mitsunori

2012-10-01

This study investigated the use of a nanometallic Ca and CaO dispersion mixture for the immobilization of heavy metals (As, Cd, Cr and Pb) in contaminated soil. Simple grinding achieved 85-90% heavy metal immobilization, but it can be enhanced further to 98-100% by addition of a nanometallic Ca/CaO dispersion mixture produced by grinding. Observations using SEM-EDS elemental maps and semi-quantitative analysis showed that the amounts of As, Cd, Cr, and Pb measurable on the soil particle surface decrease after nanometallic Ca/CaO treatment. The leachable heavy metal concentrations were reduced after nanometallic Ca/CaO treatment to concentrations lower than the Japan soil elution standard regulatory threshold: <0.01 mg L(-1) for As, Cd, and Pb; and 0.05 mg L(-1) for Cr. Effects of soil moisture and pH on heavy metal immobilization were not strongly influenced. The most probable mechanisms for the enhancement of heavy metal immobilization capacity with nanometallic Ca/CaO treatment might be due to adsorption and entrapment of heavy metals into newly formed aggregates, thereby prompting aggregation of soil particles and enclosure/binding with Ca/CaO-associated immobile salts. Results suggest that the nanometallic Ca/CaO mixture is suitable for use in immobilization of heavy-metal-contaminated soil under normal moisture conditions. Copyright © 2012 Elsevier Ltd. All rights reserved.

Enhancement of Surfactin and Fengycin Production by Bacillus mojavensis A21: Application for Diesel Biodegradation.

PubMed

Hmidet, Noomen; Ben Ayed, Hanen; Jacques, Philippe; Nasri, Moncef

2017-01-01

This work concerns the study of the enhancement of surfactin and fengycin production by B. mojavensis A21 and application of the produced product in diesel biodegradation. The influences of the culture medium and cells immobilization were studied. The highest lipopeptides production was achieved after 72 hours of incubation in a culture medium containing 30 g/L glucose as carbon source and a combination of yeast extract (1 g/L) and glutamic acid (5 g/L) as nitrogen sources with initial pH 7.0 at 30°C and 90% volumetric aeration. The study of primary metabolites production showed mainly the production of acetoin, with a maximum production after 24 h of strain growth. The use of immobilized cells seemed to be a promising method for improving lipopeptides productivity. In fact, the synthesis of both lipopeptides, mainly fengycin, was greatly enhanced by the immobilization of A21 cells. An increase of diesel degradation capacity of approximately 20, 27, and 40% in the presence of 0.5, 1, and 2 g/L of produced lipopeptides, respectively, was observed. Considering these properties, B. mojavensis A21 strain producing a lipopeptide mixture, containing both surfactin and fengycin, may be considered as a potential candidate for future use in bioremediation and crop protection.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vienna, John D.; Todd, Terry A.; Gray, Kimberly D.

The U.S. Department of Energy, Office of Nuclear Energy has chartered an effort to develop technologies to enable safe and cost effective recycle of commercial used nuclear fuel (UNF) in the U.S. Part of this effort includes the evaluation of exiting waste management technologies for effective treatment of wastes in the context of current U.S. regulations and development of waste forms and processes with significant cost and/or performance benefits over those existing. This study summarizes the results of these ongoing efforts with a focus on the highly radioactive primary waste streams. The primary streams considered and the recommended waste formsmore » include: •Tritium separated from either a low volume gas stream or a high volume water stream. The recommended waste form is low-water cement in high integrity containers. •Iodine-129 separated from off-gas streams in aqueous processing. There are a range of potentially suitable waste forms. As a reference case, a glass composite material (GCM) formed by the encapsulation of the silver Mordenite (AgZ) getter material in a low-temperature glass is assumed. A number of alternatives with distinct advantages are also considered including a fused silica waste form with encapsulated nano-sized AgI crystals. •Carbon-14 separated from LWR fuel treatment off-gases and immobilized as a CaCO3 in a cement waste form. •Krypton-85 separated from LWR and SFR fuel treatment off-gases and stored as a compressed gas. •An aqueous reprocessing high-level waste (HLW) raffinate waste which is immobilized by the vitrification process in one of three forms: a single phase borosilicate glass, a borosilicate based glass ceramic, or a multi-phased titanate ceramic [e.g., synthetic rock (Synroc)]. •An undissolved solids (UDS) fraction from aqueous reprocessing of LWR fuel that is either included in the borosilicate HLW glass or is immobilized in the form of a metal alloy in the case of glass ceramics or titanate ceramics. •Zirconium-based LWR fuel cladding hulls and stainless steel (SS) fuel assembly hardware that are washed and super-compacted for disposal or as an alternative Zr purification and reuse (or disposal as low-level waste, LLW) by reactive gas separations. •Electrochemical process salt HLW which is immobilized in a glass bonded Sodalite waste form known as the ceramic waste form (CWF). •Electrochemical process UDS and SS cladding hulls which are melted into an iron based alloy waste form. Mass and volume estimates for each of the recommended waste forms based on the source terms from a representative flowsheet are reported.« less

Evaluation of protein immobilization capacity on various carbon nanotube embedded hydrogel biomaterials.

PubMed

Derkus, Burak; Emregul, Kaan Cebesoy; Emregul, Emel

2015-11-01

This study investigates effective immobilization of proteins, an important procedure in many fields of bioengineering and medicine, using various biomaterials. Gelatin, alginate and chitosan were chosen as polymeric carriers, and applied in both their composites and nanocomposite forms in combination with carbon nanotubes (CNTs). The prepared nano/composite structures were characterized using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), thermal gravimetric analysis (TG) and contact angle analysis (CA). Electrochemical impedance spectroscopy analysis revealed gelatin composites in general to exhibit better immobilization performance relative to the native gelatin which can be attributed to enhanced film morphologies of the composite structures. Moreover, superior immobilization efficiencies were obtained with the addition of carbon nanotubes, due to their conducting and surface enhancement features, especially in the gelatin-chitosan structures due to the presence of structural active groups. Copyright © 2015 Elsevier B.V. All rights reserved.

Surface Immobilization of Molecular Electrocatalysts for Energy Conversion

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bullock, R. Morris; Das, Atanu K.; Appel, Aaron M.

2017-03-22

Electrocatalysts are critically important for a secure energy future, as they facilitate the conversion between electrical energy and chemical energy. Molecular catalysts offer precise control of their structure, and the ability to modify the substituents to understand structure-reactivity relationships that are more difficult to achieve with heterogeneous catalysts. Molecular electrocatalysts can be immobilized on surfaces by covalent bonds or through non-covalent interactions. Advantages of surface immobilization include the need for less catalyst, avoidance of bimolecular decomposition pathways, and easier determination of catalyst lifetime. Copper-catalyzed click reactions are often used to form covalent bonds to surfaces, and pi-pi stacking of pyrenemore » substituents appended to the ligand of a molecular complex is a frequently used method to achieve non-covalent surface immobilization. This mini-review highlights surface confinement of molecular electrocatalysts for reduction of O2, oxidation of H2O, production of H2, and reduction of CO2.« less

Fiber reinforced concrete: An advanced technology for LL/ML radwaste conditioning and disposal

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tchemitcheff, E.; Verdier, A.

Radioactive waste immobilization is an integral part of operations in nuclear facilities. The goal of immobilization is to contain radioactive materials in a waste form which can maintain its integrity over very long periods of time, thus effectively isolating the materials from the environment and hence from the public. This is true regardless of the activity of the waste, including low-, and medium-level waste (LLW, MLW). A multiple-year research effort by Cogema culminated in the development of a new process to immobilize nuclear waste in concrete containers reinforced with metal fibers. The fiber concrete containers satisfy all French safety requirementsmore » relating to waste immobilization and disposal, and have been certified by ANDRA, the national radioactive waste management agency. The fiber concrete containers have been fabricated on a production scale since July 1990 by Sogefibre, a jointly-owned subsidiary of SGN and Compagnie Generale des Eaux.« less

Immobilization of pectinase and lipase on macroporous resin coated with chitosan for treatment of whitewater from papermaking.

PubMed

Liu, Kai; Zhao, Guanglei; He, Beihai; Chen, Lihui; Huang, Liulian

2012-11-01

Anionic residues and pitch deposits in whitewater negatively impact the operation of paper-forming equipment. In order to remove these substances, a macroporous resin based on a methyl acrylate matrix was synthesized and coated with chitosan of various molecular weights through glutaraldehyde cross-linking. Pectinase from Bacillus licheniformis and lipase from Thermomyces lanuginosus were immobilized on the resin coated with chitosan by a Schiff base reaction. The highest hydrolysis activities of the immobilized enzymes were achieved by using chitosan with 10×10(5)DaMW for coating and 0.0025% glutaraldehyde for cross-linking chitosan. The cationic demand and pitch deposits in whitewater were reduced by 58% and 74%, respectively, when treating whitewater with immobilized dual-enzymes for 15min at 55°C and pH 7.5. This method is useful for treatment of whitewater in the papermaking industry. Copyright © 2012 Elsevier Ltd. All rights reserved.

Process for immobilizing plutonium into vitreous ceramic waste forms

DOEpatents

Feng, Xiangdong; Einziger, Robert E.

1997-01-01

Disclosed is a method for converting spent nuclear fuel and surplus plutonium into a vitreous ceramic final waste form wherein spent nuclear fuel is bound in a crystalline matrix which is in turn bound within glass.

Process for immobilizing plutonium into vitreous ceramic waste forms

DOEpatents

Feng, X.; Einziger, R.E.

1997-08-12

Disclosed is a method for converting spent nuclear fuel and surplus plutonium into a vitreous ceramic final waste form wherein spent nuclear fuel is bound in a crystalline matrix which is in turn bound within glass.

Process for immobilizing plutonium into vitreous ceramic waste forms

DOEpatents

Feng, X.; Einziger, R.E.

1997-01-28

Disclosed is a method for converting spent nuclear fuel and surplus plutonium into a vitreous ceramic final waste form wherein spent nuclear fuel is bound in a crystalline matrix which is in turn bound within glass.

Surface modification of silk fibroin fabric using layer-by-layer polyelectrolyte deposition and heparin immobilization for small-diameter vascular prostheses.

PubMed

Elahi, M Fazley; Guan, Guoping; Wang, Lu; Zhao, Xinzhe; Wang, Fujun; King, Martin W

2015-03-03

There is an urgent need to develop a biologically active implantable small-diameter vascular prosthesis with long-term patency. Silk-fibroin-based small-diameter vascular prosthesis is a promising candidate having higher patency rate; however, the surface modification is indeed required to improve its further hemocompatibility. In this study, silk fibroin fabric was modified by a two-stage process. First, the surface of silk fibroin fabric was coated using a layer-by-layer polyelectrolyte deposition technique by stepwise dipping the silk fibroin fabric into a solution of cationic poly(allylamine hydrochloride) (PAH) and anionic poly(acrylic acid) (PAA) solution. The dipping procedure was repeated to obtain the PAH/PAA multilayers deposited on the silk fibroin fabrics. Second, the polyelectrolyte-deposited silk fibroin fabrics were treated in EDC/NHS-activated low-molecular-weight heparin (LMWH) solution at 4 °C for 24 h, resulting in immobilization of LMWH on the silk fibroin fabrics surface. Scanning electron microscopy, atomic force microscopy, and energy-dispersive X-ray data revealed the accomplishment of LMWH immobilization on the polyelectrolyte-deposited silk fibroin fabric surface. The higher the number of PAH/PAA coating layers on the silk fibroin fabric, the more surface hydrophilicity could be obtained, resulting in a higher fetal bovine serum protein and platelets adhesion resistance properties when tested in vitro. In addition, compared with untreated sample, the surface-modified silk fibroin fabrics showed negligible loss of bursting strength and thus reveal the acceptability of polyelectrolytes deposition and heparin immobilization approach for silk-fibroin-based small-diameter vascular prostheses modification.

Self-immobilization and/or thermal treatment for preparing silica-poly(methyloctylsiloxane) stationary phases.

PubMed

Bottoli, Carla B G; Vigna, Camila R M; Fischer, Gerd; Albert, Klaus; Collins, Kenneth E; Collins, Carol H

2004-03-19

Batches of poly(methyloctylsiloxane) (PMOS)-loaded silica were prepared by the deposition of PMOS, into the pores of HPLC silica. Portions of PMOS-loaded silica were allowed to remain at ambient temperature, without further treatment for 2, 9, 20, 31, 51, 105 and 184 days after preparation to undergo self-immobilization (irreversible adsorption of a layer of polymer on silica at ambient temperature in the absence of initiators). Other portions were subjected to a thermal treatment (100 degrees C for 4h) after 1, 2, 5, 7, 9, 15, 20, 25, 70, 111 and 184 days. Self-immobilized and thermally treated samples were characterized by % C, 29Si cross-polarization magic angle spinning (CP/MAS) NMR spectroscopy and reversed-phase column performance. The results show that thermal immobilization accelerates the distribution and rearrangement of the polymer on the silica surface. However, from the time that a monolayer has been formed by self-immobilization (approximately 100 days for PMOS on Kromasil silica), the thermal treatment does not alter this configuration and, thus, does not change the resulting chromatographic parameters.

Synthesis of galactooligosaccharides by CBD fusion β-galactosidase immobilized on cellulose.

PubMed

Lu, Lili; Xu, Shuze; Zhao, Renfei; Zhang, Dayu; Li, Zhengyi; Li, Yumei; Xiao, Min

2012-07-01

The β-galactosidase gene (bgaL3) was cloned from Lactobacillus bulgaricus L3 and fused with cellulose binding domain (CBD) using pET-35b (+) vector in Escherichia coli. The resulting fusion protein (CBD-BgaL3) was directly adsorbed onto microcrystalline cellulose with a high immobilization efficiency of 61%. A gram of cellulose was found to absorb 97.6 U of enzyme in the solution containing 100mM NaCl (pH 5.8) at room temperature for 20 min. The enzymatic and transglycosylation characteristics of the immobilized CBD-BgaL3 were similar to the free form. Using the immobilized enzyme as the catalyst, the yield of galactooligosaccharides (GOS) reached a maximum of 49% (w/w) from 400 g/L lactose (pH 7.6) at 45 °C for 75 min, with a high productivity of 156.8 g/L/h. Reusability assay was subsequently performed under the same reaction conditions. The immobilized enzyme could retain over 85% activity after twenty batches with the GOS yields all above 40%. Copyright © 2012 Elsevier Ltd. All rights reserved.

SOLIDIFICATION OF THE HANFORD LAW WASTE STREAM PRODUCED AS A RESULT OF NEAR-TANK CONTINUOUS SLUDGE LEACHING AND SODIUM HYDROXIDE RECOVERY

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reigel, M.; Johnson, F.; Crawford, C.

2011-09-20

The U.S. Department of Energy (DOE), Office of River Protection (ORP), is responsible for the remediation and stabilization of the Hanford Site tank farms, including 53 million gallons of highly radioactive mixed wasted waste contained in 177 underground tanks. The plan calls for all waste retrieved from the tanks to be transferred to the Waste Treatment Plant (WTP). The WTP will consist of three primary facilities including pretreatment facilities for Low Activity Waste (LAW) to remove aluminum, chromium and other solids and radioisotopes that are undesirable in the High Level Waste (HLW) stream. Removal of aluminum from HLW sludge canmore » be accomplished through continuous sludge leaching of the aluminum from the HLW sludge as sodium aluminate; however, this process will introduce a significant amount of sodium hydroxide into the waste stream and consequently will increase the volume of waste to be dispositioned. A sodium recovery process is needed to remove the sodium hydroxide and recycle it back to the aluminum dissolution process. The resulting LAW waste stream has a high concentration of aluminum and sodium and will require alternative immobilization methods. Five waste forms were evaluated for immobilization of LAW at Hanford after the sodium recovery process. The waste forms considered for these two waste streams include low temperature processes (Saltstone/Cast stone and geopolymers), intermediate temperature processes (steam reforming and phosphate glasses) and high temperature processes (vitrification). These immobilization methods and the waste forms produced were evaluated for (1) compliance with the Performance Assessment (PA) requirements for disposal at the IDF, (2) waste form volume (waste loading), and (3) compatibility with the tank farms and systems. The iron phosphate glasses tested using the product consistency test had normalized release rates lower than the waste form requirements although the CCC glasses had higher release rates than the quenched glasses. However, the waste form failed to meet the vapor hydration test criteria listed in the WTP contract. In addition, the waste loading in the phosphate glasses were not as high as other candidate waste forms. Vitrification of HLW waste as borosilicate glass is a proven process; however the HLW and LAW streams at Hanford can vary significantly from waste currently being immobilized. The ccc glasses show lower release rates for B and Na than the quenched glasses and all glasses meet the acceptance criterion of < 4 g/L. Glass samples spiked with Re{sub 2}O{sub 7} also passed the PCT test. However, further vapor hydration testing must be performed since all the samples cracked and the test could not be performed. The waste loading of the iron phosphate and borosilicate glasses are approximately 20 and 25% respectively. The steam reforming process produced the predicted waste form for both the high and low aluminate waste streams. The predicted waste loadings for the monolithic samples is approximately 39%, which is higher than the glass waste forms; however, at the time of this report, no monolithic samples were made and therefore compliance with the PA cannot be determined. The waste loading in the geopolymer is approximately 40% but can vary with the sodium hydroxide content in the waste stream. Initial geopolymer mixes revealed compressive strengths that are greater than 500 psi for the low aluminate mixes and less than 500 psi for the high aluminate mixes. Further work testing needs to be performed to formulate a geopolymer waste form made using a high aluminate salt solution. A cementitious waste form has the advantage that the process is performed at ambient conditions and is a proven process currently in use for LAW disposal. The Saltstone/Cast Stone formulated using low and high aluminate salt solutions retained at least 97% of the Re that was added to the mix as a dopant. While this data is promising, additional leaching testing must be performed to show compliance with the PA. Compressive strength tests must also be performed on the Cast Stone monoliths to verify PA compliance. Based on testing performed for this report, the borosilicate glass and Cast Stone are the recommended waste forms for further testing. Both are proven technologies for radioactive waste disposal and the initial testing using simulated Hanford LAW waste shows compliance with the PA. Both are resistant to leaching and have greater than 25% waste loading.« less

Arsenite and Ferrous Iron Oxidation Linked to Chemolithotrophic Denitrification for the Immobilization of Arsenic in Anoxic Environments

PubMed Central

Sun, Wenjie; Sierra-Alvarez, Reyes; Milner, Lily; Oremland, Ron; Field, Jim A.

2014-01-01

The objective of this study was to explore a bioremediation strategy based on injecting NO3− to support the anoxic oxidation of ferrous iron (Fe(II)) and arsenite (As(III)) in the subsurface as a means to immobilize As in the form of arsenate (As(V)) adsorbed onto biogenic ferric (Fe(III)) (hydr)oxides. Continuous flow sand filled columns were used to simulate a natural anaerobic groundwater and sediment system with co-occurring As(III) and Fe(II) in the presence (SF1) or absence (SF2) of nitrate, respectively. During operation for 250 days, the average influent arsenic concentration of 567 µg l−1 was reduced to 10.6 (±9.6) µg l−1 in the effluent of column SF1. The cumulative removal of Fe(II) and As(III) in SF1 was 6.5–10-fold higher than that in SF2. Extraction and measurement of the mass of iron and arsenic immobilized on the sand packing of the columns was close to the iron and arsenic removed from the aqueous phase during column operation. The dominant speciation of the immobilized iron and arsenic was Fe(III) and As(V) in SF1, compared with Fe(II) and As(III) in SF2. The speciation was confirmed by XRD and XPS. The results indicate that microbial oxidation of As(III) and Fe(II) linked to denitrification resulted in the enhanced immobilization of aqueous arsenic in anaerobic environments by forming Fe(III) (hydr)oxides coated sands with adsorbed As(V). PMID:19764221

Secondary Waste Form Down Selection Data Package – Ceramicrete

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cantrell, Kirk J.; Westsik, Joseph H.

2011-08-31

As part of high-level waste pretreatment and immobilized low activity waste processing, liquid secondary wastes will be generated that will be transferred to the Effluent Treatment Facility on the Hanford Site for further treatment. These liquid secondary wastes will be converted to stable solid waste forms that will be disposed in the Integrated Disposal Facility. Currently, four waste forms are being considered for stabilization and solidification of the liquid secondary wastes. These waste forms are Cast Stone, Ceramicrete, DuraLith, and Fluidized Bed Steam Reformer. The preferred alternative will be down selected from these four waste forms. Pacific Northwest National Laboratorymore » is developing data packages to support the down selection process. The objective of the data packages is to identify, evaluate, and summarize the existing information on the four waste forms being considered for stabilization and solidification of the liquid secondary wastes. The information included will be based on information available in the open literature and from data obtained from testing currently underway. This data package is for the Ceramicrete waste form. Ceramicrete is a relatively new engineering material developed at Argonne National Laboratory to treat radioactive and hazardous waste streams (e.g., Wagh 2004; Wagh et al. 1999a, 2003; Singh et al. 2000). This cement-like waste form can be used to treat solids, liquids, and sludges by chemical immobilization, microencapsulation, and/or macroencapsulation. The Ceramicrete technology is based on chemical reaction between phosphate anions and metal cations to form a strong, dense, durable, low porosity matrix that immobilizes hazardous and radioactive contaminants as insoluble phosphates and microencapsulates insoluble radioactive components and other constituents that do not form phosphates. Ceramicrete is a type of phosphate-bonded ceramic, which are also known as chemically bonded phosphate ceramics. The Ceramicrete binder is formed through an acid-base reaction between calcined magnesium oxide (MgO; a base) and potassium hydrogen phosphate (KH{sub 2}PO{sub 4}; an acid) in aqueous solution. The reaction product sets at room temperature to form a highly crystalline material. During the reaction, the hazardous and radioactive contaminants also react with KH{sub 2}PO{sub 4} to form highly insoluble phosphates. In this data package, physical property and waste acceptance data for Ceramicrete waste forms fabricated with wastes having compositions that were similar to those expected for secondary waste effluents, as well as secondary waste effluent simulants from the Hanford Tank Waste Treatment and Immobilization Plant were reviewed. With the exception of one secondary waste form formulation (25FA+25 W+1B.A. fabricated with the mixed simulant did not meet the compressive strength requirement), all the Ceramicrete waste forms that were reviewed met or exceeded Integrated Disposal Facility waste acceptance criteria.« less

Immobilization of antibacterial metallic cations (Ga3+, Zn2+ and Co2+) in a polypyrrole coating formed on Nitinol.

PubMed

Saugo, M; Brugnoni, L I; Flamini, D O; Saidman, S B

2018-05-01

Gallium, zinc and cobalt species were immobilized in hollow rectangular-sectioned microtubes of polypyrrole (PPy) electrosynthesized on Nitinol (NiTi) alloy by means of two different methods. One of them involved the immobilization after the PPy electropolymerization and the other one during the electrosynthesis process. The antibacterial activity of the coating against Escherichia coli (E. coli) was evaluated and the best results were obtained with gallium species. Characterization results demonstrated that gallium is incorporated into the PPy matrix as Ga 3+ ions. The PPy film with gallium species incorporated during the electropolymerization exhibited a good corrosion protection performance. Copyright © 2018 Elsevier B.V. All rights reserved.

Novel microfabrication stage allowing for one-photon and multi-photon light assisted molecular immobilization and for multi-photon microscope

NASA Astrophysics Data System (ADS)

Gonçalves, Odete; Snider, Scott; Zadoyan, Ruben; Nguyen, Quoc-Thang; Vorum, Henrik; Petersen, Steffen B.; Neves-Petersen, Maria Teresa

2017-02-01

Light Assisted Molecular Immobilization (LAMI) results in spatially oriented and localized covalent coupling of biomolecules onto thiol reactive surfaces. LAMI is possible due to the conserved spatial proximity between aromatic residues and disulfide bridges in proteins. When aromatic residues are excited with UV light (275-295nm), disulphide bridges are disrupted and the formed thiol groups covalently bind to surfaces. Immobilization hereby reported is achieved in a microfabrication stage coupled to a fs-laser, through one- or multi-photon excitation. The fundamental 840nm output is tripled to 280nm and focused onto the sample, leading to one-photon excitation and molecular immobilization. The sample rests on a xyz-stage with micrometer step resolution and is illuminated according to a pattern uploaded to the software controlling the stage and the shutter. Molecules are immobilized according to such pattern, with micrometer spatial resolution. Spatial masks inserted in the light path lead to light diffraction patterns used to immobilize biomolecules with submicrometer spatial resolution. Light diffraction patterns are imaged by an inbuilt microscope. Two-photon microscopy and imaging of the fluorescent microbeads is shown. Immobilization of proteins, e.g. C-reactive protein, and of an engineered molecular beacon has been successfully achieved. The beacon was coupled to a peptide containing a disulfide bridge neighboring a tryptophan residue, being this way possible to immobilize the beacon on a surface using one-photon LAMI. This technology is being implemented in the creation of point-of-care biosensors aiming at the detection of cancer and cardiovascular disease markers.

Anti-infiltration for fabrication of a suspended nanoparticle layer on porous close-packed colloidal arrays.

PubMed

Teh, Lay K; Yan, Qingfeng; Wong, Chee C

2009-04-01

We develop a new method to fabricate suspended sheets of nanocrystals (NCs) on porous surfaces. The method relies on the resistance of an aqueous suspension droplet to infiltrate a porous network; hence, the method is named anti-infiltration. The process works by combining fluid dynamics of a liquid droplet during impact/absorption onto a porous surface with the convective self-assembly of NCs. The immobilization of the liquid droplet edge due to the self-assembly of NCs at the meniscus is harnessed to halt the lateral spreading of the droplet and, consequently, the capillary penetration of the liquid immediately after droplet impact. Further capillary penetration of the liquid is drastically reduced because of the competition between capillary forces and convective losses as well as the rapid occlusion of the pores as soon as a continuous NC film has formed upon evaporation of the suspension. This method holds promise for a wide variety of optoelectronic, sensing, and separation membrane applications. As an example, we demonstrate that these suspended NC layers are suitable candidates as planar defects embedded within a colloidal photonic crystal.

Immobilized humic substances and immobilized aggregates of humic substances as sorbent for solid phase extraction.

PubMed

Erny, Guillaume L; Gonçalves, Bruna M; Esteves, Valdemar I

2013-09-06

In this work, humic substances (HS) immobilized, as a thin layer or as aggregates, on silica gel were tested as material for solid phase extraction. Some triazines (simazine, atrazine, therbutylazine, atrazine-desethyl-desisopropyl-2-hydroxy, ametryn and terbutryn), have been selected as test analytes due to their environmental importance and to span a large range of solubility and octanol/water partition coefficient (logP). The sorbent was obtained immobilizing a thin layer of HS via physisorption on a pre-coated silica gel with a cationic polymer (polybrene). While the sorbent could be used as it is, it was demonstrated that additional HS could be immobilized, via weak interactions, to form stable humic aggregates. However, while a higher quantity of HS could be immobilized, no significant differences were observed in the sorption parameters. This sorbent have been tested for solid phase extraction to concentrate triazines from aqueous matrixes. The sorbent demonstrated performances equivalent to commercial alternatives as a concentration factor between 50 and 200, depending on the type of triazines, was obtained. Moreover the low cost and the high flow rate of sample through the column allowed using high quantity of sorbent. The analytical procedure was tested with different matrixes including tap water, river water and estuarine water. Copyright © 2013 Elsevier B.V. All rights reserved.

Optimization and characterization of biomolecule immobilization on silicon substrates using (3-aminopropyl)triethoxysilane (APTES) and glutaraldehyde linker

NASA Astrophysics Data System (ADS)

Gunda, Naga Siva Kumar; Singh, Minashree; Norman, Lana; Kaur, Kamaljit; Mitra, Sushanta K.

2014-06-01

In the present work, we developed and optimized a technique to produce a thin, stable silane layer on silicon substrate in a controlled environment using (3-aminopropyl)triethoxysilane (APTES). The effect of APTES concentration and silanization time on the formation of silane layer is studied using spectroscopic ellipsometry and Fourier transform infrared spectroscopy (FTIR). Biomolecules of interest are immobilized on optimized silane layer formed silicon substrates using glutaraldehyde linker. Surface analytical techniques such as ellipsometry, FTIR, contact angle measurement system, and atomic force microscopy are employed to characterize the bio-chemically modified silicon surfaces at each step of the biomolecule immobilization process. It is observed that a uniform, homogenous and highly dense layer of biomolecules are immobilized with optimized silane layer on the silicon substrate. The developed immobilization method is successfully implemented on different silicon substrates (flat and pillar). Also, different types of biomolecules such as anti-human IgG (rabbit monoclonal to human IgG), Listeria monocytogenes, myoglobin and dengue capture antibodies were successfully immobilized. Further, standard sandwich immunoassay (antibody-antigen-antibody) is employed on respective capture antibody coated silicon substrates. Fluorescence microscopy is used to detect the respective FITC tagged detection antibodies bound to the surface after immunoassay.

Indirect immobilized Jagged1 suppresses cell cycle progression and induces odonto/osteogenic differentiation in human dental pulp cells.

PubMed

Manokawinchoke, Jeeranan; Nattasit, Praphawi; Thongngam, Tanutchaporn; Pavasant, Prasit; Tompkins, Kevin A; Egusa, Hiroshi; Osathanon, Thanaphum

2017-08-31

Notch signaling regulates diverse biological processes in dental pulp tissue. The present study investigated the response of human dental pulp cells (hDPs) to the indirect immobilized Notch ligand Jagged1 in vitro. The indirect immobilized Jagged1 effectively activated Notch signaling in hDPs as confirmed by the upregulation of HES1 and HEY1 expression. Differential gene expression profiling using an RNA sequencing technique revealed that the indirect immobilized Jagged1 upregulated genes were mainly involved in extracellular matrix organization, disease, and signal transduction. Downregulated genes predominantly participated in the cell cycle, DNA replication, and DNA repair. Indirect immobilized Jagged1 significantly reduced cell proliferation, colony forming unit ability, and the number of cells in S phase. Jagged1 treated hDPs exhibited significantly higher ALP enzymatic activity, osteogenic marker gene expression, and mineralization compared with control. Pretreatment with a γ-secretase inhibitor attenuated the Jagged1-induced ALP activity and mineral deposition. NOTCH2 shRNA reduced the Jagged1-induced osteogenic marker gene expression, ALP enzymatic activity, and mineral deposition. In conclusion, indirect immobilized Jagged1 suppresses cell cycle progression and induces the odonto/osteogenic differentiation of hDPs via the canonical Notch signaling pathway.

Optimal immobilization of β-galactosidase onto κ-carrageenan gel beads using response surface methodology and its applications.

PubMed

Elnashar, Magdy M; Awad, Ghada E; Hassan, Mohamed E; Mohy Eldin, Mohamed S; Haroun, Bakry M; El-Diwany, Ahmed I

2014-01-01

β-Galactosidase (β-gal) was immobilized by covalent binding on novel κ-carrageenan gel beads activated by two-step method; the gel beads were soaked in polyethyleneimine followed by glutaraldehyde. 2(2) full-factorial central composite experiment designs were employed to optimize the conditions for the maximum enzyme loading efficiency. 11.443 U of enzyme/g gel beads was achieved by soaking 40 units of enzyme with the gel beads for eight hours. Immobilization process increased the pH from 4.5 to 5.5 and operational temperature from 50 to 55 °C compared to the free enzyme. The apparent K(m) after immobilization was 61.6 mM compared to 22.9 mM for free enzyme. Maximum velocity Vmax was 131.2 μ mol · min(-1) while it was 177.1 μ mol · min(-1) for free enzyme. The full conversion experiment showed that the immobilized enzyme form is active as that of the free enzyme as both of them reached their maximum 100% relative hydrolysis at 4 h. The reusability test proved the durability of the κ-carrageenan beads loaded with β -galactosidase for 20 cycles with retention of 60% of the immobilized enzyme activity to be more convenient for industrial uses.

Aroma Release in Wine Using Co-Immobilized Enzyme Aggregates.

PubMed

Ahumada, Katherine; Martínez-Gil, Ana; Moreno-Simunovic, Yerko; Illanes, Andrés; Wilson, Lorena

2016-11-08

Aroma is a remarkable factor of quality and consumer preference in wine, representing a distinctive feature of the product. Most aromatic compounds in varietals are in the form of glycosidic precursors, which are constituted by a volatile aglycone moiety linked to a glucose residue by an O -glycosidic bond; glucose is often linked to another sugar (arabinose, rhamnose or apiose). The use of soluble β-glycosidases for aroma liberation implies the addition of a precipitating agent to remove it from the product and precludes its reuse after one batch. An attractive option from a technological perspective that will aid in removing such constraints is the use of immobilized glycosidases. Immobilization by aggregation and crosslinking is a simple strategy producing enzyme catalysts of very high specific activity, being an attractive option to conventional immobilization to solid inert supports. The purpose of this work was the evaluation of co-immobilized β-glycosidases crosslinked aggregates produced from the commercial preparation AR2000, which contains the enzymes involved in the release of aromatic terpenes in Muscat wine (α-l-arabinofuranosidase and β-d-glucopyranosidase). To do so, experiments were conducted with co-immobilized crosslinked enzyme aggregates (combi-CLEAs), and with the soluble enzymes, using an experiment without enzyme addition as control. Stability of the enzymes at the conditions of winemaking was assessed and the volatiles composition of wine was determined by SPE-GC-MS. Stability of enzymes in combi-CLEAs was much higher than in soluble form, 80% of the initial activity remaining after 60 days in contact with the wine; at the same conditions, the soluble enzymes had lost 80% of their initial activities after 20 days. Such higher stabilities will allow prolonged use of the enzyme catalyst reducing its impact in the cost of winemaking. Wine treated with combi-CLEAs was the one exhibiting the highest concentration of total terpenes (18% higher than the control) and the highest concentrations of linalool (20% higher), nerol (20% higher) and geraniol (100% higher), which are the most important terpenes in determining Muscat typicity. Co-immobilized enzymes were highly stable at winemaking conditions, so their reutilization is possible and technologically attractive by reducing the impact of enzyme cost on winemaking cost.

Transport of fine sediment over a coarse, immobile riverbed

USGS Publications Warehouse

Grams, Paul E.; Wilcock, Peter R.

2014-01-01

Sediment transport in cobble-boulder rivers consists mostly of fine sediment moving over a coarse, immobile bed. Transport rate depends on several interrelated factors: boundary shear stress, the grain size and volume of fine sediment, and the configuration of fine sediment into interstitial deposits and bed forms. Existing models do not incorporate all of these factors. Approaches that partition stress face a daunting challenge because most of the boundary shear is exerted on immobile grains. We present an alternative approach that divides the bed into sand patches and interstitial deposits and is well constrained by two clear end-member cases: full sand cover and absence of sand. Entrainment from sand patches is a function of their aerial coverage. Entrainment from interstices among immobile grains is a function of sand elevation relative to the size of the immobile grains. The bed-sand coverage function is used to predict the ratio of the rate of entrainment from a partially covered bed to the rate of entrainment from a completely sand-covered bed, which is determined using a standard sand transport model. We implement the bed-sand coverage function in a morphodynamic routing model and test it against observations of sand bed elevation and suspended sand concentration for conditions of nonuniform fine sediment transport in a large flume with steady uniform flow over immobile hemispheres. The results suggest that this approach may provide a simple and robust method for predicting the transport and migration of fine sediment through rivers with coarse, immobile beds.

Antimicrobial and cell viability measurement of bovine serum albumin capped silver nanoparticles (Ag/BSA) loaded collagen immobilized poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) film.

PubMed

Bakare, Rotimi; Hawthrone, Samantha; Vails, Carmen; Gugssa, Ayele; Karim, Alamgir; Stubbs, John; Raghavan, Dharmaraj

2016-03-01

Bacterial infection of orthopedic devices has been a major concern in joint replacement procedures. Therefore, this study is aimed at formulating collagen immobilized poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) film loaded with bovine serum albumin capped silver nanoparticles (Ag/BSA NPs) to inhibit bacterial growth while retaining/promoting osteoblast cells viability. The nanoparticles loaded collagen immobilized PHBV film was characterized for its composition by X-ray Photoelectron Spectroscopy and Anodic Stripping Voltammetry. The extent of loading of Ag/BSA NPs on collagen immobilized PHBV film was found to depend on the chemistry of the functionalized PHBV film and the concentration of Ag/BSA NPs solution used for loading nanoparticles. Our results showed that more Ag/BSA NPs were loaded on higher molecular weight collagen immobilized PHEMA-g-PHBV film. Maximum loading of Ag/BSA NPs on collagen immobilized PHBV film was observed when 16ppm solution was used for adsorption studies. Colony forming unit and optical density measurements showed broad antimicrobial activity towards Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa at significantly lower concentration i.e., 0.19 and 0.31μg/disc, compared to gentamicin and sulfamethoxazole trimethoprim while MTT assay showed that released nanoparticles from Ag/BSA NPs loaded collagen immobilized PHBV film has no impact on MCTC3-E1 cells viability. Copyright © 2015 Elsevier Inc. All rights reserved.

Method to synthesize dense crystallized sodalite pellet for immobilizing halide salt radioactive waste

DOEpatents

Koyama, Tadafumi

1994-01-01

A method for immobilizing waste chloride salts containing radionuclides such as cesium and strontium and hazardous materials such as barium. A sodalite intermediate is prepared by mixing appropriate amounts of silica, alumina and sodium hydroxide with respect to sodalite and heating the mixture to form the sodalite intermediate and water. Heating is continued to drive off the water to form a water-free intermediate. The water-free intermediate is mixed with either waste salt or waste salt which has been contacted with zeolite to concentrate the radionuclides and hazardous material. The waste salt-intermediate mixture is then compacted and heated under conditions of heat and pressure to form sodalite with the waste salt, radionuclides and hazardous material trapped within the sodalite cage structure. This provides a final product having excellent leach resistant capabilities.

Method to synthesize dense crystallized sodalite pellet for immobilizing halide salt radioactive waste

DOEpatents

Koyama, Tadafumi.

1994-08-23

A method is described for immobilizing waste chloride salts containing radionuclides such as cesium and strontium and hazardous materials such as barium. A sodalite intermediate is prepared by mixing appropriate amounts of silica, alumina and sodium hydroxide with respect to sodalite and heating the mixture to form the sodalite intermediate and water. Heating is continued to drive off the water to form a water-free intermediate. The water-free intermediate is mixed with either waste salt or waste salt which has been contacted with zeolite to concentrate the radionuclides and hazardous material. The waste salt-intermediate mixture is then compacted and heated under conditions of heat and pressure to form sodalite with the waste salt, radionuclides and hazardous material trapped within the sodalite cage structure. This provides a final product having excellent leach resistant capabilities.

Method to synthesize dense crystallized sodalite pellet for immobilizing halide salt radioactive waste

DOEpatents

Koyama, T.

1992-01-01

This report describes a method for immobilizing waste chloride salts containing radionuclides such as cesium and strontium and hazardous materials such as barium. A sodalite intermediate is prepared by mixing appropriate amounts of silica, alumina and sodium hydroxide with respect to sodalite and heating the mixture to form the sodalite intermediate and water. Heating is continued to drive off the water to form a water-free intermediate. The water-free intermediate is mixed with either waste salt or waste salt which has been contacted with zeolite to concentrate the radionuclides and hazardous material. The waste salt-intermediate mixture is then compacted and heated under conditions of heat and pressure to form sodalite with the waste salt, radionuclides and hazardous material trapped within the sodalite cage structure. This provides a final product having excellent leach resistant capabilities.

Retinal detachment and cataract, facial dysmorphism, generalized osteoporosis, immobile spine and platyspondyly in a consanguinous kindred--a possible new syndrome.

PubMed

Schmidt, H; Rudolph, G; Hergersberg, M; Schneider, K; Moradi, S; Meitinger, T

2001-02-01

We report on a consanguineous family with 6 children (out of 7) affected by a spondylo-ocular syndrome. Clinical features include cataract, loss of vision due to retinal detachment, facial dysmorphism, facial hypotonia, normal height with disproportional short trunk, immobile spine with thorakal kyphosis and reduced lumbal lordosis. On ophthalmological examination of the index patient, a dense cataract and complete retinal detachment could be detected on the right eye. On the left eye, an absent lens nucleus was found, but no retinal detachment. On radiological examination, there was generalized moderate osteoporosis; the spine showed marked platyspondyly and the bone age was advanced. On laboratory investigations, a normal excretion of amino acids, mucopolysaccharides and oligosaccharides could be found. The phenotypical spectrum observed in the 6 affected individuals was rather uniform. The karyotype was normal in all affected children. This hitherto undescribed combination of oculo-skeletal symptoms shows most resemblance with connective tissue disorders, suggesting a range of candidate genes for mutation analysis.

Effects of temperature and relative humidity on the stability of paper-immobilized antibodies.

PubMed

Wang, Jingyun; Yiu, Brian; Obermeyer, Jaclyn; Filipe, Carlos D M; Brennan, John D; Pelton, Robert

2012-02-13

The stability of a paper-immobilized antibody was investigated over a range of temperatures (40-140 °C) and relative humidities (RH, 30-90%) using both unmodified filter paper and the same paper impregnated with polyamide-epichlorohydrin (PAE) as supports. Antibody stability decreased with increasing temperature, as expected, but also decreased with increasing RH. At 40 °C, the half-life was more than 10 days, with little dependence on RH. However, at 80 °C, the half-life varied from ~3 days at low RH to less than half an hour at 90% RH, demonstrating that hydration of the antibody promotes unfolding. Antibody stability was not influenced by the PAE paper surface treatment. This work shows that antibodies are good candidates for development of bioactive paper as they have sufficient stability at high temperature to withstand printing and other roll-to-roll processing steps, and sufficient low temperature stability to allow long-term storage of bioactive paper materials.

Poly High Internal Phase Emulsion for the Immobilization of Chemical Warfare Agents.

PubMed

Wright, Alexander J; Main, Marcus J; Cooper, Nicholas J; Blight, Barry A; Holder, Simon J

2017-09-20

We report a facile method for the absorption (characterized by the weight/weight swelling degree, Q) of a variety of chemical warfare agents (CWAs); including sulfur mustard (HD) (Q = 40) and V-series (VM, VX, i-Bu-VX, n-Bu-VX) of nerve agents (Q ≥ 45) and a simulant, methyl benzoate (Q = 55), through the use of a poly(styrene-co-vinyl benzyl chloride-co-divinylbenzene) lightly cross-linked poly high internal phase emulsion (polyHIPE). By varying the vinyl benzyl chloride (VBC) content and the volume of the internal phase of the precursor emulsion it is demonstrated that absorption is facilitated both by the swelling of the polymer and the uptake of liquid in the pores. In particular the sample prepared from a 95% internal emulsion water content showed rapid swelling (<5 min to total absorption) and the ability to swell both from a monolithic state and from a compressed state, making these systems ideal practical candidates for the rapid immobilization of CWAs.

Bioethanol production from the nutrient stress-induced microalga Chlorella vulgaris by enzymatic hydrolysis and immobilized yeast fermentation.

PubMed

Kim, Kyoung Hyoun; Choi, In Seong; Kim, Ho Myeong; Wi, Seung Gon; Bae, Hyeun-Jong

2014-02-01

The microalga Chlorella vulgaris is a potential feedstock for bioenergy due to its rapid growth, carbon dioxide fixation efficiency, and high accumulation of lipids and carbohydrates. In particular, the carbohydrates in microalgae make them a candidate for bioethanol feedstock. In this study, nutrient stress cultivation was employed to enhance the carbohydrate content of C. vulgaris. Nitrogen limitation increased the carbohydrate content to 22.4% from the normal content of 16.0% on dry weight basis. In addition, several pretreatment methods and enzymes were investigated to increase saccharification yields. Bead-beating pretreatment increased hydrolysis by 25% compared with the processes lacking pretreatment. In the enzymatic hydrolysis process, the pectinase enzyme group was superior for releasing fermentable sugars from carbohydrates in microalgae. In particular, pectinase from Aspergillus aculeatus displayed a 79% saccharification yield after 72h at 50°C. Using continuous immobilized yeast fermentation, microalgal hydrolysate was converted into ethanol at a yield of 89%. Copyright © 2013 Elsevier Ltd. All rights reserved.

ZnS nanoparticles electrodeposited onto ITO electrode as a platform for fabrication of enzyme-based biosensors of glucose.

PubMed

Du, Jian; Yu, Xiuping; Wu, Ying; Di, Junwei

2013-05-01

The electrochemical and photoelectrochemical biosensors based on glucose oxidase (GOD) and ZnS nanoparticles modified indium tin oxide (ITO) electrode were investigated. The ZnS nanoparticles were electrodeposited directly on the surface of ITO electrode. The enzyme was immobilized on ZnS/ITO electrode surface by sol-gel method to fabricate glucose biosensor. GOD could electrocatalyze the reduction of dissolved oxygen, which resulted in a great increase of the reduction peak current. The reduction peak current decreased linearly with the addition of glucose, which could be used for glucose detection. Moreover, ZnS nanoparticles deposited on ITO electrode surface showed good photocurrent response under illumination. A photoelectrochemical biosensor for the detection of glucose was also developed by monitoring the decreases in the cathodic peak photocurrent. The results indicated that ZnS nanoparticles deposited on ITO substrate were a good candidate material for the immobilization of enzyme in glucose biosensor construction. Copyright © 2013 Elsevier B.V. All rights reserved.

Oxidative particle mixtures for groundwater treatment

DOEpatents

Siegrist, Robert L.; Murdoch, Lawrence C.

2000-01-01

The invention is a method and a composition of a mixture for degradation and immobilization of contaminants in soil and groundwater. The oxidative particle mixture and method includes providing a material having a minimal volume of free water, mixing at least one inorganic oxidative chemical in a granular form with a carrier fluid containing a fine grained inorganic hydrophilic compound and injecting the resulting mixture into the subsurface. The granular form of the inorganic oxidative chemical dissolves within the areas of injection, and the oxidative ions move by diffusion and/or advection, therefore extending the treatment zone over a wider area than the injection area. The organic contaminants in the soil and groundwater are degraded by the oxidative ions, which form solid byproducts that can sorb significant amounts of inorganic contaminants, metals, and radionuclides for in situ treatment and immobilization of contaminants. The method and composition of the oxidative particle mixture for long-term treatment and immobilization of contaminants in soil and groundwater provides for a reduction in toxicity of contaminants in a subsurface area of contamination without the need for continued injection of treatment material, or for movement of the contaminants, or without the need for continuous pumping of groundwater through the treatment zone, or removal of groundwater from the subsurface area of contamination.

Supported liquid inorganic membranes for nuclear waste separation

DOEpatents

Bhave, Ramesh R; DeBusk, Melanie M; DelCul, Guillermo D; Delmau, Laetitia H; Narula, Chaitanya K

2015-04-07

A system and method for the extraction of americium from radioactive waste solutions. The method includes the transfer of highly oxidized americium from an acidic aqueous feed solution through an immobilized liquid membrane to an organic receiving solvent, for example tributyl phosphate. The immobilized liquid membrane includes porous support and separating layers loaded with tributyl phosphate. The extracted solution is subsequently stripped of americium and recycled at the immobilized liquid membrane as neat tributyl phosphate for the continuous extraction of americium. The sequestered americium can be used as a nuclear fuel, a nuclear fuel component or a radiation source, and the remaining constituent elements in the aqueous feed solution can be stored in glassified waste forms substantially free of americium.

Tyrosinase-catalyzed site-specific immobilization of engineered C-phycocyanin to surface

PubMed Central

Faccio, Greta; Kämpf, Michael M.; Piatti, Chiara; Thöny-Meyer, Linda; Richter, Michael

2014-01-01

Enzymatic crosslinking of proteins is often limited by the steric availability of the target residues, as of tyrosyl side chains in the case of tyrosinase. Carrying an N-terminal peptide-tag containing two tyrosine residues, the fluorescent protein C-phycocyanin HisCPC from Synechocystis sp. PCC6803 was crosslinked to fluorescent high-molecular weight forms with tyrosinase. Crosslinking with tyrosinase in the presence of L-tyrosine produced non fluorescent high-molecular weight products. Incubated in the presence of tyrosinase, HisCPC could also be immobilized to amino-modified polystyrene beads thus conferring a blue fluorescence. Crosslinking and immobilization were site-specific as both processes required the presence of the N-terminal peptide in HisCPC. PMID:24947668

Potential value of phosphate compounds in enhancing immobilization and reducing bioavailability of mixed heavy metal contaminants in shooting range soil.

PubMed

Seshadri, B; Bolan, N S; Choppala, G; Kunhikrishnan, A; Sanderson, P; Wang, H; Currie, L D; Tsang, Daniel C W; Ok, Y S; Kim, G

2017-10-01

Shooting range soils contain mixed heavy metal contaminants including lead (Pb), cadmium (Cd), and zinc (Zn). Phosphate (P) compounds have been used to immobilize these metals, particularly Pb, thereby reducing their bioavailability. However, research on immobilization of Pb's co-contaminants showed the relative importance of soluble and insoluble P compounds, which is critical in evaluating the overall success of in situ stabilization practice in the sustainable remediation of mixed heavy metal contaminated soils. Soluble synthetic P fertilizer (diammonium phosphate; DAP) and reactive (Sechura; SPR) and unreactive (Christmas Island; CPR) natural phosphate rocks (PR) were tested for Cd, Pb and Zn immobilization and later their mobility and bioavailability in a shooting range soil. The addition of P compounds resulted in the immobilization of Cd, Pb and Zn by 1.56-76.2%, 3.21-83.56%, and 2.31-74.6%, respectively. The reactive SPR significantly reduced Cd, Pb and Zn leaching while soluble DAP increased their leachate concentrations. The SPR reduced the bioaccumulation of Cd, Pb and Zn in earthworms by 7.13-23.4% and 14.3-54.6% in comparison with earthworms in the DAP and control treatment, respectively. Bioaccessible Cd, Pb and Zn concentrations as determined using a simplified bioaccessibility extraction test showed higher long-term stability of P-immobilized Pb and Zn than Cd. The differential effect of P-induced immobilization between P compounds and metals is due to the variation in the solubility characteristics of P compounds and nature of metal phosphate compounds formed. Therefore, Pb and Zn immobilization by P compounds is an effective long-term remediation strategy for mixed heavy metal contaminated soils. Copyright © 2017 Elsevier Ltd. All rights reserved.

EPS production by Propionibacterium freudenreichii facilitates its immobilization for propionic acid production.

PubMed

Belgrano, F D S; Verçoza, B R F; Rodrigues, J C F; Hatti-Kaul, R; Pereira, N

2018-04-28

Immobilization of microbial cells is a useful strategy for developing high cell density bioreactors with improved stability and productivity for production of different chemicals. Functionalization of the immobilization matrix or biofilm forming property of some strains has been utilized for achieving cell attachment. The aim of the present study was to investigate the production of exopolysaccharide (EPS) by Propionibacterium freudenreichii C.I.P 59.32 and utilize this feature for immobilization of the cells on porous glass beads for production of propionic acid. Propionibacterium freudenreichii was shown to produce both capsular and excreted EPS during batch cultivations using glucose as carbon source. Different electron microscopy techniques confirmed the secretion of EPS and formation of cellular aggregates. The excreted EPS was mainly composed of mannose and glucose in a 5·3 : 1 g g -1 ratio. Immobilization of the cells on untreated and polyethyleneimine (PEI)-treated Poraver beads in a bioreactor was evaluated. Higher productivity and yield of propionic acid (0·566 g l -1  h -1 and 0·314 g g -1 , respectively) was achieved using cells immobilized to untreated beads and EPS production reached 617·5 mg l -1 after 48 h. These results suggest an important role of EPS-producing strains for improving cell immobilization and propionic acid production. This study demonstrates the EPS-producing microbe to be easily immobilized on a solid matrix and to be used in a bioprocess. Such a system could be optimized for achieving high cell density in fermentations without the need for functionalization of the matrix. © 2018 The Society for Applied Microbiology.

Arsenite and ferrous iron oxidation linked to chemolithotrophic denitrification for the immobilization of arsenic in anoxic environments

USGS Publications Warehouse

Sun, W.; Sierra-Alvarez, R.; Milner, L.; Oremland, R.; Field, J.A.

2009-01-01

The objective of this study was to explore a bioremediation strategy based on injecting NO3- to support the anoxic oxidation of ferrous iron (Fe(II)) and arsenite (As(III)) in the subsurface as a means to immobilize As in the form of arsenate (As(V)) adsorbed onto biogenic ferric (Fe(III)) (hydr)oxides. Continuous flows and filled columns were used to simulate a natural anaerobic groundwater and sediment system with co-occurring As(III) and Fe(II) in the presence (column SF1) or absence (column SF2) of nitrate, respectively. During operation for 250 days, the average influent arsenic concentration of 567 ??g L-1 was reduced to 10.6 (??9.6) ??g L-1 in the effluent of column SF1. The cumulative removal of Fe(II) and As(III) in SF1 was 6.5 to 10-fold higher than that in SF2. Extraction and measurement of the mass of iron and arsenic immobilized on the sand packing of the columns were close to the iron and arsenic removed from the aqueous phase during column operation. The dominant speciation of the immobilized iron and arsenic was Fe(III) and As(V) in SF1, compared with Fe(II) and As(III) in SF2. The speciation was confirmed by X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The results indicate that microbial oxidation of As(III) and Fe(II) linked to denitrification resulted in the enhanced immobilization of aqueous arsenic in anaerobic environments by forming Fe(III) (hydr)oxide coated sands with adsorbed As(V). ?? 2009 American Chemical Society.

Characterization and Leaching Tests of the Fluidized Bed Steam Reforming (FBSR) Waste Form for LAW Immobilization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Neeway, James J.; Qafoku, Nikolla; Brown, Christopher F.

2013-10-01

Several supplemental technologies for treating and immobilizing Hanford low activity waste (LAW) have been evaluated. One such immobilization technology is the Fluidized Bed Steam Reforming (FBSR) granular product. The FBSR granular product is composed of insoluble sodium aluminosilicate (NAS) feldspathoid minerals. Production of the FBSR mineral product has been demonstrated both at the industrial and laboratory scale. Pacific Northwest National Laboratory (PNNL) was involved in an extensive characterization campaign. This goal of this campaign was study the durability of the FBSR mineral product and the mineral product encapsulated in a monolith to meet compressive strength requirements. This paper gives anmore » overview of results obtained using the ASTM C 1285 Product Consistency Test (PCT), the EPA Test Method 1311 Toxicity Characteristic Leaching Procedure (TCLP), and the ASTMC 1662 Single-Pass Flow-Through (SPFT) test. Along with these durability tests an overview of the characteristics of the waste form has been collected using Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), microwave digestions for chemical composition, and surface area from Brunauer, Emmett, and Teller (BET) theory.« less

Characterization and Leaching Tests of the Fluidized Bed Steam Reforming (FBSR) Waste Form for LAW Immobilization - 13400

DOE Office of Scientific and Technical Information (OSTI.GOV)

Neeway, James J.; Qafoku, Nikolla P.; Peterson, Reid A.

2013-07-01

Several supplemental technologies for treating and immobilizing Hanford low activity waste (LAW) have been evaluated. One such immobilization technology is the Fluidized Bed Steam Reforming (FBSR) granular product. The FBSR granular product is composed of insoluble sodium aluminosilicate (NAS) feldspathoid minerals. Production of the FBSR mineral product has been demonstrated both at the industrial and laboratory scale. Pacific Northwest National Laboratory (PNNL) was involved in an extensive characterization campaign. The goal of this campaign was to study the durability of the FBSR mineral product and the encapsulated FBSR product in a geo-polymer monolith. This paper gives an overview of resultsmore » obtained using the ASTM C 1285 Product Consistency Test (PCT), the EPA Test Method 1311 Toxicity Characteristic Leaching Procedure (TCLP), and the ASTMC 1662 Single-Pass Flow-Through (SPFT) test. Along with these durability tests an overview of the characteristics of the waste form has been collected using Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), microwave digestions for chemical composition, and surface area from Brunauer, Emmett, and Teller (BET) theory. (authors)« less

DNA-directed trypsin immobilization on a polyamidoamine dendrimer-modified capillary to form a renewable immobilized enzyme microreactor.

PubMed

Wu, Nan; Wang, Siming; Yang, Ye; Song, Jiayi; Su, Ping; Yang, Yi

2018-07-01

A novel type of trypsin capillary microreactor was developed based on a DNA-directed immobilization (DDI) technique applied to a fused-silica capillary modified with polyamidoamine (PAMAM) dendrimers. Trypsin binding to the inner wall of the capillary was confirmed by confocal laser scanning microscopy. The properties of the trypsin-DNA conjugated, PAMAM-modified capillary microreactor were investigated by monitoring hydrolysis of Nα-benzoyl- L -arginine ethyl ester. Through the hybridization and dehybridization of the DNA, the inner wall of the capillary functionalized with trypsin can be regenerated, thus indicating the renewability of this enzyme microreactor. In addition, these results demonstrated that introduction of PAMAM enabled higher amounts of trypsin to be immobilized, markedly improving the enzymolysis efficiency, compared with traditional modified capillaries. The digestion performance of the trypsin capillary microreactor was further evaluated by digesting cytochrome C, and a peptide numbers of 8, and a sequence coverage of 59% were obtained. This renewable and efficient immobilized trypsin capillary microreactor combines advantages of both DDI technology and PAMAM, and is potentially adaptable to high-throughput enzyme assays in biochemical and clinical research. Copyright © 2018. Published by Elsevier B.V.

Immobilization-stabilization of proteins on nanofibrillated cellulose derivatives and their bioactive film formation.

PubMed

Arola, Suvi; Tammelin, Tekla; Setälä, Harri; Tullila, Antti; Linder, Markus B

2012-03-12

In a number of different applications for enzymes and specific binding proteins a key technology is the immobilization of these proteins to different types of supports. In this work we describe a concept for protein immobilization that is based on nanofibrillated cellulose (NFC). NFC is a form of cellulose where fibers have been disintegrated into fibrils that are only a few nanometers in diameter and have a very large aspect ratio. Proteins were conjugated through three different strategies using amine, epoxy, and carboxylic acid functionalized NFC. The conjugation chemistries were chosen according to the reactive groups on the NFC derivatives; epoxy amination, heterobifunctional modification of amino groups, and EDC/s-NHS activation of carboxylic acid groups. The conjugation reactions were performed in solution and immobilization was performed by spin coating the protein-NCF conjugates. The structure of NFC was shown to be advantageous for both protein performance and stability. The use of NFC allows all covalent chemistry to be performed in solution, while the immobilization is achieved by a simple spin coating or spreading of the protein-NFC conjugates on a support. This allows more scalable methods and better control of conditions compared to the traditional methods that depend on surface reactions.

From protein engineering to immobilization: promising strategies for the upgrade of industrial enzymes.

PubMed

Singh, Raushan Kumar; Tiwari, Manish Kumar; Singh, Ranjitha; Lee, Jung-Kul

2013-01-10

Enzymes found in nature have been exploited in industry due to their inherent catalytic properties in complex chemical processes under mild experimental and environmental conditions. The desired industrial goal is often difficult to achieve using the native form of the enzyme. Recent developments in protein engineering have revolutionized the development of commercially available enzymes into better industrial catalysts. Protein engineering aims at modifying the sequence of a protein, and hence its structure, to create enzymes with improved functional properties such as stability, specific activity, inhibition by reaction products, and selectivity towards non-natural substrates. Soluble enzymes are often immobilized onto solid insoluble supports to be reused in continuous processes and to facilitate the economical recovery of the enzyme after the reaction without any significant loss to its biochemical properties. Immobilization confers considerable stability towards temperature variations and organic solvents. Multipoint and multisubunit covalent attachments of enzymes on appropriately functionalized supports via linkers provide rigidity to the immobilized enzyme structure, ultimately resulting in improved enzyme stability. Protein engineering and immobilization techniques are sequential and compatible approaches for the improvement of enzyme properties. The present review highlights and summarizes various studies that have aimed to improve the biochemical properties of industrially significant enzymes.

Fabrication of an electrochemical platform based on the self-assembly of graphene oxide-multiwall carbon nanotube nanocomposite and horseradish peroxidase: direct electrochemistry and electrocatalysis

NASA Astrophysics Data System (ADS)

Zhang, Qian; Yang, Shaojun; Zhang, Jing; Zhang, Ling; Kang, Pingli; Li, Jinghong; Xu, Jingwei; Zhou, Hua; Song, Xi-Ming

2011-12-01

A novel hybrid nanomaterial (GO-MWNTs) was explored based on the self-assembly of multiwall carbon nanotubes (MWNTs) and graphene oxide (GO). Compared with pristine MWNTs, such a nanocomposite could be well dispersed in aqueous solution and exhibit a negative charge. Driven by the electrostatic interaction, positively charged horseradish peroxidase (HRP) could then be immobilized onto GO-MWNTs at the surface of a glassy carbon (GC) electrode to form a HRP/GO-MWNT/GC electrode under mild conditions. TEM was used to characterize the morphology of the GO-MWNT nanocomposite. UV-vis and FTIR spectra suggested that HRP was immobilized onto the hybrid matrix without denaturation. Furthermore, the immobilized HRP showed enhanced direct electron transfer for the HRP-Fe(III)/Fe(II) redox center. Based on the direct electron transfer of the immobilized HRP, the HRP/GO-MWNT/GC electrode exhibited excellent electrocatalytic behavior to the reduction of H2O2 and NaNO2, respectively. Therefore, GO-MWNTs could provide a novel and efficient platform for the immobilization and biosensing of redox enzymes, and thus may find wide potential applications in the fabrication of biosensors, biomedical devices, and bioelectronics.

Biosynthesis of Thermoresponsive Magnetic Nanoparticles by Magnetosome Display System.

PubMed

Yoshino, Tomoko; Shimada, Takumi; Ito, Yasuhito; Honda, Toru; Maeda, Yoshiaki; Matsunaga, Tadashi; Tanaka, Tsuyoshi

2018-05-16

Thermoresponsive magnetic nanoparticles (MNPs) were synthesized using a magnetosome display system. An elastin-like polypeptide decamer of VPGVG (ELP 10 ), which is hydrophobic above the transition temperature ( T t ) and can form an insoluble aggregation, was immobilized on biogenic MNPs in the magnetotactic bacterium, Magnetospirillum magneticum AMB-1. It was suggested that hydrophobicity of the MNP surface increased at 60 °C compared with 20 °C by the immobilization of ELP 10 . Size distribution analysis indicated that the immobilization of ELP 10 onto MNPs induced the increased hydrophobicity with increasing temperatures up to 60 °C, promoting aggregation of the particles by hydrophobic and magnetic interactions. These results suggest that the acceleration of magnetic collection at 60 °C was caused by particle aggregation promoted by hydrophobic interaction between ELP-MNPs. Furthermore, the immobilization of ELP on MNPs gave a quick magnetic collection at 60 °C by external magnetic field. The thermoresponsive properties will further expand the utility of biotechnological applications of biogenic MNPs.

Immobilization of Chloroperoxidase on Aminopropyl-Glass

PubMed Central

Kadima, Tenshuk A.; Pickard, Michael A.

1990-01-01

Chloroperoxidase (CPO) purified from Caldariomyces fumago CMI 89362 was covalently bound to aminopropyl-glass by using a modification of an established method. Acid-washed glass was derivatized by using aminopropyltriethoxysilane, and the enzyme was ionically bound at low ionic strength. Further treatment with glutaraldehyde covalently linked the enzyme to the glass beads in an active form. No elution of bound activity from glass beads could be detected with a variety of washings. The loading of enzyme protein to the glass beads was highest, 100 mg of CPO per g of glass, at high reaction ratios of CPO to glass, but the specific activity of the immobilized enzyme was highest, 36% of theoretical, at low enzyme-to-carrier ratios. No differences in the properties of the soluble and immobilized enzymes could be detected by a number of criteria: their pH-activity and pH-stability profiles were similar, as were their thermal stabilities. After five uses, the immobilized enzyme retained full activity between pH 6.0 and 6.7. PMID:16348352

Pore Structure and Fluoride Ion Adsorption Characteristics of Zr (IV) Surface-Immobilized Resin Prepared Using Polystyrene as a Porogen

NASA Astrophysics Data System (ADS)

Mizuki, Hidenobu; Ito, Yudai; Harada, Hisashi; Uezu, Kazuya

Zr(IV) surface-immobilized resins for removal of fluoride ion were prepared by surface template polymerization using polystyrene as a porogen. At polymerization, polystyrene was added in order to increase mesopores (2-50 nm) and macropore (>50 nm) with large macropores (around 300 nm) formed with internal aqueous phase of W⁄O emulsion. The pore structure of Zr(IV) surface-immobilized resins was evaluated by measuring specific surface area, pore volume, and pore size distribution with volumetric adsorption measurement instrument and mercury porosimeter. The adsorption isotherms were well fitted by Langmuir equation. The removal of fluoride was also carried out with column method. Zr(IV) surface-immobilized resins, using 10 g⁄L polystyrene in toluene at polymerization, possessed higher volume of not only mesopores and macropores but also large macropores. Furethermore, by adding the polystyrene with smaller molecular size, the pore volume of mesopores, macropores and large macropores was significantly increased, and the fluoride ion adsorption capacity and the column utilization also increased.

Methane storage in metal-organic frameworks.

PubMed

He, Yabing; Zhou, Wei; Qian, Guodong; Chen, Banglin

2014-08-21

Natural gas (NG), whose main component is methane, is an attractive fuel for vehicular applications. Realization of safe, cheap and convenient means and materials for high-capacity methane storage can significantly facilitate the implementation of natural gas fuelled vehicles. The physisorption based process involving porous materials offers an efficient storage methodology and the emerging porous metal-organic frameworks have been explored as potential candidates because of their extraordinarily high porosities, tunable pore/cage sizes and easily immobilized functional sites. In this view, we provide an overview of the current status of metal-organic frameworks for methane storage.

[Biosensor development in clinical analysis].

PubMed

Boitieux, J L; Desmet, G; Thomas, D

1985-01-01

The use of enzymes immobilized or as markers formed the subject of more than thousand publications in the field of industry or biomedical applications, during the last five years. Recently, some authors published works concerning immobilization of total microorganisms for catalytic purposes, others use the enzymatic activity for marking molecules involved in immunological analysis processes. Together industrial biotechnology and medical analysis laboratory are interested with the evolution of these procedures involving the activity of immobilized enzymes. Enzyme immobilization allowed the lowering of analysis costs for, in this case, the enzyme can be used several times. We take account of the two main cases which are encountered during utilization of immobilized enzymes of analytical purposes. The enzyme is used directly for the catalysed reaction or it is used as enzymatic marker. These both aspects are developed mainly for the elaboration of enzymatic and immunoenzymatic electrodes and the realization of automatic computerized devices allowing continuous estimation of numerous biological blood parameters. From these two precise examples, glucose and antigen determination, the authors show the evolution of these technologies in the field of immobilized enzymes or captors and the analysis of signals given by these electrodes requiring a computerized treatment. This new technology opens to important potentialities in the analytical field. The automatization of these devices allowing the control in real time, will probably make easier the optimization steps of procedures actually used in the biomedical sphere.

Contribution of Extracellular Polymeric Substances from Shewanella sp. HRCR-1 Biofilms to U(VI) Immobilization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cao, Bin; Ahmed, B.; Kennedy, David W.

2011-06-05

The goal of this study was to quantify the contribution of extracellular polymeric substances (EPS) in U(VI) immobilization by Shewanella sp. HRCR-1. Through comparison of U(VI) immobilization using cells with bound EPS (bEPS) and cells without EPS, we showed that i) bEPS from Shewanella sp. HRCR-1 biofilms contributed significantly to U(VI) immobilization, especially at low initial U(VI) concentrations, through both sorption and reduction; ii) bEPS could be considered as a functional extension of the cells for U(VI) immobilization and they likely play more important roles at initial U(VI) concentrations; and iii) U(VI) reduction efficiency was found to be dependent uponmore » initial U(VI) concentration and the efficiency decreased at lower concentrations. To quantify relative contribution of sorption and reduction in U(VI) immobilization by EPS fractions, we isolated loosely associated EPS (laEPS) and bEPS from Shewanella sp. HRCR-1 biofilms grown in a hollow fiber membrane biofilm reactor and tested their reactivity with U(V). We found that, when in reduced form, the isolated cell-free EPS fractions could reduce U(VI). Polysaccharides in the EPS likely contributed to U(VI) sorption and dominated reactivity of laEPS while redox active components (e.g., outer membrane c-type cytochromes), especially in bEPS, might facilitate U(VI) reduction.« less

ABIOTIC REDUCTION AND DETOXIFICATION OF CHROMATE PRESENT IN SOILS

EPA Science Inventory

Theoretical and experimental research has supported the assertion that Cr(III) species are the most stable, immobile, and nontoxic forms of chromium and that they may form rapidly when adequate reducing agents are introduced in an oxidized soil environment. The objective of this ...

Immobilization of Lipase Inhibitor on the Biopolymers from Agaricus bisporus Cell Walls

PubMed Central

2017-01-01

Summary One of the methods for curing obesity is the inclusion of some substances with the antilipase activity in the diet and thus reducing the uptake of fat components from food. The aim of this research is to provide a stabilized form of lipase inhibitor by immobilization of enzyme on the biopolymers from Agaricus bisporus cell walls. The phenolic compounds extracted from the rapeseed were considered as the lipase inhibitor. The activity of the inhibitor was considerably reduced in the gastric juice, as well as at temperatures above 37 °C and during its storage, which determined the suitability of the inhibitor for stabilization on the matrix. The effectiveness of the phenolic compound stabilization was investigated by means of immobilization on the biopolymers from Agaricus bisporus cell wall matrix. The biopolymers used were β-glucan, chitin, melanin and proteins. A number of samples, which differed both in the content of the inhibitor (from 1 to 16%) and in the ratio of biopolymers in the matrix composition, was obtained. The conditions of immobilization (temperature, duration of the process) were also varied. The expediency of obtaining the sample with the inhibitor content of 12% and matrix containing 47.9% of glucan, 18.8% of chitin, 18.8% of melanin and 11.1% of proteins was shown. The best immobilization was carried out at 20–25 °C for 30 min. Thermal analysis and infrared spectroscopy data confirmed that immobilization of the lipase inhibitor on the matrix was due to the hydrogen bonds. The immobilized inhibitor had higher pH stability and higher thermal stability than the original one. The remaining activity of the immobilized inhibitor was higher than the original one after incubation in the gastric acid and bile. The immobilized inhibitor was characterized by a low loss of activity after 12 months of storage. PMID:29540987

Enhancement of Surfactin and Fengycin Production by Bacillus mojavensis A21: Application for Diesel Biodegradation

PubMed Central

Ben Ayed, Hanen; Jacques, Philippe; Nasri, Moncef

2017-01-01

This work concerns the study of the enhancement of surfactin and fengycin production by B. mojavensis A21 and application of the produced product in diesel biodegradation. The influences of the culture medium and cells immobilization were studied. The highest lipopeptides production was achieved after 72 hours of incubation in a culture medium containing 30 g/L glucose as carbon source and a combination of yeast extract (1 g/L) and glutamic acid (5 g/L) as nitrogen sources with initial pH 7.0 at 30°C and 90% volumetric aeration. The study of primary metabolites production showed mainly the production of acetoin, with a maximum production after 24 h of strain growth. The use of immobilized cells seemed to be a promising method for improving lipopeptides productivity. In fact, the synthesis of both lipopeptides, mainly fengycin, was greatly enhanced by the immobilization of A21 cells. An increase of diesel degradation capacity of approximately 20, 27, and 40% in the presence of 0.5, 1, and 2 g/L of produced lipopeptides, respectively, was observed. Considering these properties, B. mojavensis A21 strain producing a lipopeptide mixture, containing both surfactin and fengycin, may be considered as a potential candidate for future use in bioremediation and crop protection. PMID:29082251

Screening of bacterial strains isolated from uranium mill tailings porewaters for bioremediation purposes.

PubMed

Sánchez-Castro, Iván; Amador-García, Ahinara; Moreno-Romero, Cristina; López-Fernández, Margarita; Phrommavanh, Vannapha; Nos, Jeremy; Descostes, Michael; Merroun, Mohamed L

2017-01-01

The present work characterizes at different levels a number of bacterial strains isolated from porewaters sampled in the vicinity of two French uranium tailing repositories. The 16S rRNA gene from 33 bacterial isolates, corresponding to the different morphotypes recovered, was almost fully sequenced. The resulting sequences belonged to 13 bacterial genera comprised in the phyla Firmicutes, Actinobacteria and Proteobacteria. Further characterization at physiological level and metals/metalloid tolerance provided evidences for an appropriate selection of bacterial strains potentially useful for immobilization of uranium and other common contaminants. By using High Resolution Transmission Electron Microscope (HRTEM), this potential ability to immobilize uranium as U phosphate mineral phases was confirmed for the bacterial strains Br3 and Br5 corresponding to Arthrobacter sp. and Microbacterium oxydans, respectively. Scanning Transmission Electron Microscope- High-Angle Annular Dark-Field (STEM-HAADF) analysis showed U accumulates on the surface and within bacterial cytoplasm, in addition to the extracellular space. Energy Dispersive X-ray (EDX) element-distribution maps demonstrated the presence of U and P within these accumulates. These results indicate the potential of certain bacterial strains isolated from porewaters of U mill tailings for immobilizing uranium, likely as uranium phosphates. Some of these bacterial isolates might be considered as promising candidates in the design of uranium bioremediation strategies. Copyright © 2016 Elsevier Ltd. All rights reserved.

Selective, Specific, and Versatile Personal Biosensors to Organophosphate Chemical Toxins Composed of Polyurethane Immobilized Enzymes

DTIC Science & Technology

2002-01-01

cuvette containing a stir bar and the biosensor. RESULTS ACh ChE Acetate + Choline Betaine aldehyde+H2O2 Choline oxidase O2+ Choline oxidase...O2+ H2O2 Betaine + 2H2O2 carbon electrode electrochemical detection luminol fluorescent adducts visible adducts chemiluminescence fluorescence (CPM...soluble BChE and biosensor BChE at different tem- peratures relative to 30oC. Figure 7. Top: Immobilized choline oxidase and soluble form of the

Immobilized Enzymes/Bacteria for Naval Applications - Initial Data Base.

DTIC Science & Technology

1981-05-31

system was chosen as the primary organ - izational reference. Thus, the tables attached are listed in their sequence by enzyme number according to the...abstracts presented in section 4.1 are organized in tabu- lar form. The IUB number, formal name, and common name(s) and reaction(s) catalyzed are shown...antigen control 1.1.6.9 Purification of biochemicals 1.1.6.10 Artificial organs using immobilized enzymes 1.1.7 Pharmaceuticals 1.1.7.1 Amino acid

Hydrophilic porous magnetic poly(GMA-MBAA-NVP) composite microspheres containing oxirane groups: An efficient carrier for immobilizing penicillin G acylase

NASA Astrophysics Data System (ADS)

Xue, Ping; Su, Weiguang; Gu, Yaohua; Liu, Haifeng; Wang, Julan

2015-03-01

Magnetic hydrophilic polymeric microspheres containing oxirane groups were prepared by inverse suspension polymerization of glycidyl methacrylate (GMA), N, N‧-methylene bisacrylamide (MBAA) and N-vinyl pyrrolidone (NVP) in the existence of formamide, which were denoted as magnetic poly(GMA-MBAA-NVP) microspheres. The magnetic poly(GMA-MBAA-NVP) microspheres were characterized by scanning electron microscopy (SEM), FT-IR spectroscopy, X-ray diffraction (XRD), vibrating sample magnetometer (VSM) and so on. The results showed that poly(GMA-MBAA-NVP) microspheres possessed well spherical shape, narrow size distribution, abundant porous structure, reactive oxirane groups and superparamagnetic properties. Formamide used in the present work served as a modifier, a dispersant and a porogen to form final porous polymer microspheres. The penicillin G acylase (PGA) was covalently immobilized onto the magnetic microspheres through the reaction between the amino groups of enzyme and the oxirane groups on the microspheres for producing 6-aminopenicillanic acid (6-APA). The effects of GMA/NVP ratio and crosslink density on the activity of immobilized PGA were investigated. The highest apparent activity, enzyme loading and coupling yield of immobilized PGA were 821 IU/g, 65.3 mg/g and 42.3% respectively when the mass ratio of GMA/NVP was 1:1 and crosslink density was 60%. Compared with the free PGA, immobilized PGA showed a wider range of pH value and reaction temperature. The relative activity and reaction rate of immobilized PGA remained almost constant after 20 recycles. The magnetic poly(GMA-MBAA-NVP) microspheres would be very promising carriers for immobilizing enzymes in industrial application.

THE EFFECT OF TEMPERATURE AND UNIAXIAL PRESSURE ON THE DENSIFICATION BEHAVIOR OF SILICA AEROGEL GRANULES

DOE Office of Scientific and Technical Information (OSTI.GOV)

Matyas, Josef; Robinson, Matthew J.; Fryxell, Glen E.

Materials are being developed in U.S. for the removal and immobilization of iodine from gaseous products of nuclear fuel reprocessing in support of the Fuel Cycle Technology Separations and Waste Forms Campaign. The silver-functionalized silica aerogel proved to be an excellent candidate for this treatment because of its high selectivity and sorption capacity for radioiodine and its possible conversion to a durable silica-based waste form. The present study investigated with nitrogen sorption and helium pycnometry the effect of pressureless isothermal sintering at temperatures of 900-1400°C for 2.5-90 min or isothermal hot-pressing at 1200°C for 2.5 min on densification of rawmore » and silver-functionalized silica aerogel granules. Rapid sintering was observed at 1050 and 1200°C. Only 15 min of pressureless sintering at 1200°C resulted in almost complete densification. The macropores disappeared, surface area decreased from 1114 m2/g to 25 m2/g, pore volume from 7.41 cm3/g to 0.09 cm3/g, and adsorption pore size from 18.7 to 7 nm. The skeletal density of sintered granules was similar to the bulk density of amorphous silica (2.2 g/cm3). The hot-pressing accelerated the sintering process, decreasing significantly the pore size and volume.« less

Secondary Waste Form Down-Selection Data Package—Fluidized Bed Steam Reforming Waste Form

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qafoku, Nikolla; Westsik, Joseph H.; Strachan, Denis M.

2011-09-12

The Hanford Site in southeast Washington State has 56 million gallons of radioactive and chemically hazardous wastes stored in 177 underground tanks (ORP 2010). The U.S. Department of Energy (DOE), Office of River Protection (ORP), through its contractors, is constructing the Hanford Tank Waste Treatment and Immobilization Plant (WTP) to convert the radioactive and hazardous wastes into stable glass waste forms for disposal. Within the WTP, the pretreatment facility will receive the retrieved waste from the tank farms and separate it into two treated process streams. These waste streams will be vitrified, and the resulting waste canisters will be sentmore » to offsite (high-level waste [HLW]) and onsite (immobilized low-activity waste [ILAW]) repositories. As part of the pretreatment and ILAW processing, liquid secondary wastes will be generated that will be transferred to the Effluent Treatment Facility (ETF) on the Hanford Site for further treatment. These liquid secondary wastes will be converted to stable solid waste forms that will be disposed of in the Integrated Disposal Facility (IDF). To support the selection of a waste form for the liquid secondary wastes from WTP, Washington River Protection Solutions (WRPS) has initiated secondary waste form testing work at Pacific Northwest National Laboratory (PNNL). In anticipation of a down-selection process for a waste form for the Solidification Treatment Unit to be added to the ETF, PNNL is developing data packages to support that down-selection. The objective of the data packages is to identify, evaluate, and summarize the existing information on the four waste forms being considered for stabilizing and solidifying the liquid secondary wastes. At the Hanford Site, the FBSR process is being evaluated as a supplemental technology for treating and immobilizing Hanford LAW radioactive tank waste and for treating secondary wastes from the WTP pretreatment and LAW vitrification processes.« less

Immobilization of oxalate-degrading enzymes into p(HEMA) for inhibiting encrustation on ureteral stents

NASA Astrophysics Data System (ADS)

Mellman, James Kenneth

Ureteral stents develop calcium-bearing deposits, called encrustation, that diminish their biocompatibility due to complications, such as chronic abrasion to the lumen of the ureter wall and subsequent infection. A reduction of encrustation, namely calcium oxalate, will improve the lifetime, health care costs, and infection resistance of such devices. The purpose of this research project is to study oxalate-degrading enzymes entrapped into a coating material that will control the interface to the urinary environment for ureteral stents. The coating material was a lightly crosslinked poly(2-hydroxyethyl methacrylate) (p(HEMA)) matrix in which the active enzymes were entrapped within the bulk material's free volume. The swelling of p(HEMA) films was comparable in ddH2O and urine. This hydrophilic matrix allows oxalate anions to diffuse into the bulk so that enzyme activity against oxalate can lower its local concentration, and thereby reduce the supersaturation of calcium oxalate. Oxalate oxidase (OxO) and oxalate decarboxylase (OxDc) were the oxalate-degrading enzymes examined herein. Michaelis Menten kinetic models were applied to free and immobilized enzyme activity. A substrate inhibition model was applied to OxO. The free form of OxO had a Vmax of 1.8 +/- 0.1 muM/min-mug, a km of 1.8 +/- 0.1 mM, and a ks of 35.4 +/- 3.7 mM while the immobilized form had a Vmax of 1.2 +/- 0.2 muM/min-mug, a km of 4.1 +/- 0.6 mM, and a ks of 660 +/- 140 mM. The free form of OxDc had a Vmax of 23.5 +/- 1.4 muM/min-mug and a km of 0.5 +/- 0.1 mM while the immobilized form had a Vmax of 5.0 +/- 1.9 muM/min-mug and km of 23.2 +/- 9.1 mM. The enzyme activity was measured to indicate viable application conditions for the coating, such as storing the films in urine over time. The maximum activity was shown at pH 4.2 to 4.5 and activity drops to be negligible by pH 7.0. Storing the enzyme at pH 6.1 exhibited a larger retained activity than storing at pH 4.2, yet storing in urine showed the highest retention. In a six moth trial period in urine, immobilized OxO lost 30% activity to 0.7 muM/min-mug, whereas the activity for immobilized OxDc fell 50% from about 5.9 to 2.9 muM/min-mug. Coating p(HEMA) onto polyurethane ureteral stents was applied by dip coating into a monomer-based coating solution. To achieve successful coatings, the viscosity of the coating solution and adhesion to the stent were optimized through a series of experiments with glycerol and superglue to form a primer of p(HEMA). The enzymes were applied to the primer through successive layers without the use of glycerol or superglue. The enzyme activity was used to compare various processing routes, such as dip time, dip cycles, and the use of Triton X-100. An encrustation model was established using artificial and real urine, and an antibiotic/antimycotic solution was added to prevent infection. The solutions were spiked with 0.5 mM oxalate to optimize encrustation conditions. The encrustation study was conducted up to two months in these solutions, and samples were analyzed using polarized light microscopy. Immobilized OxDc inhibited crystal growth up to two-months, although OxO developed encrustation to a similar extent of the control group. This opens the possibility of utilizing the immobilized enzyme as a therapy for degrading oxalate concentrations in urine, which can be employed as a coating on ureteral stents.

Excess Weapons Plutonium Immobilization in Russia

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jardine, L.; Borisov, G.B.

2000-04-15

The joint goal of the Russian work is to establish a full-scale plutonium immobilization facility at a Russian industrial site by 2005. To achieve this requires that the necessary engineering and technical basis be developed in these Russian projects and the needed Russian approvals be obtained to conduct industrial-scale immobilization of plutonium-containing materials at a Russian industrial site by the 2005 date. This meeting and future work will provide the basis for joint decisions. Supporting R&D projects are being carried out at Russian Institutes that directly support the technical needs of Russian industrial sites to immobilize plutonium-containing materials. Special R&Dmore » on plutonium materials is also being carried out to support excess weapons disposition in Russia and the US, including nonproliferation studies of plutonium recovery from immobilization forms and accelerated radiation damage studies of the US-specified plutonium ceramic for immobilizing plutonium. This intriguing and extraordinary cooperation on certain aspects of the weapons plutonium problem is now progressing well and much work with plutonium has been completed in the past two years. Because much excellent and unique scientific and engineering technical work has now been completed in Russia in many aspects of plutonium immobilization, this meeting in St. Petersburg was both timely and necessary to summarize, review, and discuss these efforts among those who performed the actual work. The results of this meeting will help the US and Russia jointly define the future direction of the Russian plutonium immobilization program, and make it an even stronger and more integrated Russian program. The two objectives for the meeting were to: (1) Bring together the Russian organizations, experts, and managers performing the work into one place for four days to review and discuss their work with each other; and (2) Publish a meeting summary and a proceedings to compile reports of all the excellent Russian plutonium immobilization contract work. This proceedings document presents the wide extent of Russian immobilization activities, provides a reference for their work, and makes it available to others.« less

Demonstration of lysosomal localization for the mammalian ependymin-related protein using classical approaches combined with a novel density shift method.

PubMed

Della Valle, Maria Cecilia; Sleat, David E; Sohar, Istvan; Wen, Ting; Pintar, John E; Jadot, Michel; Lobel, Peter

2006-11-17

Most newly synthesized soluble lysosomal proteins are delivered to the lysosome via the mannose 6-phosphate (Man-6-P)-targeting pathway. The presence of the Man-6-P post-translational modification allows these proteins to be affinity-purified on immobilized Man-6-P receptors. This approach has formed the basis for a number of proteomic studies that identified multiple as yet uncharacterized Man-6-P glycoproteins that may represent new lysosomal proteins. Although the presence of Man-6-P is suggestive of lysosomal function, the subcellular localization of such candidates requires experimental verification. Here, we have investigated one such candidate, ependymin-related protein (EPDR). EPDR is a protein of unknown function with some sequence similarity to ependymin, a fish protein thought to play a role in memory consolidation and learning. Using classical subcellular fractionation on rat brain, EPDR co-distributes with lysosomal proteins, but there is significant overlap between lysosomal and mitochondrial markers. For more definitive localization, we have developed a novel approach based upon a selective buoyant density shift of the brain lysosomes in a mutant mouse lacking NPC2, a lysosomal protein involved in lipid transport. EPDR, in parallel with lysosomal markers, shows this density shift in gradient centrifugation experiments comparing mutant and wild type mice. This approach, combined with morphological analyses, demonstrates that EPDR resides in the lysosome. In addition, the lipidosis-induced density shift approach represents a valuable tool for identification and validation of both luminal and membrane lysosomal proteins that should be applicable to high throughput proteomic studies.

Immobilization of Polyoxometalates on Tailored Polymeric Surfaces.

PubMed

Aguado-Ureta, Saioa; Rodríguez-Hernández, Juan; Del Campo, Adolfo; Perez-Álvarez, Leyre; Ruiz-Rubio, Leire; Vilas, José Luis; Artetxe, Beñat; Reinoso, Santiago; Gutiérrez-Zorrilla, Juan M

2018-03-02

Herein we describe the preparation of hybrid polymer-inorganic interfaces by the immobilization of polyoxometalate nanoclusters on functionalized polymer surfaces. The polymeric surfaces were made of polystyrene- b -poly(acrylic acid)/polystyrene (PS- b -PAA/PS) blends by spin coating on a silicon wafer. The functionalization of the polymer film was obtained by interfacial migration of the amphiphilic block copolymer toward the interface upon water vapor annealing. The carboxylic acid functional groups contained in the PAA block were then employed to anchor the [Ln III (α-SiW 11 O 39 )] 5- polyoxometalates (Ln: Ce, Er). This purpose was achieved by immersing the films in aqueous solutions of the in situ-formed inorganic nanoclusters. X-ray photoelectron and confocal Raman spectroscopies, together with atomic force microscopy, confirmed the immobilization of the inorganic species at the interface.

Covalent immobilization of lipase onto chitosan-mesoporous silica hybrid nanomaterials by carboxyl functionalized ionic liquids as the coupling agent.

PubMed

Xiang, Xinran; Suo, Hongbo; Xu, Chao; Hu, Yi

2018-05-01

Chitosan-mesoporous silica SBA-15 hybrid nanomaterials (CTS-SBA-15) were synthesized by means of carboxyl functionalized ionic liquids as the coupling agent. The as-prepared CTS-SBA-15 support was characterized by TEM, FTIR, TG and nitrogen adsorption-desorption techniques. Porcine pancreas lipase (PPL) was then bound to the hybrid nanomaterials by using the cross-linking reagent glutaraldehyde (GA). Further, the parameters like cross-linking concentration, time and ratio of supports to enzyme were optimized. The property of immobilized lipase were tested in detail by enzyme activity assays. The results indicated that the hybrid nanomaterials could form three-dimensional (3D) structure with homogeneous mesoporous structures and immobilized PPL revealed excellent enzymatic performance. Copyright © 2018 Elsevier B.V. All rights reserved.

Immobilized metal ion affinity electrophoresis. A study with several model proteins containing histidine.

PubMed

Goubran-Botros, H; Nanak, E; Abdul Nour, J; Birkenmeir, G; Vijayalakshmi, M A

1992-04-24

Immobilized metal ion affinity electrophoresis (IMA-Elec) is one among the many methods derived from the immobilized metal ion affinity chromatography. Two approaches for incorporating the metal ligand, were studied. One was in the form of insoluble particulate material based on Sepharose 6B and the other in the form of soluble polymer based on polyethylene glycol (PEG) 5000. Both the polymers coupled with iminodiacetate and metallized with copper or zinc were used as ligands, incorporated into soluble agarose as the electrophoretic gel. Several histidine-containing model proteins were studied with both the systems and their metal binding strengths were determined as the dissociation constants, Kd. The results clearly demonstrated that the mechanism of protein recognition by immobilized copper or zinc via the accessible histidyl residues was maintained in the IMA-Elec system. Proteins with increasing numbers of histidine residues showed increasing binding strength (lower Kd values). While this basic mechanism was conserved, the supporting polymers (Sepharose 6B and the PEG 5000) showed significant differences in the metal binding to the protein. The polysaccharide Sepharose 6B enhanced the binding strength compared with PEG 5000. The optimum electrophoretic parameters were determined to be current intensities up to 20 mA and pH ca. 7.0. At pH greater than 8.0, a significant decrease in the affinity was observed, this decrease being greater with PEG 5000 than Sepharose 6B as supporting material.

Surface plasmon resonance label-free monitoring of antibody antigen interactions in real time

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kausaite, A.; van Dijk, M.; Castrop, J.

2007-01-01

Detection of biologically active compounds is one of the most important topics in molecular biology and biochemistry. One of the most promising detection methods is based on the application of surface plasmon resonance for label-free detection of biologically active compounds. This method allows one to monitor binding events in real time without labeling. The system can therefore be used to determine both affinity and rate constants for interactions between various types of molecules. Here, we describe the application of a surface plasmon resonance biosensor for label-free investigation of the interaction between an immobilized antigen bovine serum albumin (BSA) and antibodymore » rabbit anti-cow albumin IgG1 (anti-BSA). The formation of a self-assembled monolayer (SAM) over a gold surface is introduced into this laboratory training protocol as an effective immobilization method, which is very promising in biosensing systems based on detection of affinity interactions. In the next step, covalent attachment via artificially formed amide bonds is applied for the immobilization of proteins on the formed SAM surface. These experiments provide suitable experience for postgraduate students to help them understand immobilization of biologically active materials via SAMs, fundamentals of surface plasmon resonance biosensor applications, and determination of non-covalent biomolecular interactions. The experiment is designed for master and/or Ph.D. students. In some particular cases, this protocol might be adoptable for bachelor students that already have completed an extended biochemistry program that included a background in immunology.« less

Analysis of nucleotides and oligonucleotides immobilized as self-assembled monolayers by static secondary ion mass spectrometry.

PubMed

Patrick, J S; Cooks, R G; Pachuta, S J

1994-11-01

Nucleic acid constituents can be bound to a metal surface in the form of self-assembled monolayers. Binding is achieved either through ionic interactions with a self-assembled 2-aminoethanethiol monolayer or by direct covalent binding of a dithiophosphate oligonucleotide to a metal surface through a sulfur-metal bond. Nucleotides, polynucleotides (both normal and a dithiophosphate analog) and double-stranded DNA have all been bound to surfaces. When the surfaces are interrogated using static secondary ion mass spectrometry (SIMS), the surface-bound nucleic acid constituents are observed in the form of the characteristic protonated nucleic acid base ions (BH2+). While a silver foil substrate was found to provide the highest absolute signal, vapor-deposited gold yields the best signal-to-noise ratio for ionically bound deoxyguanosine monophosphate. Under comparable conditions, a Cs+ projectile produces a 10-fold increase in the secondary ion signal relative to a Ga+ projectile. The experiment has been extended to a triple-quadrupole instrument where tandem mass spectrometric experiments on ionically immobilized dGMP showed the characteristic loss of ammonia from the released BH2+ ion. When a 'biomimetic' surface formed by ionically immobilizing double-stranded DNA is exposed to a solution containing ethidium bromide, ions corresponding to the non-covalent adduct are readily detectable using SIMS. This adduct and the nucleic acid constituents can be monitored at levels below 10 fmol.

Epsilon Metal Waste Form for Immobilization of Noble Metals from Used Nuclear Fuel

DOE Office of Scientific and Technical Information (OSTI.GOV)

Crum, Jarrod V.; Strachan, Denis M.; Rohatgi, Aashish

2013-10-01

Epsilon metal (ε-metal), an alloy of Mo, Pd, Rh, Ru, and Tc, is being developed as a waste form to treat and immobilize the undissolved solids and dissolved noble metals from aqueous reprocessing of commercial used nuclear fuel. Epsilon metal is an attractive waste form for several reasons: increased durability relative to borosilicate glass, it can be fabricated without additives (100% waste loading), and in addition it also benefits borosilicate glass waste loading by eliminating noble metals from the glass and thus the processing problems related there insolubility in glass. This work focused on the processing aspects of the epsilonmore » metal waste form development. Epsilon metal is comprised of refractory metals resulting in high reaction temperatures to form the alloy, expected to be 1500 - 2000°C making it a non-trivial phase to fabricate by traditional methods. Three commercially available advanced technologies were identified: spark-plasma sintering, microwave sintering, and hot isostatic pressing, and investigated as potential methods to fabricate this waste form. Results of these investigations are reported and compared in terms of bulk density, phase assemblage (X-ray diffraction and elemental analysis), and microstructure (scanning electron microscopy).« less

Enhanced uranium immobilization and reduction by Geobacter sulfurreducens biofilms.

PubMed

Cologgi, Dena L; Speers, Allison M; Bullard, Blair A; Kelly, Shelly D; Reguera, Gemma

2014-11-01

Biofilms formed by dissimilatory metal reducers are of interest to develop permeable biobarriers for the immobilization of soluble contaminants such as uranium. Here we show that biofilms of the model uranium-reducing bacterium Geobacter sulfurreducens immobilized substantially more U(VI) than planktonic cells and did so for longer periods of time, reductively precipitating it to a mononuclear U(IV) phase involving carbon ligands. The biofilms also tolerated high and otherwise toxic concentrations (up to 5 mM) of uranium, consistent with a respiratory strategy that also protected the cells from uranium toxicity. The enhanced ability of the biofilms to immobilize uranium correlated only partially with the biofilm biomass and thickness and depended greatly on the area of the biofilm exposed to the soluble contaminant. In contrast, uranium reduction depended on the expression of Geobacter conductive pili and, to a lesser extent, on the presence of the c cytochrome OmcZ in the biofilm matrix. The results support a model in which the electroactive biofilm matrix immobilizes and reduces the uranium in the top stratum. This mechanism prevents the permeation and mineralization of uranium in the cell envelope, thereby preserving essential cellular functions and enhancing the catalytic capacity of Geobacter cells to reduce uranium. Hence, the biofilms provide cells with a physically and chemically protected environment for the sustained immobilization and reduction of uranium that is of interest for the development of improved strategies for the in situ bioremediation of environments impacted by uranium contamination. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Enhanced Uranium Immobilization and Reduction by Geobacter sulfurreducens Biofilms

PubMed Central

Cologgi, Dena L.; Speers, Allison M.; Bullard, Blair A.; Kelly, Shelly D.

2014-01-01

Biofilms formed by dissimilatory metal reducers are of interest to develop permeable biobarriers for the immobilization of soluble contaminants such as uranium. Here we show that biofilms of the model uranium-reducing bacterium Geobacter sulfurreducens immobilized substantially more U(VI) than planktonic cells and did so for longer periods of time, reductively precipitating it to a mononuclear U(IV) phase involving carbon ligands. The biofilms also tolerated high and otherwise toxic concentrations (up to 5 mM) of uranium, consistent with a respiratory strategy that also protected the cells from uranium toxicity. The enhanced ability of the biofilms to immobilize uranium correlated only partially with the biofilm biomass and thickness and depended greatly on the area of the biofilm exposed to the soluble contaminant. In contrast, uranium reduction depended on the expression of Geobacter conductive pili and, to a lesser extent, on the presence of the c cytochrome OmcZ in the biofilm matrix. The results support a model in which the electroactive biofilm matrix immobilizes and reduces the uranium in the top stratum. This mechanism prevents the permeation and mineralization of uranium in the cell envelope, thereby preserving essential cellular functions and enhancing the catalytic capacity of Geobacter cells to reduce uranium. Hence, the biofilms provide cells with a physically and chemically protected environment for the sustained immobilization and reduction of uranium that is of interest for the development of improved strategies for the in situ bioremediation of environments impacted by uranium contamination. PMID:25128347

A novel step towards immobilization of biocatalyst using agro waste and its application for ester synthesis.

PubMed

Tomke, Prerana D; Rathod, Virendra K

2018-05-04

This work explains the utilization of agro-waste (coconut and peanut shell) to produce mesoporous activated carbon which further utilized as a support material for lipase immobilization (Candida antarctica B, CALB). Various parameters affecting the binding of enzyme to activated carbon with high surface area (1603 m 2  g -1 ) were optimized. Maximum 200 μg g -1 CALB has been loaded at 40 °C and pH 6.8 in 12 h by using glutaraldehyde as a cross-linker. The operational parameters such as pH (5.8-8.8) and temperature (30-70 °C) were optimized for free and immobilized form of lipase. In thermal stability (50-70 °C) study, immobilization of enzyme showed 2.35 folds increased half-life with respect to free enzyme. The samples, before and after immobilization, were characterized by specific surface area, FT-IR, SEM, XRD. This immobilized lipase was successfully used for the synthesis of cinnamyl acetate by transesterification reaction producing 94% conversion in 60 min. Catalytic efficiency (58 ± 1.08) was seen to be retained for more than five consecutive cycles of chemical reaction for repeated applications. Sequential results towards activity retention were obtained upto 30 days of storability study. In the context, this process constitutes a clean route for the development of sustainable biocatalysts from agro waste, capable of applications in various area. Copyright © 2017. Published by Elsevier B.V.

Immobilization of Pb and Cu in polluted soil by superphosphate, multi-walled carbon nanotube, rice straw and its derived biochar.

PubMed

Rizwan, Muhammad Shahid; Imtiaz, Muhammad; Huang, Guoyong; Chhajro, Muhammad Afzal; Liu, Yonghong; Fu, Qingling; Zhu, Jun; Ashraf, Muhammad; Zafar, Mohsin; Bashir, Saqib; Hu, Hongqing

2016-08-01

Lead (Pb) and copper (Cu) contamination in croplands pose severe health hazards and environmental concerns throughout soil-food chain transfer. In the present study, BCR, TCLP, CaCl2, and SBET techniques were employed to evaluate the simultaneous effectiveness of rice straw (RS) and its derived biochar (BC), multiwall carbon nanotube (MWCNT), and single superphosphate (SSP) to immobilize the Pb and Cu in co-contaminated soil. The BCR sequential extraction results suggested that with increasing BC and SSP amount, the acid-soluble fractions decreased while oxidizable and residual proportions of Pb and Cu were increased significantly. Compared to SSP, the application of BC amendment substantially modified partitioning of Cu from easily exchangeable phase to less bioavailable residual bound fraction. The immobilized Pb and Cu were mainly transformed to reducible forms. The TCLP and CaCl2-extracted Pb and Cu were reduced significantly by the addition of BC compared to RS and MWCNT, whereas the bio-accessibility of Pb significantly reduced with RS addition. SSP showed better results for Pb immobilization while marginal for Cu in co-contaminated soil. Overall, the addition of BC offered the best results and could be effective in both Pb and Cu immobilization thereby reducing their mobility and bioavailability in the co-contaminated soil.

Oriented antibody immobilization to polystyrene macrocarriers for immunoassay modified with hydrazide derivatives of poly(meth)acrylic acid

PubMed Central

Shmanai, Vadim V; Nikolayeva, Tamara A; Vinokurova, Ludmila G; Litoshka, Anatoli A

2001-01-01

Background Hydrophobic polystyrene is the most common material for solid phase immunoassay. Proteins are immobilized on polystyrene by passive adsorption, which often causes considerable denaturation. Biological macromolecules were found to better retain their functional activity when immobilized on hydrophilic materials. Polyacrylamide is a common material for solid-phase carriers of biological macromolecules, including immunoreagents used in affinity chromatography. New macroformats for immunoassay modified with activated polyacrylamide derivatives seem to be promising. Results New polymeric matrices for immunoassay in the form of 0.63-cm balls which contain hydrazide functional groups on hydrophilic polymer spacer arms at their surface shell are synthesized by modification of aldehyde-containing polystyrene balls with hydrazide derivatives of poly(meth)acrylic acid. The beads contain up to 0.31 μmol/cm2 active hydrazide groups accessible for covalent reaction with periodate-oxidized antibodies. The matrices obtained allow carrying out the oriented antibody immobilization, which increases the functional activity of immunosorbents. Conclusions An efficient site-directed antibody immobilization on a macrosupport is realized. The polymer hydrophilic spacer arms are the most convenient and effective tools for oriented antibody coupling with molded materials. The suggested scheme can be used for the modification of any other solid supports containing electrophilic groups reacting with hydrazides. PMID:11545680

Sequential application of waste whey as a medium component for Kluyveromyces lactis cultivation and a co-feeder for lipase immobilization by CLEA method.

PubMed

Veteikytė, Aušra; Šiekštelė, Rimantas; Tvaska, Bronius; Matijošytė, Inga

2017-05-01

Currently, much attention is paid to technologies which can be drivers of the circular economy across different sectors, in particular, to develop technologies for utilization or reusability of biocompatible materials from industrial waste. One of such is the milk whey, which is a cheap biobased raw material, the disposal of which is a major problem for the dairy industry. Our proposed and investigated technology is based on a continuous exploitation of the whey combining microbiology and biotechnology. Primarily, whey was used as a nutrition source for the cultivation of Kluyveromyces lactis with the aim to produce the targeted biocatalyst-lipase. During cultivation, the whey was transformed into the hydrolyzed form, which was further successfully applied as a protein feeder (external linker) for immobilization of lipase by cross-linked enzyme aggregate (CLEA) method. The first time use of whey as a co-feeder for immobilization of enzymes by CLEA method has shown promising results and increased the stability of lipases for temperature and organic solvents. Hydrolysis of rapeseed oil catalyzed with immobilized derivatives was obtained with 45-96% efficiency at non-optimized conditions. Additionally, the determined kinetic parameters indicated that the rate of p-nitrophenyl palmitate hydrolysis was not changed drastically after immobilization.

Immobilization and direct electrochemistry of glucose oxidase on a tetragonal pyramid-shaped porous ZnO nanostructure for a glucose biosensor.

PubMed

Dai, Zhihui; Shao, Guojian; Hong, Jianmin; Bao, Jianchun; Shen, Jian

2009-01-01

A tetragonal pyramid-shaped porous ZnO (TPSP-ZnO) nanostructure is used for the immobilization, direct electrochemistry and biosensing of proteins. The prepared ZnO has a large surface area and good biocompatibility. Using glucose oxidase (GOD) as a model, this shaped ZnO is tested for immobilization of proteins and the construction of electrochemical biosensors with good electrochemical performances. The interaction between GOD and TPSP-ZnO is examined by using AFM, N(2) adsorption isotherms and electrochemical methods. The immobilized GOD at a TPSP-ZnO-modified glassy carbon electrode shows a good direct electrochemical behavior, which depends on the properties of the TPSP-ZnO. Based on a decrease of the electrocatalytic response of the reduced form of GOD to dissolved oxygen, the proposed biosensor exhibits a linear response to glucose concentrations ranging from 0.05 to 8.2mM with a detection limit of 0.01mM at an applied potential of -0.50V which has better biosensing properties than those from other morphological ZnO nanoparticles. The biosensor shows good stability, reproducibility, low interferences and can diagnose diabetes very fast and sensitively. Such the TPSP-ZnO nanostructure provides a good matrix for protein immobilization and biosensor preparation.

Production of L-glutamic Acid with Corynebacterium glutamicum (NCIM 2168) and Pseudomonas reptilivora (NCIM 2598): A Study on Immobilization and Reusability

PubMed Central

Shyamkumar, Rajaram; Moorthy, Innasi Muthu Ganesh; Ponmurugan, Karuppiah; Baskar, Rajoo

2014-01-01

Background L-glutamic acid is one of the major amino acids that is present in a wide variety of foods. It is mainly used as a food additive and flavor enhancer in the form of sodium salt. Corynebacterium glutamicum (C. glutamicum) is one of the major organisms widely used for glutamic acid production. Methods The study was dealing with immobilization of C. glutamicum and mixed culture of C. glutamicum and Pseudomonas reptilivora (P. reptilivora) for L-glutamic acid production using submerged fermentation. 2, 3 and 5% sodium alginate concentrations were used for production and reusability of immobilized cells for 5 more trials. Results The results revealed that 2% sodium alginate concentration produced the highest yield (13.026±0.247 g/l by C. glutamicum and 16.026±0.475 g/l by mixed immobilized culture). Moreover, reusability of immobilized cells was evaluated in 2% concentration with 5 more trials. However, when the number of cycles increased, the production of L-glutamic acid decreased. Conclusion Production of glutamic acid using optimized medium minimizes the time needed for designing the medium composition. It also minimizes external contamination. Glutamic acid production gradually decreased due to multiple uses of beads and consequently it reduces the shelf life. PMID:25215180

Production of L-glutamic Acid with Corynebacterium glutamicum (NCIM 2168) and Pseudomonas reptilivora (NCIM 2598): A Study on Immobilization and Reusability.

PubMed

Shyamkumar, Rajaram; Moorthy, Innasi Muthu Ganesh; Ponmurugan, Karuppiah; Baskar, Rajoo

2014-07-01

L-glutamic acid is one of the major amino acids that is present in a wide variety of foods. It is mainly used as a food additive and flavor enhancer in the form of sodium salt. Corynebacterium glutamicum (C. glutamicum) is one of the major organisms widely used for glutamic acid production. The study was dealing with immobilization of C. glutamicum and mixed culture of C. glutamicum and Pseudomonas reptilivora (P. reptilivora) for L-glutamic acid production using submerged fermentation. 2, 3 and 5% sodium alginate concentrations were used for production and reusability of immobilized cells for 5 more trials. The results revealed that 2% sodium alginate concentration produced the highest yield (13.026±0.247 g/l by C. glutamicum and 16.026±0.475 g/l by mixed immobilized culture). Moreover, reusability of immobilized cells was evaluated in 2% concentration with 5 more trials. However, when the number of cycles increased, the production of L-glutamic acid decreased. Production of glutamic acid using optimized medium minimizes the time needed for designing the medium composition. It also minimizes external contamination. Glutamic acid production gradually decreased due to multiple uses of beads and consequently it reduces the shelf life.

Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation.

PubMed

Hoesli, Corinne A; Raghuram, Kamini; Kiang, Roger L J; Mocinecová, Dušana; Hu, Xiaoke; Johnson, James D; Lacík, Igor; Kieffer, Timothy J; Piret, James M

2011-02-01

Alginate has been used to protect transplanted pancreatic islets from immune rejection and as a matrix to increase the insulin content of islet progenitor cells. The throughput of alginate bead generation by the standard extrusion and external gelation method is limited by the rate of droplet formation from nozzles. Alginate bead generation by emulsion and internal gelation is a scaleable alternative that has been used with biological molecules and microbial cells, but not mammalian cells. We describe the novel adaptation of this process to mammalian cell immobilization. After optimization, the emulsion process yielded 90 ± 2% mouse insulinoma 6 (MIN6) cell survival, similar to the extrusion process. The MIN6 cells expanded at the same rate in both bead types to form pseudo-islets with increased glucose stimulation index compared to cells in suspension. The emulsion process was suitable for primary pancreatic exocrine cell immobilization, leading to 67 ± 32 fold increased insulin expression after 10 days of immobilized culture. Due to the scaleability and broad availability of stirred mixers, the emulsion process represents an attractive option for laboratories that are not equipped with extrusion-based cell encapsulators, as well as for the production of immobilized or encapsulated cellular therapeutics on a clinical scale. © 2010 Wiley Periodicals, Inc.

DNA surface hybridization regimes

PubMed Central

Gong, Ping; Levicky, Rastislav

2008-01-01

Surface hybridization reactions, in which sequence-specific recognition occurs between immobilized and solution nucleic acids, are routinely carried out to quantify and interpret genomic information. Although hybridization is fairly well understood in bulk solution, the greater complexity of an interfacial environment presents new challenges to a fundamental understanding, and hence application, of these assays. At a surface, molecular interactions are amplified by the two-dimensional nature of the immobilized layer, which focuses the nucleic acid charge and concentration to levels not encountered in solution, and which impacts the hybridization behavior in unique ways. This study finds that, at low ionic strengths, an electrostatic balance between the concentration of immobilized oligonucleotide charge and solution ionic strength governs the onset of hybridization. As ionic strength increases, the importance of electrostatics diminishes and the hybridization behavior becomes more complex. Suppression of hybridization affinity constants relative to solution values, and their weakened dependence on the concentration of DNA counterions, indicate that the immobilized strands form complexes that compete with hybridization to analyte strands. Moreover, an unusual regime is observed in which the surface coverage of immobilized oligonucleotides does not significantly influence the hybridization behavior, despite physical closeness and hence compulsory interactions between sites. These results are interpreted and summarized in a diagram of hybridization regimes that maps specific behaviors to experimental ranges of ionic strength and probe coverage. PMID:18381819

Photonic activation of disulfide bridges achieves oriented protein immobilization on biosensor surfaces.

PubMed

Neves-Petersen, Maria Teresa; Snabe, Torben; Klitgaard, Søren; Duroux, Meg; Petersen, Steffen B

2006-02-01

Photonic induced immobilization is a novel technology that results in spatially oriented and spatially localized covalent coupling of biomolecules onto thiol-reactive surfaces. Immobilization using this technology has been achieved for a wide selection of proteins, such as hydrolytic enzymes (lipases/esterases, lysozyme), proteases (human plasminogen), alkaline phosphatase, immunoglobulins' Fab fragment (e.g., antibody against PSA [prostate specific antigen]), Major Histocompability Complex class I protein, pepsin, and trypsin. The reaction mechanism behind the reported new technology involves "photonic activation of disulfide bridges," i.e., light-induced breakage of disulfide bridges in proteins upon UV illumination of nearby aromatic amino acids, resulting in the formation of free, reactive thiol groups that will form covalent bonds with thiol-reactive surfaces (see Fig. 1). Interestingly, the spatial proximity of aromatic residues and disulfide bridges in proteins has been preserved throughout molecular evolution. The new photonic-induced method for immobilization of proteins preserves the native structural and functional properties of the immobilized protein, avoiding the use of one or more chemical/thermal steps. This technology allows for the creation of spatially oriented as well as spatially defined multiprotein/DNA high-density sensor arrays with spot size of 1 microm or less, and has clear potential for biomedical, bioelectronic, nanotechnology, and therapeutic applications.

From Protein Engineering to Immobilization: Promising Strategies for the Upgrade of Industrial Enzymes

PubMed Central

Singh, Raushan Kumar; Tiwari, Manish Kumar; Singh, Ranjitha; Lee, Jung-Kul

2013-01-01

Enzymes found in nature have been exploited in industry due to their inherent catalytic properties in complex chemical processes under mild experimental and environmental conditions. The desired industrial goal is often difficult to achieve using the native form of the enzyme. Recent developments in protein engineering have revolutionized the development of commercially available enzymes into better industrial catalysts. Protein engineering aims at modifying the sequence of a protein, and hence its structure, to create enzymes with improved functional properties such as stability, specific activity, inhibition by reaction products, and selectivity towards non-natural substrates. Soluble enzymes are often immobilized onto solid insoluble supports to be reused in continuous processes and to facilitate the economical recovery of the enzyme after the reaction without any significant loss to its biochemical properties. Immobilization confers considerable stability towards temperature variations and organic solvents. Multipoint and multisubunit covalent attachments of enzymes on appropriately functionalized supports via linkers provide rigidity to the immobilized enzyme structure, ultimately resulting in improved enzyme stability. Protein engineering and immobilization techniques are sequential and compatible approaches for the improvement of enzyme properties. The present review highlights and summarizes various studies that have aimed to improve the biochemical properties of industrially significant enzymes. PMID:23306150

Carbon fibers with a nano-hydroxyapatite coating as an excellent biofilm support for bioreactors

NASA Astrophysics Data System (ADS)

Liu, Qijie; Zhang, Chao; Bao, Yanling; Dai, Guangze

2018-06-01

A biofilm support with high biocompatibility is needed for bioreactors. A nano-hydroxyapatite (HA) coating on carbon fibers (CFs) was prepared by electrochemical deposition (ECD). The sludge immobilization assays, bacterial cells adhesion assays and Derjaguin-Landau-Verwey-Overbeek (DLVO) theory were used to evaluate the capacity of CF supports to immobilize activated sludge and bacterial cells. The sludge immobilization and bacterial cells adhesion assays illustrated that HA coating could enhance the capacity of CFs to immobilize microorganisms. SEM images showed that HA and bacterial cells formed a dense film on CFs surface. In addition, HA, acting as a glue, could combine CFs with bacterial cells or between cells, which helped CFs capture more bacterial cells. DLVO theory illustrated that CFs with HA coating had a lower total interaction energy than CFs without handling, explaining the higher capacity of CFs with HA coating to immobilize bacterial cells. This result was owning to the less negative zeta potential and higher hydrophilicity of CFs with HA coating, and the hydrophilicity made a greater contribution to the lower total interaction energy. Experiments and theory reveal that HA coating could enhance the biocompatibility of CFs, and CFs with HA coating could be used as an excellent biofilm support for bioreactors.

Recoverable immobilization of transuranic elements in sulfate ash

DOEpatents

Greenhalgh, Wilbur O.

1985-01-01

Disclosed is a method of reversibly immobilizing sulfate ash at least about 20% of which is sulfates of transuranic elements. The ash is mixed with a metal which can be aluminum, cerium, samarium, europium, or a mixture thereof, in amounts sufficient to form an alloy with the transuranic elements, plus an additional amount to reduce the transuranic element sulfates to elemental form. Also added to the ash is a fluxing agent in an amount sufficient to lower the percentage of the transuranic element sulfates to about 1% to about 10%. The mixture of the ash, metal, and fluxing agent is heated to a temperature sufficient to melt the fluxing agent and the metal. The mixture is then cooled and the alloy is separated from the remainder of the mixture.

Tellurite glasses for vitrification of technetium-99 from pyrochemical processing

NASA Astrophysics Data System (ADS)

Pyo, Jae-Young; Lee, Cheong Won; Park, Hwan-Seo; Yang, Jae Hwan; Um, Wooyong; Heo, Jong

2017-09-01

A new alkali-alumino tellurite glass composition was developed to immobilize highly-volatile technetium (Tc) wastes generated from the pyrochemical processing technology. Tellurite glass can incorporate up to 7 mass% of rhenium (Re, used as a surrogate for Tc) with an average retention of 86%. Normalized elemental releases evaluated by seven-day product consistency test (PCT) satisfied the immobilized low activity waste requirements of United States when concentration of Ca(ReO4)2 in the glass was <12 mass%. Re ions form Re7+ and are coordinated with four oxygens to form ReO4- tetrahedra. These tetrahedra bond to modifiers such as Ca2+ or Na+ that are further connected to the tellurite glass network by Ca2+ (or Na+) - non-bridging oxygen bonds.

Method and article for primary containment of cesium wastes. [DOE patent application

DOEpatents

Angelini, P.; Lackey, W.J.; Stinton, D.P.; Blanco, R.E.; Bond, W.D.; Arnold, W.D. Jr.

1981-09-03

A method for producing a cesium-retentive waste form, characterized by a high degree of compositional stability and mechanical integrity, is provided by subjecting a cesium-loaded zeolite to heat under conditions suitable for stabilizing the zeolite and immobilizing the cesium, and coating said zeolite for sufficient duration within a suitable environment with at least one dense layer of pyrolytic carbon to seal therein said cesium to produce a final, cesium-bearing waste form. Typically, the zolite is stabilized and the cesium immobilized in less than four hours by confinement within an air environment maintained at about 600/sup 0/C. Coatings are thereafter applied by confining the calcined zeolite within a coating environment comprising inert fluidizing and carbon donor gases maintained at 1000/sup 0/C for a suitable duration.

Method for primary containment of cesium wastes

DOEpatents

Angelini, Peter; Lackey, Walter J.; Stinton, David P.; Blanco, Raymond E.; Bond, Walter D.; Arnold, Jr., Wesley D.

1983-01-01

A method for producing a cesium-retentive waste form, characterized by a high degree of compositional stability and mechanical integrity, is provided by subjecting a cesium-loaded zeolite to heat under conditions suitable for stabilizing the zeolite and immobilizing the cesium, and coating said zeolite for sufficient duration within a suitable environment with at least one dense layer of pyrolytic carbon to seal therein said cesium to produce a final, cesium-bearing waste form. Typically, the zeolite is stabilized and the cesium immobilized in less than four hours by confinement within an air environment maintained at about 600.degree. C. Coatings are thereafter applied by confining the calcined zeolite within a coating environment comprising inert fluidizing and carbon donor gases maintained at 1,000.degree. C. for a suitable duration.

Phase 2 testing results of immobilization of WTP effluent management facility vaporator bottoms simulant

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reigel, M.; Cozzi, A.; McCabe, D.

2017-09-08

The Hanford Waste Treatment and Immobilization Plant (WTP) Low Activity Waste (LAW) vitrification facility will generate an aqueous condensate recycle stream (LAW Melter Off-Gas Condensate) from the primary off-gas system. This work examined three waste form formulations based on previous testing with related simulants: 8 wt% ordinary portland cement (OPC), 47 wt% blast furnace slag (BFS), 45 wt% fly ash (FA) known as Cast Stone formulation; 20 wt% Aquaset® II-GH and 80 wt% BFS; 20 wt% OPC and 80 wt% BFS. These tests successfully produced one waste form that set within five days (Cast Stone formulation); however the other twomore » formulations, Aquaset® II-GH/BFS and OPC/BFS, took approximately eight and fourteen days to set, respectively.« less

Immobilization of acetylcholinesterase in lipid membranes deposited on self-assembled monolayers.

PubMed

Milkani, Eftim; Khaing, Aung M; Huang, Fei; Gibson, Daniel G; Gridley, Scott; Garceau, Norman; Lambert, Christopher R; McGimpsey, W Grant

2010-12-21

Human red blood cell acetylcholinesterase was incorporated into planar lipid membranes deposited on alkanethiol self-assembled monolayers (SAMs) on gold substrates. Activity of the protein in the membrane was detected with a standard photometric assay and was determined to be similar to the protein in detergent solution or incorporated in lipid vesicles. Monolayer and bilayer lipid membranes were generated by fusing liposomes to hydrophobic and hydrophilic SAMs, respectively. Liposomes were formed by the injection method using the lipid dimyristoylphosphatidylcholine (DMPC). The formation of alkanethiol SAMs and lipid monolayers on SAMs was confirmed by sessile drop goniometry, ellipsometry, and electrochemical impedance spectroscopy. In this work, we report acetylcholinesterase immobilization in lipid membranes deposited on SAMs formed on the gold surface and compare its activity to enzyme in solution.

Simultaneous Alcoholic and Malolactic Fermentations by Saccharomyces cerevisiae and Oenococcus oeni Cells Co-immobilized in Alginate Beads

PubMed Central

Bleve, Gianluca; Tufariello, Maria; Vetrano, Cosimo; Mita, Giovanni; Grieco, Francesco

2016-01-01

Malolactic fermentation (MLF) usually takes place after the end of alcoholic fermentation (AF). However, the inoculation of lactic acid bacteria together with yeast starter cultures is a promising system to enhance the quality and safety of wine. In recent years, the use of immobilized cell systems has been investigated, with interesting results, for the production of different fermented foods and beverages. In this study we have carried out the simultaneous immobilization of Saccharomyces cerevisiae and Oenococcus oeni in alginate beads and used them in microvinifications tests to produce Negroamaro wine. The process was monitored by chemical and sensorial analyses and dominance of starters and cell leaking from beads were also checked. Co-immobilization of S. cerevisiae and O. oeni allowed to perform an efficient fermentation process, producing low volatile acidity levels and ethanol and glycerol concentrations comparable with those obtained by cell sequential inoculum and co-inoculum of yeast and bacteria cells in free form. More importantly, co-immobilization strategy produced a significant decrease of the time requested to complete AF and MLF. The immobilized cells could be efficiently reused for the wine fermentation at least three times without any apparent loss of cell metabolic activities. This integrated biocatalytic system is able to perform simultaneously AF and MLF, producing wines similar in organoleptic traits in comparison with wines fermented following traditional sequential AF and MLF with free cell starters. The immobilized-cell system, that we here describe for the first time in our knowledge, offers many advantages over conventional free cell fermentations, including: (i) elimination of non-productive cell growth phases; (ii) feasibility of continuous processing; (iii) re-use of the biocatalyst. PMID:27379072

Binary Colloidal Crystal Layers as Platforms for Surface Patterning of Puroindoline-Based Antimicrobial Peptides.

PubMed

Boden, Andrew; Bhave, Mrinal; Wang, Peng-Yuan; Jadhav, Snehal; Kingshott, Peter

2018-01-24

The ability of bacteria to form biofilms and the emergence of antibiotic-resistant strains have prompted the need to develop the next generation of antibacterial coatings. Antimicrobial peptides (AMPs) are showing promise as molecules that can address these issues, especially if used when immobilized as a surface coating. We present a method that explores how surface patterns together with the selective immobilization of an AMP called PuroA (FPVTWRWWKWWKG-NH 2 ) can be used to both kill bacteria and also as a tool to study bacterial attachment mechanisms. Surface patterning is achieved using stabilized self-assembled binary colloidal crystal (BCC) layers, allowing selective PuroA immobilization to carboxylated particles using N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide (EDC) hydrochloride/N-hydroxysuccinimide (NHS) coupling chemistry. Covalent immobilization of PuroA was compared with physical adsorption (i.e., without the addition of EDC/NHS). The AMP-functionalized colloids and BCC layers were characterized by X-ray photoelectron spectroscopy, ζ potentials, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Surface antimicrobial activity was assessed by viability assays using Escherichia coli. MALDI-TOF MS analysis revealed that although not all of PuroA was successfully covalently immobilized, a relatively low density of PuroA (1.93 × 10 13 molecules/cm 2 and 7.14 × 10 12 molecules/cm 2 for covalent and physical immobilization, respectively) was found to be sufficient at significantly decreasing the viability of E. coli by 70% when compared to that of control samples. The findings provide a proof of concept that BCC layers are a suitable platform for the patterned immobilization of AMPs and the importance of ascertaining the success of small-molecule grafting reactions using surface-MALDI, something that is often assumed to be successful in the field.

A-site compositional effects in Ga-doped hollandite materials of the form BaxCsyGa2x+yTi8−2x−yO16: implications for Cs immobilization in crystalline ceramic waste forms

PubMed Central

Xu, Yun; Wen, Yi; Grote, Rob; Amoroso, Jake; Shuller Nickles, Lindsay; Brinkman, Kyle S.

2016-01-01

The hollandite structure is a promising crystalline host for Cs immobilization. A series of Ga-doped hollandite BaxCsyGa2x+yTi8−2x−yO16 (x = 0, 0.667, 1.04, 1.33; y = 1.33, 0.667, 0.24, 0) was synthesized through a solid oxide reaction method resulting in a tetragonal hollandite structure (space group I4/m). The lattice parameter associated with the tunnel dimension was found to increases as Cs substitution in the tunnel increased. A direct investigation of cation mobility in tunnels using electrochemical impedance spectroscopy was conducted to evaluate the ability of the hollandite structure to immobilize cations over a wide compositional range. Hollandite with the largest tunnel size and highest aspect ratio grain morphology resulting in rod-like microstructural features exhibited the highest ionic conductivity. The results indicate that grain size and optimized Cs stoichiometry control cation motion and by extension, the propensity for Cs release from hollandite. PMID:27273791

Final Technical Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sobecky, Patricia A; Taillefert, Martial

This final technical report describes results and findings from a research project to examine the role of microbial phosphohydrolase enzymes in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of the radionuclide uranium through the production of insoluble uranium phosphate minerals. The research project investigated the microbial mechanisms and the physical and chemical processes promoting uranium biomineralization and sequestration in oxygenated subsurface soils. Uranium biomineralization under aerobic conditions can provide a secondary biobarrier strategy to immobilize radionuclides should the metal precipitates formed by microbial dissimilatory mechanisms remobilize due to a change in redox state.

Enhancement of algicidal activity by immobilization of algicidal bacteria antagonistic to Stephanodiscus hantzschii (Bacillariophyceae).

PubMed

Kang, Y-H; Kim, B-R; Choi, H J; Seo, J G; Kim, B-H; Han, M-S

2007-11-01

Enhancement of algicidal activity by immobilization of algicidal bacteria antagonistic to Stephanodiscus hantzschii. In laboratory studies, A diatom-lysing bacterium, Pseudomonas fluorescens HYK0210-SK09 showed strong algicidal activity against S. hantzschii, but a natural mesocosm study revealed that this bacterium failed to fully control natural blooms of Stephanodiscus at the low water temperatures that favour these blooms. Here, we sought to develop an effective immobilization strategy for enhancing the algicidal activity of HYK0210-SK09 in the natural setting. Bacterium HYK0210-SK09 was immobilized with various carriers including agar, alginate, polyurethane and cellulose sponge. The bacterial cells immobilized with cellulose sponge (CIS) induced more rapid and complete lysis of S. hantzschii than other carriers, and had a higher packing ability than polyurethane. Furthermore, CIS-immobilized cells showed higher lysis of S. hantzschii at the same concentrations as that of free cells (< or =1 x 10(7) cells ml(-1)), and had especially strong algicidal activity at the low temperatures (<10 degrees C). Based on these laboratory studies, we assessed the possible application of HYK0210-SK09 cells in the field by performing a mesocosm study during the winter season. The CIS-immobilized cells with species-specific activity towards the genera Stephanodiscus showed extremely high algicidal activity (up to 95%) against a bloom of Stephanodiscus hantzschii even at low water temperatures, because of high cell packing and subsequent cell protection against low temperatures and predators, whereas free cells showed negligible algicidal activities under these conditions. Immobilizing cells of HYK0210-SK09 in CIS foam, rather than in the other matrices tested, could achieve more efficient control of Stephanodiscus blooms and showed a significant algicidal activity on in vitro and in vivo blooms, even at low water temperature. Collectively, these results indicate that CIS of algicidal bacteria may form an important strategy for effective management of Stephanodiscus blooms at low water temperatures.

Virus immobilization on biomaterial scaffolds through biotin-avidin interaction for improving bone regeneration.

PubMed

Hu, Wei-Wen; Wang, Zhuo; Krebsbach, Paul H

2016-02-01

To spatially control therapeutic gene delivery for potential tissue engineering applications, a biotin-avidin interaction strategy was applied to immobilize viral vectors on biomaterial scaffolds. Both adenoviral vectors and gelatin sponges were biotinylated and avidin was applied to link them in a virus-biotin-avidin-biotin-material (VBABM) arrangement. The tethered viral particles were stably maintained within scaffolds and SEM images illustrated that viral particles were evenly distributed in three-dimensional (3D) gelatin sponges. An in vivo study demonstrated that transgene expression was restricted to the implant sites only and transduction efficiency was improved using this conjugation method. For an orthotopic bone regeneration model, adenovirus encoding BMP-2 (AdBMP2) was immobilized to gelatin sponges before implanting into critical-sized bone defects in rat calvaria. Compared to gelatin sponges with AdBMP2 loaded in a freely suspended form, the VBABM method enhanced gene transfer and bone regeneration was significantly improved. These results suggest that biotin-avidin immobilization of viral vectors to biomaterial scaffolds may be an effective strategy to facilitate tissue regeneration. Copyright © 2013 John Wiley & Sons, Ltd.

Experimental study on cesium immobilization in struvite structures.

PubMed

Wagh, Arun S; Sayenko, S Y; Shkuropatenko, V A; Tarasov, R V; Dykiy, M P; Svitlychniy, Y O; Virych, V D; Ulybkina, Е А

2016-01-25

Ceramicrete, a chemically bonded phosphate ceramic, was developed for nuclear waste immobilization and nuclear radiation shielding. Ceramicrete products are fabricated by an acid-base reaction between magnesium oxide and mono potassium phosphate that has a struvite-K mineral structure. In this study, we demonstrate that this crystalline structure is ideal for incorporating radioactive Cs into a Ceramicrete matrix. This is accomplished by partially replacing K by Cs in the struvite-K structure, thus forming struvite-(K, Cs) mineral. X-ray diffraction and thermo-gravimetric analyses are used to confirm such a replacement. The resulting product is non-leachable and stable at high temperatures, and hence it is an ideal matrix for immobilizing Cs found in high-activity nuclear waste streams. The product can also be used for immobilizing secondary waste streams generated during glass vitrification of spent fuel, or the method described in this article can be used as a pretreatment method during glass vitrification of high level radioactive waste streams. Furthermore, it suggests a method of producing safe commercial radioactive Cs sources. Copyright © 2015 Elsevier B.V. All rights reserved.

Fermentative hydrogen production from molasses wastewater in a continuous mixed immobilized sludge reactor.

PubMed

Han, Wei; Wang, Bing; Zhou, Yan; Wang, De-Xin; Wang, Yan; Yue, Li-Ran; Li, Yong-Feng; Ren, Nan-Qi

2012-04-01

A novel continuous mixed immobilized sludge reactor (CMISR) containing activated carbon as support carrier was used for fermentative hydrogen production from molasses wastewater. When the CMISR system operated at the conditions of influent COD of 2000-6000mg/L, hydraulic retention time (HRT) of 6h and temperature of 35°C, stable ethanol type fermentation was formed after 40days operation. The H(2) content in biogas and chemical oxygen demand (COD) removal were estimated to be 46.6% and 13%, respectively. The effects of organic loading rates (OLRs) on the CMISR hydrogen production system were also investigated. It was found that the maximum hydrogen production rate of 12.51mmol/hL was obtained at OLR of 32kg/m(3)d and the maximum hydrogen yield by substrate consumed of 130.57mmol/mol happened at OLR of 16kg/m(3)d. Therefore, the continuous mixed immobilized sludge reactor (CMISR) could be a promising immobilized system for fermentative hydrogen production. Copyright © 2012 Elsevier Ltd. All rights reserved.

Facile preparation of magnetic carbon nanotubes-immobilized lipase for highly efficient synthesis of 1,3-dioleoyl-2-palmitoylglycerol-rich human milk fat substitutes.

PubMed

Zheng, Mingming; Wang, Shi; Xiang, Xia; Shi, Jie; Huang, Juan; Deng, Qianchun; Huang, Fenghong; Xiao, Junyong

2017-08-01

In this study, Candida lipolytica lipase (CLL) was immobilized on magnetic multi-walled carbon nanotubes (mMWCNTs) via hydrophobic and cation-exchange interaction. The resultant immobilized CLL showed much better thermal stability, biocatalyst activity and easier recycling than the free form. A method for efficient enzymatic acidolysis of tripalmitin (PPP) with oleic acid (OA), to produce OPO-rich TAGs, was developed, using the immobilized CLL as the biocatalyst. Under optimized conditions (2% water, 20mg/ml enzyme, 1:6 PPP/OA, 50°C, 2h), the content of OPO in the final product reached 46.5%. CLL@mMWCNTs had a better activity and manipulative stability than commercial lipases. More importantly, the feasibility of CLL@mMWCNTs was also validated in the practical production of OPO-rich TAGs, using lard and restructured palm oil as the raw material. These results suggest that CLL@mMWCNTs is a promising biocatalyst for the OPO-rich TAGs production and will be helpful for the infant formula industry. Copyright © 2017 Elsevier Ltd. All rights reserved.

Applications of fiber reinforced concrete containers in France and in Slovakia

DOE Office of Scientific and Technical Information (OSTI.GOV)

Verdier, A.; Delgrande, J.; Remias, V.

Radioactive waste immobilization is an integral part of operations in nuclear facilities. The goal of immobilization is to contain radioactive materials in a waste form which can maintain its integrity over very long periods of time, thus effectively isolating the materials from the environment and hence from the public. This is true regardless of the activity of the waste, including low-, and medium-level waste (LLW, MLW). A multiple-year research effort by COGEMA culminated in the development of a new process to immobilize nuclear waste in concrete containers reinforced with metal fibers. The fiber reinforced concrete containers satisfy all French safetymore » requirements relating to waste immobilization and disposal, and have been certified by ANDRA, the national radioactive waste management agency. The fiber reinforced concrete containers have been fabricated on a production scale since July 1990 by Sogefibre, a jointly-owned subsidiary of SGN and Campaign Generale des Eaux. This technology is being transferred to Slovenske Elektrarne (Slovak Power Plant) to intern the waste produced by Bohunice and Mochovce power plants in cubical fiber reinforced concrete containers.« less

Immobilization of Polyoxometalates on Tailored Polymeric Surfaces

PubMed Central

Aguado-Ureta, Saioa; Rodríguez-Hernández, Juan; del Campo, Adolfo; Perez-Álvarez, Leyre

2018-01-01

Herein we describe the preparation of hybrid polymer–inorganic interfaces by the immobilization of polyoxometalate nanoclusters on functionalized polymer surfaces. The polymeric surfaces were made of polystyrene-b-poly(acrylic acid)/polystyrene (PS-b-PAA/PS) blends by spin coating on a silicon wafer. The functionalization of the polymer film was obtained by interfacial migration of the amphiphilic block copolymer toward the interface upon water vapor annealing. The carboxylic acid functional groups contained in the PAA block were then employed to anchor the [LnIII(α-SiW11O39)]5− polyoxometalates (Ln: Ce, Er). This purpose was achieved by immersing the films in aqueous solutions of the in situ-formed inorganic nanoclusters. X-ray photoelectron and confocal Raman spectroscopies, together with atomic force microscopy, confirmed the immobilization of the inorganic species at the interface. PMID:29498656

Method for carbon dioxide sequestration

DOEpatents

Wang, Yifeng; Bryan, Charles R.; Dewers, Thomas; Heath, Jason E.

2015-09-22

A method for geo-sequestration of a carbon dioxide includes selection of a target water-laden geological formation with low-permeability interbeds, providing an injection well into the formation and injecting supercritical carbon dioxide (SC--CO.sub.2) into the injection well under conditions of temperature, pressure and density selected to cause the fluid to enter the formation and splinter and/or form immobilized ganglia within the formation. This process allows for the immobilization of the injected SC--CO.sub.2 for very long times. The dispersal of scCO2 into small ganglia is accomplished by alternating injection of SC--CO.sub.2 and water. The injection rate is required to be high enough to ensure the SC--CO.sub.2 at the advancing front to be broken into pieces and small enough for immobilization through viscous instability.

Mold Pectinase Modified with Dialdehyde Derivatives of Dextran and Cellulose.

PubMed

Kobayashi, M; Chiba, Y; Funane, K; Ohya, S; Kato, Y

1996-01-01

Chemical modification of mold pectinase with dextran- and cellulose-dialdehydes was examined to improve the enzyme characteristics. The modified pectinase with dextran-dialdehyde retained about 50% of the original activity, and more than 80% of the total amino groups were modified. HPLC gel filtration analysis showed an increase in molecular weight of the reaction product. Reaction with cellulose-dialdehyde provided an immobilized form of pectinase. The immobilized pectinase was resistant to both acidic and alkaline pHs, and also acquired heat stability at 60°C. The optimum pH of the modified enzyme shifted from pH 4.5 to 5.0-5.5, and this enzyme had higher activity at neutral pH regions than the native enzyme. A rather low recovery of immobilized enzyme (14.5%) should be improved by the combination with various methods hitherto established.

Proteomic Analysis of an α7 Nicotinic Acetylcholine Receptor Interactome

PubMed Central

Paulo, Joao A.; Brucker, William J.; Hawrot, Edward

2009-01-01

The α7 nicotinic acetylcholine receptor (nAChR) is well established as the principal high-affinity α-bungarotoxin-binding protein in the mammalian brain. We isolated carbachol-sensitive α-bungarotoxin-binding complexes from total mouse brain tissue by affinity immobilization followed by selective elution, and these proteins were fractionated by SDS-PAGE. The proteins in subdivided gel lane segments were tryptically digested, and the resulting peptides were analyzed by standard mass spectrometry. We identified 55 proteins in wild-type samples that were not present in comparable brain samples from α7 nAChR knockout mice that had been processed in a parallel fashion. Many of these 55 proteins are novel proteomic candidates for interaction partners of the α7 nAChR, and many are associated with multiple signaling pathways that may be implicated in α7 function in the central nervous system. The newly identified potential protein interactions, together with the general methodology that we introduce for α-bungarotoxin-binding protein complexes, form a new platform for many interesting follow-up studies aimed at elucidating the physiological role of neuronal α7 nAChRs. PMID:19714875

Palladium nanoparticles deposited on graphene and its electrochemical performance for glucose sensing

NASA Astrophysics Data System (ADS)

Mijowska, Ewa; Onyszko, Magdalena; Urbas, Karolina; Aleksandrzak, Malgorzata; Shi, Xiaoze; Moszyński, Dariusz; Penkala, Krzysztof; Podolski, Jacek; El Fray, Mirosława

2015-11-01

This paper reports on the fabrication and characterization of glucose oxidase (GOx) immobilized onto a glassy carbon electrode (GCE) modified with reduced graphene oxide/palladium nanocomposite (RGO-Pd). Characterization tools showed well dispersed uniform Pd nanoparticles on a partly reduced graphene oxide surface. Cyclic voltammetry demonstrated successful immobilization of GOx on RGO-Pd modified GCE (GCE-RGO-Pd) using covalent bonding of GOx with RGO-Pd (RGO-Pd-GOx). Therefore, it was used as an electrochemical biosensor of glucose. RGO-Pd-GOx exhibited good electrocatalysis toward glucose in different glucose concentrations (from 2 to 10 mM, which includes the blood glucose levels of both normal and diabetic persons) with O2 saturated phosphate buffer solution (PBS) at pH 7.4. The system showed a linear increase in current at potential -0.085 V in the concentration range examined, with a correlation coefficient of 0.996. The sensitivity of the biosensor was 41.3 μA cm-2 mM-1, suggesting that RGO-Pd-GOx-modified GCE could be a potential candidate as a glucose sensor.

Apatite and sodalite based glass-bonded waste forms for immobilization of 129I and mixed halide radioactive wastes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Goel, Ashutosh; McCloy, John S.; Riley, Brian J.

The goal of the project was to utilize the knowledge accumulated by the team, in working with minerals for chloride wastes and biological apatites, toward the development of advanced waste forms for immobilizing 129I and mixed-halide wastes. Based on our knowledge, experience, and thorough literature review, we had selected two minerals with different crystal structures and potential for high chemical durability, sodalite and CaP/PbV-apatite, to form the basis of this project. The focus of the proposed effort was towards: (i) low temperature synthesis of proposed minerals (iodine containing sodalite and apatite) leading to the development of monolithic waste forms, (ii)more » development of a fundamental understanding of the atomic-scale to meso-scale mechanisms of radionuclide incorporation in them, and (iii) understanding of the mechanism of their chemical corrosion, alteration mechanism, and rates. The proposed work was divided into four broad sections. deliverables. 1. Synthesis of materials 2. Materials structural and thermal characterization 3. Design of glass compositions and synthesis glass-bonded minerals, and 4. Chemical durability testing of materials.« less

Rational Design of Peptide-Functionalized Surface Plasmon Resonance Sensor for Specific Detection of TNT Explosive.

PubMed

Wang, Jin; Muto, Masaki; Yatabe, Rui; Onodera, Takeshi; Tanaka, Masayoshi; Okochi, Mina; Toko, Kiyoshi

2017-09-30

In this study, a rationally-designed 2,4,6-trinitrotoluene (TNT) binding peptide derived from an amino acid sequence of the complementarity-determining region (CDR) of an anti-TNT monoclonal antibody was used for TNT detection based on a maleimide-functionalized surface plasmon resonance (SPR) sensor. By antigen-docking simulation and screening, the TNT binding candidate peptides were obtained as TNTHCDR1 derived from the heavy chain of CDR1, TNTHCDR2 derived from CDR2, and TNTHCDR3 from CDR3 of an anti-TNT antibody. The binding events between candidate peptides and TNT were evaluated using the SPR sensor by direct determination based on the 3-aminopropyltriethoxysilane (APTES) surface. The TNT binding peptide was directly immobilized on the maleimide-functionalized sensor chip surface from N-γ-maleimidobutyryl-oxysuccinimide ester (GMBS). The results demonstrated that peptide TNTHCDR3 was identified and selected as a TNT binding peptide among the other two candidate peptides. Five kinds of TNT analogues were also investigated to testify the selectivity of TNT binding peptide TNTHCDR3. Furthermore, the results indicated that the APTES-GMBS-based SPR sensor chip procedure featured a great potential application for the direct detection of TNT.

Rational Design of Peptide-Functionalized Surface Plasmon Resonance Sensor for Specific Detection of TNT Explosive

PubMed Central

Wang, Jin; Muto, Masaki; Yatabe, Rui; Onodera, Takeshi; Okochi, Mina; Toko, Kiyoshi

2017-01-01

In this study, a rationally-designed 2,4,6-trinitrotoluene (TNT) binding peptide derived from an amino acid sequence of the complementarity-determining region (CDR) of an anti-TNT monoclonal antibody was used for TNT detection based on a maleimide-functionalized surface plasmon resonance (SPR) sensor. By antigen-docking simulation and screening, the TNT binding candidate peptides were obtained as TNTHCDR1 derived from the heavy chain of CDR1, TNTHCDR2 derived from CDR2, and TNTHCDR3 from CDR3 of an anti-TNT antibody. The binding events between candidate peptides and TNT were evaluated using the SPR sensor by direct determination based on the 3-aminopropyltriethoxysilane (APTES) surface. The TNT binding peptide was directly immobilized on the maleimide-functionalized sensor chip surface from N-γ-maleimidobutyryl-oxysuccinimide ester (GMBS). The results demonstrated that peptide TNTHCDR3 was identified and selected as a TNT binding peptide among the other two candidate peptides. Five kinds of TNT analogues were also investigated to testify the selectivity of TNT binding peptide TNTHCDR3. Furthermore, the results indicated that the APTES-GMBS-based SPR sensor chip procedure featured a great potential application for the direct detection of TNT. PMID:28973962

Epsilon metal waste form for immobilization of noble metals from used nuclear fuel

NASA Astrophysics Data System (ADS)

Crum, Jarrod V.; Strachan, Denis; Rohatgi, Aashish; Zumhoff, Mac

2013-10-01

Epsilon metal (ɛ-metal), an alloy of Mo, Pd, Rh, Ru, and Tc, is being developed as a waste form to treat and immobilize the undissolved solids and dissolved noble metals from aqueous reprocessing of commercial used nuclear fuel. Epsilon metal is an attractive waste form for several reasons: increased durability relative to borosilicate glass, it can be fabricated without additives (100% waste loading), and in addition it also benefits borosilicate glass waste loading by eliminating noble metals from the glass, thus the processing problems related to their insolubility in glass. This work focused on the processing aspects of the epsilon metal waste form development. Epsilon metal is comprised of refractory metals resulting in high alloying temperatures, expected to be 1500-2000 °C, making it a non-trivial phase to fabricate by traditional methods. Three commercially available advanced technologies were identified: spark-plasma sintering, microwave sintering, and hot isostatic pressing, and investigated as potential methods to fabricate this waste form. Results of these investigations are reported and compared in terms of bulk density, phase assemblage (X-ray diffraction and elemental analysis), and microstructure (scanning electron microscopy).

Radioactive Demonstrations Of Fluidized Bed Steam Reforming (FBSR) With Hanford Low Activity Wastes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jantzen, C. M.; Crawford, C. L.; Burket, P. R.

Several supplemental technologies for treating and immobilizing Hanford low activity waste (LAW) are being evaluated. One immobilization technology being considered is Fluidized Bed Steam Reforming (FBSR) which offers a low temperature (700-750?C) continuous method by which wastes high in organics, nitrates, sulfates/sulfides, or other aqueous components may be processed into a crystalline ceramic (mineral) waste form. The granular waste form produced by co-processing the waste with kaolin clay has been shown to be as durable as LAW glass. The FBSR granular product will be monolithed into a final waste form. The granular component is composed of insoluble sodium aluminosilicate (NAS)more » feldspathoid minerals such as sodalite. Production of the FBSR mineral product has been demonstrated both at the industrial, engineering, pilot, and laboratory scales on simulants. Radioactive testing at SRNL commenced in late 2010 to demonstrate the technology on radioactive LAW streams which is the focus of this study.« less

Microbial-enhanced lindane removal by sugarcane (Saccharum officinarum) in doped soil-applications in phytoremediation and bioaugmentation.

PubMed

Salam, Jaseetha Abdul; Hatha, Mohammed A A; Das, Nilanjana

2017-05-15

The aim of this study was to examine the effect of lindane-degrading yeast on the growth and lindane uptake by Saccharum sp., in doped garden soils. The rhizosphere of Saccharum plant was amended with yeast Candida VITJzN04 by root-inoculation. The bio-augment yeast was applied in two different forms viz., planktonic form and cells immobilized on sugarcane-bagasse, in the pot experiments. Garden soils (lindane∼100 mg/kg) exposed to various treatments were monitored for a period of 30 days, for residual lindane by gas-chromatography analysis. The lindane-removal rates in soil were expressed in terms of half-life period and were recorded as 13.3 days (yeast), 43.3 days (Saccharum), 9.8 days (free yeast-plant) and 7.1 days (immobilized yeast-plant). Additionally, Candida sp., was also identified as a plant growth promoting yeast due to its ability to produce growth hormone and solubilize insoluble phosphates in the soil for better uptake by the plant species. Bio-stimulation of the soil with yeast immobilized on sugarcane bagasse further enhanced the total yeast activity in the soil which in turn had a positive influence on lindane-removal. Combined treatment with bagasse immobilized yeast and plant showed the best lindane degradation. Results suggested that the synergistic activity of plant and yeast resulted in fast and efficient degradation of lindane. Thus, it can be concluded that Saccharum plant in combination with Candida VITJzN04 is an effective alternative for the conventional remediation strategies. Copyright © 2017 Elsevier Ltd. All rights reserved.

Assessment of waste oyster shells and coal mine drainage sludge for the stabilization of As-, Pb-, and Cu-contaminated soil.

PubMed

Moon, Deok Hyun; Cheong, Kyung Hoon; Koutsospyros, Agamemnon; Chang, Yoon-Young; Hyun, Seunghun; Ok, Yong Sik; Park, Jeong-Hun

2016-02-01

A novel treatment mix was designed for the simultaneous immobilization of As, Cu, and Pb in contaminated soils using natural (waste oyster shells (WOS)) and industrial (coal mine drainage sludge (CMDS)) waste materials. The treatments were conducted using the standard U.S. sieve size no. 20 (0.85 mm) calcined oyster shells (COS) and CMDS materials with a curing time of 1 and 28 days. The As immobilization treatments were evaluated using the 1-N HCl extraction fluid, whereas the Pb and Cu immobilization treatments were evaluated using the 0.1-N HCl extraction fluid based on the Korean leaching standards. The treatment results showed that the immobilization of As, Cu, and Pb was best achieved using a combination mix of 10 wt% COS and 10 wt% CMDS. This treatment mix was highly effective leading to superior leachability reductions for all three target contaminants (>93 % for As and >99 % for Cu and Pb) for a curing period of 28 days. The X-ray absorption near-edge structure (XANES) results showed that As was present in the form of As(V) in the control sample and that no changes in As speciation were observed following the COS-CMDS treatments. The scanning electron microscopy (SEM)-energy dispersive X-ray spectroscopy (EDX) sample treated with 10 wt% COS and 10 wt% CMDS indicated that As immobilization may be associated with the formation of Ca-As and Fe-As precipitates while Pb and Cu immobilization was most probably linked to calcium silicate hydrates (CSHs) and calcium aluminum hydrates (CAHs).

Features of the supercritical CO2-assisted immobilization of fluorinated tetraphenylporphyrins into tetrafluoroethylene copolymers

NASA Astrophysics Data System (ADS)

Shershnev, I. V.; Cherkasova, A. V.; Kopylov, A. S.; Glagolev, N. N.; Bragina, N. A.; Solov'eva, A. B.

2017-07-01

The immobilization of fluorinated tetraphenylporphyrins (FTPPs) into tetrafluoroethylene copolymers (fluoroplast F-42 and MF-4SK, a perfluorinated sulfonic acid cation exchanger in H+-form) is conducted in supercritical CO2 (scCO2). The effects the conditions of immobilization (the temperature and pressure of scCO2, reaction time, and the addition of cosolvents) and the structure of the carrier polymer have on the content of porphyrin in these polymers is studied. The porphyrin-loaded polymer systems are shown to exhibit photosensitizing activity in anthracene and cholesterol oxidation in scCO2. Under conditions of photocatalysis, chemical and functional stability is a feature of only MF-4SK polymer systems; this is attributed to the formation of protonated forms of the porphyrins and their interaction with SO3 --groups of the polymer (an ion exchange process), which prevents leaching of the FTPP from the polymer matrix. The photocatalytic process actually occurs inside the matrix of the perfluorinated copolymer, with the protonated form of the porphyrin acting as a photosensitizer. The rate constant of anthracene photooxidation in the presence of FTPP-loaded MF-4SK films in scCO2 is found to pass through a maximum as a function of the porphyrin content and the polymer film thickness. The use of such catalytic systems for cholesterol photooxidation in scCO2 is shown to produce a virtual monoproduct (yield, 10%): 6-formyl-B-norcholestane-3,5-diol, a compound with high biological activity.

The effect of operational parameters on the biodegradation of bisphenols by Trametes versicolor laccase immobilized on Hippospongia communis spongin scaffolds.

PubMed

Zdarta, Jakub; Antecka, Katarzyna; Frankowski, Robert; Zgoła-Grześkowiak, Agnieszka; Ehrlich, Hermann; Jesionowski, Teofil

2018-02-15

Due to the rapid growth in quantities of phenolic compounds in wastewater, the development of efficient and environmentally friendly methods for their removal becomes a necessity. Thus, in a presented work, for the first time, a novel material, Hippospongia communis spongin-based scaffold, was used as a biopolymeric support for the immobilization of laccase from Trametes versicolor. The resulting biocatalytic systems were used for the biodegradation of three bisphenols: bisphenol A (BPA), bisphenol F (BPF) and bioremoval-resistant bisphenol S (BPS). Optimization of the immobilization and biodegradation methodologies was performed to increase bisphenols removal. The effect of temperature, pH and initial pollutant concentration was evaluated. It was shown that under optimal conditions, almost 100% of BPA (pH5, 30°C) and BPF (pH5, 40°C), and over 40% of BPS (pH4, 30°C) was removed from the solution at a concentration of 2mg/mL. Furthermore, the immobilized laccase exhibited good reusability and storage stability, retaining over 80% of its initial activity after 50days of storage. In addition, the main biodegradation products of BPA and BPF were identified. It was shown that mainly dimers and trimers were formed following the oxidation of bisphenols by the immobilized laccase. Copyright © 2017 Elsevier B.V. All rights reserved.

Evaluation of Phosphate Fertilizers for the Immobilization of Cd in Contaminated Soils

PubMed Central

Yan, Yin; Zhou, Yi Qun; Liang, Cheng Hua

2015-01-01

A laboratory investigation was conducted to evaluate the efficiency of four phosphate fertilizers, including diammonium phosphate (DAP), potassium phosphate monobasic (MPP), calcium superphosphateon (SSP), and calcium phosphate tribasic (TCP), in terms of the toxicity and bioavailability of Cd in contaminated soils. The efficiency of immobilization was evaluated on the basis of two criteria: (a) the reduction of extractable Cd concentration below the TCLP regulatory level and (b) the Cd changes associated with specific operational soil fractions on the basis of sequential extraction data. Results showed that after 50 d immobilization, the extractable concentrations of Cd in DAP, MPP, SSP, and TCP treated soils decreased from 42.64 mg/kg (in the control) to 23.86, 21.86, 33.89, and 35.59 mg/kg, respectively, with immobilization efficiency in the order of MPP > DAP > SSP > TCP. Results from the assessment of Cd speciation via the sequential extraction procedure revealed that the soluble exchangeable fraction of Cd in soils treated with phosphate fertilizers, especially TCP, was considerably reduced. In addition, the reduction was correspondingly related to the increase in the more stable forms of Cd, that is, the metal bound to manganese oxides and the metal bound to crystalline iron oxides. Treatment efficiency increased as the phosphate dose (according to the molar ratio of PO4/Cd) increased. Immobilization was the most effective under the molar ratio of PO4/Cd at 4:1. PMID:25915051

Bioconversion of Airborne Methylamine by Immobilized Recombinant Amine Oxidase from the Thermotolerant Yeast Hansenula polymorpha

PubMed Central

Sigawi, Sasi; Nitzan, Yeshayahu

2014-01-01

Aliphatic amines, including methylamine, are air-pollutants, due to their intensive use in industry and the natural degradation of proteins, amino acids, and other nitrogen-containing compounds in biological samples. It is necessary to develop systems for removal of methylamine from the air, since airborne methylamine has a negative effect on human health. The primary amine oxidase (primary amine : oxygen oxidoreductase (deaminating) or amine oxidase, AMO; EC 1.4.3.21), a copper-containing enzyme from the thermotolerant yeast Hansenula polymorpha which was overexpressed in baker's yeast Saccharomyces cerevisiae, was tested for its ability to oxidize airborne methylamine. A continuous fluidized bed bioreactor (CFBR) was designed to enable bioconversion of airborne methylamine by AMO immobilized in calcium alginate (CA) beads. The results demonstrated that the bioreactor with immobilized AMO eliminates nearly 97% of the airborne methylamine. However, the enzymatic activity of AMO causes formation of formaldehyde. A two-step bioconversion process was therefore proposed. In the first step, airborne methylamine was fed into a CFBR which contained immobilized AMO. In the second step, the gas flow was passed through another CFBR, with alcohol oxidase from the yeast H. polymorpha immobilized in CA, in order to decompose the formaldehyde formed in the first step. The proposed system provided almost total elimination of the airborne methylamine and the formaldehyde. PMID:24672387

Design of a Papain Immobilized Antimicrobial Food Package with Curcumin as a Crosslinker

PubMed Central

Sivakumar, Ponnurengam Malliappan; Doble, Mukesh

2015-01-01

Contamination of food products by spoilage and pathogenic microorganisms during post process handling is one of the major causes for food spoilage and food borne illnesses. The present green sustainable approach describes the covalent immobilization of papain to LDPE (low density polyethylene), HDPE (high density polyethylene), LLDPE (linear low density polyethylene) and PCL (polycaprolactam) with curcumin as the photocrosslinker. About 50% of curcumin and 82-92% of papain were successfully immobilized on these polymers. After 30 days, the free enzyme retained 87% of its original activity, while the immobilized enzyme retained more than 90% of its activity on these polymers. Papain crosslinked to LLDPE exhibited the best antibiofilm properties against Acinetobacter sp. KC119137.1 and Staphylococcus aureus NCIM 5021 when compared to the other three polymers, because of the highest amount of enzyme immobilized on this surface. Papain acts by damaging the cell membrane. The enzyme is able to reduce the amount of carbohydrate and protein contents in the biofilms formed by these organisms. Meat wrapped with the modified LDPE and stored at 4°C showed 9 log reduction of these organisms at the end of the seventh day when compared to samples wrapped with the bare polymer. This method of crosslinking can be used on polymers with or without functional groups and can be adopted to bind any type of antimicrobial agent. PMID:25906061

Chelation by collagen in the immobilization of Aspergillus oryzae β-galactosidase: A potential biocatalyst to hydrolyze lactose by batch processes.

PubMed

Gennari, Adriano; Mobayed, Francielle Herrmann; Volpato, Giandra; de Souza, Claucia Fernanda Volken

2018-04-01

This work is the first study of the immobilization of Aspergillus oryzae β-galactosidase (Gal) on powdered collagen (Col) that had formed a chelate with aluminum (Col-Al-Gal). Other collagen treatments, including those with acetic acid, glutaraldehyde, and a combination of aluminum and glutaraldehyde (Col-Al-Glu-Gal), were also tested. High-yield (superior to 80%) and high-efficiency (superior to 99%) immobilization was obtained for the derivatives Col-Al-Gal and Col-Al-Glu-Gal, even at high protein loads (500-1,000 mg g -1 of support). The storage stability of Gal immobilized on Col-Al and Col-Al-Glu resulted in Gal retaining approximately 60% of its initial activity after 90 days at 4 °C. The half-life values of derivatives Col-Al-Gal and Col-Al-Glu-Gal were higher than those of soluble enzyme at 65, 68, 70, and 73 °C. The derivatives Col-Al-Gal and Col-Al-Glu-Gal retained high enzyme activity in batch hydrolysis of lactose in permeate and lactose solutions for 50 and 60 cycles, respectively. Our results suggest that powdered collagen treated with aluminum, a low-cost support, is a promising support for the immobilization of β-galactosidase. Copyright © 2017 Elsevier B.V. All rights reserved.

Biodegradation of phenol in synthetic and industrial wastewater by Rhodococcus erythropolis UPV-1 immobilized in an air-stirred reactor with clarifier.

PubMed

Prieto, M B; Hidalgo, A; Rodríguez-Fernández, C; Serra, J L; Llama, M J

2002-05-01

Phenol biodegradation by suspended and immobilized cells of Rhodococcus erythropolis UPV-1 was studied in discontinuous and continuous mode under optimum culture conditions. Phenol-acclimated cells were adsorbed on diatomaceous earth, where they grew actively forming a biofilm of short filaments. Immobilization protected cells against phenol and resulted in a remarkable enhancement of their respiratory activity and a shorter lag phase preceding active phenol degradation. Under optimum operation conditions in a laboratory-scale air-stirred reactor, the immobilized cells were able to completely degrade phenol in synthetic wastewater at a volumetric productivity of 11.5 kg phenol m(-3) day(-1). Phenol biodegradation was also tested in two different industrial wastewaters (WW1 and WW2) obtained from local resin manufacturing companies, which contained both phenols and formaldehyde. In this case, after wastewater conditioning (i.e., dilution, pH, nitrogen and phosphorous sources and micronutrient amendments) the immobilized cells were able to completely remove the formaldehyde present in both waters. Moreover, they biodegraded phenols completely at a rate of 0.5 kg phenol m(-3) day(-1) in the case of WW1 and partially (but at concentrations lower than 50 mg l(-1)) at 0.1 and 1.0 kg phenol m(-3) day(-1) in the cases of WW2 and WW1, respectively.

Thermostable trypsin conjugates immobilized to biogenic magnetite show a high operational stability and remarkable reusability for protein digestion

NASA Astrophysics Data System (ADS)

Pečová, M.; Šebela, M.; Marková, Z.; Poláková, K.; Čuda, J.; Šafářová, K.; Zbořil, R.

2013-03-01

In this work, magnetosomes produced by microorganisms were chosen as a suitable magnetic carrier for covalent immobilization of thermostable trypsin conjugates with an expected applicability for efficient and rapid digestion of proteins at elevated temperatures. First, a biogenic magnetite was isolated from Magnetospirillum gryphiswaldense and its free surface was coated with the natural polysaccharide chitosan containing free amino and hydroxy groups. Prior to covalent immobilization, bovine trypsin was modified by conjugating with α-, β- and γ-cyclodextrin. Modified trypsin was bound to the magnetic carriers via amino groups using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide as coupling reagents. The magnetic biomaterial was characterized by magnetometric analysis and electron microscopy. With regard to their biochemical properties, the immobilized trypsin conjugates showed an increased resistance to elevated temperatures, eliminated autolysis, had an unchanged pH optimum and a significant storage stability and reusability. Considering these parameters, the presented enzymatic system exhibits properties that are superior to those of trypsin forms obtained by other frequently used approaches. The proteolytic performance was demonstrated during in-solution digestion of model proteins (horseradish peroxidase, bovine serum albumin and hen egg white lysozyme) followed by mass spectrometry. It is shown that both magnetic immobilization and chemical modification enhance the characteristics of trypsin making it a promising tool for protein digestion.

Immobilization of methotrexate anticancer drug onto the graphene surface and interaction with calf thymus DNA and 4T1 cancer cells.

PubMed

Karimi Shervedani, Reza; Mirhosseini, Hadiseh; Samiei Foroushani, Marzieh; Torabi, Mostafa; Rahsepar, Fatemeh Rahnemaye; Norouzi-Barough, Leila

2018-02-01

Immobilization of methotrexate (MTX) anticancer drug onto the graphene surface is reported through three methods, including either covalent linkage via (a) EDC/NHS organic activators and (b) electrografting of MTX diazonium salt, or (c) noncovalent bonding, resulting in three different systems. To evaluate the interaction ability of the immobilized MTX with biological species, calf thymus DNA (ctDNA), mouse 4T1 breast tumor, and Human foreskin fibroblast (hFF) cells as models of the primary intracellular target of anticancer drugs, cancer and normal cells, respectively, are examined. The features of the constructed systems and their interactions with ctDNA are followed by surface analysis techniques and electrochemical methods. The results indicate that (i) the amount of the immobilized MTX on the graphene surface is affected by type of the immobilization method; and a maximum value of (Γ=9.3±0.9pmolcm -2 ) is found via electrografting method, (ii) graphene-modified-MTX has high affinity for ctDNA in a wide dynamic range of concentrations, and (iii) the nature of the interaction is of electrostatic and/or hydrogen bonding type, formed most probably between OH, NH and CO groups of MTX and different DNA functions. Finally, electrochemical impedance spectroscopy results approved the high affinity of the systems for 4T1 cancer cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Immobilization of concanavalin A receptors during differentiation of neuroblastoma cells.

PubMed

Fishman, M C; Dragsten, P R; Spector, I

1981-04-30

Neuroblastoma cells serve as a useful model of neuronal development because compounds such as dimethyl sulphoxide (DMSO) and dibutyryl cyclic AMP cause them to undergo a process of controlled differentiation in tissue culture, during which they can extend long processes, develop characteristic excitability mechanisms, synthesize neurotransmitters and form synapses. We have used the technique of fluorescence photobleaching recovery to study the lateral mobility of cell-surface constituents during the differentiation of neuroblastoma clone N1E-115 cells. The concanavalin A (Con A) binding sites appear as discrete patches distributed over the entire cell surface and exhibit lateral mobility in undifferentiated cells comparable with that of surface glycoproteins of other cells. After induction of differentiation, however, the vast majority of Con A binding sites become immobilized, and we present data which suggest that the mechanism of this immobilization may involve linkage to the internal actin network.

Membrane Compartmentalization Reducing the Mobility of Lipids and Proteins within a Model Plasma Membrane.

PubMed

Koldsø, Heidi; Reddy, Tyler; Fowler, Philip W; Duncan, Anna L; Sansom, Mark S P

2016-09-01

The cytoskeleton underlying cell membranes may influence the dynamic organization of proteins and lipids within the bilayer by immobilizing certain transmembrane (TM) proteins and forming corrals within the membrane. Here, we present coarse-grained resolution simulations of a biologically realistic membrane model of asymmetrically organized lipids and TM proteins. We determine the effects of a model of cytoskeletal immobilization of selected membrane proteins using long time scale coarse-grained molecular dynamics simulations. By introducing compartments with varying degrees of restraints within the membrane models, we are able to reveal how compartmentalization caused by cytoskeletal immobilization leads to reduced and anomalous diffusional mobility of both proteins and lipids. This in turn results in a reduced rate of protein dimerization within the membrane and of hopping of membrane proteins between compartments. These simulations provide a molecular realization of hierarchical models often invoked to explain single-molecule imaging studies of membrane proteins.

Surface immobilization of heparin on functional polyisobutylene-based thermoplastic elastomer as a potential artificial vascular graft

NASA Astrophysics Data System (ADS)

Wu, Yi-Bo; Li, Kang; Xiang, Dong; Zhang, Min; Yang, Dan; Zhang, Jin-Han; Mao, Jing; Wang, Hao; Guo, Wen-Li

2018-07-01

Polyisobutylene-based thermoplastic elastomer (TPE) is a new soft biomaterial. Hydroxyl functional dendritic polyisobutylene-based TPEs (arb-SIBS-OH), which satisfy the design requirements for small-diameter vascular substitutes, were synthesized by controlled carbocationic polymerization. Creep property, which is the destructive weakness of polyisobutylene-based TPEs, was significantly improved with the formation of a "double network" promoted by branched structure and microphase separation. Compatibility of arb-SIBS-OH with rabbit blood was markedly enhanced by modifying heparin grafted from these hydroxyl functional groups. Application of "click chemistry" to immobilize heparin on arb-SIBS-OH surface was apparently effective in enhancing the bioactivity of heparin. Immobilized heparin, which directly bonded by ester bonds, was more likely to form multi-point binding on arb-SIBS-OH surface. This process hindered the accessibility of the heparin active sequence to antithrombin.

Synthesis of magnetic thermosensitive microcontainers for enzyme immobilization

NASA Astrophysics Data System (ADS)

Wang, Jianzhi; Zhao, Guanghui; Wang, Xinyu; Peng, Xiaomen; Li, Yanfeng

2015-05-01

We present a new approach for the fabrication of magnetic thermoresponsive polymer microcapsules with mobile magnetic spherical cores. The microcontainers form fried-egg-like structures with a polymer shell layer of 50 nm due to the existence of hollow cavities. The microcontainers undergo a temperature-induced volume phase transition upon changing the temperature and present an impressive magnetic response. The magnetic saturation of these smart microcontainers (42 emu/g) is high enough to meet most requirements of bioapplications. To further investigate the potential application of these smart microcontainers in biotechnology, Candida rugosa lipase was selected for the enzyme immobilization process. The immobilized lipase exhibited excellent thermal stability and reusability in comparison with the free enzyme. The adsorption/release of the lipase from the microcontainers can be controlled by the environmental temperature and magnetic force, thus, offering new potential applications such as in controlled drug delivery, bioseparation, and catalysis.

Immobilization of Trametes versicolor cultures for improving laccase production in bubble column reactor intensified by sonication.

PubMed

Wang, Feng; Guo, Chen; Liu, Chun-Zhao

2013-01-01

The mycelia of Trametes versicolor immobilized in alginate beads provided higher laccase production than that in pelleted form. An efficient ultrasonic treatment enhanced laccase production from the immobilized T. versicolor cultures. The optimized treatment process consisted of exposing 36-h-old bead cultures to 7-min ultrasonic treatments twice with a 12-h interval using a fixed ultrasonic power and frequency (120 W, 40 kHz). Using the intensification strategy with sonication, laccase production increased by more than 2.1-fold greater than the untreated control in both flasks and bubble column reactors. The enhancement of laccase production by ultrasonic treatment is related to the improved mass transfer of nutrients and product between the liquid medium and the gel matrix. These results provide a basis for the large-scale and highly-efficient production of laccase using sonobioreactors.

Construction of titanium dioxide nanorod/graphite microfiber hybrid electrodes for a high performance electrochemical glucose biosensor

NASA Astrophysics Data System (ADS)

Zhang, Jian; Yu, Xin; Guo, Weibo; Qiu, Jichuan; Mou, Xiaoning; Li, Aixue; Liu, Hong

2016-04-01

The demand for a highly sensitive and selective glucose biosensor which can be used for implantable or on-time monitoring is constantly increasing. In this work, TiO2 nanorods were synthesized in situ on the surface of graphite microfibers to yield TiO2 nanorod/graphite microfiber hybrid electrodes. The TiO2 nanorods not only retain the high activity of the immobilized glucose molecule, but also promote the direct electron transfer process on the electrode surface. As a working electrode in an electrochemical glucose biosensor in a flowing system, the microfiber hybrid electrodes exhibit high sensitivity, selectivity and stability. Due to its simplicity, low cost, high stability, and unique morphology, the TiO2 nanorod/graphite microfiber hybrid electrode is expected to be an excellent candidate for an implantable biosensor or for in situ flow monitoring.The demand for a highly sensitive and selective glucose biosensor which can be used for implantable or on-time monitoring is constantly increasing. In this work, TiO2 nanorods were synthesized in situ on the surface of graphite microfibers to yield TiO2 nanorod/graphite microfiber hybrid electrodes. The TiO2 nanorods not only retain the high activity of the immobilized glucose molecule, but also promote the direct electron transfer process on the electrode surface. As a working electrode in an electrochemical glucose biosensor in a flowing system, the microfiber hybrid electrodes exhibit high sensitivity, selectivity and stability. Due to its simplicity, low cost, high stability, and unique morphology, the TiO2 nanorod/graphite microfiber hybrid electrode is expected to be an excellent candidate for an implantable biosensor or for in situ flow monitoring. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr01360k

Evaluation of free or anchored antimicrobial peptides as candidates for the prevention of orthopaedic device-related infections.

PubMed

D'Este, Francesca; Oro, Debora; Boix-Lemonche, Gerard; Tossi, Alessandro; Skerlavaj, Barbara

2017-10-01

The prevention of implant-associated infection, one the most feared complications in orthopaedic surgery, remains a major clinical challenge and urges development of effective methods to prevent bacterial colonization of implanted devices. Alpha-helical antimicrobial peptides (AMPs) may be promising candidates in this respect due to their potent and broad-spectrum antimicrobial activity, their low tendency to elicit resistance and possible retention of efficacy in the immobilized state. The aim of this study was to evaluate the potential of five different helical AMPs, the cathelicidins BMAP-27 and BMAP-28, their (1-18) fragments and the rationally designed, artificial P19(9/G7) peptide, for the prevention of orthopaedic implant infections. Peptides were effective at micromolar concentrations against 22 Staphylococcus and Streptococcus isolates from orthopaedic infections, while only BMAP-28 and to a lesser extent BMAP-27 were active against Enterococcus faecalis. Peptides in solution showed activities comparable to those of cefazolin and linezolid, on a molar basis, and also a variable capacity to neutralize bacterial lipopolysaccharide, while devoid of adverse effects on MG-63 osteoblast cells at concentrations corresponding to the MIC. The (1-18) BMAP fragments and P19(9/G7) were selected for further examination, based on better selectivity indices, and showed effectiveness in the presence of hyaluronic acid and in synovial fluid, while human serum affected their activity to variable extents, with BMAP-27(1-18) best retaining activity. This peptide was immobilized on streptavidin-resin beads and retained activity against reference Staphylococcus epidermidis and Staphylococcus aureus strains, with negligible toxicity towards osteoblasts, underlining its potential for the development of infection-resistant biomaterials for orthopaedic application. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

Test plan for formulation and evaluation of grouted waste forms with shine process wastes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ebert, W. L.; Jerden, J. L.

2015-09-01

The objective of this experimental project is to demonstrate that waste streams generated during the production of Mo99 by the SHINE Medical Technologies (SHINE) process can be immobilized in cement-based grouted waste forms having physical, chemical, and radiological stabilities that meet regulatory requirements for handling, storage, transport, and disposal.

Use of a novel immobilization yeast system for winemaking.

PubMed

Peinado, Rafael A; Moreno, Juan J; Maestre, Oscar; Mauricio, Juan C

2005-09-01

Penicillium was used to immobilize Saccharomyces cerevisiae, without using physico-chemical external supports, to form yeast biocapsules. The biocapsules, once the Penicillium was killed by the ethanol produced, were used in a grape must fermentation. Must fermentation was carried out for 160 h with the biocapsules and for 300 h with free yeast cells. Acetaldehyde (84 vs. 63 mg/l), isobutanol (217 vs. 194 mg/l), L: -proline (7.7 vs. 6.5 mM: ) and aspartic acid (0.42 vs. 0 mM: ) in final wine were higher with the biocapsules than with free cells.

Experimental gastric ulcers induced by immobilization and electric shock of rats and their pharmacotherapy

NASA Technical Reports Server (NTRS)

Zabrodin, O. N.

1980-01-01

The mechanism of development of experimental gastric ulcers, induced in rats by combined immobilization and electric shock, was analyzed pharmacologically with peripheral neurotropic agents. It is concluded that: (1) The most marked preventive effect in the development of the experimentally induced gastric ulcers was displayed by agents capable of blocking the ascending activation system of the reticular formation. (2) Sympathetic fibers, which disrupt the trophism of the gastric wall, form the efferent portion of the reflex arc. (3) Gastric secretion does not appear to be the primary cause of ulceration.

Solid electrolytes

DOEpatents

Abraham, Kuzhikalail M.; Alamgir, Mohamed

1993-06-15

This invention pertains to Li ion (Li.sup.+) conductive solid polymer electrolytes composed of solvates of Li salts immobilized (encapsulated) in a solid organic polymer matrix. In particular, this invention relates to solid polymer electrolytes derived by immobilizing complexes (solvates) formed between a Li salt such as LiAsF.sub.6, LiCF.sub.3 SO.sub.3 or LiClO.sub.4 and a mixture of aprotic organic solvents having high dielectric constants such as ethylene carbonate (EC) (dielectric constant=89.6) and propylene carbonate (PC) (dielectric constant=64.4) in a polymer matrix such as polyacrylonitrile, poly(tetraethylene glycol diacrylate), or poly(vinyl pyrrolidinone).

Development of a paper-based carbon nanotube sensing microfluidic device for biological detection.

PubMed

Yang, Shih-I; Lei, Kin Fong; Tsai, Shiao-Wen; Hsu, Hsiao-Ting

2013-01-01

Carbon nanotube (CNT) has been utilized for the biological detection due to its extremely sensitive to biological molecules. A paper-based CNT sensing microfluidic device has been developed for the detection of protein, i.e., biotin-avidin, binding. We have developed a fabrication method that allows controlled deposition of bundled CNTs with well-defined dimensions to form sensors on paper. Then, polydimethyl siloxane (PDMS) was used to pattern the hydrophobic boundary on paper to form the reaction sites. The proposed fabrication method is based on vacuum filtration process with a metal mask covering on a filter paper for the definition of the dimension of sensor. The length, width, and thickness of the CNT-based sensors are readily controlled by the metal mask and the weight of the CNT powder used during the filtration process, respectively. Homogeneous deposition of CNTs with well-defined dimensions can be achieved. The CNT-based sensor on paper has been demonstrated on the detection of the protein binding. Biotin was first immobilized on the CNT's sidewall and avidin suspended solution was applied to the site. The result of the biotin-avidin binding was measured by the resistance change of the sensor, which is a label-free detection method. It showed the CNT is sensitive to the biological molecules and the proposed paper-based CNT sensing device is a possible candidate for point-of-care biosensors. Thus, electrical bio-assays on paper-based microfluidics can be realized to develop low cost, sensitive, and specific diagnostic devices.

76 FR 82313 - Intent To Request Renewal From OMB of One Current Public Collection of Information...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-12-30

... Security Officer (TSO) to ensure their qualifications to perform TSO duties pursuant to 49 U.S.C 44965. In certain cases, TSO candidates' health care providers may be asked to complete supplemental forms. DATES... candidates and, in certain cases, supplemental forms completed by TSO candidates' health care providers. The...

Biomineralization of U(VI) phosphate promoted by microbially-mediated phytate hydrolysis in contaminated soils

NASA Astrophysics Data System (ADS)

Salome, Kathleen R.; Beazley, Melanie J.; Webb, Samuel M.; Sobecky, Patricia A.; Taillefert, Martial

2017-01-01

The bioreduction of uranium may immobilize a significant fraction of this toxic contaminant in reduced environments at circumneutral pH. In oxic and low pH environments, however, the low solubility of U(VI)-phosphate minerals also makes them good candidates for the immobilization of U(VI) in the solid phase. As inorganic phosphate is generally scarce in soils, the biomineralization of U(VI)-phosphate minerals via microbially-mediated organophosphate hydrolysis may represent the main immobilization process of uranium in these environments. In this study, contaminated sediments were incubated aerobically in two pH conditions to examine whether phytate, a naturally-occurring and abundant organophosphate in soils, could represent a potential phosphorous source to promote U(VI)-phosphate biomineralization by natural microbial communities. While phytate hydrolysis was not evident at pH 7.0, nearly complete hydrolysis was observed both with and without electron donor at pH 5.5, suggesting indigenous microorganisms express acidic phytases in these sediments. While the rate of hydrolysis of phytate generally increased in the presence of uranium, the net rate of inorganic phosphate production in solution was decreased and inositol phosphate intermediates were generated in contrast to similar incubations conducted without uranium. These findings suggest uranium stress enhanced the phytate-metabolism of the microbial community, while simultaneously inhibiting phosphatase production and/or activity by the indigenous population. Finally, phytate hydrolysis drastically decreased uranium solubility, likely due to formation of ternary sorption complexes, U(VI)-phytate precipitates, and U(VI)-phosphate minerals. Overall, the results of this study provide evidence for the ability of natural microbial communities to liberate phosphate from phytate in acidic sediments, possibly as a detoxification mechanism, and demonstrate the potential utility of phytate-promoted uranium immobilization in subsurface environments. These processes should be investigated in more detail with pure cultures isolated from these sediments.

Setting and stiffening of cementitious components in Cast Stone waste form for disposal of secondary wastes from the Hanford waste treatment and immobilization plant

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chung, Chul-Woo; Chun, Jaehun, E-mail: jaehun.chun@pnnl.gov; Um, Wooyong

2013-04-01

Cast Stone is a cementitious waste form, a viable option to immobilize secondary nuclear liquid wastes generated from the Hanford Waste Treatment and Immobilization Plant. However, no study has been performed to understand the flow and stiffening behavior, which is essential to ensure proper workability and is important to safety in a nuclear waste field-scale application. X-ray diffraction, rheology, and ultrasonic wave reflection methods were used to understand the specific phase formation and stiffening of Cast Stone. Our results showed a good correlation between rheological properties of the fresh mixture and phase formation in Cast Stone. Secondary gypsum formation wasmore » observed with low concentration simulants, and the formation of gypsum was suppressed in high concentration simulants. A threshold concentration for the drastic change in stiffening was found at 1.56 M Na concentration. It was found that the stiffening of Cast Stone was strongly dependent on the concentration of simulant. Highlights: • A combination of XRD, UWR, and rheology gives a better understanding of Cast Stone. • Stiffening of Cast Stone was strongly dependent on the concentration of simulant. • A drastic change in stiffening of Cast Stone was found at 1.56 M Na concentration.« less

Preparation and characterization compatible pellets for immobilization of colloidal sulphur nanoparticles

NASA Astrophysics Data System (ADS)

Adlim, M.; Zarlaida, F.; Khaldun, I.; Dewi, R.; Jamilah, M.

2018-03-01

Mercury pollution in atmosphere is dominated by mercury vapour release from coal burning and gold-amalgam separation in gold mining. The initial steps in formulating a compatible mercury absorbent for mercury stabilization was fabrication of pellet supported colloidal sulphur. Sulphur is used to stabilize mercury vapour by formation of metacinnabar that has much lower toxicity. The sulphur reactivity toward mercury vapour can be enhanced by using colloidal sulphur nanoparticles immobilized on compatible pellets. Clay pellets would have heat resistance but in fact, they were less stable in aqueous solution although their stability increased with inclusion of rice husk ash and sawdust or pineapple leaf fibre in the composite. Pellets made of rice husk ash and polyvinyl acetate were stable in water at least for 24 hours. Sulphur from thiosulfate precursor that immobilized onto surface of pellet using chitosan as the stabilizer and the binding agent gave lower sulphur content compared to sulphur from other precursors (sulphur powder and sulphur-CS2). Sulphur from thiosulfate precursor was in form of colloid, has nanosize, and disperse particles on the surface of rice husk ash pellets. Sulphur immobilization methods affect on sulphur particles exposure on the pellet surface.

Method for immobilizing particulate materials in a packed bed

DOEpatents

Even, W.R. Jr.; Guthrie, S.E.; Raber, T.N.; Wally, K.; Whinnery, L.L.; Zifer, T.

1999-02-02

The present invention pertains generally to immobilizing particulate matter contained in a packed bed reactor so as to prevent powder migration, compaction, coalescence, or the like. More specifically, this invention relates to a technique for immobilizing particulate materials using a microporous foam-like polymer such that (a) the particulate retains its essential chemical nature, (b) the local movement of the particulate particles is not unduly restricted, (c) bulk powder migration and is prevented, (d) physical and chemical access to the particulate is unchanged over time, and (e) very high particulate densities are achieved. The immobilized bed of the present invention comprises a vessel for holding particulate matter, inlet and an outlet ports or fittings, a loosely packed bed of particulate material contained within the vessel, and a three dimensional porous matrix for surrounding and confining the particles thereby fixing the movement of an individual particle to a limited local position. The established matrix is composed of a series of cells or chambers comprising walls surrounding void space, each wall forming the wall of an adjacent cell; each wall containing many holes penetrating through the wall yielding an overall porous structure and allowing useful levels of gas transport. 4 figs.

The influence of Pb addition on the properties of fly ash-based geopolymers.

PubMed

Nikolić, Violeta; Komljenović, Miroslav; Džunuzović, Nataša; Miladinović, Zoran

2018-05-15

Preventing or reducing negative effects on the environment from the waste landfilling is the main goal defined by the European Landfill Directive. Generally geopolymers can be considered as sustainable binders for immobilization of hazardous wastes containing different toxic elements. In this paper the influence of addition of high amount of lead on structure, strength, and leaching behavior (the effectiveness of Pb immobilization) of fly ash-based geopolymers depending on the geopolymer curing conditions was investigated. Lead was added during the synthesis of geopolymers in the form of highly soluble salt - lead-nitrate. Structural changes of geopolymers as a result of lead addition/immobilization were assessed by means of XRD, SEM/EDS, and 29 Si MAS NMR analysis. Investigated curing conditions significantly influenced structure, strength and leaching behavior of geopolymers. High addition of lead caused a sizeable decrease in compressive strength of geopolymers and promoted formation of aluminum-deficient aluminosilicate gel (depolymerization of aluminosilicate gel), regardless of the curing conditions investigated. According to the EUWAC limitations, 4% of lead was successfully immobilized by fly ash-based geopolymers cured for 28 days in a humid chamber at room temperature. Copyright © 2018 Elsevier B.V. All rights reserved.

Biochemical surface modification of Co-Cr-Mo.

PubMed

Puleo, D A

1996-01-01

Because of the limited mechanical properties of tissue substitutes formed by culturing cells on polymeric scaffolds, other approaches to tissue engineering must be explored for applications that require complete and immediate ability to bear weight, e.g. total joint replacements. Biochemical surface modification offers a way to partially regulate events at the bone-implant interface to obtain preferred tissue responses. Tresyl chloride, gamma-aminopropyltriethoxysilane (APS) and p-nitrophenyl chloroformate (p-NPC) immobilization schemes were used to couple a model enzyme, trypsin, on bulk samples of Co-Cr-Mo. For comparison, samples were simply adsorbed with protein. The three derivatization schemes resulted in different patterns and levels of activity. Tresyl chloride was not effective in immobilizing active enzyme on Co-Cr-Mo. Aqueous silanization with 12.5% APS resulted in optimal immobilized activity. Activity on samples derivatized with 0.65 mg p-NPC cm-2 was four to five times greater than that on samples simple adsorbed with enzyme or optimally derivatized with APS and was about eight times that on tresylated samples. This work demonstrates that, although different methods have different effectiveness, chemical derivatization can be used to alter the amount and/or stability of biomolecules immobilized on the surface of Co-Cr-Mo.

The enhancement of the hydrolysis of bamboo biomass in ionic liquid with chitosan-based solid acid catalysts immobilized with metal ions.

PubMed

Cheng, Jie; Wang, Nan; Zhao, Dezhou; Qin, Dandan; Si, Wenqing; Tan, Yunfei; Wei, Shun'an; Wang, Dan

2016-11-01

Three kinds of sulfonated cross-linked chitosan (SCCR) immobilized with metal ions of Cu(2+), Fe(3+) and Zn(2+) individually were synthesized and firstly used as solid acid catalysts in the hydrolysis of bamboo biomass. FTIR spectra showed that metal ions had been introduced into SCCR and the N-metal ions coordinate bound was formed. The particle sizes of these catalysts were about 500-1000μm with a pore size of 50-160μm. All of the three kinds of catalysts performed well for bamboo hydrolysis with 1-butyl-3-methyl-imidazolium chloride used as solvent. The most effective one was sulfonated cross-linked chitosan immobilized with Fe(3+) (Fe(3+)-SCCR). TRS yields were up to 73.42% for hydrolysis of bamboo powder in [C4mim]Cl with Fe(3+)-SCCR at 120°C and 20RPM after 24h. These novel chitosan-based metal ions immobilized solid acid catalysts with ionic liquids as the solvent might be promising to facilitate cost-efficient conversion of biomass into biofuels and bioproducts. Copyright © 2016 Elsevier Ltd. All rights reserved.

Mercury bioremediation by mercury accumulating Enterobacter sp. cells and its alginate immobilized application.

PubMed

Sinha, Arvind; Khare, Sunil Kumar

2012-02-01

The effective microbial remediation of the mercury necessitates the mercury to be trapped within the cells without being recycled back to the environment. The study describes a mercury bioaccumulating strain of Enterobacter sp., which remediated mercury from the medium simultaneous to its growth. The transmission electron micrographs and electron dispersive X-ray analysis revealed the accumulation of remediated mercury as nano-size particles in the cytoplasm as well as on the cell wall. The Enterobacter sp. in the present work was able to accumulate mercury, without being engineered in its native form. The possibility of recovering the accumulated mercury from the cells is also indicated. The applicability of the alginate immobilized cells in removing mercury from synthetic and complex industrial effluent in a batch mode was amply demonstrated. The initial load of 7.3 mg l(-1) mercury in the industrial effluent was completely removed in 72 h. The cells immobilized in calcium alginate were similarly effective in the complete removal of 5 mg l(-1) HgCl(2) of mercury from the synthetic effluent in less than 72 h. The immobilized cells could be reused for multiple cycles.

Porous immobilized C coated N doped TiO2 containing in-situ generated polyenes for enhanced visible light photocatalytic activity

NASA Astrophysics Data System (ADS)

Sabri, N. A.; Nawi, M. A.; Nawawi, W. I.

2015-10-01

Carbon coated nitrogen-doped Degussa P25TiO2 (or C,N-P25TiO2) was successfully immobilized on a glass plate using epoxidized natural rubber (ENR-50) and polyvinyl chloride (PVC) as the organic binders. Photo-etching of the fabricated system for 10 h oxidized its PVC binder into polyenes as well as forming a highly porous surface. The band gap energy (Eg) of the photo-etched immobilized photocatalyst system (C,N-P25TiO2/ENR/PVC-10 h) was reduced from 2.91 to 2.86 eV. Its photocatalytic activity was studied via photocatalytic degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) under a 45 W visible light fluorescent lamp. C,N-P25TiO2/ENR/PVC-10 h with polyenes performed better than its slurry counterpart under visible light irradiation where the conjugated double bonds acted as photo sensitizers. The immobilized C,N-P25TiO2/ENR/PVC-10 h has excellent reusability and sustainable with an average k value of 0.056 ± 0.011 min-1 and average percent removal of 99.18 ± 0.54%.

Method for immobilizing particulate materials in a packed bed

DOEpatents

Even, Jr., William R.; Guthrie, Stephen E.; Raber, Thomas N.; Wally, Karl; Whinnery, LeRoy L.; Zifer, Thomas

1999-01-01

The present invention pertains generally to immobilizing particulate matter contained in a "packed" bed reactor so as to prevent powder migration, compaction, coalescence, or the like. More specifically, this invention relates to a technique for immobilizing particulate materials using a microporous foam-like polymer such that a) the particulate retains its essential chemical nature, b) the local movement of the particulate particles is not unduly restricted, c) bulk powder migration and is prevented, d) physical and chemical access to the particulate is unchanged over time, and e) very high particulate densities are achieved. The immobilized bed of the present invention comprises a vessel for holding particulate matter, inlet and an outlet ports or fittings, a loosely packed bed of particulate material contained within the vessel, and a three dimensional porous matrix for surrounding and confining the particles thereby fixing the movement of individual particle to a limited local position. The established matrix is composed of a series of cells or chambers comprising walls surrounding void space, each wall forming the wall of an adjacent cell; each wall containing many holes penetrating through the wall yielding an overall porous structure and allowing useful levels of gas transport.

Increasing binding density of yeast cells by control of surface charge with allylamine grafting to ion modified polymer surfaces.

PubMed

Tran, Clara T H; Kondyurin, Alexey; Chrzanowski, Wojciech; Bilek, Marcela M M; McKenzie, David R

2014-10-01

Plasma immersion ion implantation (PIII) treatment of polymers creates a biointerface capable of direct covalent immobilization of biomolecules. The immobilization of protein molecules is achieved by covalent bonds formed between embedded radicals on the treated surface and amino acid side chains and cells can be immobilized through cell-wall proteins. The attachment density of negatively charged entities on a PIII treated surface is inhibited by its negative surface charge at neutral pH. To reduce the negative charge of PIII treated surfaces in phosphate buffer (pH 7.4, 11mM), we develop an effective approach of grafting allylamine monomers onto the treated surface. The results reveal reactions between allylamine and radicals on the PIII treated surface. One of these triggers polymerization, increasing the number of amine groups grafted. As a consequence, the PIII treated polystyrene surface after allylamine exposure becomes more hydrophobic and less negatively charged in phosphate buffer. Using yeast cells as an example, we have shown a significant improvement (6-15 times) of cell density immobilized on the PIII treated surface after exposure to allylamine. Copyright © 2014 Elsevier B.V. All rights reserved.

Bubble Formation and Lattice Parameter Changes Resulting from He Irradiation of Defect-Fluorite Gd2Zr2O7

DOE Office of Scientific and Technical Information (OSTI.GOV)

Taylor, Caitlin A.; Patel, Maulik K.; Aguiar, Jeffery A.

2016-08-15

Pyrochlores have long been considered as potential candidates for advanced ceramic waste-forms for the immobilization of radioactive waste nuclides. This work provides evidence that Gd2Zr2O7, often considered the most radiation tolerant pyrochlore, could be susceptible to radiation damage in the form of bubble nucleation at the highest He doses expected over geological time. Ion irradiations were utilized to experimentally simulate the radiation damage and He accumulation produced by ..alpha..-decay. Samples were pre-damaged using 7 MeV Au3+ to induce the pyrochlore to defect-fluorite phase transformation, which would occur due to ..alpha..-recoil damage within several hundred years of storage in a Gd2Zr2O7more » waste-form. These samples were then implanted to various He concentrations in order to study the long-term effects of He accumulation. Helium bubbles 1-3 nm in diameter were observed in TEM at a concentration of 4.6 at.% He. Some bubbles remained isolated, while others formed chains 10-30 nm in length parallel to the surface. GIXRD measurements showed lattice swelling after irradiating pristine Gd2Zr2O7 with 7 MeV Au3+ to a fluence of 2.2 x 1015 Au/cm2. An increase in lattice swelling was also measured after 2.2 x 1015 Au/cm2 + 2 x 1015 He/cm2 and 2.2 x 1015 Au/cm2 + 2 x 1016 He/cm2. A decrease in lattice swelling was measured after irradiation with 2.2 x 1015 Au/cm2 + 2 x 1017 He/cm2, the fluence where bubbles and bubble chains were observed in TEM. Bubble chains are thought to form in order to reduce lattice strain normal to the surface, which is produced by the Au and He irradiation damage.« less

[Phase transfer catalyzed bioconversion of penicillin G to 6-APA by immobilized penicillin acylase in recyclable aqueous two-phase systems with light/pH sensitive copolymers].

PubMed

Jin, Ke-ming; Cao, Xue-jun; Su, Jin; Ma, Li; Zhuang, Ying-ping; Chu, Ju; Zhang, Si-liang

2008-03-01

Immobilized penicillin acylase was used for bioconversion of penicillin PG into 6-APA in aqueous two-phase systems consisting of a light-sensitive polymer PNBC and a pH-sensitive polymer PADB. Partition coefficients of 6-APA was found to be about 5.78 in the presence of 1% NaCl. Enzyme kinetics showed that the reaction reached equilibrium at roughly 7 h. The 6-APA mole yields were 85.3% (pH 7.8, 20 degrees C), with about 20% increment as compared with the reaction of single aqueous phase buffer. The partition coefficient of PG (Na) varied scarcely, while that of the product, 6-APA and phenylacetic acid (PA) significantly varied due to Donnan effect of the phase systems and hydrophobicity of the products. The variation of the partition coefficients of the products also affected the bioconversion yield of the products. In the aqueous two-phase systems, the substrate, PG, the products of 6-APA and PA were biased in the top phase, while immobilized penicillin acylase at completely partitioned at the bottom. The substrate and PG entered the bottom phase, where it was catalyzed into 6-APA and PA and entered the top phase. Inhibition of the substrate and products was removed to result in improvement of the product yield, and the immobilized enzyme showed higher efficiency than the immobilized cells and occupied smaller volume. Compared with the free enzyme, immobilized enzyme had greater stability, longer life-time, and was completely partitioned in the bottom phase and recycle. Bioconversion in two-phase systems using immobilized penicillin acylase showed outstanding advantage. The light-sensitive copolymer forming aqueous two-phase systems could be recovered by laser radiation at 488 nm or filtered 450 nm light, while pH-sensitive polymer PADB could be recovered at the isoelectric point (pH 4.1). The recovery of the two copolymers was between 95% and 99%.

Effective L-Tyrosine Hydroxylation by Native and Immobilized Tyrosinase

PubMed Central

Lewańczuk, Marcin; Koźlecki, Tomasz; Liesiene, Jolanta; Bryjak, Jolanta

2016-01-01

Hydroxylation of L-tyrosine to 3,4-dihydroxyphenylalanine (L-DOPA) by immobilized tyrosinase in the presence of ascorbic acid (AH2), which reduces DOPA-quinone to L-DOPA, is characterized by low reaction yields that are mainly caused by the suicide inactivation of tyrosinase by L-DOPA and AH2. The main aim of this work was to compare processes with native and immobilized tyrosinase to identify the conditions that limit suicide inactivation and produce substrate conversions to L-DOPA of above 50% using HPLC analysis. It was shown that immobilized tyrosinase does not suffer from partitioning and diffusion effects, allowing a direct comparison of the reactions performed with both forms of the enzyme. In typical processes, additional aeration was applied and boron ions to produce the L-DOPA and AH2 complex and hydroxylamine to close the cycle of enzyme active center transformations. It was shown that the commonly used pH 9 buffer increased enzyme stability, with concomitant reduced reactivity of 76%, and that under these conditions, the maximal substrate conversion was approximately 25 (native) to 30% (immobilized enzyme). To increase reaction yield, the pH of the reaction mixture was reduced to 8 and 7, producing L-DOPA yields of approximately 95% (native enzyme) and 70% (immobilized). A three-fold increase in the bound enzyme load achieved 95% conversion in two successive runs, but in the third one, tyrosinase lost its activity due to strong suicide inactivation caused by L-DOPA processing. In this case, the cost of the immobilized enzyme preparation is not overcome by its reuse over time, and native tyrosinase may be more economically feasible for a single use in L-DOPA production. The practical importance of the obtained results is that highly efficient hydroxylation of monophenols by tyrosinase can be obtained by selecting the proper reaction pH and is a compromise between complexation and enzyme reactivity. PMID:27711193

Immobilization of Cd in landfill-leachate-contaminated soil with cow manure compost as soil conditioners: A laboratory study.

PubMed

Liao, Zhuwei; Wang, Jia; Wan, Rui; Xi, Shuang; Chen, Zhuqi; Chen, Zhulei; Yu, Yingjian; Long, Sijie; Wang, Huabin

2016-12-01

Introducing cow manure compost as an amendment in landfill-leachate-contaminated soils is proved to be an effective technique for the immobilization of Cd in this study. Landfill-leachate-contaminated soil was collected from an unlined landfill in China and amended with a different blending quantity of cow manure compost (0, 12, 24, 36, and 48 g per 200 g soil), which was made by mixing cow manure and chaff at a ratio of 1/1 and maturing for 6 months. pH values of five different blending quantity mixtures increased by 0.2-0.4, and the organic matter levels increased by 2.5-7%, during a remediation period of 5 weeks. Four fractions of Cd named exchangeable Cd, reducible Cd, oxidizable Cd, and residual Cd in soil were respectively analyzed by a sequential extraction procedure. Introducing the cow manure compost application resulted in more than 40% lower exchangeable Cd but a higher concentration of oxidizable Cd in soils, and mass balance results showed nearly no Cd absorption by applied material, indicating that transformation of exchangeable Cd into oxidization forms was the main mechanism of Cd immobilization when cow manure compost was used as an amendment. The Pearson correlation showed that increasing of pH values significantly improved the efficiency of Cd immobilization, with a correlation coefficiency of 0.940 (p < 0.05). This is the first attempt at heavy metal immobilization in landfill-leachate-contaminated soil by cow manure compost, and findings of this work can be integrated to guide the application. Addition of cow manure compost (CMC) was effective in reducing exchangeable Cd in landfill-leachate-contaminated soils (LLCS). The immobilization effect of Cd was mainly assigned to the redistribution of labile soil Cd. Organic matter (OM) and pH value increased with CMC application. The pH values were more sensitive to Cd immobilization efficiency. It was proved that CMC can be safely and effectively used for the restoration of LLCS.

Immobilization of Lecitase® Ultra onto a novel polystyrene DA-201 resin: characterization and biochemical properties.

PubMed

Liu, Ning; Fu, Min; Wang, Yong; Zhao, Qiangzhong; Sun, Weizheng; Zhao, Mouming

2012-11-01

A simple, rapid, and economic method of enzyme immobilization was developed for phospholipase Lecitase® ultra (LU) via interfacial adsorption. The effect of nature of the polystyrene supports and the kinetic behavior and stability of immobilized lecitase® ultra (IM-LU) were evaluated. Six macroporous resins (AB-8, X-5, DA-201, NKA-9, D101, D4006) and two anion resins (D318 and D201) were studied as the supports. DA-201 resin was selected because of its best immobilization effect for LU. Immobilization conditions were investigated, including immobilization time, pH, and enzyme concentration. IM-LU with a lipase activity of 1,652.4 ± 8.6 U/g was obtained. The adsorption process was modeled by Langmuir and Freundlich equations, and the experimental data were better fit for the former one. The kinetic constant (K (m)) values were found to be 192.7 ± 2.2 mM for the free LU and 249.3 ± 5.4 mM for the IM-LU, respectively. The V (max) value of free LU (169.5 ± 4.3 mM/min) was higher than that of the IM-LU (53.8 ± 1.5 mM/min). Combined strategies of scanning electron micrograph, thermogravimetric analysis, and Fourier transform infrared (FTIR) spectroscopy were employed to characterize the IM-LU. FTIR spectroscopy showed that the secondary conformation of the enzyme had changed after immobilization, through which a decrease of α-helix content and an increase of β-sheet content were observed. The IM-LU possessed an improved thermal stability as well as metal ionic tolerance when compared with its free form. The reusability of IM-LU was also evaluated through catalyzing esterification reaction between oleic acid and glycerol. It exhibited approximately 70 % of relative esterification efficiency after six successive cycles. This immobilized enzyme on hydrophobic support may well be used for the synthesis of structural lipids in lipid area.

From Evaluation to Collaborative Reflection: Teacher Candidate Perceptions of a Digital Learner-Centered Classroom Observation Form

ERIC Educational Resources Information Center

Flores, Raymond; Krutka, Daniel G.; Mason, Katherine; Bergman, Daniel J.

2014-01-01

In efforts to promote a more learner-centered approach to supervision, faculty members at a Midwestern U.S. university piloted a new digital classroom observation form. Participants included middle/secondary teacher candidates (N = 28) in their final semester of their programs. The overall goal of this study was to gather teacher candidates'…

The Challenges of PFAS Remediation

EPA Science Inventory

Many military bases and their surrounding communities are impacted by contamination with per- and polyfluoroalkyl substances (PFAS) from Aqueous Film-Forming Foams (AFFFs). Soil sorption technologies provide a promising solution to immobilize PFAS in the soil and prevent groundwa...

Removal of caffeine from industrial wastewater using Trichosporon asahii.

PubMed

Lakshmi, V; Das, Nilanjana

2013-07-01

Caffeine (1,3,7-trimethylxanthine), a natural alkaloid present mainly in tea and coffee products has been suggested as an environmental pollutant. Decaffeination is an important process for the removal of caffeine from coffee industrial wastes. In the present study, caffeine removal (through degradation) by yeast isolate, Trichosporon asahii immobilized on various conventional matrices (sodium alginate, carboxymethyl cellulose, chitosan, agar and agarose) was investigated using the method of entrapment. The biofilm forming ability of T. asahii was monitored by atomic force microscopy and scanning electron microscopy. Exopolysaccharide produced by T asahii biofilm was characterized by FT-IR spectroscopy and HPLC analysis. Caffeine removal from coffee processing industrial effluent was found to be 75 and 80 % by alginate immobilized yeast and yeast biofilm formed on gravels over a period of 48 hr in batch mode. Effectiveness of the process was also tested involving the continuous--flow column studies.

Kinetics during the redox biotransformation of pollutants mediated by immobilized and soluble humic acids.

PubMed

Cervantes, Francisco J; Martínez, Claudia M; Gonzalez-Estrella, Jorge; Márquez, Arturo; Arriaga, Sonia

2013-03-01

The aim of this study was to elucidate the kinetic constraints during the redox biotransformation of the azo dye, Reactive Red 2 (RR2), and carbon tetrachloride (CT) mediated by soluble humic acids (HAs) and immobilized humic acids (HAi), as well as by the quinoid model compounds, anthraquinone-2,6-disulfonate (AQDS) and 1,2-naphthoquinone-4-sulfonate (NQS). The microbial reduction of both HAs and HAi by anaerobic granular sludge (AGS) was the rate-limiting step during decolorization of RR2 since the reduction of RR2 by reduced HAi proceeded at more than three orders of magnitute faster than the electron-transferring rate observed during the microbial reduction of HAi by AGS. Similarly, the reduction of RR2 by reduced AQDS proceeded 1.6- and 1.9-fold faster than the microbial reduction of AQDS by AGS when this redox mediator (RM) was supplied in soluble and immobilized form, respectively. In contrast, the reduction of NQS by AGS occurred 1.6- and 19.2-fold faster than the chemical reduction of RR2 by reduced NQS when this RM was supplied in soluble and immobilized form, respectively. The microbial reduction of HAs and HAi by a humus-reducing consortium proceeded 1,400- and 790-fold faster than the transfer of electrons from reduced HAs and HAi, respectively, to achieve the reductive dechlorination of CT to chloroform. Overall, the present study provides elucidation on the rate-limiting steps involved in the redox biotransformation of priority pollutants mediated by both HAs and HAi and offers technical suggestions to overcome the kinetic restrictions identified in the redox reactions evaluated.

Simultaneous immobilization of lead and atrazine in contaminated soils using dairy-manure biochar.

PubMed

Cao, Xinde; Ma, Lena; Liang, Yuan; Gao, Bin; Harris, Willie

2011-06-01

Biochar produced from waste biomass is increasingly being recognized as a green, cost-effective amendment for environmental remediation. This work was to determine the ability of biochar to immobilize heavy metal Pb and organic pesticide atrazine in contaminated soils. Biochar prepared from dairy manure was incubated with contaminated soils at rates of 0, 2.5, and 5.0% by weight for 210 d. A commercial activated carbon (AC) was included as a comparison. The AC was effective in immobilizing atrazine, but was ineffective for Pb. However, biochar was effective in immobilizing both atrazine and Pb and the effectiveness was enhanced with increasing incubation time and biochar rates. After 210 d, soils treated with the highest rate of 5.0% biochar showed more than 57% and 66% reduction in Pb and atrazine concentrations in 0.01 M CaCl(2) extraction, respectively. Lead and atrazine concentrations in the toxicity characteristic leaching procedure solutions were reduced by 70-89% and 53-77%, respectively. Uptake of Pb and atrazine by earthworms (Eisenia fetida) was reduced by up to 79% and 73%. Phosphorus originally contained in biochar reacted with soil Pb to form insoluble hydroxypyromorphite Pb(5)(PO(4))(3)(OH), as determined by X-ray diffraction, which was presumably responsible for soil Pb immobilization, whereas atrazine stabilization may result from its adsorption by biochar demonstrated by the significant exponential decrease of extractable atrazine with increasing organic C in biochar (r(2) > 0.97, p < 0.05). The results highlighted the potential of dairy-manure biochar as a unique amendment for immobilization of both heavy metal and organic contaminants in cocontaminated soils.

Metasomatic tourmalinite formation along basement-cover decollements, Orobic Alps, Italy

USGS Publications Warehouse

Slack, J.F.; Passchier, C.W.; Zhang, J.S.

1996-01-01

Cryptocrystalline tourmalinites that occur discontinuously for ???30 km along basement-cover de??collements of the Orohic Alps (Italy) formed by the metasomatism of aluminous cataclasites derived from Permian conglomerates and/or feldspathic sandstones. Using Al as an immobile element monitor, calculations show that the majority of tourmalinites in the region formed through the addition of moderate to significant amounts of B, Mg, Na, Sr, and Be, and the loss of moderate to significant Mn, Ca, K, P, Rb, Ba, and Cr; minor Si, Ti, V, light REE, and Eu also were lost. Data tor relatively immobile Al, Zr, Th, Sc, Nb, and heavy REE indicate that, on average, these tourmalinites formed through ???12% net mass loss assuming an original conglomerate protolith, or through ???7% net mass loss assuming a sandstone protolith. The B and other introduced constituents in the tourmalinites were deposited by hydrothermal fluids focused along and near basement-cover de??collements. These fluids, believed to be associated with late Hercynian felsic magmatism, probably are related to fluids that formed the tourmaline-rich U-Mo-Zn deposits at the nearby Novazza mine and the U-Zn deposits at the nearby Val Vedello mine.

Coupling of 2,4,6-trinitrotoluene (TNT) metabolites onto humic monomers by a new laccase from Trametes modesta.

PubMed

Nyanhongo, Gibson S; Couto, Susana Rodríguez; Guebitz, Georg M

2006-06-01

During degradation of trinitrotoluene (TNT) by Trametes modesta, addition of humic monomers prevented the accumulation of all major stable TNT metabolites (aminodinitrotoluenes [AMDNT]) by at least 92% in the presence of 200 mM ferulic acid and guaiacol. Acute toxicity tests with individual TNT metabolites and in T. modesta cultures supplemented with 200 microM TNT demonstrated that the TNT biodegradation process lead to less toxic metabolites. Toxicity decreased in the order TNT>4-HADNT (4-hydroxylaminodinitrotoluene)>2-HADNT>2,6-DNT (2,6-dinitrotoluene)>2',2',6,6-azoxytetranitrotoluene>4-AMDNT>2-AMDNT>2,4-diamninonitrotoluene (2,4-DAMNT) while 2,4-DNT and 2,6-DAMNT were the least toxic. Ferulic acid is the best candidate for immobilization TNT biodegradation metabolites since it prevented the accumulation of AMDNTs in cultures during TNT biodegradation and its products were less toxic. All humic monomers were very effective in immobilizing 2-HADNT [100%], 4-HADNT [100%] and 2,2,6,6-azoxytetranitrotoluene [100%]. Two distinct laccase isoenzymes (LTM1 and LTM2) potentially involved in immobilization of TNT degradation products were purified to electrophoretic homogeneity. LTM1 and LTM2 have molecular weights of 77.6 and 52.5 kDa, are 18% and 24% glycosylated, have pI values of 3.6 and 4.2, respectively. Both enzymes oxidized all the typical laccase substrates tested. LTM1 showed highest kinetic constants (K(m)=0.03 microM; K(cat)=8.8 4x 10(7)s(-1)) with syringaldazine as substrate.

Identification, characterization, immobilization, and mutational analysis of a novel acetylesterase with industrial potential (LaAcE) from Lactobacillus acidophilus.

PubMed

Wang, Ying; Ryu, Bum Han; Yoo, Wanki; Lee, Chang Woo; Kim, Kyeong Kyu; Lee, Jun Hyuck; Kim, T Doohun

2018-01-01

Lactic acid bacteria, which are involved in the fermentation of vegetables, meats, and dairy products, are widely used for the productions of small organic molecules and bioactive peptides. Here, a novel acetylesterase (LaAcE) from Lactobacillus acidophilus NCFM was identified, functionally characterized, immobilized, and subjected to site-directed mutagenesis for biotechnological applications. The enzymatic properties of LaAcE were investigated using biochemical and biophysical methods including native polyacrylamide gel electrophoresis, acetic acid release, biochemical assays, enzyme kinetics, and spectroscopic methods. Interestingly, LaAcE exhibited the ability to act on a broad range of substrates including glucose pentaacetate, glyceryl tributyrate, fish oil, and fermentation-related compounds. Furthermore, immobilization of LaAcE showed good recycling ability and high thermal stability compared with free LaAcE. A structural model of LaAcE was used to guide mutational analysis of hydrophobic substrate-binding region, which was composed of Leu 156 , Phe 164 , and Val 204 . Five mutants (L156A, F164A, V204A, L156A/F164A, and L156A/V204A) were generated and investigated to elucidate the roles of these hydrophobic residues in substrate specificity. This work provided valuable insights into the properties of LaAcE, and demonstrated that LaAcE could be used as a model enzyme of acetylesterase in lactic acid bacteria, making LaAcE a great candidate for industrial applications. Copyright © 2017 Elsevier B.V. All rights reserved.

Site-specific fab fragment biotinylation at the conserved nucleotide binding site for enhanced Ebola detection.

PubMed

Mustafaoglu, Nur; Alves, Nathan J; Bilgicer, Basar

2015-07-01

The nucleotide binding site (NBS) is a highly conserved region between the variable light and heavy chains at the Fab domains of all antibodies, and a small molecule that we identified, indole-3-butyric acid (IBA), binds specifically to this site. Fab fragment, with its small size and simple production methods compared to intact antibody, is good candidate for use in miniaturized diagnostic devices and targeted therapeutic applications. However, commonly used modification techniques are not well suited for Fab fragments as they are often more delicate than intact antibodies. Fab fragments are of particular interest for sensor surface functionalization but immobilization results in damage to the antigen binding site and greatly reduced activity due to their truncated size that allows only a small area that can bind to surfaces without impeding antigen binding. In this study, we describe an NBS-UV photocrosslinking functionalization method (UV-NBS(Biotin) in which a Fab fragment is site-specifically biotinylated with an IBA-EG11-Biotin linker via UV energy exposure (1 J/cm(2)) without affecting its antigen binding activity. This study demonstrates successful immobilization of biotinylated Ebola detecting Fab fragment (KZ52 Fab fragment) via the UV-NBS(Biotin) method yielding 1031-fold and 2-fold better antigen detection sensitivity compared to commonly used immobilization methods: direct physical adsorption and NHS-Biotin functionalization, respectively. Utilization of the UV-NBS(Biotin) method for site-specific conjugation to Fab fragment represents a proof of concept use of Fab fragment for various diagnostic and therapeutic applications with numerous fluorescent probes, affinity molecules and peptides. © 2015 Wiley Periodicals, Inc.

Isolation of phosphate solubilizing bacteria and their potential for lead immobilization in soil.

PubMed

Park, Jin Hee; Bolan, Nanthi; Megharaj, Mallavarapu; Naidu, Ravi

2011-01-30

Lead (Pb), a highly toxic heavy metal forms stable compounds with phosphate (P). The potential of phosphate solubilizing bacteria (PSB) to immobilize Pb by enhancing solubilization of insoluble P compounds was tested in this research. Eighteen different PSB strains isolated from P amended and Pb contaminated soils were screened for their efficiency in P solubilization. The PSB isolated from P amended soils solubilized 217-479 mg/L of P while the PSB from Pb contaminated soil solubilized 31-293 mg/L of P. Stepwise multiple regression analysis and P solubility kinetics indicated that the major mechanism of P solubilization by PSB is the pH reduction through the release of organic acids. From the isolated bacteria, two PSB were chosen for Pb immobilization and these bacteria were identified as Pantoea sp. and Enterobacter sp., respectively. The PSB significantly increased P solubilization by 25.0% and 49.9% in the case of Pantoea sp., and 63.3% and 88.6% in the case of Enterobacter sp. for 200 and 800 mg/kg of rock phosphate (RP) addition, respectively, thereby enhancing the immobilization of Pb by 8.25-13.7% in the case of Pantoea sp. and 14.7-26.4% in the case of Enterobacter sp. The ability of PSB to solubilize P, promote plant growth, and immobilize Pb can be used for phytostabilization of Pb contaminated soils. Copyright © 2010 Elsevier B.V. All rights reserved.

Immobilization of FLAG-Tagged Recombinant Adeno-Associated Virus 2 onto Tissue Engineering Scaffolds for the Improvement of Transgene Delivery in Cell Transplants.

PubMed

Li, Hua; Zhang, Feng-Lan; Shi, Wen-Jie; Bai, Xue-Jia; Jia, Shu-Qin; Zhang, Chen-Guang; Ding, Wei

2015-01-01

The technology of virus-based genetic modification in tissue engineering has provided the opportunity to produce more flexible and versatile biomaterials for transplantation. Localizing the transgene expression with increased efficiency is critical for tissue engineering as well as a challenge for virus-based gene delivery. In this study, we tagged the VP2 protein of type 2 adeno-associated virus (AAV) with a 3×FLAG plasmid at the N-terminus and packaged a FLAG-tagged recombinant AAV2 chimeric mutant. The mutant AAVs were immobilized onto the tissue engineering scaffolds with crosslinked anti-FLAG antibodies by N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP). Cultured cells were seeded to scaffolds to form 3D transplants, and then tested for viral transduction both in vitro and in vivo. The results showed that our FLAG-tagged AAV2 exerted similar transduction efficiency compared with the wild type AAV2 when infected cultured cells. Following immobilization onto the scaffolds of PLGA or gelatin sponge with anti-FLAG antibodies, the viral mediated transgene expression was significantly improved and more localized. Our data demonstrated that the mutation of AAV capsid targeted for antibody-based immobilization could be a practical approach for more efficient and precise transgene delivery. It was also suggested that the immobilization of AAV might have attractive potentials in applications of tissue engineering involving the targeted gene manipulation in 3D tissue cultures.

Optical Sensors for Biomolecules Using Nanoporous Sol-Gel Materials

NASA Technical Reports Server (NTRS)

Fang, Jonathan; Zhou, Jing C.; Lan, Esther H.; Dunn, Bruce; Gillman, Patricia L.; Smith, Scott M.

2004-01-01

An important consideration for space missions to Mars is the ability to detect biosignatures. Solid-state sensing elements for optical detection of biological entities are possible using sol-gel based biologically active materials. We have used these materials as optical sensing elements in a variety of bioassays, including immunoassays and enzyme assays. By immobilizing an appropriate biomolecule in the sol-gel sensing element, we have successfully detected analytes such as amino acids and hormones. In the case of the amino acid glutamate, the enzyme glutamate dehydrogenase was the immobilized molecule, whereas in the case of the hormone cortisol, an anti-cortisol antibody was immobilized in the sensing element. In this previous work with immobilized enzymes and antibodies, excellent sensitivity and specificity were demonstrated in a variety of formats including bulk materials, thin films and fibers. We believe that the sol-gel approach is an attractive platform for bioastronautics sensing applications because of the ability to detect a wide range of entities such as amino acids, fatty acids, hopanes, porphyrins, etc. The sol-gel approach produces an optically transparent 3D silica matrix that forms around the biomolecule of interest, thus stabilizing its structure and functionality while allowing for optical detection. This encapsulation process protects the biomolecule and leads to a more "rugged" sensor. The nanoporous structure of the sol-gel matrix allows diffusion of small target molecules but keeps larger, biomolecules immobilized in the pores. We are currently developing these biologically active sol-gel materials into small portable devices for on-orbit cortisol detection

Using dual-polarization interferometry to study surface-initiated DNA hybridization chain reactions in real time.

PubMed

Huang, Fujian; Xu, Pingping; Liang, Haojun

2014-01-15

In this study we used dual-polarization interferometry to investigate DNA hybridization chain reactions (HCRs) at solid-liquid interfaces. We monitored the effects of variations in mass, thickness, and density of the immobilized initiator on the subsequent HCRs at various salt concentrations. At low salt concentrations, the single-stranded DNA (ssDNA) initiator was attached uniformly to the chip surface. At high salt concentrations, it lay on the surface at the onset of the immobilization process, but the approaching ssDNA forced the pre-immobilized ssDNA strands to extend into solution as a result of increased electrostatic repulsion between the pre-adsorbed and approaching ssDNA chains. Injection of a mixture of H1 and H2 increased the mass and thickness of the films initially, but thereafter the thickness decreased. These changes indicate that the long double-stranded DNA that formed lay on the surface, rather than extended into the solution, thereby suppressing the subsequent initiation activity of the released single-strand parts of H1 and H2. Increasing the salt concentration increased the HCR efficiency and reaction rate. The HCR efficiency of the initiator ssDNA immobilized on its 5' end was higher than that immobilized on its 3' end, suggesting that the released single-strand parts of H1 and H2 close to the chip surface decreased the initiation activity relative to those of the ones extending into solution. © 2013 Elsevier B.V. All rights reserved.

Immobilization of iron- and manganese-oxidizing bacteria with a biofilm-forming bacterium for the effective removal of iron and manganese from groundwater.

PubMed

Li, Chunyan; Wang, Shuting; Du, Xiaopeng; Cheng, Xiaosong; Fu, Meng; Hou, Ning; Li, Dapeng

2016-11-01

In this study, three bacteria with high Fe- and Mn-oxidizing capabilities were isolated from groundwater well sludge and identified as Acinetobacter sp., Bacillus megaterium and Sphingobacterium sp. The maximum removal ratios of Fe and Mn (99.75% and 96.69%) were obtained by an optimal combination of the bacteria at a temperature of 20.15°C, pH 7.09 and an inoculum size of 2.08%. Four lab-scale biofilters were tested in parallel for the removal of iron and manganese ions from groundwater. The results indicated that the Fe/Mn removal ratios of biofilter R4, which was inoculated with iron- and manganese-oxidizing bacteria and a biofilm-forming bacterium, were approximately 95% for each metal during continuous operation and were better than the other biofilters. This study demonstrated that the biofilm-forming bacterium could promote the immobilization of the iron- and manganese-oxidizing bacteria on the biofilters and enhance the removal efficiency of iron and manganese ions from groundwater. Copyright © 2016 Elsevier Ltd. All rights reserved.

Liquidus temperature and chemical durability of selected glasses to immobilize rare earth oxides waste

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mohd Fadzil, Syazwani Binti; Hrma, Pavel R.; Schweiger, Michael J.

Pyroprocessing is a reprocessing method for managing and reusing used nuclear fuel (UNF) by dissolving it in an electrorefiner with a molten alkali or alkaline earth chloride salt mixture while avoiding wet reprocessing. Pyroprocessing UNF with a LiCl-KCl eutectic salt releases the fission products from the fuel and generates a variety of metallic and salt-based species, including rare earth (RE) chlorides. If the RE-chlorides are converted to oxides, borosilicate glass is a prime candidate for their immobilization because of its durability and ability to dissolve almost any RE waste component into the matrix at high loadings. Crystallization that occurs inmore » waste glasses as the waste loading increases may complicate glass processing and affect the product quality. This work compares three types of borosilicate glasses in terms of liquidus temperature (TL): the International Simple Glass designed by the International Working Group, sodium borosilicate glass developed by Korea Hydro and Nuclear Power, and the lanthanide aluminoborosilicate (LABS) glass established in the United States. The LABS glass allows the highest waste loadings (over 50 mass% RE2O3) while possessing an acceptable chemical durability.« less

Liquidus temperature and chemical durability of selected glasses to immobilize rare earth oxides waste

NASA Astrophysics Data System (ADS)

Mohd Fadzil, Syazwani; Hrma, Pavel; Schweiger, Michael J.; Riley, Brian J.

2015-10-01

Pyroprocessing is are processing method for managing and reusing used nuclear fuel (UNF) by dissolving it in an electrorefiner with a molten alkali or alkaline earth chloride salt mixture while avoiding wet reprocessing. Pyroprocessing UNF with a LiCl-KCl eutectic salt releases the fission products from the fuel and generates a variety of metallic and salt-based species, including rare earth (RE) chlorides. If the RE-chlorides are converted to oxides, borosilicate glass is a prime candidate for their immobilization because of its durability and ability to dissolve almost any RE waste component into the glass matrix at high loadings. Crystallization that occurs in waste glasses as the waste loading increases may complicate glass processing and affect the product quality. This work compares three types of borosilicate glasses in terms of liquidus temperature (TL): the International Simple Glass designed by the International Working Group, sodium borosilicate glass developed by Korea Hydro and Nuclear Power, and the lanthanide aluminoborosilicate (LABS) glass established in the United States. The LABS glass allows the highest waste loadings (over 50 mass% RE2O3) while possessing an acceptable chemical durability.

Advances in immobilized artificial membrane (IAM) chromatography for novel drug discovery.

PubMed

Tsopelas, Fotios; Vallianatou, Theodosia; Tsantili-Kakoulidou, Anna

2016-01-01

The development of immobilized artificial membrane (IAM) chromatography has unfolded new perspectives for the use of chromatographic techniques in drug discovery, combining simulation of the environment of cell membranes with rapid measurements. The present review describes the characteristics of phosphatidylcholine-based stationary phases and analyses the molecular factors governing IAM retention in comparison to n-octanol-water and liposomes partitioning systems as well as to reversed phase chromatography. Other biomimetic stationary phases are also briefly discussed. The potential of IAM chromatography to model permeability through the main physiological barriers and drug membrane interactions is outlined. Further applications to calculate complex pharmacokinetic properties, related to tissue binding, and to screen drug candidates for phospholipidosis, as well as to estimate cell accumulation/retention are surveyed. The ambivalent nature of IAM chromatography, as a border case between passive diffusion and binding, defines its multiple potential applications. However, despite its successful performance in many permeability and drug-membrane interactions studies, IAM chromatography is still used as a supportive and not a stand-alone technique. Further studies looking at IAM chromatography in different biological processes are still required if this technique is to have a more focused and consistent application in drug discovery.

Advances in identification and validation of protein targets of natural products without chemical modification.

PubMed

Chang, J; Kim, Y; Kwon, H J

2016-05-04

Covering: up to February 2016Identification of the target proteins of natural products is pivotal to understanding the mechanisms of action to develop natural products for use as molecular probes and potential therapeutic drugs. Affinity chromatography of immobilized natural products has been conventionally used to identify target proteins, and has yielded good results. However, this method has limitations, in that labeling or tagging for immobilization and affinity purification often result in reduced or altered activity of the natural product. New strategies have recently been developed and applied to identify the target proteins of natural products and synthetic small molecules without chemical modification of the natural product. These direct and indirect methods for target identification of label-free natural products include drug affinity responsive target stability (DARTS), stability of proteins from rates of oxidation (SPROX), cellular thermal shift assay (CETSA), thermal proteome profiling (TPP), and bioinformatics-based analysis of connectivity. This review focuses on and reports case studies of the latest advances in target protein identification methods for label-free natural products. The integration of newly developed technologies will provide new insights and highlight the value of natural products for use as biological probes and new drug candidates.

Ethanol Production from Wet-Exploded Wheat Straw Hydrolysate by Thermophilic Anaerobic Bacterium Thermoanaerobacter BG1L1 in a Continuous Immobilized Reactor

NASA Astrophysics Data System (ADS)

Georgieva, Tania I.; Mikkelsen, Marie J.; Ahring, Birgitte K.

Thermophilic ethanol fermentation of wet-exploded wheat straw hydrolysate was investigated in a continuous immobilized reactor system. The experiments were carried out in a lab-scale fluidized bed reactor (FBR) at 70°C. Undetoxified wheat straw hydrolysate was used (3-12% dry matter), corresponding to sugar mixtures of glucose and xylose ranging from 12 to 41 g/1. The organism, thermophilic anaerobic bacterium Thermoanaerobacter BG1L1, exhibited significant resistance to high levels of acetic acid (up to 10 g/1) and other metabolic inhibitors present in the hydrolysate. Although the hydrolysate was not detoxified, ethanol yield in a range of 0.39-0.42 g/g was obtained. Overall, sugar efficiency to ethanol was 68-76%. The reactor was operated continuously for approximately 143 days, and no contamination was seen without the use of any agent for preventing bacterial infections. The tested microorganism has considerable potential to be a novel candidate for lignocellulose bioconversion into ethanol. The work reported here also demonstrates that the use of FBR configuration might be a viable approach for thermophilic anaerobic ethanol fermentation.

75 FR 69221 - Endangered and Threatened Wildlife and Plants; Review of Native Species That Are Candidates for...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-10

...In this Candidate Notice of Review (CNOR), we, the U.S. Fish and Wildlife Service (Service), present an updated list of plant and animal species native to the United States that we regard as candidates for or have proposed for addition to the Lists of Endangered and Threatened Wildlife and Plants under the Endangered Species Act of 1973, as amended. Identification of candidate species can assist environmental planning efforts by providing advance notice of potential listings, allowing landowners and resource managers to alleviate threats and thereby possibly remove the need to list species as endangered or threatened. Even if we subsequently list a candidate species, the early notice provided here could result in more options for species management and recovery by prompting candidate conservation measures to alleviate threats to the species. The CNOR summarizes the status and threats that we evaluated in order to determine that species qualify as candidates and to assign a listing priority number (LPN) to each species or to determine that species should be removed from candidate status. Additional material that we relied on is available in the Species Assessment and Listing Priority Assignment Forms (species assessment forms, previously called candidate forms) for each candidate species. Overall, this CNOR recognizes five new candidates, changes the LPN for four candidates, and removes one species from candidate status. Combined with other decisions for individual species that were published separately from this CNOR in the past year, the current number of species that are candidates for listing is 251. This document also includes our findings on resubmitted petitions and describes our progress in revising the Lists of Endangered and Threatened Wildlife and Plants during the period October 1, 2009, through September 30, 2010. We request additional status information that may be available for the 251 candidate species identified in this CNOR.

Presidential Search: Selecting the Top Candidates.

ERIC Educational Resources Information Center

Stead, Ronald S.

1988-01-01

The winnowing of a large number of presidential candidates to a smaller, more manageable group is discussed. Initial screening, candidate information, evaluation form, reference checking, and selecting interviewees are described. (MLW)

Green synthesis and characterization of novel gold nanocomposites for electrochemical sensing applications.

PubMed

Tanwar, Shivani; Ho, Ja-an Annie; Magi, Emanuele

2013-12-15

Synthesis, characterization and application of Au-PANI-Calix and Au-PANI-Nap nanocomposites, is reported herein. An easy template free green synthesis is proposed and discussed for easy expediency. A variety of analytical techniques were used to characterize the nanocomposites: UV-vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, Dynamic light scattering (DLS), X-ray diffraction (XRD), Energy-dispersive X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS) were used to characterize the nanocomposites. Surface morphology was studied by transmission electron microscopy (TEM). The nanocomposites were immobilized on screen-printed electrode and showed electroactivity in neutral pH, making them promising candidates for various analytical applications. A sensitive and selective detection of Cu(2+) was perceived on the Au-PANI-Calix modified electrode with no interference from ions K(+), Ni(2+), Co(2+), Pb(2+), Cr(3+) with a detection limit of 10nM. The copper detection is facilitated for accessible ligation with 4-sulfocalix[4]arene, so as the Cu(II)-Calix complex formed. The electrode modified with Au-PANI-Nap showed sensing application towards H2O2 with a detection limit of 1 μM. The modified electrodes were reproducible and stable for 2 months. © 2013 Elsevier B.V. All rights reserved.

Separating and stabilizing phosphate from high-level radioactive waste: process development and spectroscopic monitoring.

PubMed

Lumetta, Gregg J; Braley, Jenifer C; Peterson, James M; Bryan, Samuel A; Levitskaia, Tatiana G

2012-06-05

Removing phosphate from alkaline high-level waste sludges at the Department of Energy's Hanford Site in Washington State is necessary to increase the waste loading in the borosilicate glass waste form that will be used to immobilize the highly radioactive fraction of these wastes. We are developing a process which first leaches phosphate from the high-level waste solids with aqueous sodium hydroxide, and then isolates the phosphate by precipitation with calcium oxide. Tests with actual tank waste confirmed that this process is an effective method of phosphate removal from the sludge and offers an additional option for managing the phosphorus in the Hanford tank waste solids. The presence of vibrationally active species, such as nitrate and phosphate ions, in the tank waste processing streams makes the phosphate removal process an ideal candidate for monitoring by Raman or infrared spectroscopic means. As a proof-of-principle demonstration, Raman and Fourier transform infrared (FTIR) spectra were acquired for all phases during a test of the process with actual tank waste. Quantitative determination of phosphate, nitrate, and sulfate in the liquid phases was achieved by Raman spectroscopy, demonstrating the applicability of Raman spectroscopy for the monitoring of these species in the tank waste process streams.

Similar interaction chromatography of proteins: A cross interaction chromatographic approach to estimate the osmotic second virial coefficient.

PubMed

Quigley, A; Williams, D R

2016-08-12

Self-interaction chromatography (SIC) has established itself as an important experimental technique for the measurement of the second osmotic virial coefficients B22. B22 data are critical for understanding a range of protein solution phenomena, particularly aggregation and crystallisation. A key limitation to the more extensive use of SIC is the need to develop a method for immobilising each specific protein of interest onto a chromatographic support. This requirement is both a time and protein consuming constraint, which means that SIC cannot be used as a high throughput method for screening a wide range of proteins and their variants. Here an experimental framework is presented for estimating B22 values using Similar Interaction Chromatography (SimIC). This work uses experimental B23 and B32 data for lysozyme, lactoferrin, catalase and concanavalin A to reliably estimate B22 using arithmetic mean field approximations and is demonstrated to give good agreement with SIC measurements of B22 for the same proteins. SimIC could form the basis of a rapid protein variant screening methods to assess the developability of protein therapeutic candidates for industrial and academic researchers with respect to aggregation behaviour by eluting target proteins through a series of well-characterised protein immobilized reference columns. Copyright © 2016. Published by Elsevier B.V.

MUC1-Targeted Cancer Cell Photothermal Ablation Using Bioinspired Gold Nanorods.

PubMed

Zelasko-Leon, Daria C; Fuentes, Christina M; Messersmith, Phillip B

2015-01-01

Recent studies have highlighted the overexpression of mucin 1 (MUC1) in various epithelial carcinomas and its role in tumorigenesis. These mucins present a novel targeting opportunity for nanoparticle-mediated photothermal cancer treatments due to their unique antenna-like extracellular extension. In this study, MUC1 antibodies and albumin were immobilized onto the surface of gold nanorods using a "primer" of polydopamine (PD), a molecular mimic of catechol- and amine-rich mussel adhesive proteins. PD forms an adhesive platform for the deposition of albumin and MUC1 antibodies, achieving a surface that is stable, bioinert and biofunctional. Two-photon luminescence confocal and darkfield scattering imaging revealed targeting of MUC1-BSA-PD-NRs to MUC1+ MCF-7 breast cancer and SCC-15 squamous cell carcinoma cells lines. Treated cells were exposed to a laser encompassing the near-infrared AuNR longitudinal surface plasmon and assessed for photothermal ablation. MUC1-BSA-PD-NRs substantially decreased cell viability in photoirradiated MCF-7 cell lines vs. MUC1- MDA-MB-231 breast cancer cells (p < 0.005). Agents exhibited no cytotoxicity in the absence of photothermal treatment. The facile nature of the coating method, combined with targeting and photoablation efficacy, are attractive features of these candidate cancer nanotherapeutics.

Paraformaldehyde fixation of neutrophils for immunolabeling of granule antigens in cryoultrasections.

PubMed

Elliott, E; Dennison, C; Fortgens, P H; Travis, J

1995-10-01

Paraformaldehyde (PFA) fixation was optimized to facilitate the immobilization and labeling of multiple granule antigens, using short fixation regimens and cryoultramicrotomy of unembedded neutrophils (PMNs). In the optimal protocol, extraction of azurophil granule antigens (especially of the abundant elastase) was obviated by manipulating the polymeric state of PFA, and hence its rate of cross-linking, by altering its concentration and pH in a multistep process. Primary fixation conditions used (4% PFA, pH 8.0, 5 min) favor fixative penetration and rapid cross-linking. Stable cross-linking of the antigen was achieved in a secondary fixation step using conditions that favor larger, more cross-linking polymeric forms of PFA (8% PFA, pH 7.2, 15 min). Immobilization of granule antigens was enhanced by flotation of cut sections on fixative (8% PFA, pH 8.0) before labeling and by using post-labeling fixation with 1% glutaraldehyde. The optimized protocol facilitated immobilization and immunolabeling of elastase, myeloperoxidase, lactoferrin, and cathepsin D in highly hydrated, unembedded PMNs.

A Sortase A-Immobilized Mesoporous Hollow Carbon Sphere-Based Biosensor for Detection of Gram-Positive Bacteria

NASA Astrophysics Data System (ADS)

Wang, Hongsu; Luo, Ruiping; Chen, Yang; Si, Qi; Niu, Xiaodi

2018-05-01

A sensor based on mesoporous carbon materials immobilized with sortase A (SrtA) for determination of Staphylococcus aureus (S. aureus) is reported. To prepare the biosensor, we first synthesized carboxyl-functionalized mesoporous hollow carbon spheres, then applied them as carriers for immobilization of SrtA. Based on the catalytic mechanism of SrtA, a highly sensitive, inexpensive, and rapid method was developed for S. aureus detection. The sensor showed a linear response in the bacterial concentration range of 0.125 × 102 colony-forming units (CFU) mL-1 to 2.5 × 102 CFU mL-1, with detection limit as low as 9.0 CFU mL-1. The method was successfully used for quantitative detection of S. aureus in whole milk samples, giving results similar to experimental results obtained from the plate counting method. This biosensor could also be used to detect other Gram-positive bacteria that secrete SrtA.

Micrometer sized immobilization of protein molecules onto quartz, silicium and gold.

NASA Astrophysics Data System (ADS)

Petersen, Steffen B.; Neves-Petersen, Maria Teresa; Klitgaard, Søren; Duroux, Meg Crookshanks

2006-02-01

We demonstrate that ultraviolet light can be used to make sterically oriented covalent immobilization of a large variety of protein molecules onto either gold or thiolated quartz or silicium. The reaction mechanism behind the reported new technology involves light induced breakage of disulphide bridges in proteins upon UV illumination of nearby aromatic amino acids, resulting in the formation of free, reactive thiol groups that will form covalent bonds with thiol reactive surfaces. The protein molecules in general retain their function. The size of the immobilization spot is determined by the dimension of the UV beam. In principle, the spot size may be as small as 1 micrometer or less. We have developed the necessary technology for preparing large protein arrays of enzymes and fragments of monoclonal antibodies. Dedicated Image Processing Software has been developed for making quality assessment of the protein arrays. A multitude of important application areas such as drug carriers and drug delivery, bioelectronics, carbon nanotubes, nanoparticles as well as protein glue are discussed.

Osteoinductive activity of insulin-functionalized cell culture surfaces obtained using diazonium chemistry

NASA Astrophysics Data System (ADS)

Mikulska, Anna; Filipowska, Joanna; Osyczka, Anna; Nowakowska, Maria; Szczubiałka, Krzysztof

2014-12-01

Polymeric surfaces suitable for cell culture (DR/Pec) were constructed from diazoresin (DR) and pectin (Pec) in a form of ultrathin films using the layer-by-layer (LbL) technique. The surfaces were functionalized with insulin using diazonium chemistry. Such functionalized surfaces were used to culture human mesenchymal stem cells (hMSCs) to assess their suitability for bone tissue engineering and regeneration. The activity of insulin immobilized on the surfaces (DR/Pec/Ins) was compared to that of insulin dissolved in the culture medium. Human MSC grown on insulin-immobilized DR/Pec surfaces displayed increased proliferation and higher osteogenic activity. The latter was determined by means of alkaline phosphatase (ALP) activity, which increases at early stages of osteoblasts differentiation. Insulin dissolved in the culture medium did not stimulate cell proliferation and its osteogenic activity was significantly lower. Addition of recombinant human bone morphogenetic protein 2 (rhBMP-2) to the culture medium further increased ALP activity in hMSCs indicating additive osteogenic action of immobilized insulin and rhBMP-2

Microbially-induced Carbonate Precipitation for Immobilization of Toxic Metals.

PubMed

Kumari, Deepika; Qian, Xin-Yi; Pan, Xiangliang; Achal, Varenyam; Li, Qianwei; Gadd, Geoffrey Michael

2016-01-01

Rapid urbanization and industrialization resulting from growing populations contribute to environmental pollution by toxic metals and radionuclides which pose a threat to the environment and to human health. To combat this threat, it is important to develop remediation technologies based on natural processes that are sustainable. In recent years, a biomineralization process involving ureolytic microorganisms that leads to calcium carbonate precipitation has been found to be effective in immobilizing toxic metal pollutants. The advantage of using ureolytic organisms for bioremediating metal pollution in soil is their ability to immobilize toxic metals efficiently by precipitation or coprecipitation, independent of metal valence state and toxicity and the redox potential. This review summarizes current understanding of the ability of ureolytic microorganisms for carbonate biomineralization and applications of this process for toxic metal bioremediation. Microbial metal carbonate precipitation may also be relevant to detoxification of contaminated process streams and effluents as well as the production of novel carbonate biominerals and biorecovery of metals and radionuclides that form insoluble carbonates. Copyright © 2016 Elsevier Inc. All rights reserved.

Enzymatic synthesis of eugenol benzoate by immobilized Staphylococcus aureus lipase: optimization using response surface methodology and determination of antioxidant activity.

PubMed

Horchani, Habib; Ben Salem, Nadia; Zarai, Zied; Sayari, Adel; Gargouri, Youssef; Chaâbouni, Moncef

2010-04-01

The ability of a non-commercial immobilized Staphylococcus aureus lipase to catalyze the esterification of eugenol with benzoic acid was checked and the antioxidant power of the ester formed was evaluated. Response surface methodology based on four variables (the reaction temperature, the amount of lipase, the benzoic acid/eugenol molar ratio and the volume of solvent) was used to optimize the experimental conditions of eugenol benzoate synthesis. The maximum conversion yield (75%) was obtained using 240 IU of immobilized lipase, a benzoic acid/eugenol molar ratio of 1.22 dissolved in 4.6 ml chloroform at 41 degrees Celsius. The antioxidant activities of eugenol and its ester were evaluated. Compared to BHT, used as a model synthetic antioxidant, the eugenol benzoate showed a higher antioxidative activity. The IC(50) value for 1,1-diphenyl-2-picrylhydrazyl was found to be 18.2 microg/ml versus 20.2 microg/ml for eugenol and eugenol benzoate. Copyright 2009 Elsevier Ltd. All rights reserved.

Selective retention of basic compounds by metal aquo-ion affinity chromatography.

PubMed

Asakawa, Yoshiki; Yamamoto, Eiichi; Asakawa, Naoki

2014-10-01

A novel metal aquo-ion affinity chromatography has been developed for the analysis of basic compounds using heat-treated silica gel containing hydrated metal cations (metal aquo-ions) as the packing material. The packing materials of the metal aquo-ion affinity chromatography were prepared by the immobilization of a single metal component such as Fe(III), Al(III), Ag(I), and Ni(II) on silica gel followed by extensive heat treatment. The immobilized metals form aquo-ions to present cation-exchange ability for basic analytes and the cation-exchange ability for basic analytes depends on pKa of the immobilized metal species. In the present study, to evaluate the retention characteristics of metal aquo-ion affinity chromatography, the on-line solid-phase extraction of drugs was investigated. Obtained data clearly evidence the selective retention capability of metal aquo-ion affinity chromatography for basic analytes with sufficient capacity. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Effect of hydrothermal carbonization on migration and environmental risk of heavy metals in sewage sludge during pyrolysis.

PubMed

Liu, Tingting; Liu, Zhengang; Zheng, Qingfu; Lang, Qianqian; Xia, Yu; Peng, Nana; Gai, Chao

2018-01-01

The heavy metals distribution during hydrothermal carbonization (HTC) of sewage sludge, and pyrolysis of the resultant hydrochar was investigated and compared with raw sludge pyrolysis. The results showed that HTC reduced exchangeable/acid-soluble and reducible fraction of heavy metals and lowered the potential risk of heavy metals in sewage sludge. The pyrolysis favored the transformation of extracted/mobile fraction of heavy metals to residual form especially at high temperature, immobilizing heavy metals in the chars. Compared to the chars from raw sludge pyrolysis, the chars derived from hydrochar pyrolysis was more alkaline and had lower risk and less leachable heavy metals, indicating that pyrolysis imposed more positive effect on immobilization of heavy metals for the hydrochar than for sewage sludge. The present study demonstrated that HTC is a promising pretreatment prior to pyrolysis from the perspective of immobilization of heavy metals in sewage sludge. Copyright © 2017 Elsevier Ltd. All rights reserved.

Study on the spectrophotometric detection of free fatty acids in palm oil utilizing enzymatic reactions.

PubMed

Azeman, Nur Hidayah; Yusof, Nor Azah; Abdullah, Jaafar; Yunus, Robiah; Hamidon, Mohd Nizar; Hajian, Reza

2015-07-07

In this paper, a comprehensive study has been made on the detection of free fatty acids (FFAs) in palm oil via an optical technique based on enzymatic aminolysis reactions. FFAs in crude palm oil (CPO) were converted into fatty hydroxamic acids (FHAs) in a biphasic lipid/aqueous medium in the presence of immobilized lipase. The colored compound formed after complexation between FHA and vanadium (V) ion solution was proportional to the FFA content in the CPO samples and was analyzed using a spectrophotometric method. In order to develop a rapid detection system, the parameters involved in the aminolysis process were studied. The utilization of immobilized lipase as catalyst during the aminolysis process offers simplicity in the product isolation and the possibility of conducting the process under extreme reaction conditions. A good agreement was found between the developed method using immobilized Thermomyces lanuginose lipase as catalyst for the aminolysis process and the Malaysian Palm Oil Board (MPOB) standard titration method (R2 = 0.9453).

Aminosilane multilayer formed on a single-crystalline diamond surface with controlled nanoscopic hardness and bioactivity by a wet process.

PubMed

Amemiya, Yosuke; Hatakeyama, Akiko; Shimamoto, Nobuo

2009-01-06

Diamond could be an excellent support for nanodevices utilizing biomolecules if it is covered with a polymer layer immobilizing a variety of biomolecules. We report a wet method to form a 3-aminopropyltriethoxysilane (APTES) multilayer with a controlled hardness, roughness, and capacity for immobilizing protein. The method is feasible in typical biochemical laboratories where biomolecules are prepared. Atomic force microscopy (AFM) revealed that the surface geometries and nanoscopic hardness of the multilayers on an oxygen-terminated single-crystalline diamond surface depended on the dielectric constant of the solvent; the smaller the constant, the harder the layer. The hard multilayers had holes and APTES aggregates on the surfaces, while less hard ones had homogeneous surfaces with rare holes and little aggregates. The secondary deposition of APTES in a solvent with a large dielectric constant on a hard multilayer removed the holes, and further treatment of the multilayer in acidic ethanol solution diminished the aggregates. Such a surface can immobilize streptavidin with enough specificity against nonspecific adsorption using a combination of polyethylene glycol reagents. The results of a scratching test and nanoindentation test with AFM provided consistent results, suggesting some universality of the scratching test independent of the tip structure of the cantilever. The mechanism of formation of multilayers on the diamond surface and their binding to it is discussed.

In Situ Immobilization of Selenium in Sediment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moore, Robert C.; Stewart, Thomas Austin

2014-09-01

This project focused on the use of a sorbent, carbonated apatite, to immobilize selenium in the environment. It is know that apatite will sorb selenium and based on the mechanism of sorption it is theorized that carbonated apatite will be more effective that pure apatite. Immobilization of selenium in the environment is through the use of a sorbent in a permeable reactive barrier (PRB). A PRB can be constructed by trenching and backfill with the sorbent or in the case of apatite as the sorbent formed in situ using the apatite forming solution of Moore (2003, 2004). There is verymore » little data on selenium sorption by carbonated apatite in the literature. Therefore, in this work, the basic sorptive properties of carbonated apatite were investigated. Carbonated apatite was synthesized by a precipitation method and characterized. Batch selenium kinetic and equilibrium experiments were performed. The results indicate the carbonated apatite contained 9.4% carbonate and uptake of selenium as selenite was rapid; 5 hours for complete uptake of selenium vs. more than 100 hours for pure hydroxyapatite reported in the literature. Additionally, the carbonated apatite exhibited significantly higher distribution coefficients in equilibrium experiments than pure apatite under similar experimental conditions. The next phase of this work will be to seek additional funds to continue the research with the goal of eventually demonstrating the technology in a field application.« less

Cellulase and alcohol dehydrogenase immobilized in Langmuir and Langmuir-Blodgett films and their molecular-level effects upon contact with cellulose and ethanol.

PubMed

Rodrigues, Dilmer; Camilo, Fernanda Ferraz; Caseli, Luciano

2014-02-25

The key challenges for producing devices based on nanostructured films with control over the molecular architecture are to preserve the catalytic activity of the immobilized biomolecules and to provide a reliable method for determining the intermolecular interactions and the accommodation of molecules at very small scales. In this work, the enzymes cellulase and alcohol dehydrogenase (ADH) were coimmobilized with dipalmitoylphosphatidylcholine (DPPC) as Langmuir-Blodgett (LB) films, and their biological activities were assayed by accommodating the structure formed in contact with cellulose. For this purpose, the polysaccharide was dissolved in an ionic liquid, 1-buthyl-3-methylimidazolium chloride (BMImCl), and dropped on the top of the hybrid cellulase-ADH-DPPC LB film. The interactions between cellulose and ethanol, which are the catalytic substrates of the enzymes as well as important elements in the production of second-generation fuels, were then investigated using polarization-modulation infrared reflection-absorption spectroscopy (PM-IRRAS). Investigation of the secondary structures of the enzymes was performed using PM-IRRAS, through which the presence of ethanol and cellulose was observed to highly affect the structures of ADH and cellulase, respectively. The detection of products formed from the catalyzed reactions as well as the changes of secondary structure of the enzymes immobilization could be carried out, which opens the possibility to produce a means for producing second-generation ethanol using nanoscale arrangements.

A rapid method for detection of genetically modified organisms based on magnetic separation and surface-enhanced Raman scattering.

PubMed

Guven, Burcu; Boyacı, İsmail Hakkı; Tamer, Ugur; Çalık, Pınar

2012-01-07

In this study, a new method combining magnetic separation (MS) and surface-enhanced Raman scattering (SERS) was developed to detect genetically modified organisms (GMOs). An oligonucleotide probe which is specific for 35 S DNA target was immobilized onto gold coated magnetic nanospheres to form oligonucleotide-coated nanoparticles. A self assembled monolayer was formed on gold nanorods using 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) and the second probe of the 35 S DNA target was immobilized on the activated nanorod surfaces. Probes on the nanoparticles were hybridized with the target oligonucleotide. Optimization parameters for hybridization were investigated by high performance liquid chromatography. Optimum hybridization parameters were determined as: 4 μM probe concentration, 20 min immobilization time, 30 min hybridization time, 55 °C hybridization temperature, 750 mM buffer salt concentration and pH: 7.4. Quantification of the target concentration was performed via SERS spectra of DTNB on the nanorods. The correlation between the target concentration and the SERS signal was found to be linear within the range of 25-100 nM. The analyses were performed with only one hybridization step in 40 min. Real sample analysis was conducted using Bt-176 maize sample. The results showed that the developed MS-SERS assay is capable of detecting GMOs in a rapid and selective manner. This journal is © The Royal Society of Chemistry 2012

Biogeochemical Gradients in Wetland Sediments and their Effect on the Fate Trace Metals

NASA Astrophysics Data System (ADS)

Jaffe, P. R.; Choi, J.; Xu, S.

2005-12-01

The interactions between sediment biogeochemistry processes and higher plants play a major role on trace metal mobility in wetlands. Most wetland sediments are characterized by steep redox gradients, resulting from the sequential utilization of different electron acceptors during the degradation of organic matter provided by leaf litter and root turnover. Metals in wetland sediments may be immobilized due to precipitation or adsorption to different organic and inorganic sediment constituents. Adsorption onto iron, and manganese oxides, are important in the rhizosphere where iron oxyhydroxide plaques may form on the surface of roots. As the sediments becomes more reduced, bioavailable iron and manganese oxides are used as electron acceptor and are gradually depleted, resulting in the mobilization of some adsorbed species (i.e., As(V), phosphate, etc.), the reduction of some trace metals such as Cr(VI) (which is then immobilized as Cr(III)), and for more reduced conditions the immobilization of trace metals (i.e., Cd, Pb, Zn) as sulfides. Results from numerical simulations, laboratory experiments, and field measurements will be presented, showing how redox gradients and hence, trace-metal immobilization, in wetlands respond to external forcing functions such as changes in nutrient loading, plant distribution, seasonal and diurnal plant activity (specifically evapotranspiration and oxygen release), and temporal or spatial changes in the profile of iron and manganese oxides.

Immobilization of tetracyclines in manure and manure-amended soils using aluminum-based drinking water treatment residuals.

PubMed

Punamiya, Pravin; Sarkar, Dibyendu; Rakshit, Sudipta; Elzinga, Evert J; Datta, Rupali

2016-02-01

Veterinary antibiotics (VAs) are emerging contaminants of concern in the environment, mainly due to the potential for development of antibiotic-resistant bacteria and effect on microbiota that could interfere with crucial ecosystem functions such as nutrient cycling and decomposition. High levels of VAs such as tetracyclines (TCs) have been reported in agricultural soils amended with manure, which also has the potential to cause surface and groundwater contamination. Several recent studies have focused on developing methods to immobilize VAs such as composting with straw, hardwood chips, commercial biochar, aeration, mixing, heat treatment, etc. The major shortcomings of these methods include high cost and limited effectiveness. In the current study, we assessed the effectiveness of aluminum-based drinking water treatment residuals (Al-WTR) as a "green" sorbent to immobilize TCs in manure and manure-applied soils with varying physicochemical properties by laboratory incubation study. Results show that Al-WTR is very effective in immobilizing tetracycline (TTC) and oxytetracycline (OTC). The presence of phosphate resulted in significant (p < 0.01) decrease in TTC/OTC sorption by Al-WTR, but the presence of sulfate did not. attenuated total reflection (ATR)-FTIR spectroscopy indicate that TTC and OTC likely forming surface complexes via inner-sphere-type bonds in soils, manure, and manure-applied soils amended with Al-WTR.

Redox transformation and reductive immobilization of Cr(VI) in the Columbia River hyporheic zone sediments

NASA Astrophysics Data System (ADS)

Xu, Fen; Liu, Yuanyuan; Zachara, John; Bowden, Mark; Kennedy, David; Plymale, Andrew E.; Liu, Chongxuan

2017-12-01

An experimental and modeling study was conducted to investigate the redox transformation and reductive immobilization of groundwater contaminant Cr in hyporheic zone (HZ) sediments from U.S. DOE's Hanford Site, where groundwater Cr(VI) is migrating and discharging to the nearby Columbia River. Experimental results revealed that Cr(VI) can be reduced and immobilized by the HZ sediments in the presence/absence of O2. Anaerobic pre-incubation of the sediments increased the effective rate of Cr reduction that was correlated with the increase in HCl-extractable Fe(II) content in the sediments. The reduced Cr was stable when exposed to O2 under field-relevant pH (7.5) with and without dissolved Mn(II), which might be oxidized to form Mn(III/IV) oxides that may oxidize reduced Cr. The Cr(VI) reduction rate showed a multi-rate behavior, apparently reflecting the presence of reductants with different reactivity in the sediments. The results from this study indicated that the HZ sediments can reductively immobilize Cr and the sediment redox capacity can be recharged through microbial activities. The results implied that HZ can play a role as a natural permeable redox barrier for removing groundwater Cr before it discharges into a river system.

Electrostatic immobilization of antimicrobial peptides on polyethylenimine and their antibacterial effect against Staphylococcus epidermidis.

PubMed

Hernandez-Montelongo, J; Corrales Ureña, Y R; Machado, D; Lancelloti, M; Pinheiro, M P; Rischka, K; Lisboa-Filho, P N; Cotta, M A

2018-04-01

Staphylococcus epidermidis is a gram-positive bacterium, and one of the most prevalent causes of nosocomial infections due to its strong ability to form biofilms on catheters and surgical implants. Here we explore the antimicrobial properties of Tet-124 peptides, which are part of the innate defense against different multicellular organisms in nature. Two different Tet-124 peptides were immobilized on a polyethylenimine (PEI) film to determine their impact on the antimicrobial properties: KLWWMIRRW (Tet-124), which contains only natural amino acids, and KLWWMIRRWG-(F-Br)-G (F-Br = 4-Bromophenylalanine), a modified Tet-124 sequence with the addition of an unnatural amino acid. The immobilization was obtained as a result of the electrostatic interaction between PEI amino groups and the C-terminal carboxylic groups of tryptophan and glycine amino acids of Tet-124 and Tet-124-Br peptides, respectively. The process was monitored and studied by water contact angle, Atomic Force Microscopy (AFM), X-ray Photoelectron Spectroscopy (XPS) and Quartz Crystal Microbalance with Dissipation (QCM-D) measurements. The antibacterial effect of our samples against S. epidermis was evaluated by the spread plate counting method, and cytotoxicity was tested using fibroblast cultures. Our results indicate the feasibility to immobilize electrostatically both Tet-124 peptides for biomedical applications. Copyright © 2018 Elsevier B.V. All rights reserved.

Induction of myofibroblastic differentiation in vitro by covalently immobilized transforming growth factor-beta(1).

PubMed

Metzger, Wolfgang; Grenner, Nadine; Motsch, Sandra E; Strehlow, Rothin; Pohlemann, Tim; Oberringer, Martin

2007-11-01

Growth factors are an important tool in tissue engineering. Bone morphogenetic protein-2 and transforming growth factor-beta(1) (TGF-beta(1)) are used to provide bioactivity to surgical implants and tissue substitute materials. Mostly growth factors are used in soluble or adsorbed form. However, simple adsorption of proteins to surfaces is always accompanied by reduced stability and undefined pharmacokinetics. This study aims to prove that TGF-beta(1) can be covalently immobilized to functionalized surfaces, maintaining its ability to induce myofibroblastic differentiation of normal human dermal fibroblasts. In vivo, fibroblasts differentiate to myofibroblasts (MFs) during soft tissue healing by the action of TGF-beta(1). As surfaces for our experiments, we used slides bearing aldehyde, epoxy, or amino groups. For our in vitro cell culture experiments, we used the expression of alpha-smooth muscle actin as a marker for MFs after immunochemical staining. Using the aldehyde and the epoxy slides, we were able to demonstrate the activity of immobilized TGF-beta(1) through a significant increase in MF differentiation rate. A simple immunological test was established to detect TGF-beta(1) on the surfaces. This technology enables the creation of molecular "landscapes" consisting of several factors arranged in a distinct spatial pattern and immobilized on appropriate surfaces.

DNA microdevice for electrochemical detection of Escherichia coli 0157:H7 molecular markers.

PubMed

Berganza, J; Olabarria, G; García, R; Verdoy, D; Rebollo, A; Arana, S

2007-04-15

An electrochemical DNA sensor based on the hybridization recognition of a single-stranded DNA (ssDNA) probe immobilized onto a gold electrode to its complementary ssDNA is presented. The DNA probe is bound on gold surface electrode by using self-assembled monolayer (SAM) technology. An optimized mixed SAM with a blocking molecule preventing the nonspecific adsorption on the electrode surface has been prepared. In this paper, a DNA biosensor is designed by means of the immobilization of a single stranded DNA probe on an electrochemical transducer surface to recognize specifically Escherichia coli (E. coli) 0157:H7 complementary target DNA sequence via cyclic voltammetry experiments. The 21 mer DNA probe including a C6 alkanethiol group at the 5' phosphate end has been synthesized to form the SAM onto the gold surface through the gold sulfur bond. The goal of this paper has been to design, characterise and optimise an electrochemical DNA sensor. In order to investigate the oligonucleotide probe immobilization and the hybridization detection, experiments with different concentration of DNA and mismatch sequences have been performed. This microdevice has demonstrated the suitability of oligonucleotide Self-assembled monolayers (SAMs) on gold as immobilization method. The DNA probes deposited on gold surface have been functional and able to detect changes in bases sequence in a 21-mer oligonucleotide.

Diazonium-protein adducts for graphite electrode microarrays modification: direct and addressed electrochemical immobilization.

PubMed

Corgier, Benjamin P; Marquette, Christophe A; Blum, Loïc J

2005-12-28

Diazonium cation electrodeposition was investigated for the direct and electro-addressed immobilization of proteins. For the first time, this reaction was triggered directly onto diazonium-modified proteins. Screen-printed (SP) graphite electrode microarrays were studied as active support for this immobilization. A 10-microelectrode (eight working electrodes, 0.2 mm2 each; one reference; and one auxiliary) setup was used to study the addressing possibilities of the method. These electrode microarrays were shown to be able to covalently graft diazonium cations through electrochemical reduction. Cyclic voltammetry and X-ray photoelectron spectroscopy were used to characterize the electrochemical grafting onto our SP graphite surface and suggested that a diazonium monolayer was deposited. Rabbit and human immunoglobulins (IgGs) were then chemically coupled to an aniline derivative (4-carboxymethylaniline), followed by diazotation to form an aryl diazonium function available for the electrodeposition. These modified proteins were both successfully electro-addressed at the surface of the graphite electrodes without cross-talk or interference. The immuno-biochip obtained using this novel approach enabled the specific detection of anti-rabbit IgG antibodies with a detection limit of 50 fmol of protein. A promising strategy to immobilize markedly different biological entities was then presented, providing an excellent spatial specificity of the electro-addressing.

Nursing leadership succession planning in Veterans Health Administration: creating a useful database.

PubMed

Weiss, Lizabeth M; Drake, Audrey

2007-01-01

An electronic database was developed for succession planning and placement of nursing leaders interested and ready, willing, and able to accept an assignment in a nursing leadership position. The tool is a 1-page form used to identify candidates for nursing leadership assignments. This tool has been deployed nationally, with access to the database restricted to nurse executives at every Veterans Health Administration facility for the purpose of entering the names of developed nurse leaders ready for a leadership assignment. The tool is easily accessed through the Veterans Health Administration Office of Nursing Service, and by limiting access to the nurse executive group, ensures candidates identified are qualified. Demographic information included on the survey tool includes the candidate's demographic information and other certifications/credentials. This completed information form is entered into a database from which a report can be generated, resulting in a listing of potential candidates to contact to supplement a local or Veterans Integrated Service Network wide position announcement. The data forms can be sorted by positions, areas of clinical or functional experience, training programs completed, and geographic preference. The forms can be edited or updated and/or added or deleted in the system as the need is identified. This tool allows facilities with limited internal candidates to have a resource with Department of Veterans Affairs prepared staff in which to seek additional candidates. It also provides a way for interested candidates to be considered for positions outside of their local geographic area.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cozzi, A.; Crawford, C.; Fox, K.

More than 56 million gallons of radioactive and hazardous waste are stored in 177 underground storage tanks at the U.S. Department of Energy’s (DOE’s) Hanford Site in Washington State. The HLW will be vitrified in the HLW facility for ultimate disposal at an offsite federal repository. A portion (~35%) of the LAW will be vitrified in the LAW vitrification facility for disposal onsite at the Integrated Disposal Facility (IDF). The pretreatment and HLW vitrification facilities will have the capacity to treat and immobilize all of the wastes destined for those facilities. However, a second facility will be needed for themore » expected volume of LAW requiring immobilization. Cast Stone, a cementitious waste form, is being considered to provide the required additional LAW immobilization capacity. The Cast Stone waste form must be acceptable for disposal in the IDF. The Cast Stone waste form and immobilization process must be tested to demonstrate that the final Cast Stone waste form can comply with the waste acceptance criteria for the disposal facility and that the immobilization processes can be controlled to consistently provide an acceptable waste form product. A testing program was developed in fiscal year (FY) 2012 describing in detail the work needed to develop and qualify Cast Stone as a waste form for the solidification of Hanford LAW. A statistically designed test matrix was used to evaluate the effects of key parameters on the properties of the Cast Stone as it is initially prepared and after curing. For the processing properties, the water-to-dry-blend mix ratio was the most significant parameter in affecting the range of values observed for each property. The single shell tank (SST) Blend simulant also showed differences in measured properties compared to the other three simulants tested. A review of the testing matrix and results indicated that an additional set of tests would be beneficial to improve the understanding of the impacts noted in the Screening Matrix tests. A set of Cast Stone formulations were devised to augment the original screening test matrix and focus on the range of the test conditions. Fly ash and blast furnace slag were limited to either northwest or southeast and the salt solutions were narrowed to the Average and the SST Blend at the 7.8M Na concentration. To fill in the matrix, a mix ratio of 0.5 was added. In addition, two admixtures, Xypex Admix C-500 and Rheomac SF100 (silica fume), were added as an additional dry material binder in select compositions. As in the Screening Matrix, both fresh and cured properties were evaluated for the formulations. In this study, properties that were influenced by the W/DM ratio in the Screening Matrix; flow diameter, plastic viscosity, density, and compressive strength, showed consistent behavior with respect to W/DM. The leach index for highly soluble components, sodium and nitrate, were not influenced by changes in formulation or the admixtures. The leach index for both iodine and Tc-99 show an influence from the addition of the admixture, Xypex Admix C-500. Additional testing should be performed to further evaluate the influence of Xypex Admix C-500 on the leach index over a range of admixture concentrations, Cast Stone formulations, and curing and storage conditions.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Asmussen, Robert M.; Lawter, Amanda R.; Stephenson, John R.

Washington River Protection Solutions (WRPS) is collecting relevant available data on waste forms for use as a supplemental immobilization technology, to provide the additional capacity needed to treat low-activity waste (LAW) in Hanford Site tanks and complete the tank waste cleanup mission in a timely and cost-effective manner. One candidate supplemental waste form, fabricated using a low-temperature process, is a cementitious grout called Cast Stone. Cast Stone has been under investigation for this application at Pacific Northwest National Laboratory (PNNL) since initial screening tests in FY13. This report is the culmination of work to lower the diffusivities of Tc andmore » I from Cast Stone using getters. Getters are compounds added to a system designed to selectively sequester a species of interest to provide increased stability to the species. The work contained within this report is related to waste form development and testing, and does not directly support the 2017 integrated disposal facility (IDF) performance assessment. However, this work contains valuable information which may be used in performance assessment maintenance past FY17, and in future waste form development. This report on performance characterization of Tc and I getters in Cast Stone fabricated with simulated LAW covers several areas of interest and major findings to WRPS: investigating performance of potassium metal sulfide (KMS-2-SS) and tin (II) apatite (Sn-A) as Tc getters when incorporated into Cast Stone; investigating performance of silver exchanged zeolite (Ag-Z) and argentite (Arg) as I getters when incorporated into Cast Stone; utilizing sequential addition of Tc and I getters to overcome any deleterious interactions between the getters in solution; determining, for the first time, Tc distribution within the cured Cast Stone and its evolution during leaching; and performing solid state characterization of getters and Cast Stone samples to support leach test findings and develop a mechanistic understanding of the processes that control Tc and I release into solution.« less

Uranium speciation and stability after reductive immobilization in aquifer sediments

NASA Astrophysics Data System (ADS)

Sharp, Jonathan O.; Lezama-Pacheco, Juan S.; Schofield, Eleanor J.; Junier, Pilar; Ulrich, Kai-Uwe; Chinni, Satya; Veeramani, Harish; Margot-Roquier, Camille; Webb, Samuel M.; Tebo, Bradley M.; Giammar, Daniel E.; Bargar, John R.; Bernier-Latmani, Rizlan

2011-11-01

It has generally been assumed that the bioreduction of hexavalent uranium in groundwater systems will result in the precipitation of immobile uraninite (UO 2). In order to explore the form and stability of uranium immobilized under these conditions, we introduced lactate (15 mM for 3 months) into flow-through columns containing sediments derived from a former uranium-processing site at Old Rifle, CO. This resulted in metal-reducing conditions as evidenced by concurrent uranium uptake and iron release. Despite initial augmentation with Shewanella oneidensis, bacteria belonging to the phylum Firmicutes dominated the biostimulated columns. The immobilization of uranium (˜1 mmol U per kg sediment) enabled analysis by X-ray absorption spectroscopy (XAS). Tetravalent uranium associated with these sediments did not have spectroscopic signatures representative of U-U shells or crystalline UO 2. Analysis by microfocused XAS revealed concentrated micrometer regions of solid U(IV) that had spectroscopic signatures consistent with bulk analyses and a poor proximal correlation (μm scale resolution) between U and Fe. A plausible explanation, supported by biogeochemical conditions and spectral interpretations, is uranium association with phosphoryl moieties found in biomass; hence implicating direct enzymatic uranium reduction. After the immobilization phase, two months of in situ exposure to oxic influent did not result in substantial uranium remobilization. Ex situ flow-through experiments demonstrated more rapid uranium mobilization than observed in column oxidation studies and indicated that sediment-associated U(IV) is more mobile than biogenic UO 2. This work suggests that in situ uranium bioimmobilization studies and subsurface modeling parameters should be expanded to account for non-uraninite U(IV) species associated with biomass.

Immobilization of hexavalent chromium in contaminated soils using biochar supported nanoscale iron sulfide composite.

PubMed

Lyu, Honghong; Zhao, Hang; Tang, Jingchun; Gong, Yanyan; Huang, Yao; Wu, Qihang; Gao, Bin

2018-03-01

Biochar supported carboxymethyl cellulose (CMC)-stabilized nanoscale iron sulfide (FeS) composite (CMC-FeS@biochar) was prepared and tested for immobilization of hexavalent chromium Cr(VI) in soil. Results of UV-vis and transmission electron microscopy (TEM) showed that the backbone of biochar suppressed the aggregation of FeS, resulting in smaller particle size and more sorption sites than bare FeS. The composite at a dosage of 2.5 mg per gram soil displayed an enhanced Cr(VI) immobilization efficiency (a 94.7% reduction in the toxicity characteristic leaching procedure (TCLP) based leachability and a 95.6% reduction in the CaCl 2 extraction) compared to plain biochar and bare FeS. Sequential extraction procedure (SEP) and X-ray photoelectron spectroscopy (XPS) analysis suggested that CMC-FeS@biochar promoted the conversion of more accessible Cr (exchangeable and carbonate-bound fractions) into the less accessible forms (iron-manganese oxides-bound, organic material-bound, and residual fractions) to reduce the toxicity of Cr(VI) and that surface sorption and reduction were dominant mechanisms for Cr(VI) immobilization. CMC-FeS@biochar greatly reduced the bioavailability of Cr(VI) to wheat and earthworms (Eisenia fetida). Moreover, the application of CMC-FeS@biochar enhanced soil organic matter content and microbial activity. This work highlighted the potential of CMC-FeS@biochar composite as a low-cost, "green", and effective amendment for immobilizing Cr(VI) in contaminated soils and improving soil properties. Copyright © 2017 Elsevier Ltd. All rights reserved.

Quartz crystal microbalance (QCM) with immobilized protein receptors: comparison of response to ligand binding for direct protein immobilization and protein attachment via disulfide linker.

PubMed

Baltus, Ruth E; Carmon, Kendra S; Luck, Linda A

2007-03-27

Results from an investigation of the frequency response resulting from ligand binding for a genetically engineered hormone-binding domain of the alpha-estrogen receptor immobilized to a piezoelectric quartz crystal are reported. Two different approaches were used to attach a genetically altered receptor to the gold electrode on the quartz surface: (1) the mutant receptor containing a single solvent-exposed cysteine was directly attached to the crystal via a sulfur to gold covalent bond, forming a self-assembled protein monolayer, and (2) the N-terminal histidine-tagged end was utilized to attach the receptor via a 3,3-dithiobis[N-(5-amino-5-carboxypentyl)propionamide-N',N'-diacetic acid] linker complexed with nickel. Previous studies have shown that these engineered constructs bind 17beta-estradiol and are fully functional. Exposure of the receptor directly attached to the piezoelectric crystal to the known ligand 17beta-estradiol resulted in a measurable frequency response, consistent with a change in conformation of the receptor with ligand binding. However, no response was observed when the receptor immobilized via the linker was exposed to the same ligand. The presence of the linker between the quartz surface and the protein receptor does not allow the crystal to sense the conformational change in the receptor that occurs with ligand binding. These results illustrate that the immobilization strategy used to bind the receptor to the sensor platform is key to eliciting an appropriate response from this biosensor. This study has important implications for the development of QCM-based sensors using protein receptors.

The on-bead digestion of protein corona on nanoparticles by trypsin immobilized on the magnetic nanoparticle.

PubMed

Hu, Zhengyan; Zhao, Liang; Zhang, Hongyan; Zhang, Yi; Wu, Ren'an; Zou, Hanfa

2014-03-21

Proteins interacting with nanoparticles would form the protein coronas on the surface of nanoparticles in biological systems, which would critically impact the biological identities of nanoparticles and/or result in the physiological and pathological consequences. The enzymatic digestion of protein corona was the primary step to achieve the identification of protein components of the protein corona for the bottom-up proteomic approaches. In this study, the investigation on the tryptic digestion of protein corona by the immobilized trypsin on a magnetic nanoparticle was carried out for the first time. As a comparison with the usual overnight long-time digestion and the severe self-digestion of free trypsin, the on-bead digestion of protein corona by the immobilized trypsin could be accomplished within 1h, along with the significantly reduced self-digestion of trypsin and the improved reproducibility on the identification of proteins by the mass spectrometry-based proteomic approach. It showed that the number of identified bovine serum (BS) proteins on the commercial Fe3O4 nanoparticles was increased by 13% for the immobilized trypsin with 1h digestion as compared to that of using free trypsin with even overnight digestion. In addition, the on-bead digestion of using the immobilized trypsin was further applied on the identification of human plasma protein corona on the commercial Fe3O4 nanoparticles, which leads the efficient digestion of the human plasma proteins and the identification of 149 human plasma proteins corresponding to putative critical pathways and biological processes. Copyright © 2014 Elsevier B.V. All rights reserved.

A novel fluorescence sensor based on covalent immobilization of 3-amino-9-ethylcarbazole by using silver nanoparticles as bridges and carriers.

PubMed

Tan, Shu-Zhen; Hu, Yan-Jun; Gong, Fu-Chun; Cao, Zhong; Xia, Jiao-Yun; Zhang, Ling

2009-03-23

A novel technique of covalent immobilization of indicator dyes in the preparation of fluorescence sensors is developed. Silver nanoparticles are used as bridges and carriers for anchoring indicator dyes. 3-amino-9-ethylcarbazole (AEC) was employed as an example of indicator dyes with terminal amino groups and covalently immobilized onto the outmost surface of a quartz glass slide. First, the glass slide was functionalized by (3-mercaptopropyl) trimethoxysilane (MPS) to form a thiol-terminated self-assembled monolayer, where silver nanoparticles were strongly bound to the surface through covalent bonding. Then, 16-mercaptohexadecanoic acid (MHDA) was self-assembled to bring carboxylic groups onto the surface of silver nanoparticles. A further activation by using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) converted the carboxylic groups into succinimide esters. Finally, the active succinimide esters on the surface of silver nanoparticles were reacted with AEC. Thus, AEC was covalently bound to the glass slide and an AEC-immobilized sensor was obtained. The sensor exhibited very satisfactory reproducibility and reversibility, rapid response and no dye-leaching. Rutin can quench the fluorescence intensity of the sensor and be measured by using the sensor. The linear response of the sensor to rutin covers the range from 2.0 x 10(-6) to 1.5 x 10(-4) molL(-1) with a detection limit of 8.0 x 10(-7) molL(-1). The proposed technique may be feasible to the covalent immobilization of other dyes with primary amino groups.

Three MOF-Templated Carbon Nanocomposites for Potential Platforms of Enzyme Immobilization with Improved Electrochemical Performance.

PubMed

Dong, Sheying; Peng, Lei; Wei, Wenbo; Huang, Tinglin

2018-05-02

An efficient and facile metal-organic framework (MOF)-template strategy for preparing carbon nanocomposites has been developed. First of all, a series of metal ions, including Fe 3+ , Zr 4+ , and La 3+ , were respectively connected with 2-aminoterephthalate (H 2 ATA) to form three metal-organic frameworks (MOFs) and then three novel MOF-derived materials were obtained by annealing them at 550 °C under N 2 atmosphere. The morphologies and microstructure results showed that they still retained the original structure of MOFs and formed carbon-supported metal oxide hybrid nanomaterials. Interestingly, it was found that La-MOF-NH 2 and its derived materials were first reported, which had wool-ball-like structure formed by many streaky-shaped particles intertwining each other. Furthermore, these MOF-derived materials were all successfully used as effective immobilization matrixes of acetylcholinesterase (AChE) to construct biosensors for the detection of methyl parathion. Especially, [La-MOF-NH 2 ] N 2 with wool-ball-like structure not only provided more active sites of multicontents to increase AChE immobilization amount but also facilitated the accessibility of electron transfer and shorten their diffusion length on the surface of electrode. Under optimal conditions, the biosensor based on [La-MOF-NH 2 ] N 2 displayed the widest linear range of 1.0 × 10 -13 -5.0 × 10 -9 g mL -1 and the lowest detection limit of 5.8 × 10 -14 g mL -1 in three biosensors. This study illustrates the feasibility and the potential of a series of MOF-derived materials for biosensors with improved electrochemical performance.

Glucose Biosensor Based on Immobilization of Glucose Oxidase in Platinum Nanoparticles/Graphene/Chitosan Nanocomposite Film

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu, Hong; Wang, Jun; Kang, Xinhuang

2009-09-01

The bionanocomposite film consisting of glucose oxidase/Pt/functional graphene sheets/chitosan (GOD/Pt/FGS/chitosan) for glucose sensing was described. With the electrocatalytic synergy of FGS and Pt nanoparticles to hydrogen peroxide, a sensitive biosensor with detection limit of 0.6 µM glucose was achieved. The biosensor also had good reproducibility, long term stability and negligible interfering signals from ascorbic acid and uric acid comparing to the response to glucose. The large surface area and good conductivity of graphene suggests that graphene is a potential candidate for sensor material. The hybrid nanocomposite glucose sensor provides new opportunity for clinical diagnosis and point-of-care applications.

Point-of-Need bioanalytics based on planar optical interferometry.

PubMed

Makarona, E; Petrou, P; Kakabakos, S; Misiakos, K; Raptis, I

2016-01-01

This review brings about a comprehensive presentation of the research on interferometric transducers, which have emerged as extremely promising candidates for viable, truly-marketable solutions for PoN applications due to the attested performance that has reached down to 10(-8) in term of effective refractive index changes. The review explores the operation of the various interferometric architectures along with their design, fabrication, and analytical performance aspects. The issues of biosensor functionalization and immobilization of receptors are also addressed. As a conclusion, the comparison among them is attempted in order to delve into and acknowledge their current limitations, and define the future trends. Copyright © 2016 Elsevier Inc. All rights reserved.

Bacterial mimetics of endocrine secretory granules as immobilized in vivo depots for functional protein drugs

PubMed Central

Céspedes, María Virtudes; Fernández, Yolanda; Unzueta, Ugutz; Mendoza, Rosa; Seras-Franzoso, Joaquin; Sánchez-Chardi, Alejando; Álamo, Patricia; Toledo-Rubio, Verónica; Ferrer-Miralles, Neus; Vázquez, Esther; Schwartz, Simó; Abasolo, Ibane; Corchero, José Luis; Mangues, Ramon; Villaverde, Antonio

2016-01-01

In the human endocrine system many protein hormones including urotensin, glucagon, obestatin, bombesin and secretin, among others, are supplied from amyloidal secretory granules. These granules form part of the so called functional amyloids, which within the whole aggregome appear to be more abundant than formerly believed. Bacterial inclusion bodies (IBs) are non-toxic, nanostructured functional amyloids whose biological fabrication can be tailored to render materials with defined biophysical properties. Since under physiological conditions they steadily release their building block protein in a soluble and functional form, IBs are considered as mimetics of endocrine secretory granules. We have explored here if the in vivo implantation of functional IBs in a given tissue would represent a stable local source of functional protein. Upon intratumoral injection of bacterial IBs formed by a potent protein ligand of CXCR4 we have observed high stability and prevalence of the material in absence of toxicity, accompanied by apoptosis of CXCR4+ cells and tumor ablation. Then, the local immobilization of bacterial amyloids formed by therapeutic proteins in tumors or other tissues might represent a promising strategy for a sustained local delivery of protein drugs by mimicking the functional amyloidal architecture of the mammals’ endocrine system. PMID:27775083

Immobilization of U(VI) from Oxic Groundwater by Hanford 300 Area Sediments and Effects of Columbia River Water

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ahmed, B.; Cao, Bin; Mishra, Bhoopesh

2012-09-23

Regions within the U.S. Department of Energy Hanford 300 Area (300 A) site experience periodic hydrologic influences from the nearby Columbia River as a result of changing river stage, which causes changes in groundwater elevation, flow direction and water chemistry. An important question is the extent to which the mixing of Columbia River water and groundwater impacts the speciation and mobility of uranium (U). In this study, we designed experiments to mimic interactions among U, oxic groundwater or Columbia River water, and 300 A sediments in the subsurface environment of Hanford 300 A. The goals were to investigate mechanisms of:more » 1) U immobilization in 300 A sediments under bulk oxic conditions and 2) U remobilization from U-immobilized 300 A sediments exposed to oxic Columbia River water. Initially, 300 A sediments in column reactors were fed with U(VI)-containing oxic 1) synthetic groundwater (SGW), 2) organic-amended SGW (OA-SGW), and 3) de-ionized (DI) water to investigate U immobilization processes. After that, the sediments were exposed to oxic Columbia River water for U remobilization studies. The results reveal that U was immobilized by 300 A sediments predominantly through reduction (80-85%) when the column reactor was fed with oxic OA-SGW. However, U was immobilized by 300 A sediments through adsorption (100%) when the column reactors were fed with oxic SGW or DI water. The reduced U in the 300 A sediments fed with OA-SGW was relatively resistant to remobilization by oxic Columbia River water. Oxic Columbia River water resulted in U remobilization (~7%) through desorption, and most of the U that remained in the 300 A sediments fed with OA-SGW (~93%) was in the form of uraninite nanoparticles. These results reveal that: 1) the reductive immobilization of U through OA-SGW stimulation of indigenous 300 A sediment microorganisms may be viable in the relatively oxic Hanford 300 A subsurface environments and 2) with the intrusion of Columbia River water, desorption may be the primary process resulting in U remobilization from OA-SGW-stimulated 300 A sediments at the subsurface of the Hanford 300 A site.« less

A reagentless amperometric biosensor for alcohol detection in column liquid chromatography based on co-immobilized peroxidase and alcohol oxidase in carbon paste.

PubMed

Johansson, K; Jönsson-Pettersson, G; Gorton, L; Marko-Varga, G; Csöregi, E

1993-12-01

A reagentless carbon paste electrode chemically modified with covalently bound alcohol oxidase and horse-radish peroxidase was examined as a selective sensor in flow injection and column liquid chromatography. A combination of carbodiimide, glutaraldehyde, and polyethyleneimine was used for immobilizing the enzymes in the paste. The surface of the electrodes was protected by first forming a layer of electropolymerized ortho-phenylenediamine followed by deposition of a cation exchange membrane (Eastman AQ 29D). The electrodes were used for detection of hydrogen peroxide, methanol, ethanol, propanol, isopropanol, and butanol. Preliminary investigations of the use of this sensor for bioprocess control are reported.

Volatile Impurities in the Plutonium Immobilization Ceramic Wasteform

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cozzi, A.D.

1999-10-15

Approximately 18 of the 50 metric tons of plutonium identified for disposition contain significant quantities of impurities. A ceramic waste form is the chosen option for immobilization of the excess plutonium. The impurities associated with the stored plutonium have been identified (CaCl2, MgF2, Pb, etc.). For this study, only volatile species are investigated. The impurities are added individually. Cerium is used as the surrogate for plutonium. Three compositions, including the baseline composition, were used to verify the ability of the ceramic wasteform to accommodate impurities. The criteria for evaluation of the effect of the impurities were the apparent porosity andmore » phase assemblage of sintered pellets.« less

A simple approach to patterned protein immobilization on silicon via electrografting from diazonium salt solutions.

PubMed

Flavel, Benjamin S; Gross, Andrew J; Garrett, David J; Nock, Volker; Downard, Alison J

2010-04-01

A highly versatile method utilizing diazonium salt chemistry has been developed for the fabrication of protein arrays. Conventional ultraviolet mask lithography was used to pattern micrometer sized regions into a commercial photoresist on a highly doped p-type silicon (100) substrate. These patterned regions were used as a template for the electrochemical grafting of the in situ generated p-aminobenzenediazonium cation to form patterns of aminophenyl film on silicon. Immobilization of biomolecules was demonstrated by coupling biotin to the aminophenyl regions followed by reaction with fluorescently labeled avidin and visualization with fluorescence microscopy. This simple patterning strategy is promising for future application in biosensor devices.

Structure and Thermochemistry of Perrhenate Sodalite and Mixed Guest Perrhenate/Pertechnetate Sodalite

DOE PAGES

Pierce, Eric M.; Lilova, Kristina; Missimer, David M.; ...

2016-12-05

Here we report that treatment and immobilization of technetium-99 ( 99Tc) contained in reprocessed nuclear waste and present in contaminated subsurface systems represents a major environmental challenge. One potential approach to managing this highly mobile and long-lived radionuclide is immobilization into micro- and meso-porous crystalline solids, specifically sodalite. We synthesized and characterized the structure of perrhenate sodalite, Na 8[AlSiO 4]6(ReO 4) 2, and the structure of a mixed guest perrhenate/pertechnetate sodalite, Na 8[AlSiO 4] 6(ReO 4) 2-x(TcO 4) x. Perrhenate was used as a chemical analogue for pertechnetate. Bulk analyses of each solid confirm a cubic sodalite-type structure (Pmore » $$\\overline{43}$$n, No. 218 space group) with rhenium and technetium in the 7+ oxidation state. High-resolution nanometer scale characterization measurements provide first-of-a-kind evidence that the ReO 4 – anions are distributed in a periodic array in the sample, nanoscale clustering is not observed, and the ReO 4 – anion occupies the center of the sodalite β-cage in Na8[AlSiO4]6(ReO4)2. We also demonstrate, for the first time, that the TcO4– anion can be incorporated into the sodalite structure. Lastly, thermochemistry measurements for the perrhenate sodalite were used to estimate the thermochemistry of pertechnetate sodalite based on a relationship between ionic potential and the enthalpy and Gibbs free energy of formation for previously measured oxyanion-bearing feldspathoid phases. The results collected in this study suggest that micro- and mesoporous crystalline solids maybe viable candidates for the treatment and immobilization of 99Tc present in reprocessed nuclear waste streams and contaminated subsurface environments.« less

Antibacterial activity and cytocompatibility of an implant coating consisting of TiO2 nanotubes combined with a GL13K antimicrobial peptide.

PubMed

Li, Tao; Wang, Na; Chen, Su; Lu, Ran; Li, Hongyi; Zhang, Zhenting

2017-01-01

Prevention of implant-associated infections at an early stage of surgery is highly desirable for the long-term efficacy of implants in dentistry and orthopedics. Infection prophylaxis using conventional antibiotics is becoming less effective due to the development of bacteria resistant to multiple antibiotics. An ideal strategy to conquer bacterial infections is the local delivery of antibacterial agents. Therefore, antimicrobial peptide (AMP) eluting coatings on implant surfaces is a promising alternative. In this study, the feasibility of utilizing TiO 2 nanotubes (TNTs), processed using anodization, as carriers to deliver a candidate AMP on titanium surfaces for the prevention of implant-associated infections is assessed. The broad-spectrum GL13K (GKIIKLKASLKLL-CONH2) AMP derived from human parotid secretory protein was selected and immobilized to TNTs using a simple soaking technique. Field emission scanning electron microscope, X-ray diffraction, Fourier transform infrared spectroscopy, and liquid chromatography-mass spectrometry analyses confirmed the successful immobilization of GL13K to anatase TNTs. The drug-loaded coatings demonstrated a sustained and slow drug release profile in vitro and eradicated the growth of Fusobacterium nucleatum and Porphyromonas gingivalis within 5 days of culture, as assessed by disk-diffusion assay. The GL13K-immobilized TNT (GL13K-TNT) coating demonstrated greater biocompatibility, compared with a coating produced by incubating TNTs with equimolar concentrations of metronidazole. GL13K-TNTs produced no observable cytotoxicity to preosteoblastic cells (MC3T3-E1). The coating may also have an immune regulatory effect, in support of rapid osseointegration around implants. Therefore, the combination of TNTs and AMP GL13K may achieve simultaneous antimicrobial and osteoconductive activities.

Toward the Reconstitution of a Two-Enzyme Cascade for Resveratrol Synthesis on Potyvirus Particles.

PubMed

Besong-Ndika, Jane; Wahlsten, Matti; Cardinale, Daniela; Pille, Jan; Walter, Jocelyne; Michon, Thierry; Mäkinen, Kristiina

2016-01-01

The highly ordered protein backbone of virus particles makes them attractive candidates for use as enzyme nano-carriers (ENCs). We have previously developed a non-covalent and versatile approach for adhesion of enzymes to virus particles. This approach makes use of z33, a peptide derived from the B-domain of Staphylococcus aureus protein A, which binds to the Fc domain of many immunoglobulins. We have demonstrated that with specific antibodies addressed against the viral capsid proteins (CPs) an 87% coverage of z33-tagged proteins can be achieved on potyvirus particles. 4-coumarate coenzyme A ligase (4CL2) and stilbene synthase (STS) catalyze consecutive steps in the resveratrol synthetic pathway. In this study, these enzymes were modified to carry an N-terminal z33 peptide and a C-terminal 6xHis tag to obtain (z)4CL2(His) and (z)STS(His), respectively. A protein chimera, (z)4CL2::STS(His), with the same modifications was also generated from the genetic fusion of both mono-enzyme encoding genes. All z33 enzymes were biologically active after expression in Escherichia coli as revealed by LC-MS analysis to identify resveratrol and assembled readily into macromolecular complexes with Potato virus A particles and α-PVA CP antibodies. To test simultaneous immobilization-purification, we applied the double antibody sandwich - ELISA protocol to capture active z33-containg mono-enzymes and protein chimera directly from clarified soluble cell lysates onto the virus particle surface. These immobilized enzymes were able to synthesize resveratrol. We present here a bottom up approach to immobilize active enzymes onto virus-based ENCs and discuss the potential to utilize this method in the purification and configuration of nano-devices.

Zwitterionic sulfobetaine polymer-immobilized surface by simple tyrosinase-mediated grafting for enhanced antifouling property.

PubMed

Kwon, Ho Joon; Lee, Yunki; Phuong, Le Thi; Seon, Gyeung Mi; Kim, Eunsuk; Park, Jong Chul; Yoon, Hyunjin; Park, Ki Dong

2017-10-01

Introducing antifouling property to biomaterial surfaces has been considered an effective method for preventing the failure of implanted devices. In order to achieve this, the immobilization of zwitterions on biomaterial surfaces has been proven to be an excellent way of improving anti-adhesive potency. In this study, poly(sulfobetaine-co-tyramine), a tyramine-conjugated sulfobetaine polymer, was synthesized and simply grafted onto the surface of polyurethane via a tyrosinase-mediated reaction. Surface characterization by water contact angle measurements, X-ray photoelectron spectroscopy and atomic force microscopy demonstrated that the zwitterionic polymer was successfully introduced onto the surface of polyurethane and remained stable for 7days. In vitro studies revealed that poly(sulfobetaine-co-tyramine)-coated surfaces dramatically reduced the adhesion of fibrinogen, platelets, fibroblasts, and S. aureus by over 90% in comparison with bare surfaces. These results proved that polyurethane surfaces grafted with poly(sulfobetaine-co-tyramine) via a tyrosinase-catalyzed reaction could be promising candidates for an implantable medical device with excellent bioinert abilities. Antifouling surface modification is one of the key strategy to prevent the thrombus formation or infection which occurs on the surface of biomaterial after transplantation. Although there are many methods to modify the surface have been reported, necessity of simple modification technique still exists to apply for practical applications. The purpose of this study is to modify the biomaterial's surface by simply immobilizing antifouling zwitterion polymer via enzyme tyrosinase-mediated reaction which could modify versatile substrates in mild aqueous condition within fast time period. After modification, pSBTA grafted surface becomes resistant to various biological factors including proteins, cells, and bacterias. This approach appears to be a promising method to impart antifouling property on biomaterial surfaces. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Effect of orientational ordering of magnetic nanoemulsions immobilized in agar gel on magnetic hyperthermia

NASA Astrophysics Data System (ADS)

Lahiri, B. B.; Ranoo, Surojit; Philip, John

2018-04-01

Magnetic nanoemulsions of droplet size ∼200 nm, loaded with single domain superparamagnetic nanoparticles (MNP), are potential candidates for multimodal hyperthermia due to availability of large loading volume and enhanced permeation and retention (EPR) in the cancerous tissues. In such nanoemulsions, radio frequency alternating magnetic field induced heating occur at two entirely different length scales, viz. Neel-Brown relaxation of the dispersed MNP and Brownian relaxation of emulsion droplets. Here we study the effects of orientation ordering or texturing of droplets, immobilized in a tissue mimicking agar matrix, on the field induced heating efficiency. A higher specific absorption rate (maximum ∼73 ± 2 W/gFe) is observed for droplets orientated parallel to the direction of the alternating magnetic field because of the enhancement of effective uniaxial anisotropy energy density and increased effective relaxation time. For identical and non-interacting MNP oriented parallel to the external DC magnetic field, a threefold increase in the effective uniaxial anisotropy energy density and ∼20-30% increased specific absorption rate are observed as compared to those oriented perpendicular to the magnetic field. Magnetic force microscopy images showed that the spherical morphology of the droplets remains intact even after orientational ordering and average topographic height of the droplets are found to be ∼220 (±17) nm, which is in good agreement with the most probable size obtained from dynamic light scattering. The residual volume magnetization of the emulsion droplets is found to be 1.1 × 10-6 emu/cc, indicating the superparamagnetic nature of the droplets in tissue equivalent environment. The observed increase in heating efficiency of the immobilized and oriented emulsion droplets shows promising applications in multimodal hyperthermia therapy because of the requirement of lower dose of MNP and shorter treatment time.

Mechanisms of iodine release from iodoapatite in aqueous solution

NASA Astrophysics Data System (ADS)

Zhang, Z.; Wang, J.

2017-12-01

Immobilization of iodine-129 with waste forms in geological setting is challenging due to its extremely long half-life and high volatility in the environment. To evaluate the long-term performance of waste form, it is imperative to determine the release mechanism of iodine hosted in the waste form materials. This study investigated the iodine released from apatite structured waste form Pb9.85 (VO4)6 I1.7 to understand how diffusion and dissolution control the durability of apatite waste form. A standard semi-dynamic leach test was adopted in this study. Samples were exposed in fresh leachant periodically and the leachant was replaced after each interval. Each experiment was carried out in cap-sealed Teflon vessels under constant temperature (e.g. 90 °C). ICP-MS analysis on the reacted leachates shows that Pb and V were released constantly and congruently with the stoichiometric ratio of Pb/V. However, iodine release is incongruent and time dependent. The iodine release rate starts significantly higher than the corresponding stoichiometric value and gradually decreases, approaching the stoichiometric value. Therefore, a dual-mode mechanism is proposed to account for the iodine release from apatite, which is dominated by short-term diffusion and long-term dissolution processes. Additional tests show that the element release rates depend on a number of test parameters, including sample surface to solution volume ratio (m-1), interval (day), temperature (°C), and solution pH. This study provides a quantitative characterization of iodine release mechanism. The activation energy of iodine leaching 21±1.6 kJ/mol was obtained by varying the test temperature. At the test conditions of to neutral pH and 90 °C, the long-term iodine release rate 3.3 mg/(m2 • day) is projected by normalizing sample surface area to solution volume ratio (S/V) to 1.0 m-1 and interval to 1 day. These findings demonstrate i) the feasibility of our approach to quantify the release mechanism and ii) the performance of iodine apatite as a favorable waste form candidate for I-129 disposal.

Versatile bio-ink for covalent immobilization of chimeric avidin on sol-gel substrates.

PubMed

Heikkinen, Jarkko J; Kivimäki, Liisa; Määttä, Juha A E; Mäkelä, Inka; Hakalahti, Leena; Takkinen, Kristiina; Kulomaa, Markku S; Hytönen, Vesa P; Hormi, Osmo E O

2011-10-15

A bio-ink for covalent deposition of thermostable, high affinity biotin-binding chimeric avidin onto sol-gel substrates was developed. The bio-ink was prepared from heterobifunctional crosslinker 6-maleimidohexanoic acid N-hydroxysuccinimide which was first reacted either with 3-aminopropyltriethoxysilane or 3-aminopropyldimethylethoxysilane to form silane linkers 6-maleimide-N-(3-(triethoxysilyl)propyl)hexanamide or -(ethoxydimethylsilyl)propyl)-hexanamide. C-terminal cysteine genetically engineered to chimeric avidin was reacted with the maleimide group of silane linker in methanol/PBS solution to form a suspension, which was printed on sol-gel modified PMMA film. Different concentrations of chimeric avidin and ratios between silane linkers were tested to find the best properties for the bio-ink to enable gravure or inkjet printing. Bio-ink prepared from 3-aminopropyltriethoxysilane was found to provide the highest amount of active immobilized chimeric avidin. The developed bio-ink was shown to be valuable for automated fabrication of avidin-functionalized polymer films. Copyright © 2011 Elsevier B.V. All rights reserved.

Atomic-Level Quality Assessment of Enzymes Encapsulated in Bioinspired Silica.

PubMed

Martelli, Tommaso; Ravera, Enrico; Louka, Alexandra; Cerofolini, Linda; Hafner, Manuel; Fragai, Marco; Becker, Christian F W; Luchinat, Claudio

2016-01-04

Among protein immobilization strategies, encapsulation in bioinspired silica is increasingly popular. Encapsulation offers high yields and the solid support is created through a protein-catalyzed polycondensation reaction that occurs under mild conditions. An integrated strategy is reported for the characterization of both the protein and bioinspired silica scaffold generated by the encapsulation of enzymes with an external silica-forming promoter or with the promoter expressed as a fusion to the enzyme. This strategy is applied to the catalytic domain of matrix metalloproteinase 12. Analysis reveals that the structure of the protein encapsulated by either method is not significantly altered with respect to the native form. The structural features of silica obtained by either strategy are also similar, but differ from those obtained by other approaches. In case of the covalently linked R5-enzyme construct, immobilization yields are higher. Encapsulation through a fusion protein, therefore, appears to be the method of choice. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fabrication of fiber-optic localized surface plasmon resonance sensor and its application to detect antibody-antigen reaction of interferon-gamma

NASA Astrophysics Data System (ADS)

Jeong, Hyeon-Ho; Erdene, Norov; Lee, Seung-Ki; Jeong, Dae-Hong; Park, Jae-Hyoung

2011-12-01

A fiber-optic localized surface plasmon (FO LSPR) sensor was fabricated by gold nanoparticles (Au NPs) immobilized on the end-face of an optical fiber. When Au NPs were formed on the end-face of an optical fiber by chemical reaction, Au NPs aggregation occurred and the Au NPs were immobilized in various forms such as monomers, dimers, trimers, etc. The component ratio of the Au NPs on the end-face of the fabricated FO LSPR sensor was slightly changed whenever the sensors were fabricated in the same condition. Including this phenomenon, the FO LSPR sensor was fabricated with high sensitivity by controlling the density of Au NPs. Also, the fabricated sensors were measured for the resonance intensity for the different optical systems and analyzed for the effect on sensitivity. Finally, for application as a biosensor, the sensor was used for detecting the antibody-antigen reaction of interferon-gamma.

Effects of supercritical carbon dioxide on immobile bound polymer chains on solid substrates

NASA Astrophysics Data System (ADS)

Sen, Mani; Asada, Mitsunori; Jiang, Naisheng; Endoh, Maya K.; Akgun, Bulent; Satija, Sushil; Koga, Tadanori

2013-03-01

Adsorbed polymer layers formed on flat solid substrates have recently been the subject of extensive studies because it is postulated to control the dynamics of technologically relevant polymer thin films, for example, in lithography. Such adsorbed layers have been reported to hinder the mobility of polymer chains in thin films even at a large length scale. Consequently, this bound layer remains immobile regardless of processing techniques (i.e. thermal annealing, solvent dissolution, etc). Here, we investigate the use of supercritical carbon dioxide (scCO2) as a novel plasticizer for bound polystyrene layers formed on silicon substrates. In-situ swelling and interdiffusion experiments using neutron reflectivity were performed. As a result, we found the anomalous plasticization effects of scCO2 on the bound polymer layers near the critical point where the anomalous adsorption of CO2 molecules in polymer thin films has been reported previously. Acknowledgement: We acknowledge the financial support from NSF Grant No. CMMI-084626.

Immobilization of Alkali Metal Fluorides via Recrystallization in a Cationic Lamellar Material, [Th(MoO4)(H2O)4Cl]Cl·H2O.

PubMed

Lin, Jian; Bao, Hongliang; Qie, Meiying; Silver, Mark A; Yue, Zenghui; Li, Xiaoyun; Zhu, Lin; Wang, Xiaomei; Zhang, Linjuan; Wang, Jian-Qiang

2018-06-05

Searching for cationic extended materials with a capacity for anion exchange resulted in a unique thorium molybdate chloride (TMC) with the formula of [Th(MoO 4 )(H 2 O) 4 Cl]Cl·H 2 O. The structure of TMC is composed of zigzagging cationic layers [Th(MoO 4 )(H 2 O) 4 Cl] + with Cl - as interlamellar charge-balancing anions. Instead of performing ion exchange, alkali thorium fluorides were formed after soaking TMC in AF (A = Na, K, and Cs) solutions. The mechanism of AF immobilization is elucidated by the combination of SEM-EDS, PXRD, FTIR, and EXAFS spectroscopy. It was observed that four water molecules coordinating with the Th 4+ center in TMC are vulnerable to competition with F - , due to the formation of more favorable Th-F bonds compared to Th-OH 2 . This leads to a single crystal-to-polycrystalline transformation via a pathway of recrystallization to form alkali thorium fluorides.

Bioresponsive Materials for Drug Delivery Based on Carboxymethyl Chitosan/Poly(γ-Glutamic Acid) Composite Microparticles

PubMed Central

Yan, Xiaoting; Tong, Zongrui; Chen, Yu; Mo, Yanghe; Feng, Huaiyu; Li, Peng; Qu, Xiaosai; Jin, Shaohua

2017-01-01

Carboxymethyl chitosan (CMCS) microparticles are a potential candidate for hemostatic wound dressing. However, its low swelling property limits its hemostatic performance. Poly(γ-glutamic acid) (PGA) is a natural polymer with excellent hydrophilicity. In the current study, a novel CMCS/PGA composite microparticles with a dual-network structure was prepared by the emulsification/internal gelation method. The structure and thermal stability of the composite were determined by Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), and thermogravimetric analysis (TGA). The effects of preparation conditions on the swelling behavior of the composite were investigated. The results indicate that the swelling property of CMCS/PGA composite microparticles is pH sensitive. Levofloxacin (LFX) was immobilized in the composite microparticles as a model drug to evaluate the drug delivery performance of the composite. The release kinetics of LFX from the composite microparticles with different structures was determined. The results suggest that the CMCS/PGA composite microparticles are an excellent candidate carrier for drug delivery. PMID:28452963

Conjugation, characterization and toxicity of lipophosphoglycan-polyacrylic acid conjugate for vaccination against leishmaniasis.

PubMed

Topuzogullari, Murat; Cakir Koc, Rabia; Dincer Isoglu, Sevil; Bagirova, Melahat; Akdeste, Zeynep; Elcicek, Serhat; Oztel, Olga N; Yesilkir Baydar, Serap; Canim Ates, Sezen; Allahverdiyev, Adil M

2013-06-03

Research on the conjugates of synthetic polyelectrolytes with antigenic molecules, such as proteins, peptides, or carbohydrates, is an attractive area due to their highly immunogenic character in comparison to classical adjuvants. For example, polyacrylic acid (PAA) is a weak polyelectrolyte and has been used in several biomedical applications such as immunological studies, drug delivery, and enzyme immobilization. However, to our knowledge, there are no studies that document immune-stimulant properties of PAA in Leishmania infection. Therefore, we aimed to develop a potential vaccine candidate against leishmaniasis by covalently conjugating PAA with an immunologically vital molecule of lipophosphoglycan (LPG) found in Leishmania parasites. In the study, LPG and PAA were conjugated by a multi-step procedure, and final products were analyzed with GPC and MALDI-TOF MS techniques. In cytotoxicity experiments, LPG-PAA conjugates did not indicate toxic effects on L929 and J774 murine macrophage cells. We assume that LPG-PAA conjugate can be a potential vaccine candidate, and will be immunologically characterized in further studies to prove its potential.

A novel first aid stretcher for immobilization and transportation of spine injured patients.

PubMed

Liu, Yan-Sheng; Feng, Ya-Ping; Xie, Jia-Xin; Luo, Zhuo-Jing; Shen, Cai-Hong; Niu, Fang; Zou, Jian; Tang, Shao-Feng; Hao, Jiang; Xu, Jia-Xiang; Xiao, Li-Ping; Xu, Xiao-Ming; Zhu, Hui

2012-01-01

Effective immobilization and transportation are vital to the life-saving acute medical care needed when treating critically injured people. However, the most common types of stretchers used today are wrought with problems that can lead to further medical complications, difficulty in employment and rescue, and ineffective transitions to hospital treatment. Here we report a novel first aid stretcher called the "emergency carpet", which solves these problems with a unique design for spine injured patients. Polyurethane composite material, obtained by a novel process of manually mixing isocyanate and additives, can be poured into a specially designed fabric bag and allowed to harden to form a rigid human-shaped stretcher. The effectiveness of the emergency carpet was examined in the pre-hospital management of victims with spinal fractures. Additionally, it was tested on flat ground and complex terrain as well as in the sea and air. We demonstrated that the emergency carpet can be assembled and solidified on the scene in 5 minutes, providing effective immobilization to the entire injured body. With the protection of the emergency carpet, none of the 20 patients, who were finally confirmed to have spinal column fracture or dislocation, had any neurological deterioration during transportation. Furthermore, the carpet can be handled and transported by multiple means under differing conditions, without compromising immobilization. Finally, the emergency carpet allows the critically injured patient to receive multiple examinations such as X-ray, CT, and MRI without being removed from the carpet. Our results demonstrate that the emergency carpet has ideal capabilities for immobilization, extrication, and transportation of the spine injured patients. Compared with other stretchers, it allows for better mobility, effective immobilization, remarkable conformity to the body, and various means for transportation. The emergency carpet is promising for its intrinsic advantages in the pre-hospital management of accident victims.

Preformulation considerations for controlled release dosage forms. Part I. Selecting candidates.

PubMed

Chrzanowski, Frank

2008-01-01

The physical-chemical properties of interest for controlled release (CR) dosage form development presented are based on the author's experience. Part I addresses selection of the final form based on a logical progression of physical-chemical properties evaluation of candidate forms and elimination of forms with undesirable properties from further evaluation in order to simplify final form selection. Several candidate forms which could include salt, free base or acid, polymorphic and amorphic forms of a new chemical entity (NCE) or existing drug substance (DS) are prepared and evaluated for critical properties in a scheme relevant to manufacturing processes, predictive of problems, requiring small amounts of test materials and simple analytical tools. A stability indicating assay is not needed to initiate the evaluation. This process is applicable to CR and immediate release (IR) dosage form development. The critical properties evaluated are melting, crystallinity, solubilities in water, 0.1 N HCl, and SIF, hygrodymamics, i.e., moisture sorption and loss at extremes of RH, and LOD at typical wet granulation drying conditions, and processability, i.e., corrosivity, and filming and/or sticking upon compression.

Immobilization of long-lived radionuclides in carbon matrices produced with the use of polyimide binders

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abdulakhatov, Murat; Bartenev, Sergey; Firsin, Nikolai

Available in abstract form only. Full text of publication follows: Conditions for immobilization of long-lived radionuclides {sup 99}Tc, {sup 129}I and {sup 241}Am in carbon matrices were investigated by using their chemical analogs. Stable isotopes of rhenium, iodine and europium were used as chemical analogs of {sup 99}Tc, {sup 129}I and {sup 241}Am, respectively. It is shown that the carbon matrices incorporating the above elements can be produced by carbonization of composites with ITA-31 polyimide binder of the following composition: equal molar ratio between dianhydride of 3,3/,4,4/-benzophenone-tetracarboxylic acid and tetraacetyl derivative of 4,4/-diaminodiphenyl ester, radionuclide being investigated or its chemicalmore » analog and carbon fabric as reinforcing component. The elements under investigation were used both in the form of salts or oxides and in the form of their complexes with ion-exchange resins. The produced composites were carbonized in inert gas (argon) or in vacuum. The physical-chemical properties of the samples were studied. It was revealed that the resultant matrices meet the requirements imposed on waste storage and final disposal. (authors)« less

Development of a glucose oxidase-based biocatalyst adopting both physical entrapment and crosslinking, and its use in biofuel cells

NASA Astrophysics Data System (ADS)

Chung, Yongjin; Ahn, Yeonjoo; Christwardana, Marcelinus; Kim, Hansung; Kwon, Yongchai

2016-04-01

New enzymatic catalysts prepared using physical entrapment and chemical bonding were used as anodic catalysts to enhance the performance of enzymatic biofuel cells (EBCs). For estimating the physical entrapment effect, the best glucose oxidase (GOx) concentration immobilized on polyethyleneimine (PEI) and carbon nanotube (CNT) (GOx/PEI/CNT) was determined, while for inspecting the chemical bonding effect, terephthalaldehyde (TPA) and glutaraldehyde (GA) crosslinkers were employed. According to the enzyme activity and XPS measurements, when the GOx concentration is 4 mg mL-1, they are most effectively immobilized (via the physical entrapment effect) and TPA-crosslinked GOx/PEI/CNT(TPA/[GOx/PEI/CNT]) forms π conjugated bonds via chemical bonding, inducing the promotion of electron transfer by delocalization of electrons. Due to the optimized GOx concentration and π conjugated bonds, TPA/[GOx/PEI/CNT], including 4 mg mL-1 GOx displays a high electron transfer rate, followed by excellent catalytic activity and EBC performance.New enzymatic catalysts prepared using physical entrapment and chemical bonding were used as anodic catalysts to enhance the performance of enzymatic biofuel cells (EBCs). For estimating the physical entrapment effect, the best glucose oxidase (GOx) concentration immobilized on polyethyleneimine (PEI) and carbon nanotube (CNT) (GOx/PEI/CNT) was determined, while for inspecting the chemical bonding effect, terephthalaldehyde (TPA) and glutaraldehyde (GA) crosslinkers were employed. According to the enzyme activity and XPS measurements, when the GOx concentration is 4 mg mL-1, they are most effectively immobilized (via the physical entrapment effect) and TPA-crosslinked GOx/PEI/CNT(TPA/[GOx/PEI/CNT]) forms π conjugated bonds via chemical bonding, inducing the promotion of electron transfer by delocalization of electrons. Due to the optimized GOx concentration and π conjugated bonds, TPA/[GOx/PEI/CNT], including 4 mg mL-1 GOx displays a high electron transfer rate, followed by excellent catalytic activity and EBC performance. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr00902f

Immobilization and Stabilization of Acylase on Carboxylated Polyaniline Nanofibers for Highly Effective Antifouling Application via Quorum Quenching.

PubMed

Lee, Jeongjoon; Lee, Inseon; Nam, Jahyun; Hwang, Dong Soo; Yeon, Kyung-Min; Kim, Jungbae

2017-05-10

Acylase (AC) was immobilized and stabilized on carboxylated polyaniline nanofibers (cPANFs) for the development of antifouling nanobiocatalysts with high enzyme loading and stability. AC was immobilized via three different approaches: covalent attachment (CA), enzyme coating (EC), and magnetically separable enzyme precipitate coating (Mag-EPC). The enzyme activity per unit weight of cPANFs with Mag-EPC was 75 and 300 times higher than that of those with CA and EC, respectively, representing improved enzyme loading in the form of Mag-EPC. After incubation under shaking at 200 rpm for 20 days, Mag-EPC maintained 55% of its initial activity, whereas CA and EC showed 3 and 16% of their initial activities, respectively. The antifouling of highly loaded and stable Mag-EPC against the biofouling/biofilm formation of Pseudomonas aeruginosa was tested under static- and continuous-flow conditions. Biofilm formation in the presence of 40 μg/mL Mag-EPC under static condition was 5 times lower than that under control condition with no addition of Mag-EPC. Under continuous membrane filtration, Mag-EPC delayed the increase of transmembrane pressure (TMP) more effectively as the concentration of added Mag-EPC increased. When separating Mag-EPC and membranes in two different vessels under internal circulation of the culture solution, Mag-EPC maintained a higher permeability than the control with no Mag-EPC addition. It was also confirmed that the addition of Mag-EPC reduced the generation of N-acyl homoserine lactone (AHL) autoinducers. This result reveals that the inhibition of biofilm formation and biofouling in the presence of Mag-EPC is due to the hydrolysis of AHL autoinducers, catalyzed by the immobilized and stabilized AC in the form of Mag-EPC. Mag-EPC of AC with high enzyme loadings and improved stability has demonstrated its great potential as an antifouling agent by reducing biofilm formation and membrane biofouling based on "enzymatic quorum quenching" of autoinducers.

Increasing the activity and enantioselectivity of lipases by sol-gel immobilization: further advancements of practical interest

NASA Astrophysics Data System (ADS)

Tielmann, Patrick; Kierkels, Hans; Zonta, Albin; Ilie, Adriana; Reetz, Manfred T.

2014-05-01

The entrapment of lipases in hydrophobic silicate matrices formed by sol-gel mediated hydrolysis of RSi(OCH3)3/Si(OCH3)4 as originally reported in 1996 has been improved over the years by a number of modifications. In the production of second-generation sol-gel lipase immobilizates, a variety of additives during the sol-gel process leads to increased activity and enhanced stereoselectivity in esterifying kinetic resolution. Recent advances in this type of lipase immobilization are reviewed here, in addition to new results regarding the sol-gel entrapment of the lipase from Burkholderia cepacia. It constitutes an excellent heterogeneous biocatalyst in the acylating kinetic resolution of two synthetically and industrially important chiral alcohols, rac-sulcatol and rac-trans-2-methoxycyclohexanol. The observation that the catalyst can be used 10 times in recycling experiments without losing its significant activity or enantioselectivity demonstrates the practical viability of the sol-gel approach.The entrapment of lipases in hydrophobic silicate matrices formed by sol-gel mediated hydrolysis of RSi(OCH3)3/Si(OCH3)4 as originally reported in 1996 has been improved over the years by a number of modifications. In the production of second-generation sol-gel lipase immobilizates, a variety of additives during the sol-gel process leads to increased activity and enhanced stereoselectivity in esterifying kinetic resolution. Recent advances in this type of lipase immobilization are reviewed here, in addition to new results regarding the sol-gel entrapment of the lipase from Burkholderia cepacia. It constitutes an excellent heterogeneous biocatalyst in the acylating kinetic resolution of two synthetically and industrially important chiral alcohols, rac-sulcatol and rac-trans-2-methoxycyclohexanol. The observation that the catalyst can be used 10 times in recycling experiments without losing its significant activity or enantioselectivity demonstrates the practical viability of the sol-gel approach. Dedicated to the pioneer of sol-gel enzyme immobilization, Professor David Avnir, on the occasion of his 65th birthday.

CASE STUDIES IN THE INTEGRATED USE OF SCALE ANALYSES TO SOLVE LEAD PROBLEMS

EPA Science Inventory

All methods of controlling lead corrosion involve immobilizing lead into relatively insoluble compounds that deposit on the interior wall of water pipes. Many different solid phases can form under the disparate conditions that exist in distribution systems, which range in how the...

Unveiling New Degradation Intermediates/Pathways from the Photocatalytic Degradation of Microcystin-LR

EPA Science Inventory

This study focuses on the identification of reaction intermediates formed during the photocatalytic degradation of the cyanotoxin microcystin-LR with immobilized TiO₂ Tphotocatalysts at neutral pH. To differentiate between impurities already existing in the MC-LR stand...

Porous solid ion exchange wafer for immobilizing biomolecules

DOEpatents

Arora, Michelle B.; Hestekin, Jamie A.; Lin, YuPo J.; St. Martin, Edward J.; Snyder, Seth W.

2007-12-11

A porous solid ion exchange wafer having a combination of a biomolecule capture-resin and an ion-exchange resin forming a charged capture resin within said wafer. Also disclosed is a porous solid ion exchange wafer having a combination of a biomolecule capture-resin and an ion-exchange resin forming a charged capture resin within said wafer containing a biomolecule with a tag. A separate bioreactor is also disclosed incorporating the wafer described above.

LKM1 autoantibodies in chronic hepatitis C infection: a case of molecular mimicry?

PubMed

Marceau, Gabriel; Lapierre, Pascal; Béland, Kathie; Soudeyns, Hugo; Alvarez, Fernando

2005-09-01

Anti-liver-kidney microsome type 1 (LKM1) autoantibodies directed against the cytochrome P450 2D6 (CYP2D6) are considered specific markers of type 2 autoimmune hepatitis, but are also found in 5% of sera from patients chronically infected by hepatitis C virus (HCV). Molecular mimicry between HCV proteins and CYP2D6 has been proposed to explain the emergence of these autoantibodies. Anti-LKM1 autoantibodies from hepatitis C-infected patients were affinity-purified against immobilized CYP2D6 protein and used to screen a phage display library. CYP2D6 conformational epitopes were identified using phage display analysis and the identification of statistically significant pairs (SSPs). Cross-reactivity between CYP2D6 and HCV protein candidates was tested by immunoprecipitation. Nineteen different clones were isolated, and their sequencing resulted in the mapping of a conformational epitope to the region of amino acids 254-288 of CYP2D6. Candidate HCV proteins for molecular mimicry included: core, E2, NS3 and NS5a. Affinity-purified autoantibodies from HCV+/LKM1+ patients immunoprecipitated either NS3, NS5a, or both, and these reactivities were specifically inhibited by immobilized CYP2D6. In conclusion, HCV+/LKM1+ sera recognize a specific conformational epitope on CYP2D6 between amino acids 254 to 288, the region that contains the major linear epitope in type 2 autoimmune hepatitis patients. Cross-reactivity due to molecular mimicry at the B-cell level was shown between the CYP2D6 and the HCV NS3 and NS5a proteins and could explain the presence of anti-LKM1 in patients chronically infected with HCV. Further investigation of the role played by this molecular mimicry in HCV-infected patients may lead to more specific strategies for diagnosis and treatment.

Immobilization of iron oxide nanoparticles within alginate nanogels for enhanced MR imaging applications.

PubMed

Sun, Wenjie; Yang, Jia; Zhu, Jianzhi; Zhou, Yiwei; Li, Jingchao; Zhu, Xiaoyue; Shen, Mingwu; Zhang, Guixiang; Shi, Xiangyang

2016-10-20

We report the design of iron oxide (Fe3O4) nanoparticle (NP)-immobilized alginate (AG) nanogels (NGs) as a novel contrast agent for enhanced magnetic resonance (MR) imaging applications. In this study, an aqueous solution of AG activated by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride was double emulsified to form NGs, followed by in situ cross-linking with polyethyleneimine (PEI)-coated Fe3O4 NPs (PEI-Fe3O4 NPs). The resultant Fe3O4 NP-immobilized AG NGs (AG/PEI-Fe3O4 NGs) were characterized via different techniques. Our results reveal that the hybrid NGs with a size of 186.1 ± 33.1 nm are water dispersible, colloidally stable, and cytocompatible in the given concentration range. Importantly, these NGs have a high r2 relaxivity (170.87 mM(-1) s(-1)) due to the high loading of Fe3O4 NPs within the NGs, and can be more significantly uptaken by cancer cells when compared with carboxylated Fe3O4 NPs. The formed AG/PEI-Fe3O4 NGs are able to be used as an effective contrast agent for the MR imaging of cancer cells in vitro and the xenografted tumor model in vivo after intravenous injection. The developed AG/PEI-Fe3O4 NGs may hold great promise for use as a novel contrast agent for the enhanced MR imaging of different biological systems.

Protein folding on Biosensor tips: Folding of Maltodextrin glucosidase monitored by its interactions with GroEL

PubMed Central

Pastor, Ashutosh; Singh, Amit K.; Fisher, Mark T.; Chaudhuri, Tapan K.

2016-01-01

Protein folding has been extensively studied for past four decades by employing solution based experiments such as solubility, enzymatic activity, secondary structure analysis, and analytical methods like FRET, NMR and HD exchange. However, for rapid analysis of the folding process, solution based approaches are often plagued with aggregation side reactions resulting in poor yields. In this work we demonstrate that a Bio-Layer Interferometry (BLI) chaperonin detection system can be potentially applied to identify superior refolding conditions for denatured proteins. The degree of immobilized protein folding as a function of time can be detected by monitoring the binding of the high-affinity nucleotide-free form of the chaperonin GroEL. GroEL preferentially interacts with proteins that have hydrophobic surfaces exposed in their unfolded or partially folded form so a decrease in GroEL binding can be correlated with burial of hydrophobic surfaces as folding progresses. The magnitude of GroEL binding to the protein immobilized on Bio-layer interferometry biosensor inversely reflects the extent of protein folding and hydrophobic residue burial. We demonstrate conditions where accelerated folding can be observed for the aggregation prone protein Maltodextrin glucosidase (MalZ). Superior immobilized folding conditions identified on the Bio-layer interferometry biosensor surface were reproduced on Ni-NTA sepharose bead surfaces and resulted in significant improvement in folding yields of released MalZ (measured by enzymatic activity) compared to bulk refolding conditions in solution. PMID:27367928

Spectroscopic evidence of uranium immobilization in acidic wetlands by natural organic matter and plant roots.

PubMed

Li, Dien; Kaplan, Daniel I; Chang, Hyun-Shik; Seaman, John C; Jaffé, Peter R; Koster van Groos, Paul; Scheckel, Kirk G; Segre, Carlo U; Chen, Ning; Jiang, De-Tong; Newville, Matthew; Lanzirotti, Antonio

2015-03-03

Biogeochemistry of uranium in wetlands plays important roles in U immobilization in storage ponds of U mining and processing facilities but has not been well understood. The objective of this work was to study molecular mechanisms responsible for high U retention by Savannah River Site (SRS) wetland sediments under varying redox and acidic (pH = 2.6-5.8) conditions using U L3-edge X-ray absorption spectroscopy. Uranium in the SRS wetland sediments existed primarily as U(VI) bonded as a bidentate to carboxylic sites (U-C bond distance at ∼2.88 Å), rather than phenolic or other sites of natural organic matter (NOM). In microcosms simulating the SRS wetland processes, U immobilization on roots was 2 orders of magnitude higher than on the adjacent brown or more distant white sands in which U was U(VI). Uranium on the roots were both U(IV) and U(VI), which were bonded as a bidentate to carbon, but the U(VI) may also form a U phosphate mineral. After 140 days of air exposure, all U(IV) was reoxidized to U(VI) but remained as a bidentate bonding to carbon. This study demonstrated NOM and plant roots can highly immobilize U(VI) in the SRS acidic sediments, which has significant implication for the long-term stewardship of U-contaminated wetlands.

Spectroscopic evidence of uranium immobilization in acidic wetlands by natural organic matter and plant roots

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Dien; Kaplan, Daniel I.; Chang, Hyun-Shik

Biogeochemistry of uranium in wetlands plays important roles in U immobilization in storage ponds of U mining and processing facilities but has not been well understood. The objective of this work was to study molecular mechanisms responsible for high U retention by Savannah River Site (SRS) wetland sediments under varying redox and acidic (pH = 2.6–5.8) conditions using U L₃-edge X-ray absorption spectroscopy. Uranium in the SRS wetland sediments existed primarily as U(VI) bonded as a bidentate to carboxylic sites (U–C bond distance at ~2.88 Å), rather than phenolic or other sites of natural organic matter (NOM). In microcosms simulatingmore » the SRS wetland processes, U immobilization on roots was two orders of magnitude higher than on the adjacent brown or more distant white sands in which U was U(VI). Uranium on the roots were both U(IV) and U(VI), which were bonded as a bidentate to carbon, but the U(VI) may also form a U phosphate mineral. After 140 days of air exposure, all U(IV) was re-oxidized to U(VI) but remained as a bidentate bonding to carbon. This study demonstrated NOM and plant roots can highly immobilize U(VI) in the SRS acidic sediments, which has significant implication for the long-term stewardship of U-contaminated wetlands.« less

Spectroscopic evidence of uranium immobilization in acidic wetlands by natural organic matter and plant roots

DOE PAGES

Li, Dien; Kaplan, Daniel I.; Chang, Hyun-Shik; ...

2015-03-03

Biogeochemistry of uranium in wetlands plays important roles in U immobilization in storage ponds of U mining and processing facilities but has not been well understood. The objective of this work was to study molecular mechanisms responsible for high U retention by Savannah River Site (SRS) wetland sediments under varying redox and acidic (pH = 2.6–5.8) conditions using U L₃-edge X-ray absorption spectroscopy. Uranium in the SRS wetland sediments existed primarily as U(VI) bonded as a bidentate to carboxylic sites (U–C bond distance at ~2.88 Å), rather than phenolic or other sites of natural organic matter (NOM). In microcosms simulatingmore » the SRS wetland processes, U immobilization on roots was two orders of magnitude higher than on the adjacent brown or more distant white sands in which U was U(VI). Uranium on the roots were both U(IV) and U(VI), which were bonded as a bidentate to carbon, but the U(VI) may also form a U phosphate mineral. After 140 days of air exposure, all U(IV) was re-oxidized to U(VI) but remained as a bidentate bonding to carbon. This study demonstrated NOM and plant roots can highly immobilize U(VI) in the SRS acidic sediments, which has significant implication for the long-term stewardship of U-contaminated wetlands.« less

A piezoelectric immunosensor for the detection of alpha-fetoprotein using an interface of gold/hydroxyapatite hybrid nanomaterial.

PubMed

Ding, Yanjun; Liu, Jia; Wang, Hua; Shen, Guoli; Yu, Ruqin

2007-04-01

The ideal immobilization methods that are suitable for binding immuno-active materials with high efficiency onto the sensing surface are the key target to pursue in the current biosensor design. In this paper, a new hybrid material formed by assembling gold nanoparticles (GNP) onto nano-sized hydroxyapatite (HA) has been employed for the interface design of piezoelectric immunosensor, on which the antibodies were bound. The detection performances of the resulting immunosensor were investigated by use of the antibody-antigen model system of alpha-Fetoprotein (AFP), an important indicator in the diagnosis of clinical cancers. The hybrid material was characterized by the UV-vis spectroscopy, the SEM and TEM measurements. The frequency and electrochemical impedance responses characteristics for the processes of immobilization and immunoreaction of anchored anti-AFP antibodies were studied in detail. The immunoresponse of the proposed immunosensor was compared with those antibodies immobilized by using HA or GNP alone. It was found that the developed sensing interface has some advantages such as the activation-free immobilization and the high antigen-binding activities of antibodies. The as-prepared immunosensor can allow for the determination of AFP in the concentration range of 15.3-600.0ngml(-1). Such an interface design with the nano-sized hybrid materials should be tailored as a new alternative used for biosensor design.

Magnetic multicore nanoparticles for hyperthermia--influence of particle immobilization in tumour tissue on magnetic properties.

PubMed

Dutz, Silvio; Kettering, Melanie; Hilger, Ingrid; Müller, Robert; Zeisberger, Matthias

2011-07-01

When using magnetic nanoparticles as a heating source for magnetic particle hyperthermia it is of particular interest to know if the particles are free to move in the interstitial fluid or are fixed to the tumour tissue. The immobilization state determines the relaxation behaviour of the administered particles and thus their specific heating power. To investigate this behaviour, magnetic multicore nanoparticles were injected into experimentally grown tumours in mice and magnetic heating treatment was carried out in an alternating magnetic field (H = 25 kA m(-1), f = 400 kHz). The tested particles were well suited for magnetic heating treatment as they heated a tumour of about 100 mg by about 22 K within the first 60 s. Upon sacrifice, histological tumour examination showed that the particles form spots in the tissue with a mainly homogeneous particle distribution in these spots. The magnetic ex vivo characterization of the removed tumour tissue gave clear evidence for the immobilization of the particles in the tumour tissue because the particles in the tumour showed the same magnetic behaviour as immobilized particles. Therefore, the particles are not able to rotate and a temperature increase due to Brown relaxation can be neglected. To accurately estimate the heating potential of magnetic materials, the respective environments influencing the nanoparticle mobility status have to be taken into account.

Laccase immobilization and insolubilization: from fundamentals to applications for the elimination of emerging contaminants in wastewater treatment.

PubMed

Ba, Sidy; Arsenault, Alexandre; Hassani, Thanina; Jones, J Peter; Cabana, Hubert

2013-12-01

Over the last few decades many attempts have been made to use biocatalysts for the biotransformation of emerging contaminants in environmental matrices. Laccase, a multicopper oxidoreductase enzyme, has shown great potential in oxidizing a large number of phenolic and non-phenolic emerging contaminants. However, laccases and more broadly enzymes in their free form are biocatalysts whose applications in solution have many drawbacks rendering them currently unsuitable for large scale use. To circumvent these limitations, the enzyme can be immobilized onto carriers or entrapped within capsules; these two immobilization techniques have the disadvantage of generating a large mass of non-catalytic product. Insolubilization of the free enzymes as cross-linked enzymes (CLEAs) is found to yield a greater volume ratio of biocatalyst while improving the characteristics of the biocatalyst. Ultimately, novel techniques of enzymes insolubilization and stabilization are feasible with the combination of cross-linked enzyme aggregates (combi-CLEAs) and enzyme polymer engineered structures (EPESs) for the elimination of emerging micropollutants in wastewater. In this review, fundamental features of laccases are provided in order to elucidate their catalytic mechanism, followed by different chemical aspects of the immobilization and insolubilization techniques applicable to laccases. Finally, kinetic and reactor design effects for enzymes in relation with the potential applications of laccases as combi-CLEAs and EPESs for the biotransformation of micropollutants in wastewater treatment are discussed.

Net-Immobilization of β-glucosidase on Nonwoven Fabrics to Lower the Cost of “Cellulosic Ethanol” and Increase Cellulose Conversions

PubMed Central

Zhu, Xing; He, Bin; Zhao, Changwen; Fan, Rong; Zhang, Lihua; Wang, Guan; Ma, Yuhong; Yang, Wantai

2016-01-01

The main limitation preventing the use of enzymatic cellulosic ethanol in industrial production is its higher cost which is mainly due to the elevated price of β-glucosidase (BG). Herein, we report on a simple strategy for the in-situ encapsulation of BG for repeated cellulosic ethanol production. In this strategy, BG was net-immobilized into a poly(ethylene glycol) (PEG) net-cloth layer on a PP nonwoven fabric by way of the visible light-induced surface controlled/living graft cross-linking polymerization. The visible light and mild reaction conditions could ensure the activity retention of BG during immobilization, while the non-swelling uniform net-mesh formed by living cross-linking polymerization could prevent the leakage of BG effectively (at the immobilization rate of more than 98.6% and the leakage rate of only 0.4%). When the BG-loaded fabric was used in combination with free cellulase (CEL), the results of the catalytic reaction demonstrated that these BG-loaded fabrics could not only give a 40% increase in cellulose conversions but also be reused for more than fifteen batches without losing the activity. These BG-loaded fabrics with characteristics including easy separation, excellent operation stability, a low cost of the polymeric matrix and a simple fabrication process are particularly interesting for a future bio-fuel production strategy. PMID:27009788

The development of flow-through bio-catalyst microreactors from silica micro structured fibers for lipid transformations.

PubMed

Anuar, Sabiqah Tuan; Villegas, Carla; Mugo, Samuel M; Curtis, Jonathan M

2011-06-01

This study demonstrates the utility of a flow-through enzyme immobilized silica microreactor for lipid transformations. A silica micro structured fiber (MSF) consisting of 168 channels of internal diameter 4-5 μm provided a large surface area for the covalent immobilization of Candida antartica lipase. The specific activity of the immobilized lipase was determined by hydrolysis of p-nitrophenyl butyrate and calculated to be 0.81 U/mg. The catalytic performance of the lipase microreactor was demonstrated by the efficient ethanolysis of canola oil. The parameters affecting the performance of the MSF microreactor, including temperature and reaction flow rate, were investigated. Characterization of the lipid products exiting the microreactor was performed by non-aqueous reversed-phased liquid chromatography (NARP-LC) with evaporative light scattering detector (ELSD) and by comprehensive two-dimensional gas chromatography (GC x GC). Under optimized conditions of 1 μL/min flow rate of 5 mg/mL trioleoylglycerol (TO) in ethanol and 50 °C reaction temperature, 2-monooleoylglycerol was the main product at > 90% reaction yield. The regioselectivity of the Candida antartica lipase immobilized MSF microreactor in the presence of ethanol was found to be comparable to that obtained under conventional conditions. The ability of these reusable flow-through microreactors to regioselectively form monoacylglycerides in high yield from triacylglycerides demonstrate their potential use in small-scale lipid transformations or analytical lipids profiling.

Pentachlorophenol (PCP) dechlorination in horizontal-flow anaerobic immobilized biomass (HAIB) reactors.

PubMed

Damianovic, M H R Z; Moraes, E M; Zaiat, M; Foresti, E

2009-10-01

This study verifies the potential applicability of horizontal-flow anaerobic immobilized biomass (HAIB) reactors to pentachlorophenol (PCP) dechlorination. Two bench-scale HAIB reactors (R1 and R2) were filled with cubic polyurethane foam matrices containing immobilized anaerobic sludge. The reactors were then continuously fed with synthetic wastewater consisting of PCP, glucose, acetic acid, and formic acid as co-substrates for PCP anaerobic degradation. Before being immobilized in polyurethane foam matrices, the biomass was exposed to wastewater containing PCP in reactors fed at a semi-continuous rate of 2.0 microg PCP g(-1) VS. The applied PCP loading rate was increased from 0.05 to 2.59 mg PCP l(-1)day(-1) for R1, and from 0.06 to 4.15 mg PCP l(-1)day(-1) for R2. The organic loading rates (OLR) were 1.1 and 1.7 kg COD m(-3)day(-1) at hydraulic retention times (HRT) of 24h for R1 and 18 h for R2. Under such conditions, chemical oxygen demand (COD) removal efficiencies of up to 98% were achieved in the HAIB reactors. Both reactors exhibited the ability to remove 97% of the loaded PCP. Dichlorophenol (DCP) was the primary chlorophenol detected in the effluent. The adsorption of PCP and metabolites formed during PCP degradation in the packed bed was negligible for PCP removal efficiency.

Synthesis of an oligonucleotide-derivatized amphipol and its use to trap and immobilize membrane proteins

PubMed Central

Bon, Christel Le; Della Pia, Eduardo Antonio; Giusti, Fabrice; Lloret, Noémie; Zoonens, Manuela; Martinez, Karen L.; Popot, Jean-Luc

2014-01-01

Amphipols (APols) are specially designed amphipathic polymers that stabilize membrane proteins (MPs) in aqueous solutions in the absence of detergent. A8–35, a polyacrylate-based APol, has been grafted with an oligodeoxynucleotide (ODN). The synthesis, purification and properties of the resulting ‘OligAPol’ have been investigated. Grafting was performed by reacting an ODN carrying an amine-terminated arm with the carboxylates of A8–35. The use of OligAPol for trapping MPs and immobilizing them onto solid supports was tested using bacteriorhodopsin (BR) and the transmembrane domain of Escherichia coli outer membrane protein A (tOmpA) as model proteins. BR and OligAPol form water-soluble complexes in which BR remains in its native conformation. Hybridization of the ODN arm with a complementary ODN was not hindered by the assembly of OligAPol into particles, nor by its association with BR. BR/OligAPol and tOmpA/OligAPol complexes could be immobilized onto either magnetic beads or gold nanoparticles grafted with the complementary ODN, as shown by spectroscopic measurements, fluorescence microscopy and the binding of anti-BR and anti-tOmpA antibodies. OligAPols provide a novel, highly versatile approach to tagging MPs, without modifying them chemically nor genetically, for specific, reversible and targetable immobilization, e.g. for nanoscale applications. PMID:24744236

Immobilization methods for the rapid total chemical synthesis of proteins on microtiter plates.

PubMed

Zitterbart, Robert; Krumrey, Michael; Seitz, Oliver

2017-07-01

The chemical synthesis of proteins typically involves the solid-phase peptide synthesis of unprotected peptide fragments that are stitched together in solution by native chemical ligation (NCL). The process is slow, and throughput is limited because of the need for repeated high performance liquid chromatography purification steps after both solid-phase peptide synthesis and NCL. With an aim to provide faster access to functional proteins and to accelerate the functional analysis of synthetic proteins by parallelization, we developed a method for the high performance liquid chromatography-free synthesis of proteins on the surface of microtiter plates. The method relies on solid-phase synthesis of unprotected peptide fragments, immobilization of the C-terminal fragment and on-surface NCL with an unprotected peptide thioester in crude form. Herein, we describe the development of a suitable immobilization chemistry. We compared (i) formation of nickel(II)-oligohistidine complexes, (ii) Cu-based [2 + 3] alkine-azide cycloaddition and (iii) hydrazone ligation. The comparative study identified the hydrazone ligation as most suitable. The sequence of immobilization via hydrazone ligation, on-surface NCL and radical desulfurization furnished the targeted SH3 domains in near quantitative yield. The synthetic proteins were functional as demonstrated by an on-surface fluorescence-based saturation binding analysis. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

Net-Immobilization of β-glucosidase on Nonwoven Fabrics to Lower the Cost of “Cellulosic Ethanol” and Increase Cellulose Conversions

NASA Astrophysics Data System (ADS)

Zhu, Xing; He, Bin; Zhao, Changwen; Fan, Rong; Zhang, Lihua; Wang, Guan; Ma, Yuhong; Yang, Wantai

2016-03-01

The main limitation preventing the use of enzymatic cellulosic ethanol in industrial production is its higher cost which is mainly due to the elevated price of β-glucosidase (BG). Herein, we report on a simple strategy for the in-situ encapsulation of BG for repeated cellulosic ethanol production. In this strategy, BG was net-immobilized into a poly(ethylene glycol) (PEG) net-cloth layer on a PP nonwoven fabric by way of the visible light-induced surface controlled/living graft cross-linking polymerization. The visible light and mild reaction conditions could ensure the activity retention of BG during immobilization, while the non-swelling uniform net-mesh formed by living cross-linking polymerization could prevent the leakage of BG effectively (at the immobilization rate of more than 98.6% and the leakage rate of only 0.4%). When the BG-loaded fabric was used in combination with free cellulase (CEL), the results of the catalytic reaction demonstrated that these BG-loaded fabrics could not only give a 40% increase in cellulose conversions but also be reused for more than fifteen batches without losing the activity. These BG-loaded fabrics with characteristics including easy separation, excellent operation stability, a low cost of the polymeric matrix and a simple fabrication process are particularly interesting for a future bio-fuel production strategy.

Effectiveness of Antimicrobial Peptide Immobilization for Preventing Perioperative Cornea Implant-Associated Bacterial Infection

PubMed Central

Tan, Xiao Wei; Goh, Tze Wei; Saraswathi, P.; Nyein, Chan Lwin; Setiawan, Melina; Riau, Andri; Lakshminarayanan, R.; Liu, Shouping; Tan, Donald; Beuerman, Roger W.

2014-01-01

Titanium (Ti) is a promising candidate biomaterial for an artificial corneal skirt. Antimicrobial peptide (AMP) immobilization may improve the bactericidal effect of the Ti substrate. In this study, we tested the bactericidal efficacy of a functionalized Ti surface in a rabbit keratitis model. A corneal stromal pocket was created by a femtosecond laser. The Ti films were then inserted into the pocket, and Staphylococcus aureus or Pseudomonas aeruginosa was inoculated into the pocket above the implant films. The corneas with Ti-AMP implants were compared with the corneas implanted with unprotected Ti by slit lamp observation and anterior segment optical coherence tomography (AS-OCT). Inflammatory responses were evaluated by bacterium counting, hematoxylin-eosin staining, and immunostaining. There was a lower incidence and a lesser extent of infection on rabbit corneas with Ti-AMP implants than on those with unprotected Ti implants. The bactericidal effect of AMP against S. aureus was comparable to that of postoperative prophylactic antibiotic treatment; hence, SESB2V AMP bound to the Ti implant provided functional activity in vivo, but its efficacy was greater against S. aureus than against P. aeruginosa. This work suggests that SESB2V AMP can be successfully functionalized in a rabbit keratitis model to prevent perioperative corneal infection. PMID:24957820

Controllable fabrication and characterization of biocompatible core-shell particles and hollow capsules as drug carrier

NASA Astrophysics Data System (ADS)

Hao, Lingyun; Gong, Xinglong; Xuan, Shouhu; Zhang, Hong; Gong, Xiuqing; Jiang, Wanquan; Chen, Zuyao

2006-10-01

SiO 2@CdSe core-shell particles were fabricated by controllable deposition CdSe nanoparticles on silica colloidal spheres. Step-wise coating process was tracked by the TEM and XRD measurements. In addition, SiO 2@CdSe/polypyrrole(PPy) multi-composite particles were synthesized based on the as-prepared SiO 2@CdSe particles by cationic polymerization. The direct electrochemistry of myoglobin (Mb) could be performed by immobilizing Mb on the surface of SiO 2@CdSe particles. Immobilized with Mb, SiO 2@CdSe/PPy-Mb also displayed good bioelectrochemical activity. It confirmed the good biocompatible property of the materials with protein. CdSe hollow capsules were further obtained as the removal of the cores of SiO 2@CdSe spheres. Hollow and porous character of CdSe sub-meter size capsules made them becoming hopeful candidates as drug carriers. Doxorubicin, a typical an antineoplastic drug, was introduced into the capsules. A good sustained drug release behavior of the loading capsules was discovered via performing a release test in the PBS buffer (pH 7.4) solution at 310 k. Furthermore, SiO 2@CdSe/PPy could be converted to various smart hollow capsules via selectively removal of their relevant components.

Albumin (BSA) adsorption onto graphite stepped surfaces

NASA Astrophysics Data System (ADS)

Rubio-Pereda, Pamela; Vilhena, J. G.; Takeuchi, Noboru; Serena, Pedro A.; Pérez, Rubén

2017-06-01

Nanomaterials are good candidates for the design of novel components with biomedical applications. For example, nano-patterned substrates may be used to immobilize protein molecules in order to integrate them in biosensing units. Here, we perform long MD simulations (up to 200 ns) using an explicit solvent and physiological ion concentrations to characterize the adsorption of bovine serum albumin (BSA) onto a nano-patterned graphite substrate. We have studied the effect of the orientation and step size on the protein adsorption and final conformation. Our results show that the protein is stable, with small changes in the protein secondary structure that are confined to the contact area and reveal the influence of nano-structuring on the spontaneous adsorption, protein-surface binding energies, and protein mobility. Although van der Waals (vdW) interactions play a dominant role, our simulations reveal the important role played by the hydrophobic lipid-binding sites of the BSA molecule in the adsorption process. The complex structure of these sites, that incorporate residues with different hydrophobic character, and their flexibility are crucial to understand the influence of the ion concentration and protein orientation in the different steps of the adsorption process. Our study provides useful information for the molecular engineering of components that require the immobilization of biomolecules and the preservation of their biological activity.

Sensitive immobilization-free electrochemical DNA sensor based on isothermal circular strand displacement polymerization reaction.

PubMed

Xuan, Feng; Luo, Xiaoteng; Hsing, I-Ming

2012-05-15

A highly sensitive electrochemical DNA sensor that requires no probe immobilization has been developed based on a target recycling mechanism utilizing a DNA polymerase with a strand displacement activity. The electrochemical detection is realized by taking advantage of the difference in diffusivity between a free ferrocene-labeled peptide nucleic acid (Fc-PNA) and a Fc-PNA hybridized with a complementary DNA, while the DNA polymerase-assisted target recycling leads to signal generation and amplification. The hybridization of the target DNA opens up a stem-loop template DNA with the Fc-PNA hybridized to its extruded 5' end and allows a DNA primer to anneal and be extended by the DNA polymerase, which results in sequential displacement of the target DNA and the Fc-PNA from the template DNA. The displaced target DNA will hybridize with another template DNA, triggering another round of primer extension and strand displacement. The released Fc-PNA, due to its neutral backbone, has much higher diffusivity towards a negatively charged electrode, compared to that when it is hybridized with a negatively charged DNA. Therefore, a significantly enhanced signal of Fc can be observed. The outstanding sensitivity and simplicity make this approach a promising candidate for next-generation electrochemical DNA sensing technologies. Copyright © 2012 Elsevier B.V. All rights reserved.

Method of preparing sodalite from chloride salt occluded zeolite

DOEpatents

Lewis, Michele A.; Pereira, Candido

1997-01-01

A method for immobilizing waste chloride salts containing radionuclides and hazardous nuclear material for permanent disposal starting with a substantially dry zeolite and sufficient glass to form leach resistant sodalite with occluded radionuclides and hazardous nuclear material. The zeolite and glass are heated to a temperature up to about 1000.degree. K. to convert the zeolite to sodalite and thereafter maintained at a pressure and temperature sufficient to form a sodalite product near theoretical density. Pressure is used on the formed sodalite to produce the required density.

Method of preparing sodalite from chloride salt occluded zeolite

DOEpatents

Lewis, M.A.; Pereira, C.

1997-03-18

A method is described for immobilizing waste chloride salts containing radionuclides and hazardous nuclear material for permanent disposal starting with a substantially dry zeolite and sufficient glass to form leach resistant sodalite with occluded radionuclides and hazardous nuclear material. The zeolite and glass are heated to a temperature up to about 1000 K to convert the zeolite to sodalite and thereafter maintained at a pressure and temperature sufficient to form a sodalite product near theoretical density. Pressure is used on the formed sodalite to produce the required density.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cozzi, Alex D.; McCabe, Daniel J.

The Hanford Waste Treatment and Immobilization Plant (WTP) Low Activity Waste (LAW) vitrification facility will generate an aqueous condensate recycle stream (LAW Melter Off-Gas Condensate) from the off-gas system. The baseline plan for disposition of this stream during full WTP operations is to send it to the WTP Pretreatment Facility, where it will be blended with LAW, concentrated by evaporation and recycled to the LAW vitrification facility. However, during the Direct Feed LAW (DFLAW) scenario, planned disposition of this stream is to evaporate it in a new evaporator in the Effluent Management Facility (EMF) and then return it to themore » LAW melter. It is important to understand the composition of the effluents from the melter and new evaporator so that the disposition of these streams can be accurately planned and accommodated. Furthermore, alternate disposition of this stream would eliminate recycling of problematic components, and would enable less integrated operation of the LAW melter and the Pretreatment Facilities. Alternate disposition would also eliminate this stream from recycling within WTP when it begins operations and would decrease the LAW vitrification mission duration and quantity of glass waste. This LAW Melter Off-Gas Condensate stream will contain components that are volatile at melter temperatures and are problematic for the glass waste form, such as halides and sulfate, along with entrained, volatile, and semi-volatile metals, such as Hg, As, and Se. Because this stream will recycle within WTP, these components accumulate in the Melter Condensate stream, exacerbating their impact on the number of LAW glass containers that must be produced. Diverting the stream reduces the halides and sulfate that get recycled to the melter, and is a key objective of this work. This overall program examines the potential treatment and immobilization of this stream to enable alternative disposal. The objective of earlier tasks was to formulate and prepare a simulant of the LAW Melter Off-gas Condensate expected during DFLAW operations and use it in evaporator testing to predict the composition of the effluents from the Effluent Management Facility (EMF) evaporator to aid in planning for their disposition. The objective of this task was to test immobilization options for this evaporator bottoms aqueous stream. This document describes the method used to formulate a simulant of this EMF evaporator bottoms stream, immobilize it, and determine if the immobilized waste forms meet disposal criteria.« less

Enzyme immobilization in novel alginate-chitosan core-shell microcapsules.

PubMed

Taqieddin, Ehab; Amiji, Mansoor

2004-05-01

Alginate-chitosan core-shell microcapsules were prepared in order to develop a biocompatible matrix for enzyme immobilization, where the protein is retained either in a liquid or solid core and the shell allows permeability control over substrates and products. The permeability coefficients of different molecular weight compounds (vitamin B2, vitamin B12, and myoglobin) were determined through sodium tripolyphosphate (Na-TPP)-crosslinked chitosan membrane. The microcapsule core was formed by crosslinking sodium alginate with either calcium or barium ions. The crosslinked alginate core was uniformly coated with a chitosan layer and crosslinked with Na-TPP. In the case of calcium alginate, the phosphate ions of Na-TPP were able to extract the calcium ions from alginate and liquefy the core. A model enzyme, beta-galactosidase, was immobilized in the alginate core and the catalytic activity was measured with o-nitrophenyl-beta-D-galactopyranoside (ONPG). Change in the activity of free and immobilized enzyme was determined at three different temperatures. Na-TPP crosslinked chitosan membranes were found to be permeable to solutes of up to 17,000Da molecular weight. The enzyme loading efficiency was higher in the barium alginate core (100%) as compared to the calcium alginate core (60%). The rate of ONPG conversion to o-nitrophenol was faster in the case of calcium alginate-chitosan microcapsules as compared to barium alginate-chitosan microcapsules. Barium alginate-chitosan microcapsules, however, did improve the stability of the enzyme at 37 degrees C relative to calcium alginate-chitosan microcapsules or free enzyme. This study illustrates a new method of enzyme immobilization for biotechnology applications using liquid or solid core and shell microcapsule technology.

Charge immobilization of the voltage sensor in domain IV is independent of sodium current inactivation.

PubMed

Sheets, Michael F; Hanck, Dorothy A

2005-02-15

Recovery from fast inactivation in voltage-dependent Na+ channels is associated with a slow component in the time course of gating charge during repolarization (i.e. charge immobilization), which results from the slow movement of the S4 segments in domains III and IV (S4-DIII and S4-DIV). Previous studies have shown that the non-specific removal of fast inactivation by the proteolytic enzyme pronase eliminated charge immobilization, while the specific removal of fast inactivation (by intracellular MTSET modification of a cysteine substituted for the phenylalanine in the IFM motif, ICMMTSET, in the inactivation particle formed by the linker between domains III and IV) only reduced the amount of charge immobilization by nearly one-half. To investigate the molecular origin of the remaining slow component of charge immobilization we studied the human cardiac Na+ channel (hH1a) in which the outermost arginine in the S4-DIV, which contributes approximately 20% to total gating charge (Qmax), was mutated to a cysteine (R1C-DIV). Gating charge could be fully restored in R1C-DIV by exposure to extracellular MTSEA, a positively charged methanethiosulphonate reagent. The RIC-DIV mutation was combined with ICMMTSET to remove fast inactivation, and the gating currents of R1C-DIV-ICM(MTSET) were recorded before and after modification with MTSEAo. Prior to MTSEAo, the time course of the gating charge during repolarization (off-charge) was best described by a single fast time constant. After MTSEA, the off-charge had both fast and slow components, with the slow component accounting for nearly 35% of Qmax. These results demonstrate that the slow movement of the S4-DIV during repolarization is not dependent upon the normal binding of the inactivation particle.

A novel immobilization multienzyme glucose fluorescence capillary biosensor.

PubMed

Li, Yong-Sheng; Du, Yun-Dong; Chen, Ting-Mei; Gao, Xiu-Feng

2010-02-15

Based on the immobilization enzyme technology and the fluorescence capillary analysis method, the authors have developed a highly sensitive fluorescence reaction system and a novel immobilization multienzyme glucose fluorescence capillary biosensor for determining glucose contents. Reaction principle of the system is that under the catalysis of glucose oxidase (GOD) and horseradish peroxidase (HRP) immobilized on inner surface of a medical capillary, beta-D-glucose reacts with dissolved oxygen to form gluconic acid-delta-lactone and hydrogen peroxide, and then the latter reacts with l-tyrosine to produce a tyrosine dimer, which has maximal excitation and emission wavelengths at 320 nm and 410 nm, respectively. Fluorescence of the dimer is proportional to the concentration of the beta-D-glucose. Optimization conditions suitable for the reaction system and the biosensor were as follows. Concentration of the L-tyrosine used as fluorescence reagent was 0.15 mol L(-1), the active concentrations of the GOD and the HRP for the immobilization were 15 kU L(-1) and 8 kU L(-1), respectively. Consumptions of the sample and reagents in one determination were 5.0 microL and 15 microL, respectively. Quantitative range of the biosensor for the glucose was in the range 1-10 micromol L(-1), its relative standard deviation was less than 4.9%, and its detection limit was 0.62 micromol L(-1). The biosensor's recovery was in the range 96-105%. Results of some serum determined with the biosensor and with a commercial glucose-kit were well coincident to each other. Accordingly, the biosensor can be applied to the determination of serum glucose contents in the diagnosis of diabetes. Copyright 2009 Elsevier B.V. All rights reserved.

Production of galacto-oligosaccharides from lactose by Aspergillus oryzae beta-galactosidase immobilized on cotton cloth.

PubMed

Albayrak, Nedim; Yang, Shang-Tian

2002-01-05

The production of galacto-oligosaccharides (GOS) from lactose by A. oryzae beta-galactosidase immobilized on cotton cloth was studied. The total amounts and types of GOS produced were mainly affected by the initial lactose concentration in the reaction media. In general, more and larger GOS can be produced with higher initial lactose concentrations. A maximum GOS production of 27% (w/w) of initial lactose was achieved at 50% lactose conversion with 500 g/L of initial lactose concentration. Tri-saccharides were the major types of GOS formed, accounting for more than 70% of the total GOS produced in the reactions. Temperature and pH affected the reaction rate, but did not result in any changes in GOS formation. The presence of galactose and glucose at the concentrations encountered near maximum GOS greatly inhibited the reactions and reduced GOS yield by as much as 15%. The cotton cloth as the support matrix for enzyme immobilization did not affect the GOS formation characteristics of the enzyme, suggesting no diffusion limitation in the enzyme carrier. The thermal stability of the enzyme increased approximately 25-fold upon immobilization on cotton cloth. The half-life for the immobilized enzyme on cotton cloth was more than 1 year at 40 degrees C and 48 days at 50 degrees C. Stable, continuous operation in a plugflow reactor was demonstrated for 2 weeks without any apparent problem. A maximum GOS production of 21 and 26% (w/w) of total sugars was attained with a feed solution containing 200 and 400 g/L of lactose, respectively, at pH 4.5 and 40 degrees C. The corresponding reactor productivities were 80 and 106 g/L/h, respectively, which are at least several-fold higher than those previously reported. Copyright 2002 John Wiley & Sons, Inc.

Fate of Uranium During Transport Across the Groundwater-Surface Water Interface

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jaffe, Peter R.; Kaplan, Daniel I.

Discharge of contaminated groundwater to surface waters is of concern at many DOE facilities. For example, at F-Area and TNX-Area on the Savannah River Site, contaminated groundwater, including uranium, is already discharging into natural wetlands. It is at this interface where contaminants come into contact with the biosphere. These this research addressed a critical knowledge gap focusing on the geochemistry of uranium (or for that matter, any redox-active contaminant) in wetland systems. Understanding the interactions between hydrological, microbial, and chemical processes will make it possible to provide a more accurate conceptual and quantitative understanding of radionuclide fate and transport undermore » these unique conditions. Understanding these processes will permit better long-term management and the necessary technical justification for invoking Monitored Natural Attenuation of contaminated wetland areas. Specifically, this research did provide new insights on how plant-induced alterations to the sediment biogeochemical processes affect the key uranium reducing microorganisms, the uranium reduction, its spatial distribution, the speciation of the immobilized uranium, and its long-term stability. This was achieved by conducting laboratory mesocosm wetland experiments as well as field measurements at the SRNL. Results have shown that uranium can be immobilized in wetland systems. To a degree some of the soluble U(VI) was reduced to insoluble U(IV), but the majority of the immobilized U was incorporated into iron oxyhydroxides that precipitated onto the root surfaces of wetland plants. This U was immobilized mostly as U(VI). Because it was immobilized in its oxidized form, results showed that dry spells, resulting in the lowering of the water table and the exposure of the U to oxic conditions, did not result in U remobilization.« less

SECONDARY WASTE MANAGEMENT FOR HANFORD EARLY LOW ACTIVITY WASTE VITRIFICATION

DOE Office of Scientific and Technical Information (OSTI.GOV)

UNTERREINER BJ

2008-07-18

More than 200 million liters (53 million gallons) of highly radioactive and hazardous waste is stored at the U.S. Department of Energy's Hanford Site in southeastern Washington State. The DOE's Hanford Site River Protection Project (RPP) mission includes tank waste retrieval, waste treatment, waste disposal, and tank farms closure activities. This mission will largely be accomplished by the construction and operation of three large treatment facilities at the Waste Treatment and Immobilization Plant (WTP): (1) a Pretreatment (PT) facility intended to separate the tank waste into High Level Waste (HLW) and Low Activity Waste (LAW); (2) a HLW vitrification facilitymore » intended to immobilize the HLW for disposal at a geologic repository in Yucca Mountain; and (3) a LAW vitrification facility intended to immobilize the LAW for shallow land burial at Hanford's Integrated Disposal Facility (IDF). The LAW facility is on target to be completed in 2014, five years prior to the completion of the rest of the WTP. In order to gain experience in the operation of the LAW vitrification facility, accelerate retrieval from single-shell tank (SST) farms, and hasten the completion of the LAW immobilization, it has been proposed to begin treatment of the low-activity waste five years before the conclusion of the WTP's construction. A challenge with this strategy is that the stream containing the LAW vitrification facility off-gas treatment condensates will not have the option of recycling back to pretreatment, and will instead be treated by the Hanford Effluent Treatment Facility (ETF). Here the off-gas condensates will be immobilized into a secondary waste form; ETF solid waste.« less

Immobilization of uranium in contaminated soil by natural apatite addition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mrdakovic Popic, Jelena; Stojanovic, Mirjana; Milosevic, Sinisa

2007-07-01

Available in abstract form only. Full text of publication follows: The goal of this study was to evaluate the effectiveness of Serbian natural mineral apatite as soil additive for reducing the migration of uranium from contaminated sediments. In laboratory study we investigated the sorption properties of domestic apatite upon different experimental conditions, such as pH, adsorbent mass, reaction period, concentration of P{sub 2}O{sub 5} in apatite, solid/liquid ratio. In second part of study, we did the quantification of uranium in soil samples, taken from uranium mine site 'Kalna', by sequential extraction method. The same procedure was, also, used for uraniummore » determination in contaminated soil samples after apatite addition, in order to determine the changes in U distribution in soil fraction. The obtained results showed the significant level of immobilization (96.7%) upon certain conditions. Increase of %P{sub 2}O{sub 5} in apatite and process of mechano-chemical activation led to increase of immobilization capacity from 17.50% till 91.64%. The best results for uranium binding were obtained at pH 5.5 and reaction period 60 days (98.04%) The sequential extraction showed the presence of uranium (48.2%) in potentially available soil fractions, but with the apatite addition uranium content in these fractions decreased (30.64%), what is considering environmental aspect significant fact. In situ immobilization of radionuclide using inexpensive sequestering agents, such as apatite, is very adequate for big contaminated areas of soil with low level of contamination. This investigation study on natural apatite from deposit 'Lisina' Serbia was the first one of this type in our country. Key words: apatite, uranium, immobilization, soil, contamination. (authors)« less

Chitosan-immobilized pectinolytics with novel catalytic features and fruit juice clarification potentialities.

PubMed

Irshad, Muhammad; Murtza, Aimen; Zafar, Muddassar; Bhatti, Khizar Hayat; Rehman, Abdul; Anwar, Zahid

2017-11-01

Biological macromolecules are primarily composed of complex polysaccharides that strengthen microbial growth for the production of industrially relevant enzymes. The presence of polysaccharides in the form of the disrupted cell wall and cell materials are among major challenges in the fruit juice industry. The breakdown of such biological macromolecules including cellulose and pectin is vital for the juices processing. In this background, pectinolytic enzymes including polygalacturonase (PG), pectin lyase (PL), and pectin methylesterase (PME) were isolated from Aspergillus ornatus, statistically optimized and purified via ammonium sulfate fractionation (ASF), dialysis, and Sephadex G-100 gel permeation chromatography. After passing through Sephadex G-100 column, PG, PL, and PME were 2.60-fold, 3.30-fold, and 4.52-fold purified with specific activities of 475.2U/mg, 557.1U/mg, and 205.7U/mg. The active PG, PL, and PME, each separately, were surface immobilized using various concentrations of chitosan and dextran polyaldehyde as a macromolecular crosslinking agent. Prior to exploit for juice clarification purposes, various parameters including pH, thermal and Michaelis-Menten kinetic constants of purified and chitosan-immobilized fractions were investigated. A considerable improvement in the pH and thermal profiles was recorded after immobilization. However, the negligible difference between the K m and V max values of purified free and chitosan-immobilized fractions revealed that the conformational flexibility of pectinolytics was retained as such. A significant color and turbidity reductions were recorded after 60min treatment with CTS-PG, followed by CTS-PME, and CTS-PL. It can be concluded that the clarification of apples, mango, peach, and apricot juices was greatly affected by CTS-PG, CTS-PME, and CTS-PL treatments rendering them as potential candidatures for food industry applications. Copyright © 2017 Elsevier B.V. All rights reserved.

An Antibody-Immobilized Silica Inverse Opal Nanostructure for Label-Free Optical Biosensors.

PubMed

Lee, Wang Sik; Kang, Taejoon; Kim, Shin-Hyun; Jeong, Jinyoung

2018-01-20

Three-dimensional SiO₂-based inverse opal (SiO₂-IO) nanostructures were prepared for use as biosensors. SiO₂-IO was fabricated by vertical deposition and calcination processes. Antibodies were immobilized on the surface of SiO₂-IO using 3-aminopropyl trimethoxysilane (APTMS), a succinimidyl-[(N-maleimidopropionamido)-tetraethyleneglycol] ester (NHS-PEG₄-maleimide) cross-linker, and protein G. The highly accessible surface and porous structure of SiO₂-IO were beneficial for capturing influenza viruses on the antibody-immobilized surfaces. Moreover, as the binding leads to the redshift of the reflectance peak, the influenza virus could be detected by simply monitoring the change in the reflectance spectrum without labeling. SiO₂-IO showed high sensitivity in the range of 10³-10⁵ plaque forming unit (PFU) and high specificity to the influenza A (H1N1) virus. Due to its structural and optical properties, SiO₂-IO is a promising material for the detection of the influenza virus. Our study provides a generalized sensing platform for biohazards as various sensing strategies can be employed through the surface functionalization of three-dimensional nanostructures.

Long-term high-level waste technology. Composite report

NASA Astrophysics Data System (ADS)

Cornman, W. R.

1981-12-01

Research and development studies on the immobilization of high-level wastes from the chemical reprocessing of nuclear reactor fuels are summarized. The reports are grouped under the following tasks: (1) program management and support; (2) waste preparation; (3) waste fixation; and (4) final handling. Some of the highlights are: leaching properties were obtained for titanate and tailored ceramic materials being developed at ICPP to immobilize zirconia calcine; comparative leach tests, hot-cell tests, and process evaluations were conducted of waste form alternatives to borosilicate glass for the immobilization of SRP high-level wastes, experiments were run at ANL to qualify neutron activation analysis and radioactive tracers for measuring leach rates from simulated waste glasses; comparative leach test samples of SYNROC D were prepared, characterized, and tested at LLNL; encapsulation of glass marbles with lead or lead alloys was demonstrated on an engineering scale at PNL; a canister for reference Commercial HLW was designed at PNL; a study of the optimization of salt-crete was completed at SRL; a risk assessment showed that an investment for tornado dampers in the interim storage building of the DWPF is unjustified.

Preparation and characterization of soy protein films with a durable water resistance-adjustable and antimicrobial surface.

PubMed

Li, Shuzhao; Donner, Elizabeth; Xiao, Huining; Thompson, Michael; Zhang, Yachuan; Rempel, Curtis; Liu, Qiang

2016-12-01

A water resistant surface was first obtained by immobilizing hydrophobic copolymers, poly (styrene-co-glycidyl methacrylate) (PSG), with functional groups on soy protein isolate (SPI) films. XPS and AFM results showed that PSG copolymers were immobilized on the film by chemical bonding, and formed a rough surface with some bumps because of the segregation of two different phases on PSG copolymers. Water resistance of the modified films could be adjusted dramatically by further immobilizing different amounts of guanidine-based antimicrobial polymers, poly (hexamethylene guanidine hydrochloride) (PHMG) on the resulting hydrophobic surface. The introduction of hydrophilic PHMG on the resulting surface generated many micropores, which potentially increased the water uptake of the modified films. Furthermore, the modified SPI films showed higher thermostability compared to native SPI film and broad-spectrum antimicrobial activity by contact killing, attributed to the presence of PHMG on the surface. The modified SPI film with a multi-functional surface showed potential for applications in the packaging and medical fields. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

Traceless Immobilization of Analytes for High-Throughput Experiments with SAMDI Mass Spectrometry.

PubMed

Helal, Kazi Y; Alamgir, Azmain; Berns, Eric J; Mrksich, Milan

2018-06-21

Label-free assays, and particularly those based on the combination of mass spectroscopy with surface chemistries, enable high-throughput experiments of a broad range of reactions. However, these methods can still require the incorporation of functional groups that allow immobilization of reactants and products to surfaces prior to analysis. In this paper, we report a traceless method for attaching molecules to a self-assembled monolayer for matrix-assisted laser desorption and ionization (SAMDI) mass spectrometry. This method uses monolayers that are functionalized with a 3-trifluoromethyl-3-phenyl-diazirine group that liberates nitrogen when irradiated and gives a carbene that inserts into a wide range of bonds to covalently immobilize molecules. Analysis of the monolayer with SAMDI then reveals peaks for each of the adducts formed from molecules in the sample. This method is applied to characterize a P450 drug metabolizing enzyme and to monitor a Suzuki-Miyaura coupling chemical reaction and is important because modification of the substrates with a functional group would alter their activities. This method will be important for high-throughput experiments in many areas, including reaction discovery and optimization.

Comparative investigation of two methods for Acetylcholinesterase enzyme immobilization on modified porous silicon

NASA Astrophysics Data System (ADS)

Khaldi, Khadidja; Sam, Sabrina; Lounas, Amel; Yaddaden, Chafiaa; Gabouze, Noure-Eddine

2017-11-01

In this work, Acetylcholinesterase enzyme (AChE) was immobilized on porous silicon (PSi) surface using two strategies. In the first method, acid chains were covalently grafted on the hydrogenated PSi by hydrosilylation reaction. The obtained acid-terminated surface was activated by a reaction with N-hydroxysuccinimide (NHS) in the presence of a peptide-coupling agent N-ethyl-N‧-(3-dimethylaminopropyl)-carbodiimide (EDC), and then reacted with the amino linker of the lysine residues AChE to anchor the enzyme by a covalent amide bond. In the second procedure, the PSi surface was first hydroxylated in piranha solution, followed by a silanization reaction with 3-aminopropyltriethoxysilane (APTES) to form amine-terminated surface. Finally, AChE was attached to the terminal amine groups by an aminolysis reaction with carboxylic acid groups of AChE in the presence of NHS/EDC mixture. Fourier transform infrared spectroscopy (FTIR) confirmed the efficiency of the surface modifications. The enzymatic activity of immobilized AChE was determined by means of a colorimetric test and was discussed according to the enzyme orientation on the surface which was revealed by contact angle measurements.

Technical Status Report: Preliminary Glass Formulation Report for INEEL HAW. Revision 1

DOE Office of Scientific and Technical Information (OSTI.GOV)

Peeler, D.; Reamer, I.; Vienna, J.

1998-03-01

Preliminary glass formulation work has been initiated at Pacific Northwest National Laboratory (PNNL) and the Savannah River Technology Center (SRTC) to support immobilization efforts of Idaho National Engineering and Environmental Laboratory (INEEL) high activity waste (HAW). Based on current pretreatment flow sheet assumptions, several glasses were fabricated and tested using an average `All Blend` waste stream composition which is dominated by the presence of ZrO{sub 2} (i.e., approximately 80 wt percent). The results of this initial work show that immobilization via vitrification is a viable option for a specific INEEL HAW waste stream. Waste loadings of at least 19 wtmore » percent can be achieved for the `All Blend` stream while maintaining targeted processing and product performance criteria. This waste loading translates into a ZrO{sub 2} content in excess of 15 wt percent in the final glass waste form. Frits developed for this work are based in the alkali borosilicate system. Although the results indicate that vitrification can be used to immobilize the `All Blend` waste stream, the glass compositions are by no means optimized.« less

Fabrication of Carbohydrate Microarrays by Boronate Formation.

PubMed

Adak, Avijit K; Lin, Ting-Wei; Li, Ben-Yuan; Lin, Chun-Cheng

2017-01-01

The interactions between soluble carbohydrates and/or surface displayed glycans and protein receptors are essential to many biological processes and cellular recognition events. Carbohydrate microarrays provide opportunities for high-throughput quantitative analysis of carbohydrate-protein interactions. Over the past decade, various techniques have been implemented for immobilizing glycans on solid surfaces in a microarray format. Herein, we describe a detailed protocol for fabricating carbohydrate microarrays that capitalizes on the intrinsic reactivity of boronic acid toward carbohydrates to form stable boronate diesters. A large variety of unprotected carbohydrates ranging in structure from simple disaccharides and trisaccharides to considerably more complex human milk and blood group (oligo)saccharides have been covalently immobilized in a single step on glass slides, which were derivatized with high-affinity boronic acid ligands. The immobilized ligands in these microarrays maintain the receptor-binding activities including those of lectins and antibodies according to the structures of their pendant carbohydrates for rapid analysis of a number of carbohydrate-recognition events within 30 h. This method facilitates the direct construction of otherwise difficult to obtain carbohydrate microarrays from underivatized glycans.

Radiation stability of resveratrol in immobilization on poly vinyl pyrrolidone hydrogel dressing for dermatological use

NASA Astrophysics Data System (ADS)

Momesso, Roberta G. R. A. P.; Moreno, Carolina S.; Rogero, Sizue O.; Rogero, José R.; Spencer, Patrick J.; Lugão, Ademar B.

2010-03-01

The polyphenol trans-resveratrol is a natural phytoalexin, which is found in red wine and in a wide variety of plant species. Resveratrol displays a wide array of biological activities, such as modulation of lipid metabolism, anti-inflammatory and antioxidant activities. This active compound immobilized in polyvinylpyrrolidone (PVP) hydrogel could be very interesting for topical administration, as a dressing form for dermatological use. However, PVP hydrogel obtained by radiation-induced crosslinking can cause undesirable hydrolysis reactions in the active compound. The aim of this work was to verify the resveratrol stability after irradiation at 0.5 and 1 kGy in the presence of ethanol, methanol or tert-butyl alcohol. The integrity of these samples was compared to unirradiated resveratrol by HPLC. The PVP hydrogel matrix was characterized by gel fraction, swelling and in vitro biocompatibility test. The results of gel fraction and swelling degree were approximately 90% and 1600%, respectively. The cytotoxicity assay showed absence of toxicity for this formulation after crosslinking and sterilization, indicating that the PVP hydrogel formulation was appropriate for resveratrol immobilization to produce a dressing for dermatological use.

Osteoinductive activity of insulin-functionalized cell culture surfaces obtained using diazonium chemistry

PubMed Central

Mikulska, Anna; Filipowska, Joanna; Osyczka, Anna M.; Nowakowska, Maria; Szczubiałka, Krzysztof

2015-01-01

Polymeric surfaces suitable for cell culture (DR/Pec) were constructed from diazoresin (DR) and pectin (Pec) in a form of ultrathin films using the layer-by-layer (LbL) technique. The surfaces were functionalized with insulin using diazonium chemistry. Such functionalized surfaces were used to culture human mesenchymal stem cells (hMSCs) to assess their suitability for bone tissue engineering and regeneration. The activity of insulin immobilized on the surfaces (DR/Pec/Ins) was compared to that of insulin dissolved in the culture medium. Human MSC grown on insulin-immobilized DR/Pec surfaces displayed increased proliferation and higher osteogenic activity. The latter was determined by means of alkaline phosphatase (ALP) activity, which increases at early stages of osteoblasts differentiation. Insulin dissolved in the culture medium did not stimulate cell proliferation and its osteogenic activity was significantly lower. Addition of recombinant human bone morphogenetic protein 2 (rhBMP-2) to the culture medium further increased ALP activity in hMSCs indicating additive osteogenic action of immobilized insulin and rhBMP-2. PMID:25629028

FORMATION OF PYROMORPHITE IN ANGLESITE-HYDROXYAPATITE SUSPENSIONS UNDER VARYING PH CONDITIONS

EPA Science Inventory

Addition of phosphate to lead [Pb(II)] contaminated soil to immobilize soil Pb by formation of pyromorphite has been proposed as an alternative remediation technique. Lead sulfate (PbSO4, anglesite), a Pb-bearing form found in contaminated soils and wastes, was reacted with a sy...

BIOSENSOR FOR DIRECT DETERMINATION OF ORGANOPHOSPHATE NERVE AGENTS. 1. POTENTIOMETRIC ENZYME ELECTRODE. (R823663)

EPA Science Inventory

A potentiometric enzyme electrode for the direct measurement of organophosphate (OP)
nerve agents was developed. The basic element of this enzyme electrode was a pH electrode
modified with an immobilized organophosphorus hydrolase (OPH) layer formed by cross-linking
OPH ...

Sand Transport, Flow Turbulence, and Bed Forms over an Immobile Gravel Bed

USDA-ARS?s Scientific Manuscript database

Channels downstream of dams often become armored because the sediment supply from upstream is cut off. Sand is generally supplied to these armored reaches intermittently from tributaries downstream of the dam or from sand bypassing. Accurate predictions of the rate of transport of sand over and th...

Synthesis, Photophysical Characterization, and Gelation Studies of a Stilbene-Cholesterol Derivative

ERIC Educational Resources Information Center

Geiger, H. Christina; Geiger, David K.; Baldwin, Christine

2006-01-01

Organogels are low molar mass organic compounds with the ability to immobilize an incredible quantity of solvent and fibrous aggregation of these compounds formed by noncovalent interaction usually involves hydrogen bonding. For stilbene-cholesterol based gelators, the driving force for molecular aggregation are weak van der Waal interactions…

Stability studies of immobilized lipase on rice husk and eggshell membrane

NASA Astrophysics Data System (ADS)

Abdulla, R.; Sanny, S. A.; Derman, E.

2017-06-01

Lipase immobilization for biodiesel production is gaining importance day by day. In this study, lipase from Burkholderia cepacia was immobilized on activated support materials namely rice husk and egg shell membrane. Both rice husk and eggshell membrane are natural wastes that holds a lot of potential as immobilization matrix. Rice husk and eggshell membrane were activated with glutaraldehyde. Lipase was immobilized on the glutaraldehyde-activated support material through adsorption. Immobilization efficiency together with enzyme activity was observed to choose the highest enzyme loading for further stability studies. Immobilization efficiency of lipase on rice husk was 81 as compared to an immobilization efficiency of 87 on eggshell membrane. Immobilized lipase on eggshell membrane exhibited higher enzyme activity as compared to immobilized lipase on rice husk. Eggshell membrane also reported higher stability than rice husk as immobilization matrix. Both types of immobilized lipase retatined its activity after ten cycles of reuse. In short, eggshell membrane showed to be a better immobilization platform for lipase as compared to rice husk. However, with further improvement in technique of immobilization, the stability of both types of immobilized lipase can be improved to a greater extent.

Immobilization and bonding scheme of radioactive iodine-129 in silver tellurite glass

NASA Astrophysics Data System (ADS)

Lee, Cheong Won; Pyo, Jae-Young; Park, Hwan-Seo; Yang, Jae Hwan; Heo, Jong

2017-08-01

Silver tellurite glasses with melting temperatures < 700 °C were prepared to immobilize the 129I that normally volatilizes during high-temperature melting. Glasses have densities of 6.31 ± 0.1 g/cm3 and glass transition temperatures of 165 ± 3 °C that provide thermal stability at the disposal site. Iodine waste loading in glasses was as high as 12.64 wt% of all metallic elements and 11.21 wt% including oxygen. Normalized elemental releases obtained from the product consistency test were well below US regulation of 2 g/m2. Iodines are surrounded by four Ag+ ions forming [Ag4I]3+ units that are further connected to tellurite network through bonds with non-bridging oxygens.

Mathematical model of the oxidation of ferrous iron by a biofilm of Thiobacillus ferrooxidans.

PubMed

Mesa, M M; Macías, M; Cantero, D

2002-01-01

Microbial oxidation of ferrous iron may be a viable alternative method of producing ferric sulfate, which is a reagent used for removal of H(2)S from biogas. The paper introduces a kinetic study of the biological oxidation of ferrous iron by Thiobacillus ferrooxidans immobilized on biomass support particles (BSP) composed of polyurethane foam. On the basis of the data obtained, a mathematical model for the bioreactor was subsequently developed. In the model described here, the microorganisms adhere by reversible physical adsorption to the ferric precipitates that are formed on the BSP. The model can also be considered as an expression for the erosion of microorganisms immobilized due to the agitation of the medium by aeration.

Oyster mushroom’s lipase enzyme entrapment on calcium alginate as biocatalyst in the synthesis of lauryl diethanolamide

NASA Astrophysics Data System (ADS)

Wijayati, N.; Masubah, K.; Supartono

2017-02-01

Lipase is an enzyme with large biotechnology applications, such as hydrolysis in the food industry, applications in chemical industry, synthesis of polymers and surfactants. Lipase was isolated from oyster mushroom with activity 0,93 U/mg and protein content 1,1234 mg/mL. Lipase was immobilized by entrapment method in a matrix of Ca-alginate. This report describes that we have developed for the synthesis of lauryl diethanolamide The result showed that the optimum condition of lipase immobilization was achieved on 3% Na-alginate solution with protein content 0,84 mg/mL and the activity 3,33 U/mg. An amide (22.911%) formed from the amidation of lauric acid and diethanolamine.

Method for synthesizing pollucite from chabazite and cesium chloride

DOEpatents

Pereira, C.

1999-02-23

A method is described for immobilizing waste chlorides salts containing radionuclides and hazardous nuclear material for permanent disposal, and in particular, a method is described for immobilizing waste chloride salts containing cesium, in a synthetic form of pollucite. The method for synthesizing pollucite from chabazite and cesium chloride includes mixing dry, non-aqueous cesium chloride with chabazite and heating the mixture to a temperature greater than the melting temperature of the cesium chloride, or above about 700 C. The method further comprises significantly improving the rate of retention of cesium in ceramic products comprised of a salt-loaded zeolite by adding about 10% chabazite by weight to the salt-loaded zeolite prior to conversion at elevated temperatures and pressures to the ceramic composite. 3 figs.

Method for synthesizing pollucite from chabazite and cesium chloride

DOEpatents

Pereira, Candido

1999-01-01

A method for immobilizing waste chlorides salts containing radionuclides and hazardous nuclear material for permanent disposal, and in particular, a method for immobilizing waste chloride salts containing cesium, in a synthetic form of pollucite. The method for synthesizing pollucite from chabazite and cesium chloride includes mixing dry, non-aqueous cesium chloride with chabazite and heating the mixture to a temperature greater than the melting temperature of the cesium chloride, or above about 700.degree. C. The method further comprises significantly improving the rate of retention of cesium in ceramic products comprised of a salt-loaded zeolite by adding about 10% chabazite by weight to the salt-loaded zeolite prior to conversion at elevated temperatures and pressures to the ceramic composite.

Peptide aptamer-assisted immobilization of green fluorescent protein for creating biomolecule-complexed carbon nanotube device

NASA Astrophysics Data System (ADS)

Nii, Daisuke; Nozawa, Yosuke; Miyachi, Mariko; Yamanoi, Yoshinori; Nishihara, Hiroshi; Tomo, Tatsuya; Shimada, Yuichiro

2017-10-01

Carbon nanotubes are a novel material for next-generation applications. In this study, we generated carbon nanotube and green fluorescent protein (GFP) conjugates using affinity binding peptides. The carbon nanotube-binding motif was introduced into the N-terminus of the GFP through molecular biology methods. Multiple GFPs were successfully aligned on a single-walled carbon nanotube via the molecular recognition function of the peptide aptamer, which was confirmed through transmission electron microscopy and optical analysis. Fluorescence spectral analysis results also suggested that the carbon nanotube-GFP complex was autonomously formed with orientation and without causing protein denaturation during immobilization. This simple process has a widespread potential for fabricating carbon nanotube-biomolecule hybrid devices.

Biotechnological storage and utilization of entrapped solar energy.

PubMed

Bhattacharya, Sumana; Schiavone, Marc; Nayak, Amiya; Bhattacharya, Sanjoy K

2005-03-01

Our laboratory has recently developed a device employing immobilized F0F1 adenosine triphosphatase (ATPase) that allows synthesis of adenosine triphosphate (ATP) from adenosine 5'-diphosphate and inorganic phosphate using solar energy. We present estimates of total solar energy received by Earth's land area and demonstrate that its efficient capture may allow conversion of solar energy and storage into bonds of biochemicals using devices harboring either immobilized ATPase or NADH dehydrogenase. Capture and storage of solar energy into biochemicals may also enable fixation of CO2 emanating from polluting units. The cofactors ATP and NADH synthesized using solar energy could be used for regeneration of acceptor D-ribulose-1,5-bisphosphate from 3-phosphoglycerate formed during CO2 fixation.

Replica amplification of nucleic acid arrays

DOEpatents

Church, George M.

2002-01-01

A method of producing a plurality of a nucleic acid array, comprising, in order, the steps of amplifying in situ nucleic acid molecules of a first randomly-patterned, immobilized nucleic acid array comprising a heterogeneous pool of nucleic acid molecules affixed to a support, transferring at least a subset of the nucleic acid molecules produced by such amplifying to a second support, and affixing the subset so transferred to the second support to form a second randomly-patterned, immobilized nucleic acid array, wherein the nucleic acid molecules of the second array occupy positions that correspond to those of the nucleic acid molecules from which they were amplified on the first array, so that the first array serves as a template to produce a plurality, is disclosed.

Immobilization of actinides in stable mineral type and ceramic materials (high temperature synthesis)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Starkov, O.; Konovalov, E.

1996-05-01

Alternative vitrification technologies are being developed in the world for the immobilization of high radioactive waste in materials with improved thermodynamic stability, as well as improved chemical and thermal stability and stability to radiation. Oxides, synthesized in the form of analogs to rock-forming minerals and ceramics, are among those materials that have highly stable properties and are compatible with the environment. In choosing the appropriate material, we need to be guided by its geometric stability, the minimal number of cations in the structure of the material and the presence of structural elements in the mineral that are isomorphs of uraniummore » and thorium, actinoids found in nature. Rare earth elements, yttrium, zirconium and calcium are therefore suitable. The minerals listed in the table (with the exception of the zircon) are pegatites by origin, i.e. they are formed towards the end of the magma crystallization of silicates form the residual melt, enriched with Ta, Nb, Ti, Zr, Ce, Y, U and Th. Uranium and thorium in the form of isomorphic admixtures form part of the lattice of the mineral. These minerals, which are rather simple in composition and structure and are formed under high temperatures, may be viewed as natural physio-chemical systems that are stable and long-lived in natural environments. The similarity of the properties of actinoids and lanthanoids plays an important role in the geochemistry of uranium and thorium; however, uranium (IV) is closer to the {open_quotes}heavy{close_quotes} group of lanthanoids (the yttrium group) while thorium (IV) is closer to the {open_quotes}light{close_quotes} group (the cerium group). That is why rare earth minerals contain uranium and thorium in the form of isomorphic admixtures.« less

Immobilization of ammonia-oxidizing bacteria by polyvinyl alcohol and sodium alginate.

PubMed

Dong, Yuwei; Zhang, Yanqiu; Tu, Baojun

Ammonia-oxidizing bacteria were immobilized by polyvinyl alcohol (PVA) and sodium alginate. The immobilization conditions and ammonia oxidation ability of the immobilized bacteria were investigated. The following immobilization conditions were observed to be optimal: PVA, 12%; sodium alginate, 1.1%; calcium chloride, 1.0%; inoculum concentration, 1.3 immobilized balls/mL of immobilized medium; pH, 10; and temperature, 30°C. The immobilized ammonia-oxidizing bacteria exhibited strong ammonia oxidation ability even after being recycled four times. The ammonia nitrogen removal rate of the immobilized ammonia-oxidizing bacteria reached 90.30% under the optimal immobilization conditions. When compared with ammonia-oxidizing bacteria immobilized by sodium alginate alone, the bacteria immobilized by PVA and sodium alginate were superior with respect to pH resistance, the number of reuses, material cost, heat resistance, and ammonia oxidation ability. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Analysis of Hanford Cast Stone Supplemental LAW using Composition Adjusted SRS Tank 50 Salt Solution

DOE Office of Scientific and Technical Information (OSTI.GOV)

Crawford, C.; Cozzi, A.; Hill, K.

Vitrification is the primary disposition path for Low Activity Waste (LAW) at the Department of Energy (DOE) Hanford Site. A cementitious waste form is one of the alternatives being considered for the supplemental immobilization of the LAW that will not be treated by the primary vitrification facility. Washington River Protection Solutions (WRPS) has been directed to generate and collect data on cementitious or pozzolanic waste forms such as Cast Stone.

Improving reliability of a residency interview process.

PubMed

Peeters, Michael J; Serres, Michelle L; Gundrum, Todd E

2013-10-14

To improve the reliability and discrimination of a pharmacy resident interview evaluation form, and thereby improve the reliability of the interview process. In phase 1 of the study, authors used a Many-Facet Rasch Measurement model to optimize an existing evaluation form for reliability and discrimination. In phase 2, interviewer pairs used the modified evaluation form within 4 separate interview stations. In phase 3, 8 interviewers individually-evaluated each candidate in one-on-one interviews. In phase 1, the evaluation form had a reliability of 0.98 with person separation of 6.56; reproducibly, the form separated applicants into 6 distinct groups. Using that form in phase 2 and 3, our largest variation source was candidates, while content specificity was the next largest variation source. The phase 2 g-coefficient was 0.787, while confirmatory phase 3 was 0.922. Process reliability improved with more stations despite fewer interviewers per station-impact of content specificity was greatly reduced with more interview stations. A more reliable, discriminating evaluation form was developed to evaluate candidates during resident interviews, and a process was designed that reduced the impact from content specificity.

Effects of immobilization by electricity and MS-222 on brown trout broodstock and their progeny

USGS Publications Warehouse

Redman, S.D.; Meinertz, J.R.; Gaikowski, M.P.

1998-01-01

To determine the effects of electrically and chemically induced immobilization on postspawn broodstock and their progeny, age-2 and age-3 female broodstock and age-2 male broodstock of brown trout Salmo trutta were immobilized with electricity or tricaine methanesulfonate (MS-222), stripped of their eggs or milt, and weighed. Eggs taken from electrically immobilized females were fertilized with milt taken from age-2 males that were immobilized with electricity, and eggs taken from females immobilized with MS-222 were fertilized with milt taken from age-2 males that were immobilized with MS-222. After spawning, the mortality and weight of broodstock were compared twice over a 6-month period. Egg viability and growth of offspring fry from each treatment group were also compared. Electricity induced complete and consistent immobilization in brown trout broodstock. Electrically immobilized fish were more easily handled than fish immobilized with MS-222; however, electrically immobilized fish survival (70%) was significantly less than fish immobilized with MS-222 (83%). Broodstock growth differences were only noted at 6 months postexposure, when the mean weight of electrically immobilized fish was slightly less than the weight of fish immobilized with MS-222. Broodstock immobilization by electricity did not reduce egg viability or fry growth.

Highly efficient method towards in situ immobilization of invertase using cryogelation.

PubMed

Olcer, Zehra; Ozmen, Mehmet Murat; Sahin, Zeynep M; Yilmaz, Faruk; Tanriseven, Aziz

2013-12-01

A novel method was developed for the immobilization of Saccharomyces cerevisiae invertase within supermacroporous polyacrylamide cryogel and was used to produce invert sugar. First, the cross-linking of invertase with soluble polyglutaraldehyde (PGA) was carried out prior to immobilization in order to increase the bulkiness of invertase and thus preventing the leakage of the cross-linked enzyme after immobilization by entrapment. And then, in situ immobilization of PGA cross-linked invertase within cryogel synthesis was achieved by free radical polymerization in semi-frozen state. The method resulted in 100 % immobilization and 74 % activity yields. The immobilized invertase retained all the initial activity for 30 days and 30 batch reactions. Immobilization had no effect on optimum temperature and it was 60 °C for both free and immobilized enzyme. However, optimum pH was affected upon immobilization. Optimum pH values for free and immobilized enzyme were 4.5 and 5.0, respectively. The immobilized enzyme was more stable than the free enzyme at high pH and temperatures. The kinetic parameters for free and immobilized invertase were also determined. The newly developed method is simple yet effective and could be used for the immobilization of some other enzymes and microorganisms.

Calcium-Citrate-Phosphate Solution Injection for In Situ Strontium-90 Immobilization

NASA Astrophysics Data System (ADS)

Fruchter, J. S.; Vermeul, V.; Szecsody, J.; Williams, M. D.; Fritz, B. G.

2010-12-01

Sr-90 present in groundwater and the vadose zone at the Hanford 100N area due to past waste disposal practices has reached the nearby Columbia River, as evidenced by Sr-90 concentrations in near river wells and aquifer tubes and near shore sediments. Sr-90 is currently being remediated by adsorption onto apatite (55 times stronger than Sr-90 adsorption to sediment), followed by incorporation of the Sr-90 into the apatite structure. If the Sr-90 can remain immobilized for 300 years (~ten 29.1-yr half-lives of Sr-90 decay), it will have decayed below regulatory limits to Y-90 and to stable Zr-90. Apatite [Ca10(PO4)6(OH)2] is being precipitated in situ by injection of an aqueous solution of Ca-citrate and Na-phosphate through a series of injection wells spaced 30 ft on center, forming a 300-ft-long permeable reactive barrier. Design criteria for the injection operations were based on 1) amendment volume and mass injected, 2) amendment arrival at adjacent wells, 3) water-level elevation during treatment, and 4) injection rate limitations associated with well plugging. An evaluation of compliance with these injection design criteria was used to assess operational performance and identify candidate wells for supplemental treatment. Injection design criteria were not fully met at 8 of the 16 injection well locations, with the primary deficiency at 4 of 8 locations being the limited vertical extent of Hanford formation treatment due to low-river-stage conditions during the injection. Wells whose extent of treatment did not meet design criteria were recommended for retreatment. Although injection design criteria were not fully met at a significant number of well locations, aqueous performance assessment monitoring data collected to date indicate good barrier performance. Aqueous Sr-90 monitoring in four compliance monitoring wells over a year following the high concentration injections indicates 84% to 95% decrease in Sr-90 concentrations (relative to the low and high end of the baseline range, respectively). In addition, post treatment sediment cores were collected to quantify the amount of apatite that was formed from the barrier-emplacement operations. Results indicate that the processes that account for the observed reduction in aqueous Sr-90 concentrations include: a) incorporation of Sr-90 into apatite (about 39.4% of the total Sr-90 mass in the core), b) ion exchange flushing due to the Ca-citrate-PO4 solution injection (about 47% of the mass), and c) a small increase in Sr-90 adsorbed to sediment and apatite precipitate.

Nanosilver–Silica Composite: Prolonged Antibacterial Effects and Bacterial Interaction Mechanisms for Wound Dressings

PubMed Central

Ge, Yanling; Palva, Airi; Nordström, Katrina

2017-01-01

Infected superficial wounds were traditionally controlled by topical antibiotics until the emergence of antibiotic-resistant bacteria. Silver (Ag) is a kernel for alternative antibacterial agents to fight this resistance quandary. The present study demonstrates a method for immobilizing small-sized (~5 nm) silver nanoparticles on silica matrix to form a nanosilver–silica (Ag–SiO2) composite and shows the prolonged antibacterial effects of the composite in vitro. The composite exhibited a rapid initial Ag release after 24 h and a slower leaching after 48 and 72 h and was effective against both methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli). Ultraviolet (UV)-irradiation was superior to filter-sterilization in retaining the antibacterial effects of the composite, through the higher remaining Ag concentration. A gauze, impregnated with the Ag–SiO2 composite, showed higher antibacterial effects against MRSA and E. coli than a commercial Ag-containing dressing, indicating a potential for the management and infection control of superficial wounds. Transmission and scanning transmission electron microscope analyses of the composite-treated MRSA revealed an interaction of the released silver ions with the bacterial cytoplasmic constituents, causing ultimately the loss of bacterial membranes. The present results indicate that the Ag–SiO2 composite, with prolonged antibacterial effects, is a promising candidate for wound dressing applications. PMID:28878170

Production of a novel camel single-domain antibody specific for the type III mutant EGFR.

PubMed

Omidfar, K; Rasaee, M J; Modjtahedi, H; Forouzandeh, M; Taghikhani, M; Golmakani, N

2004-01-01

Camelids have a unique immune system capable of producing single-domain heavy-chain antibodies. The antigen-specific domain of these heavy-chain IgGs (VHH) are the smallest binding units produced by the immune system. In this study, we report the isolation and characterization of several binders against the epidermal growth factor receptor (EGFR) vIII retrieved from immune library of camels (Camelus bactrianus and Camelus dromedarius). The EGFRvIII is a ligand-independent, constitutively active, mutated form of the wild-type EGFR. The expression of EGFRvIII has been demonstrated in a wide range of human malignancies, including gliomas, and breast, prostate, ovarian and lung cancer. Camels were immunized with a synthetic peptide corresponding to a mutated sequence and tissue homogenates. Single-domain antibodies (VHH) were directly selected by panning a phage display library on successively decreasing amounts of synthetic peptide immobilized on magnetic beads. The anti-EGFRvIII camel single-domain antibodies selectively bound to the EGFRvIII peptide and reacted specifically with the immunoaffinity-purified antigen from a non-small cell lung cancer patient. These antibodies with affinities in the nanomolar range recognized the EGFRvIII peptide and affinity-purified mutated receptor. We concluded that using the phage display technique, antigen-specific VHH antibody fragments are readily accessible from the camelids. These antibodies may be good candidates for tumor-diagnostic and therapeutic applications. Copyright 2004 S. Karger AG, Basel.

Critical Thinking Tendencies among Teacher Candidates

ERIC Educational Resources Information Center

Genc, Salih Zeki

2008-01-01

The study aims to determine critical thinking tendencies among teacher candidates. 720 students from primary school teaching department (Primary School Teaching Programme, Science Teaching Programme and Pre-School Teaching Programme) form the sample of the study. When the gender and age distributions were investigated, 253 candidates are males and…

Examining Variations in Programmatic Delivery on Teaching Candidates' Sense of Efficacy

ERIC Educational Resources Information Center

Putman, S. Michael

2012-01-01

Researchers have recommended that teacher education candidates receive multiple opportunities for systematic implementation of practices within authentic contexts. These experiences form an important foundation for candidates' beliefs about their abilities in classrooms. Yet, few researchers have examined the relationship between the context for…

Localization of heavy metals immobilized on specific organic and mineral parts of a wood-derived biochar

NASA Astrophysics Data System (ADS)

Rees, Frédéric; Watteau, Françoise; Morel, Jean-Louis

2013-04-01

Biochar has been intensively investigated over recent years, not only as a promising carbon sequestration or fertilizing agent in soils but also as a possible new sorbent to remediate contaminated soils. A few studies have revealed its high potential for heavy metals immobilization depending on the nature of biochar and trace elements. The mechanisms behind this immobilization remain however unclear: some authors have hypothesized a high sorption capacity due to biochar large surface area while others have suggested that this immobilization is mainly due to soil pH increase. In particular, the distinction between heavy metals specific sorption in biochar pores and heavy metals precipitation in or outside biochar particles is often impossible to make while it is of primary importance to evaluate biochar ability to retain these pollutants on a long-time scale. In order to evaluate the main heavy metal immobilization effects on a standard biochar and to identify the most successful biochar parts of the sample, we examined biochar particles after heavy metals immobilization in batch experiments designed to mimic real chemical processes in soils. A biochar derived from hard and soft wood and pyrolyzed at about 450°C was put in contact with relatively low concentrations of heavy metals (Pb, Cu, Cd, Zn, Ni) in an initially acidic Ca(NO3)2 solution. Following a one-week adsorption and a one-week desorption step, we recovered the biochar particles and observed them using scanning electron microscopy coupled to energy dispersive x-ray spectroscopy, focusing especially on the changes in mineral phases and the location of each of the retained heavy metals on biochar particles. We were able to distinguish different structures in the biochar samples which were linked to the degree of pyrolysis and the exact nature of the raw wood biomass. We detected the presence of concentrated metals zones (e.g. lead) in specific locations of the organic particles depending on the original plant tissues, and enlightened metal associations with newly-formed mineral phases such as calcite present on biochar surface. These observations provide new insights in the understanding of metal immobilization mechanisms on biochar such as precipitation and co-precipitation. Our findings also underline the need to consider the heterogeneity of biochar constitution for optimizing the remediation potential of biochar on contaminated sites.

Soymilk residue (okara) as a natural immobilization carrier for Lactobacillus plantarum cells enhances soymilk fermentation, glucosidic isoflavone bioconversion, and cell survival under simulated gastric and intestinal conditions

PubMed Central

Xiudong, Xia; Ying, Wang; Xiaoli, Liu; Ying, Li

2016-01-01

Cell immobilization is an alternative to microencapsulation for the maintenance of cells in a liquid medium. However, artificial immobilization carriers are expensive and pose a high safety risk. Okara, a food-grade byproduct from soymilk production, is rich in prebiotics. Lactobacilli could provide health enhancing effects to the host. This study aimed to evaluate the potential of okara as a natural immobilizer for L. plantarum 70810 cells. The study also aimed to evaluate the effects of okara-immobilized L. plantarum 70810 cells (IL) on soymilk fermentation, glucosidic isoflavone bioconversion, and cell resistance to simulated gastric and intestinal stresses. Scanning electron microscopy (SEM) was used to show cells adherence to the surface of okara. Lactic acid, acetic acid and isoflavone analyses in unfermented and fermented soymilk were performed by HPLC with UV detection. Viability and growth kinetics of immobilized and free L. plantarum 70810 cells (FL) were followed during soymilk fermentation. Moreover, changes in pH, titrable acidity and viscosity were measured by conventional methods. For in vitro testing of simulated gastrointestinal resistance, fermented soymilk was inoculated with FL or IL and an aliquot incubated into acidic MRS broth which was conveniently prepared to simulate gastric, pancreatic juices and bile salts. Survival to simulated gastric and intestinal stresses was evaluated by plate count of colony forming units on MRS agar. SEM revealed that the lactobacilli cells attached and bound to the surface of okara. Compared with FL, IL exhibited a significantly higher specific growth rate, shorter lag phase of growth, higher productions of lactic and acetic acids, a faster decrease in pH and increase in titrable acidity, and a higher soymilk viscosity. Similarly, IL in soymilk showed higher productions of daizein and genistein compared with the control. Compared with FL, IL showed reinforced resistance to simulatedgastric and intestinal stresses in vitro that included low pH, low pH plus pepsin, pancreatin, and bile salt. Our results indicate that okara is a new potential immobilization carrier to enhance the growth and glucosidic isoflavone bioconversion activities of L. plantarum in soymilk and improve cell survivability following simulated gastric and intestinal conditions. PMID:27867770

Immobilization of U(VI) from oxic groundwater by Hanford 300 Area sediments and effects of Columbia River water.

PubMed

Ahmed, Bulbul; Cao, Bin; Mishra, Bhoopesh; Boyanov, Maxim I; Kemner, Kenneth M; Fredrickson, Jim K; Beyenal, Haluk

2012-09-01

Regions within the U.S. Department of Energy Hanford 300 Area (300 A) site experience periodic hydrologic influences from the nearby Columbia River as a result of changing river stage, which causes changes in groundwater elevation, flow direction and water chemistry. An important question is the extent to which the mixing of Columbia River water and groundwater impacts the speciation and mobility of uranium (U). In this study, we designed experiments to mimic interactions among U, oxic groundwater or Columbia River water, and 300 A sediments in the subsurface environment of Hanford 300 A. The goals were to investigate mechanisms of: 1) U immobilization in 300 A sediments under bulk oxic conditions and 2) U remobilization from U-immobilized 300 A sediments exposed to oxic Columbia River water. Initially, 300 A sediments in column reactors were fed with U(VI)-containing oxic 1) synthetic groundwater (SGW), 2) organic-amended SGW (OA-SGW), and 3) de-ionized (DI) water to investigate U immobilization processes. After that, the sediments were exposed to oxic Columbia River water for U remobilization studies. The results reveal that U was immobilized by 300 A sediments predominantly through reduction (80-85%) when the column reactor was fed with oxic OA-SGW. However, U was immobilized by 300 A sediments through adsorption (100%) when the column reactors were fed with oxic SGW or DI water. The reduced U in the 300 A sediments fed with OA-SGW was relatively resistant to remobilization by oxic Columbia River water. Oxic Columbia River water resulted in U remobilization (∼7%) through desorption, and most of the U that remained in the 300 A sediments fed with OA-SGW (∼93%) was in the form of uraninite nanoparticles. These results reveal that: 1) the reductive immobilization of U through OA-SGW stimulation of indigenous 300 A sediment microorganisms may be viable in the relatively oxic Hanford 300 A subsurface environments and 2) with the intrusion of Columbia River water, desorption may be the primary process resulting in U remobilization from OA-SGW-stimulated 300 A sediments at the subsurface of the Hanford 300 A site. Copyright © 2012 Elsevier Ltd. All rights reserved.

Soymilk residue (okara) as a natural immobilization carrier for Lactobacillus plantarum cells enhances soymilk fermentation, glucosidic isoflavone bioconversion, and cell survival under simulated gastric and intestinal conditions.

PubMed

Xiudong, Xia; Ying, Wang; Xiaoli, Liu; Ying, Li; Jianzhong, Zhou

2016-01-01

Cell immobilization is an alternative to microencapsulation for the maintenance of cells in a liquid medium. However, artificial immobilization carriers are expensive and pose a high safety risk. Okara, a food-grade byproduct from soymilk production, is rich in prebiotics. Lactobacilli could provide health enhancing effects to the host. This study aimed to evaluate the potential of okara as a natural immobilizer for L. plantarum 70810 cells. The study also aimed to evaluate the effects of okara-immobilized L. plantarum 70810 cells (IL) on soymilk fermentation, glucosidic isoflavone bioconversion, and cell resistance to simulated gastric and intestinal stresses. Scanning electron microscopy (SEM) was used to show cells adherence to the surface of okara. Lactic acid, acetic acid and isoflavone analyses in unfermented and fermented soymilk were performed by HPLC with UV detection. Viability and growth kinetics of immobilized and free L. plantarum 70810 cells (FL) were followed during soymilk fermentation. Moreover, changes in pH, titrable acidity and viscosity were measured by conventional methods. For in vitro testing of simulated gastrointestinal resistance, fermented soymilk was inoculated with FL or IL and an aliquot incubated into acidic MRS broth which was conveniently prepared to simulate gastric, pancreatic juices and bile salts. Survival to simulated gastric and intestinal stresses was evaluated by plate count of colony forming units on MRS agar. SEM revealed that the lactobacilli cells attached and bound to the surface of okara. Compared with FL, IL exhibited a significantly higher specific growth rate, shorter lag phase of growth, higher productions of lactic and acetic acids, a faster decrease in pH and increase in titrable acidity, and a higher soymilk viscosity. Similarly, IL in soymilk showed higher productions of daizein and genistein compared with the control. Compared with FL, IL showed reinforced resistance to simulatedgastric and intestinal stresses in vitro that included low pH, low pH plus pepsin, pancreatin, and bile salt. Our results indicate that okara is a new potential immobilization carrier to enhance the growth and glucosidic isoflavone bioconversion activities of L. plantarum in soymilk and improve cell survivability following simulated gastric and intestinal conditions.

Synthesis and Characterization of Protein-Conjugated Silver Nanoparticles/Silver Salt Loaded Poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) Film for Prevention of Bacterial Infections and Potential Use in Bone Tissue Engineering Applications

NASA Astrophysics Data System (ADS)

Bakare, Rotimi Ayotunde

Failure of orthopedic implants due to bacterial infection has been a major concern in bone tissue engineering. To this end, we have formulated a potential orthopedic implant made of naturally occurring biodegradable polymer, i.e. poly (3-hydroxylbutyrate-co-3-hydroxylvalerate) (PHBV), modified with BSA conjugated silver nanoparticles and or silver chloride. Upon release of Ag NPs and or Ag+ in the implant region, can promote aseptic environment by inhibition of bacteria growth and also support/maintain bone cell adhesion, growth, and proliferation. For formulating nanoparticles loaded PHBV scaffold, we exploit specific interaction between bovine serum albumin (BSA) of BSA capped silver nanoparticles and collagen of collagen immobilized PHBV scaffold. Therefore, the first part of this study dealt with synthesis and characterization of collagen immobilized PHBV film for loading of BSA stabilized silver (Ag/BSA) nanoparticles. Two different approaches were used to immobilize collagen on macroporous PHBV film. First approach uses thermal radical copolymerization with 2-hydroxyethylmethacrylate (HEMA), while the second approach uses aminolysis to functionalize macroporous PHBV film. Using collagen crosslinker, type I collagen was covalently grafted to formulate collagen immobilized PHEMA-g-PHBV and collagen immobilized NH2-PHBV films, respectively. Spectroscopic (FTIR, XPS), physical (SEM), and thermal (TGA) techniques were used to characterize the functionalized PHBV films. The Ag/BSA nanoparticles were then loaded on collagen immobilized PHBV films and untreated PHBV films. The concentration of nanoparticles loaded on PHBV film was determined by atomic absorption spectrometry and fluorescence spectroscopy. The amount of nanoparticles loaded on collagen immobilized PHBV film was found to be significantly greater than that on untreated PHBV film. The amount of Ag/BSA nanoparticles loaded on collagen immobilized PHBV film was found to depend on the concentration of Ag/BSA nanoparticles solution used for loading and on the molecular weight of type I collagen used in collagen immobilization on PHBV film. At physiological pH, optimum amount of nanoparticles was retained on Type I collagen immobilized PHBV film because at pH 7.4, protonated amino groups of collagen immobilized PHBV film promote strong electrostatic interaction with the carboxylate anions of BSA stabilized silver nanoparticles. The second part of this study dealt with formulating AgCl/PHBV film that can potentially release silver ions for effective antimicrobial activity. In this study, we formulated AgCl/PHBV composite film by a salt exchange mechanism. Thermogravimetric analysis (TGA) was used to quantify the amount of NaCl present before and after salt exchange. The Na content in the pre-washed and partially washed NaCl/PHBV film was found to be 43.60% and 1.24% by mass, respectively. The AgCl/PBHV composite film was acid digested and assayed for Na+ and Ag+ content by using Atomic Absorption Spectrometry (AAS) and was found to be 2.15 and 10.25 ppm, respectively. XPS technique was used to characterize the surface elemental composition of the AgCl/PHBV composite film. The survey spectrum of AgCl/PHBV film showed emergence of Ag 3d and Cl 2s peaks compared to pure PHBV which was predominantly composed of C 1s and O 1s peaks. The release kinetics of silver ions from AgCl/PHBV composite film showed an initial burst of about 1.5 ppm of silver ions during the first day of desorption followed by a gradual release of silver ions at an average rate of 0.3 ppm per day during the span of two weeks studied. Ag/BSA nanoparticles loaded collagen immobilized PHBV films and AgCl/PHBV composite films were tested for antibacterial efficacy against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Colony forming unit and optical density measurements of Ag/BSA nanoparticles loaded collagen immobilized PHBV films showed broad antimicrobial activity at low Ag/BSA nanoparticles concentration (0.19and 0.31 microg) compared to commercially available gentamicin and sulfamethoxazole/trimethoprim which showed sometimes selective antimicrobial activity and antimicrobial activity at high concentration (10 microg and 23.75/1.25 microg/disc). Additionally, a clear zone of inhibition around AgCl/PHBV composite film was noticed on a modified Kirby-Bauer disk diffusion assay. Optical density results and colony forming unit measurements showed that AgCl/PHBV composite film exhibit broad bactericidal activity. Next, we evaluated the cytotoxicity of Ag/BSA nanoparticles loaded collagen immobilized PHBV films and AgCl/PHBV composite films towards MC3T3-E1 cells at the same concentration both films showed broad antimicrobial activity. By using MTT assay, we established that Ag/BSA nanoparticles loaded collagen immobilized PHBV film showed minimal, if any, cytotoxic effect towards MC3T3-E1 cells while AgCl/PHBV composite film showed significant cytotoxic effect compared to tissue culture polystyrene. Our research findings provide several formulations for preparation of scaffold, if properly tuned; it can be used as a potential biocompatible and biodegradable scaffold for the prevention of bacterial infections and promotion of cell attachment and proliferation in bone tissue engineering applications.

Development of iron phosphate ceramic waste form to immobilize radioactive waste solution

NASA Astrophysics Data System (ADS)

Choi, Jongkwon; Um, Wooyong; Choung, Sungwook

2014-09-01

The objective of this research was to develop an iron phosphate ceramic (IPC) waste form using converter slag obtained as a by-product of the steel industry as a source of iron instead of conventional iron oxide. Both synthetic off-gas scrubber solution containing technetium-99 (or Re as a surrogate) and LiCl-KCl eutectic salt, a final waste solution from pyrochemical processing of spent nuclear fuel, were used as radioactive waste streams. The IPC waste form was characterized for compressive strength, reduction capacity, chemical durability, and contaminant leachability. Compressive strengths of the IPC waste form prepared with different types of waste solutions were 16 MPa and 19 MPa for LiCl-KCl eutectic salt and the off-gas scrubber simulant, respectively, which meet the minimum compressive strength of 3.45 MPa (500 psi) for waste forms to be accepted into the radioactive waste repository. The reduction capacity of converter slag, a main dry ingredient used to prepare the IPC waste form, was 4136 meq/kg by the Ce(IV) method, which is much higher than those of the conventional Fe oxides used for the IPC waste form and the blast furnace slag materials. Average leachability indexes of Tc, Li, and K for the IPC waste form were higher than 6.0, and the IPC waste form demonstrated stable durability even after 63-day leaching. In addition, the Toxicity Characteristic Leach Procedure measurements of converter slag and the IPC waste form with LiCl-KCl eutectic salt met the universal treatment standard of the leachability limit for metals regulated by the Resource Conservation and Recovery Act. This study confirms the possibility of development of the IPC waste form using converter slag, showing its immobilization capability for radionuclides in both LiCl-KCl eutectic salt and off-gas scrubber solutions with significant cost savings.

Development of iron phosphate ceramic waste form to immobilize radioactive waste solution

DOE Office of Scientific and Technical Information (OSTI.GOV)

Choi, Jongkwon; Um, Wooyong; Choung, Sungwook

The objective of this research was to develop an iron phosphate ceramic (IPC) waste form using converter slag obtained as a by-product of the steel industry as a source of iron instead of conventional iron oxide. Both synthetic off-gas scrubber solution containing technetium-99 (or Re as a surrogate) and LiCl-KCl eutectic salt, a final waste solution from pyrochemical processing of spent nuclear fuel, were used as radioactive waste streams. The IPC waste form was characterized for compressive strength, reduction capacity, chemical durability, and contaminant leachability. Compressive strengths of the IPC waste form prepared with different types of waste solutions weremore » 16 MPa and 19 MPa for LiCl-KCl eutectic salt and the off-gas scrubber simulant, respectively, which meet the minimum compressive strength of 3.45 MPa (500 psi) for waste forms to be accepted into the radioactive waste repository. The reduction capacity of converter slag, a main dry ingredient used to prepare the IPC waste form, was 4,136 meq/kg by the Ce(IV) method, which is much higher than those of the conventional Fe oxides used for the IPC waste form and the blast furnace slag materials. Average leachability indexes of Tc, Li, and K for the IPC waste form were higher than 6.0, and the IPC waste form demonstrated stable durability even after 63-day leaching. In addition, the Toxicity Characteristic Leach Procedure measurements of converter slag and the IPC waste form with LiCl-KCl eutectic salt met the universal treatment standard of the leachability limit for metals regulated by the Resource Conservation and Recovery Act. This study confirms the possibility of development of the IPC waste form using converter slag, showing its immobilization capability for radionuclides in both LiCl-KCl eutectic salt and off-gas scrubber solutions with significant cost savings.« less

LC/MS/MS structure elucidation of reaction intermediates formed during the TiO2 photocatalysis of microcystin-LR

EPA Science Inventory

Microcystin-LR (MC-LR), a cyanotoxin and emerging drinking water contaminant, was treated with TiO(2) photocatalysts immobilized on stainless steel plates as an alternative to nanoparticles in slurry. The reaction intermediates of MC-LR were identified with mass spectrometry (MS)...

Ceramics in nuclear waste management

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chikalla, T D; Mendel, J E

1979-05-01

Seventy-three papers are included, arranged under the following section headings: national programs for the disposal of radioactive wastes, waste from stability and characterization, glass processing, ceramic processing, ceramic and glass processing, leaching of waste materials, properties of nuclear waste forms, and immobilization of special radioactive wastes. Separate abstracts were prepared for all the papers. (DLC)

Biocatalyst including porous enzyme cluster composite immobilized by two-step crosslinking and its utilization as enzymatic biofuel cell

NASA Astrophysics Data System (ADS)

Chung, Yongjin; Christwardana, Marcelinus; Tannia, Daniel Chris; Kim, Ki Jae; Kwon, Yongchai

2017-08-01

An enzyme cluster composite (TPA/GOx) formed from glucose oxidase (GOx) and terephthalaldehyde (TPA) that is coated onto polyethyleneimine (PEI) and carbon nanotubes (CNTs) is suggested as a new catalyst ([(TPA/GOx)/PEI]/CNT). In this catalyst, TPA promotes inter-GOx links by crosslinking to form a large and porous structure, and the TPA/GOx composite is again crosslinked with PEI/CNT to increase the amount of immobilized GOx. Such a two-step crosslinking (i) increases electron transfer because of electron delocalization by π conjugation and (ii) reduces GOx denaturation because of the formation of strong chemical bonds while its porosity facilitates mass transfer. With these features, an enzymatic biofuel cell (EBC) employing the new catalyst is fabricated and induces an excellent maximum power density (1.62 ± 0.08 mW cm-2), while the catalytic activity of the [(TPA/GOx)/PEI]/CNT catalyst is outstanding. This is clear evidence that the two-step crosslinking and porous structure caused by adoption of the TPA/GOx composite affect the performance enhancement of EBC.

Enhanced butyric acid tolerance and bioproduction by Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor.

PubMed

Jiang, Ling; Wang, Jufang; Liang, Shizhong; Cai, Jin; Xu, Zhinan; Cen, Peilin; Yang, Shangtian; Li, Shuang

2011-01-01

Repeated fed-batch fermentation of glucose by Clostridium tyrobutyricum immobilized in a fibrous bed bioreactor (FBB) was successfully employed to produce butyric acid at a high final concentration as well as to adapt a butyric-acid-tolerant strain. At the end of the eighth fed-batch fermentation, the butyric acid concentration reached 86.9 ± 2.17 g/L, which to our knowledge is the highest butyric acid concentration ever produced in the traditional fermentation process. To understand the mechanism and factors contributing to the improved butyric acid production and enhanced acid tolerance, adapted strains were harvested from the FBB and characterized for their physiological properties, including specific growth rate, acid-forming enzymes, intracellular pH, membrane-bound ATPase and cell morphology. Compared with the original culture used to seed the bioreactor, the adapted culture showed significantly reduced inhibition effects of butyric acid on specific growth rate, cellular activities of butyric-acid-forming enzyme phosphotransbutyrylase (PTB) and ATPase, together with elevated intracellular pH, and elongated rod morphology. © 2010 Wiley Periodicals, Inc.