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Sample records for cells immobilized

  1. Immobilized cells in meat fermentation.

    Science.gov (United States)

    McLoughlin, A J; Champagne, C P

    1994-01-01

    The immobilization of microbial cells can contribute to fermented meat technology at two basic levels. First, the solid/semisolid nature (low available water) of the substrate restricts the mobility of cells and results in spatial organizations based on "natural immobilization" within the fermentation matrix. The microniches formed influence the fermentation biochemistry through mass transfer limitations and the subsequent development and activity of the microflora. This form of immobilization controls the nature of competition between subpopulations within the microflora and ultimately exerts an effect on the ecological competence (ability to survive and compete) of the various cultures present. Second, immobilized cell technology (ICT) can be used to enhance the ecological competence of starter cultures added to initiate the fermentation. Immobilization matrices such as alginate can provide microniches or microenvironments that protect the culture during freezing or lyophilization, during subsequent rehydration, and when in competition with indigenous microflora. The regulated release of cells from the microenvironments can also contribute to competitive ability. The regulation of both immobilization processes can result in enhanced fermentation activity. PMID:8069934

  2. Immobilization of cells for use as biocatalysts

    Energy Technology Data Exchange (ETDEWEB)

    Vojtisek, V.; Jirku, V.; Krumphanzl, V.; Culik, K.

    1983-07-21

    Bacterial cells and cells of higher organisms are immobilized on polymers, either as whole cells, cell fragments, or subcellular components. This immobilization is used for stabilization of their various enzymic activities, which are of commercial interest, e.g. for the enzymes themselves, for alkaloid production, for hormone transformations, or for various fermentations. Thus, Sedipur CL-930 was polymerized in the presence of glutaraldehyde and the polymer was incubated with Alcaligenes metalcaligenes cells for immobilization. The nonimmobilized cells contained an aspartate ammonia-lyase activity of 550 mumol L-aspartate converted/min/g, and the immobilized cells contained an activity of 420 or 500 mumol aspartate/min/g when the polymer used was made with 2 different ratios of Sedipur to glutaraldehyde. The immobilized cell product had the form of defined platelets (lamellae) with a diameter of 100-600 mum, depending on the Sedipur/glutaraldehyde ratio. In other procedures, cells were permeabilized with tensides and/or organic solvents after the immobilization. Other cells immobilized included yeast, fungi, and plant cells. The activities which were examined included glycolytic enzymes, penicillin acylase, L-asparagine amidohydrolase and production of alkaloids and phytosterols from Solanum aviculare.

  3. Engineering aspects of nitrification with immobilized cells.

    NARCIS (Netherlands)

    Hunik, J.H.

    1993-01-01

    Several aspects of a nitrification process with artificially immobilized cells in an airlift loop reactor have been investigated and are described in this thesis. In chapter 1 an overview of immobilization methods, suitable reactors, modelling, small-scaleapplications and scale-up strategy is given.

  4. Metabolic Responses of Bacterial Cells to Immobilization

    Directory of Open Access Journals (Sweden)

    Joanna Żur

    2016-07-01

    Full Text Available In recent years immobilized cells have commonly been used for various biotechnological applications, e.g., antibiotic production, soil bioremediation, biodegradation and biotransformation of xenobiotics in wastewater treatment plants. Although the literature data on the physiological changes and behaviour of cells in the immobilized state remain fragmentary, it is well documented that in natural settings microorganisms are mainly found in association with surfaces, which results in biofilm formation. Biofilms are characterized by genetic and physiological heterogeneity and the occurrence of altered microenvironments within the matrix. Microbial cells in communities display a variety of metabolic differences as compared to their free-living counterparts. Immobilization of bacteria can occur either as a natural phenomenon or as an artificial process. The majority of changes observed in immobilized cells result from protection provided by the supports. Knowledge about the main physiological responses occurring in immobilized cells may contribute to improving the efficiency of immobilization techniques. This paper reviews the main metabolic changes exhibited by immobilized bacterial cells, including growth rate, biodegradation capabilities, biocatalytic efficiency and plasmid stability.

  5. Metabolic Responses of Bacterial Cells to Immobilization.

    Science.gov (United States)

    Żur, Joanna; Wojcieszyńska, Danuta; Guzik, Urszula

    2016-01-01

    In recent years immobilized cells have commonly been used for various biotechnological applications, e.g., antibiotic production, soil bioremediation, biodegradation and biotransformation of xenobiotics in wastewater treatment plants. Although the literature data on the physiological changes and behaviour of cells in the immobilized state remain fragmentary, it is well documented that in natural settings microorganisms are mainly found in association with surfaces, which results in biofilm formation. Biofilms are characterized by genetic and physiological heterogeneity and the occurrence of altered microenvironments within the matrix. Microbial cells in communities display a variety of metabolic differences as compared to their free-living counterparts. Immobilization of bacteria can occur either as a natural phenomenon or as an artificial process. The majority of changes observed in immobilized cells result from protection provided by the supports. Knowledge about the main physiological responses occurring in immobilized cells may contribute to improving the efficiency of immobilization techniques. This paper reviews the main metabolic changes exhibited by immobilized bacterial cells, including growth rate, biodegradation capabilities, biocatalytic efficiency and plasmid stability. PMID:27455220

  6. Surface cell immobilization within perfluoroalkoxy microchannels

    Energy Technology Data Exchange (ETDEWEB)

    Stojkovič, Gorazd; Krivec, Matic [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia); Vesel, Alenka [Jožef Stefan Institute, Jamova cesta 39, 1000 Ljubljana (Slovenia); Marinšek, Marjan [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia); Žnidaršič-Plazl, Polona, E-mail: polona.znidarsic@fkkt.uni-lj.si [Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, SI-1000 Ljubljana (Slovenia)

    2014-11-30

    Graphical abstract: - Highlights: • A very efficient approach for immobilization of cells into microreactors is presented. • It is applicable to various materials, including PFA and cyclic olefin (co)polymers. • It was used to immobilize different prokaryotic and eukaryotic microbes. • Cells were immobilized on the surface in high density and showed good stability. • Mechanisms of APTES interactions with target materials are proposed. - Abstract: Perfluoroalkoxy (PFA) is one of the most promising materials for the fabrication of cheap, solvent resistant and reusable microfluidic chips, which have been recently recognized as effective tools for biocatalytic process development. The application of biocatalysts significantly depends on efficient immobilization of enzymes or cells within the reactor enabling long-term biocatalyst use. Functionalization of PFA microchannels by 3-aminopropyltriethoxysilane (ATPES) and glutaraldehyde was used for rapid preparation of microbioreactors with surface-immobilized cells. X-ray photoelectron spectroscopy and scanning electron microscopy were used to accurately monitor individual treatment steps and to select conditions for cell immobilization. The optimized protocol for Saccharomyces cerevisiae immobilization on PFA microchannel walls comprised ethanol surface pretreatment, 4 h contacting with 10% APTES aqueous solution, 10 min treatment with 1% glutaraldehyde and 20 min contacting with cells in deionized water. The same protocol enabled also immobilization of Escherichia coli, Pseudomonas putida and Bacillus subtilis cells on PFA surface in high densities. Furthermore, the developed procedure has been proved to be very efficient also for surface immobilization of tested cells on other materials that are used for microreactor fabrication, including glass, polystyrene, poly (methyl methacrylate), polycarbonate, and two olefin-based polymers, namely Zeonor{sup ®} and Topas{sup ®}.

  7. Immobilization of microbial cells containing NAD-kinase

    Energy Technology Data Exchange (ETDEWEB)

    Hayashi, T.; Tanaka, Y.; Kawashima, K.

    1979-06-01

    Microbial cells having NAD-kinase activity, Brevibacterium ammoniagenes, were immobilized by the radiation-copolymerization method under low temperature with the activity recovery of more than 80%. Compared to the native microbial cells the immobilized cells were more stable against heat and pH change. The immobilized cells were subjected to the 5 hr reaction repeatedly 20 times without any activity loss.

  8. Ethanol tolerance of immobilized brewers' yeast cells.

    Science.gov (United States)

    Norton, S; Watson, K; D'Amore, T

    1995-04-01

    A method based on the survival of yeast cells subjected to an ethanol or heat shock was utilized to compare the stress resistance of free and carrageenan-immobilized yeast cells. Results demonstrated a significant increase of yeast survival against ethanol for immobilized cells as compared to free cells, while no marked difference in heat resistance was observed. When entrapped cells were released by mechanical disruption of the gel beads and submitted to the same ethanol stress, they exhibited a lower survival rate than entrapped cells, but a similar or slightly higher survival rate than free cells. The incidence of ethanol- or heat-induced respiratory-deficient mutants of entrapped cells was equivalent to that of control or non-stressed cells (1.3 +/- 0.5%) whereas ethanol- and heat-shocked free and released cells exhibited between 4.4% and 10.9% average incidence of respiration-deficient mutants. It was concluded that the carrageenan gel matrix provided a protection against ethanol, and that entrapped cells returned to normal physiological behaviour as soon as they were released. The cell growth rate was a significant factor in the resistance of yeast to high ethanol concentrations. The optimum conditions to obtain reliable and reproducible results involved the use of slow-growing cells after exhaustion of the sugar substrate.

  9. Aroma formation by immobilized yeast cells in fermentation processes.

    Science.gov (United States)

    Nedović, V; Gibson, B; Mantzouridou, T F; Bugarski, B; Djordjević, V; Kalušević, A; Paraskevopoulou, A; Sandell, M; Šmogrovičová, D; Yilmaztekin, M

    2015-01-01

    Immobilized cell technology has shown a significant promotional effect on the fermentation of alcoholic beverages such as beer, wine and cider. However, genetic, morphological and physiological alterations occurring in immobilized yeast cells impact on aroma formation during fermentation processes. The focus of this review is exploitation of existing knowledge on the biochemistry and the biological role of flavour production in yeast for the biotechnological production of aroma compounds of industrial importance, by means of immobilized yeast. Various types of carrier materials and immobilization methods proposed for application in beer, wine, fruit wine, cider and mead production are presented. Engineering aspects with special emphasis on immobilized cell bioreactor design, operation and scale-up potential are also discussed. Ultimately, examples of products with improved quality properties within the alcoholic beverages are addressed, together with identification and description of the future perspectives and scope for cell immobilization in fermentation processes.

  10. Immobilization of cells via activated cell walls

    Energy Technology Data Exchange (ETDEWEB)

    Markt, M.; Kas, J.; Valentova, O.; Demnerova, K.; Vodrazka, Z.

    1986-10-01

    Cell walls of Saccharomyces cerevisiae and S. uvarum were activated by periodate oxidation of vicinal diol groups in cell wall polysaccharides. The aldehyde groups thus generated allow the yeast cells to be covalently bound to modified bead cellulose or macroporous glycidyl methacrylate supports, or to enzymes such as glucose oxidase and catalase. 6 references.

  11. ENHANCED DEGRADATION OF CAPTAN BY IMMOBILIZED CELLS OF BACILLUS CIRCULANS

    Directory of Open Access Journals (Sweden)

    Veena More

    2014-10-01

    Full Text Available The possibility of using Bacillus circulans in degrading captan was evaluated by comparing the captan degradation rate by freely suspended and immobilized cells on agar, sodium alginate (SA, polyacrylamide (PA and polyurethane-foam (PUF in batch and repeated batch degradations. Under batch degradations, 50, 60, 72, and 88% of 0.1% captan was degraded by freely suspended cells, agar-, SA-, and PA-immobilized cells, respectively in 72 h; whereas 15, 47.5, 67.7 and 75% of 0.2% captan was degraded by freely suspended cells, agar-, SA-, and PA-immobilized cells, respectively in 72 h. However, 0.1 and 0.2% captan were completely degraded by PUF-immobilized cells in 48 and 72 h, respectively. Under repeated batch degradations, PUF-immobilized cells were reused more than 40 cycles for 72 h without losing the captan degradation ability, while the cells immobilized on agar, SA, and the PA could be reused for 15, 20, and 25 cycles, respectively. A significant 0.1% captan degradation by PUF-immobilized cells was observed at pH 4.0 - 10.0 and 20 - 40 ºC ranges. In contrast, freely suspended cells only degraded captan at optimum pH of 7.0 and 30 ºC. The PUF-immobilized cells were able to significantly degrade captan for 120 days at 4 ºC without losing the captan degradation ability; whereas this ability was lost in 120 days for freely suspended cells. Since the application of captan leads to pollution and reduces soil fertility, the use of immobilized cells of Bacillus circulans can thus be a better cost-effective strategy to decontaminate captan polluted sites.

  12. Smooth Muscle Cell Functionality on Collagen Immobilized Polycaprolactone Nanowire Surfaces

    Directory of Open Access Journals (Sweden)

    Victoria Leszczak

    2014-05-01

    Full Text Available Inhibition of smooth muscle cell (SMC proliferation and preservation of a differentiated state are important aspects in the management, avoidance and progression of vascular diseases. An understanding of the interaction between SMCs and the biomaterial involved is essential for a successful implant. In this study, we have developed collagen immobilized nanostructured surfaces with controlled arrays of high aspect ratio nanowires for the growth and maintenance of human aortic SMCs. The nanowire surfaces were fabricated from polycaprolactone and were immobilized with collagen. The objective of this study is to reveal how SMCs interact with collagen immobilized nanostructures. The results indicate significantly higher cellular adhesion on nanostructured and collagen immobilized surfaces; however, SMCs on nanostructured surfaces exhibit a more elongated phenotype. The reduction of MTT was significantly lower on nanowire (NW and collagen immobilized NW (colNW surfaces, suggesting that SMCs on nanostructured surfaces may be differentiated and slowly dividing. Scanning electron microscopy results reveal that SMCs on nanostructured surfaces are more elongated and that cells are interacting with the nano-features on the surface. After providing differentiation cues, heavy chain myosin and calponin, specific to a contractile SMC phenotype, are upregulated on collagen immobilized surfaces. These results suggest that nanotopography affects cell adhesion, proliferation, as well as cell elongation, while collagen immobilized surfaces greatly affect cell differentiation.

  13. Uranium uptake by immobilized cells of Pseudomonas strain EPS 5028

    International Nuclear Information System (INIS)

    Polyacrylamide-gel-immobilized cells of Pseudomonas strain EPS 5028 were effective in the removal of uranium (U) from synthetic effluents. Metal accumulation was performed in an open system in columns filled with immobilized cells that were challenged with continuous flows containing U. Possible variable of the system were studied. Uranium uptake by the immobilized cells of this microorganism was affected by pH but not by temperature or flow rate. In addition, U binding could be interpreted in terms of the Freundlich adsorption isotherm indicating single-layer adsorption. The feasibility of reusing the immobilized cells was suggested after the recovery of U with a solution of 0.1 M sodium carbonate. (orig.)

  14. Continuous culture of immobilized streptomyces cells for kasugamycin production.

    Science.gov (United States)

    Kim, C J; Chang, Y K; Chun, G T; Jeong, Y H; Lee, S J

    2001-01-01

    Continuous cultures of immobilized Streptomyces kasugaensis, a kasugamycin producer, were carried out on Celite beads. When using a prototype separator for immobilized-cell separation and recycling, the continuous operation could not be sustained for an extended period as a result of an excessive loss of immobilized cells caused by the poor performance of the separator. Accordingly, the immobilized-cell separator was revised to provide better immobilized-cell settling and thus recycling into the reactor. In a subsequent culture using the revised separator, a stable operation was maintained for over 820 h with a high kasugamycin productivity. The kasugamycin productivity ranged from 9.8 to 16.1 mg/L/h, which was about 14- to 23-fold higher than that in a batch suspended-cell culture. When the original feeding medium concentration was doubled at the end of the continuous culture, the productivity became severely impaired for several reasons, which will be discussed. An excessive formation of free cells and loss of immobilized cells through the separator were also observed. PMID:11386865

  15. Bioreduction of chromate by immobilized cells of Halomonas sp

    OpenAIRE

    S. Murugavelh, Kaustubha Mohanty

    2013-01-01

    In this work, the bioreduction of Cr(VI) by immobilized cells of Halomonas sp was reported. Ca alginate, acryl amide and agar were tested as the matrices for immobilization. Ca alginate was found to be the suitable matrix among the different matrices studied. Of the various dosages of inoculum studied 2 g/L was found to be the optimum. Glucose at 1 g/L was completely utilized by the immobilized Halomonas sp even in the presence of Cr(VI) at 40 mg/L. The optimum pH for the bioreduction of Cr(V...

  16. Fructooligosaccharides production using immobilized cells of Aspergillus japonicus

    OpenAIRE

    Mussatto, Solange I.; Dragone, Giuliano; Rodrigues, L. R.; Teixeira, J. A.

    2008-01-01

    The fructooligosaccharides (FOS) production using immobilized cells of the fungus Aspergillus japonicus ATCC 20236 was evaluated. Polyurethane foam, stainless steel sponge, vegetal fiber sponge, pumice stones, zeolites and cork were tested as immobilization carrier during the fermentation under submerged conditions. Experiments were carried out in 500 ml Erlenmeyer flasks containing 1 g of carrier and 100 ml of sucrose medium (165 g/l) enriched with nutrient sources. The flasks...

  17. Ethanol fermentation by immobilized cells of Zymomonas mobilis

    Energy Technology Data Exchange (ETDEWEB)

    Grote, W.

    1985-01-01

    Previous studies have shown that immobilized yeast cell cultures have commercial potential for fuel ethanol production. In this study the suitability of strains of Z. mobilis for whole cell immobilization was investigated. Experiments revealed that immobilization in Ca-alginate or K-carrageenan gel or use of flocculating strains was effective for ethanol production at relatively high productivities. Two laboratory size reactors were designed and constructed. These were a compartmented multiple discshaft column and a tower fermentor. Results of this work supported other studies that established that growth and fermentation could be uncoupled. The data indicated that specific metabolic rates were dependent on the nature of the fermentation media. The addition of lactobacilli to Z. mobilis continuous fermentations had only a transient effect, and was unlikely to affect an immobilized Z. mobilis process. With 150 gl/sup -1/ glucose media and a Z. mobilis ZM4 immobilized cell reactor, a maximum volumetric ethanol productivity of 55 gl/sup -1/h/sup -1/ was obtained. The fermentation of sucrose media or sucrose-based raw materials (molasses, cane juice, synthetic mill liquor) by immobilized Z. mobilis ZM4 revealed a pattern of rapid sucrose hydrolysis, preferential glucose utilization and the conversion of fructose to the undesirable by-products levan and sorbitol.

  18. Covalent immobilization of p-selectin enhances cell rolling.

    Science.gov (United States)

    Hong, Seungpyo; Lee, Dooyoung; Zhang, Huanan; Zhang, Jennifer Q; Resvick, Jennifer N; Khademhosseini, Ali; King, Michael R; Langer, Robert; Karp, Jeffrey M

    2007-11-20

    Cell rolling is an important physiological and pathological process that is used to recruit specific cells in the bloodstream to a target tissue. This process may be exploited for biomedical applications to capture and separate specific cell types. One of the most commonly studied proteins that regulate cell rolling is P-selectin. By coating surfaces with this protein, biofunctional surfaces that induce cell rolling can be prepared. Although most immobilization methods have relied on physisorption, chemical immobilization has obvious advantages, including longer functional stability and better control over ligand density and orientation. Here we describe chemical methods to immobilize P-selectin covalently on glass substrates. The chemistry was categorized on the basis of the functional groups on modified glass substrates: amine, aldehyde, and epoxy. The prepared surfaces were first tested in a flow chamber by flowing microspheres functionalized with a cell surface carbohydrate (sialyl Lewis(x)) that binds to P-selectin. Adhesion bonds between P-selectin and sialyl Lewis(x) dissociate readily under shear forces, leading to cell rolling. P-selectin immobilized on the epoxy glass surfaces exhibited enhanced long-term stability of the function and better homogeneity as compared to that for surfaces prepared by other methods and physisorbed controls. The microsphere rolling results were confirmed in vitro with isolated human neutrophils. This work is essential for the future development of devices for isolating specific cell types based on cell rolling, which may be useful for hematologic cancers and certain metastatic cancer cells that are responsive to immobilized selectins.

  19. Immobilized cell technology in beer brewing: Current experience and results

    Directory of Open Access Journals (Sweden)

    Leskošek-Čukalov Ida J.

    2005-01-01

    Full Text Available Immobilized cell technology (ICT has been attracting continual attention in the brewing industry over the past 30 years. Some of the reasons are: faster fermentation rates and increased volumetric productivity, compared to those of traditional beer production based on freely suspended cells, as well as the possibility of continuous operation. Nowadays, ICT technology is well established in secondary fermentation and alcohol- free and low-alcohol beer production. In main fermentation, the situation is more complex and this process is still under scrutiny on both the lab and pilot levels. The paper outlines the most important ICT processes developed for beer brewing and provides an overview of carrier materials, bioreactor design and examples of their industrial applications, as well as some recent results obtained by our research group. We investigated the possible applications of polyvinyl alcohol in the form of LentiKats®, as a potential porous matrices carrier for beer fermentation. Given are the results of growth studies of immobilized brewer's yeast Saccharomyces uvarum and the kinetic parameters obtained by using alginate microbeads with immobilized yeast cells and suspension of yeast cells as controls. The results indicate that the immobilization procedure in LentiKat® carriers has a negligible effect on cell viability and growth. The apparent specific growth rate of cells released in medium was comparable to that of freely suspended cells, implying preserved cell vitality. A series of batch fermentations performed in shaken flasks and an air-lift bioreactor indicated that the immobilized cells retained high fermentation activity. The full attenuation in green beer was reached after 48 hours in shaken flasks and less than 24 hours of fermentation in gas-lift bioreactors.

  20. Decolorization of reactive Brilliant Blue KN-R by immobilized cells of Aspergillus ficuum

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Aspergillus ficuum was immobilized with sodium alginate, and decolorization of Reactive Brilliant Blue KN-R was studied on immobilized and free Aspergillus ficuum. The optimal preparation condition of the strain immobilization was obtained by the orthogonal test, it is sodium alginate 3%, CaCl2 5%, wet mycelia 30 g/L, calcific time 8 h. It was found that the immobilized cells could effectively decolorize Reactive Brilliant Blue KN-R, the optimum temperature and pH were 33℃ and 5.0, respectively. The kinetics study of decolorization of immobilized cells showed that the decolorization of Aspergillus ficuum immobilized conformed to zero-order reaction model. The decolorization efficiency of immobilized cell compared with that of free cell in different physical conditions. Results showed that the decolorization of immobilized cells with mycelia had the best efficiency. The immobilized cells could be reused after the first decolorization.

  1. Immobilization of chlorine dioxide modified cells for uranium absorption

    International Nuclear Information System (INIS)

    There has been a trend towards the use of microorganisms to recover metals from industrial wastewater, for which various methods have been reported to be used to improve microorganism adsorption characteristics such as absorption capacity, tolerance and reusability. In present study, chlorine dioxide(ClO2), a high-efficiency, low toxicity and environment-benign disinfectant, was first reported to be used for microorganism surface modification. The chlorine dioxide modified cells demonstrated a 10.1% higher uranium adsorption capacity than control ones. FTIR analysis indicated that several cell surface groups are involved in the uranium adsorption and cell surface modification. The modified cells were further immobilized on a carboxymethylcellulose (CMC) matrix to improve their reusability. The cell-immobilized adsorbent could be employed either in a high concentration system to move vast UO22+ ions or in a low concentration system to purify UO22+ contaminated water thoroughly, and could be repeatedly used in multiple adsorption-desorption cycles with about 90% adsorption capacity maintained after seven cycles. - Highlights: • Chlorine dioxide was first reported to be used for microorganism surface modification. • The chlorine dioxide modified cells demonstrated a 10.1% higher uranium adsorption capacity than control ones. • The chlorine dioxide modified cells were further immobilized by carboxymethylcellulose to improve their reusability

  2. Immobilized cell technology in beer brewing: Current experience and results

    OpenAIRE

    Leskošek-Čukalov Ida J.; Nedović Viktor A.

    2005-01-01

    Immobilized cell technology (ICT) has been attracting continual attention in the brewing industry over the past 30 years. Some of the reasons are: faster fermentation rates and increased volumetric productivity, compared to those of traditional beer production based on freely suspended cells, as well as the possibility of continuous operation. Nowadays, ICT technology is well established in secondary fermentation and alcohol- free and low-alcohol beer production. In main fermentation, the sit...

  3. Bioreduction of chromate by immobilized cells of Halomonas sp

    Energy Technology Data Exchange (ETDEWEB)

    Murugavelh, S.; Mohanty, Kaustubha [Department of Chemical Engineering, Indian Institute of Technology Guwahati, Guwahati – 781039, Assam (India)

    2013-07-01

    In this work, the bioreduction of Cr(VI) by immobilized cells of Halomonas sp was reported. Ca alginate, acryl amide and agar were tested as the matrices for immobilization. Ca alginate was found to be the suitable matrix among the different matrices studied. Of the various dosages of inoculum studied 2 g/L was found to be the optimum. Glucose at 1 g L-1 was completely utilized by the immobilized Halomonas sp even in the presence of Cr(VI) at 40 mg L-1. The optimum pH for the bioreduction of Cr(VI) by immobilized Halomonas sp was found to be pH 6. The mechanical strength of the beads plays an essential role in the bioreduction process. Halomonas sp entrapped in a alginate matrix reported a maximum of 98.9 % of reduction for an initial Cr(VI) concentration of 10 mg L-1. The alginate beads can be reused for 3 times with slight drop in the percentage reduction. The presence of other metals decreased the bioreduction percentage.

  4. Bioreduction of chromate by immobilized cells of Halomonas sp

    Directory of Open Access Journals (Sweden)

    S. Murugavelh, Kaustubha Mohanty

    2013-01-01

    Full Text Available In this work, the bioreduction of Cr(VI by immobilized cells of Halomonas sp was reported. Ca alginate, acryl amide and agar were tested as the matrices for immobilization. Ca alginate was found to be the suitable matrix among the different matrices studied. Of the various dosages of inoculum studied 2 g/L was found to be the optimum. Glucose at 1 g/L was completely utilized by the immobilized Halomonas sp even in the presence of Cr(VI at 40 mg/L. The optimum pH for the bioreduction of Cr(VI by immobilized Halomonas sp was found to be pH 6. The mechanical strength of the beads plays an essential role in the bioreduction process. Halomonas sp entrapped in a alginate matrix reported a maximum of 98.9 % of reduction for an initial Cr(VI concentration of 10 mg/L. The alginate beads can be reused for 3 times with slight drop in the percentage reduction. The presence of other metals decreased the bioreduction percentage.

  5. Protein-free cell culture on an artificial substrate with covalently immobilized insulin.

    OpenAIRE

    Ito, Y.; Zheng, J.; Imanishi, Y.; Yonezawa, K; Kasuga, M.

    1996-01-01

    Insulin was immobilized on a surface-hydrolyzed poly(methyl methacrylate) film. Chinese hamster ovary cells overexpressing human insulin receptors were cultured on the film in the absence of serum or soluble proteins. Small amounts of immobilized insulin (1-10% of the required amount of free insulin) were sufficient to stimulate cell proliferation. In addition, the maximal mitogenic effect of immobilized insulin was greater than that of free insulin. Immobilized insulin activated the insulin ...

  6. Smooth Muscle Cell Functionality on Collagen Immobilized Polycaprolactone Nanowire Surfaces

    OpenAIRE

    Victoria Leszczak; Baskett, Dominique A.; Popat, Ketul C.

    2014-01-01

    Inhibition of smooth muscle cell (SMC) proliferation and preservation of a differentiated state are important aspects in the management, avoidance and progression of vascular diseases. An understanding of the interaction between SMCs and the biomaterial involved is essential for a successful implant. In this study, we have developed collagen immobilized nanostructured surfaces with controlled arrays of high aspect ratio nanowires for the growth and maintenance of human aortic SMCs. The nanow...

  7. A response calculus for immobilized T cell receptor ligands

    DEFF Research Database (Denmark)

    Andersen, P S; Menné, C; Mariuzza, R A;

    2001-01-01

    determine the level of T cell activation. When fitted to T cell responses against purified ligands immobilized on plastic surfaces, the 2D-affinity model adequately simulated changes in cellular activation as a result of varying ligand affinity and ligand density. These observations further demonstrated......To address the molecular mechanism of T cell receptor (TCR) signaling, we have formulated a model for T cell activation, termed the 2D-affinity model, in which the density of TCR on the T cell surface, the density of ligand on the presenting surface, and their corresponding two-dimensional affinity...... the importance of receptor cross-linking density in determining TCR signaling. Moreover, it was found that the functional two-dimensional affinity of TCR ligands was affected by the chemical composition of the ligand-presenting surface. This makes it possible that cell-bound TCR ligands, despite their low...

  8. Ethanol fermentation in an immobilized cell reactor using Saccharomyces cerevisiae.

    Science.gov (United States)

    Najafpour, Ghasem; Younesi, Habibollah; Syahidah Ku Ismail, Ku

    2004-05-01

    Fermentation of sugar by Saccharomyces cerevisiae, for production of ethanol in an immobilized cell reactor (ICR) was successfully carried out to improve the performance of the fermentation process. The fermentation set-up was comprised of a column packed with beads of immobilized cells. The immobilization of S. cerevisiae was simply performed by the enriched cells cultured media harvested at exponential growth phase. The fixed cell loaded ICR was carried out at initial stage of operation and the cell was entrapped by calcium alginate. The production of ethanol was steady after 24 h of operation. The concentration of ethanol was affected by the media flow rates and residence time distribution from 2 to 7 h. In addition, batch fermentation was carried out with 50 g/l glucose concentration. Subsequently, the ethanol productions and the reactor productivities of batch fermentation and immobilized cells were compared. In batch fermentation, sugar consumption and ethanol production obtained were 99.6% and 12.5% v/v after 27 h while in the ICR, 88.2% and 16.7% v/v were obtained with 6 h retention time. Nearly 5% ethanol production was achieved with high glucose concentration (150 g/l) at 6 h retention time. A yield of 38% was obtained with 150 g/l glucose. The yield was improved approximately 27% on ICR and a 24 h fermentation time was reduced to 7 h. The cell growth rate was based on the Monod rate equation. The kinetic constants (K(s) and mu(m)) of batch fermentation were 2.3 g/l and 0.35 g/lh, respectively. The maximum yield of biomass on substrate (Y(X-S)) and the maximum yield of product on substrate (Y(P-S)) in batch fermentations were 50.8% and 31.2% respectively. Productivity of the ICR were 1.3, 2.3, and 2.8 g/lh for 25, 35, 50 g/l of glucose concentration, respectively. The productivity of ethanol in batch fermentation with 50 g/l glucose was calculated as 0.29 g/lh. Maximum production of ethanol in ICR when compared to batch reactor has shown to increase

  9. The production of sorbitol by permeabilized and immobilized cells of Z. mobilis in sucrose

    OpenAIRE

    Josiane Alessandra Vignoli; Maria Antonia Colabone Celligoi; Rui Sérgio Ferreira da Silva; Márcio de Barros

    2006-01-01

    The production of sorbitol by permeabilized and immobilized cells of Zymomonas mobilis in Luffa cylindrica was investigated in sucrose medium. A full 2³ factorial design was used to verify the influence of each factor and its interactions. The cell permeabilization showed a significant and negative effect upon the production of sorbitol, while the time of cultivation and the immobilization process were significant and positive. The results demonstrated that the cell immobilization and the tim...

  10. Transient Behavior of Ethanol Fermentation in Immobilized Cell Bioreactors*

    OpenAIRE

    Tohru, KANNO; Yoshinori, FUJISHIGE; Hiroyuki, Ito; koichi, yamazaki; Masayoshi, KOBAYASHI

    1990-01-01

    The dynamic behavior of ethanol fermentation catalysed by an immobilized cell has been studied in batch and continuous stirred tank bioreactors, changing the operating conditions in a stepwise fashion. The rate of ethanol fermentation in the flow reactor reaches a new steady state within 60 min for the stepwise change in temperature or flow rate at 15〜30℃ and the residence time t_R=40 hr. The rate of fermentation obeys the Lineweaven-Burk plot and the Michaelis constant is calculated

  11. Ethanol production from concentrated food waste hydrolysates with yeast cells immobilized on corn stalk

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Shoubao [Huainan Normal Univ., Anhui (China). School of Life Science; Chen, Xiangsong; Wu, Jingyong; Wang, Pingchao [Chinese Academy of Sciences, Hefei (China). Key Lab. of Ion Beam Bio-engineering of Inst. of Plasma Physics

    2012-05-15

    The aim of the present study was to examine ethanol production from concentrated food waste hydrolysates using whole cells of S. cerevisiae immobilized on corn stalks. In order to improve cell immobilization efficiency, biological modification of the carrier was carried out by cellulase hydrolysis. The results show that proper modification of the carrier with cellulase hydrolysis was suitable for cell immobilization. The mechanism proposed, cellulase hydrolysis, not only increased the immobilized cell concentration, but also disrupted the sleek surface to become rough and porous, which enhanced ethanol production. In batch fermentation with an initial reducing sugar concentration of 202.64 {+-} 1.86 g/l, an optimal ethanol concentration of 87.91 {+-} 1.98 g/l was obtained using a modified corn stalk-immobilized cell system. The ethanol concentration produced by the immobilized cells was 6.9% higher than that produced by the free cells. Ethanol production in the 14th cycle repeated batch fermentation demonstrated the enhanced stability of the immobilized yeast cells. Under continuous fermentation in an immobilized cell reactor, the maximum ethanol concentration of 84.85 g/l, and the highest ethanol yield of 0.43 g/g (of reducing sugar) were achieved at hydraulic retention time (HRT) of 3.10 h, whereas the maximum volumetric ethanol productivity of 43.54 g/l/h was observed at a HRT of 1.55 h. (orig.)

  12. MC3T3-E1 Cells on Titanium Surfaces with Nanometer Smoothness and Fibronectin Immobilization

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    Tohru Hayakawa

    2012-01-01

    Full Text Available The present study was aimed to evaluate the viability and total protein contents of osteoblast-like cells on the titanium surface with different surface mechanical treatment, namely, nanometer smoothing (Ra: approximately 2.0 nm and sandblasting (Ra: approximately 1.0 μm, and biochemical treatment, namely, with or without fibronectin immobilization. Fibronectin could be easily immobilized by tresyl chloride-activation technique. MC3T3-E1 cells were seeded on the different titanium surfaces. Cell viability was determined by MTT assay. At 1 day of cell culture, there were no significant differences in cell viability among four different titanium surfaces. At 11 days, sandblasted titanium surface with fibronectin immobilization showed the significantly highest cell viability than other titanium surface. No significant differences existed for total protein contents among four different titanium surfaces at 11 days of cell culture. Scanning electron microscopy observation revealed that smoothness of titanium surface produced more spread cell morphologies, but that fibronectin immobilization did not cause any changes of the morphologies of attached cells. Fibronectin immobilization provided greater amount of the number of attached cells and better arrangement of attached cells. In conclusion, the combination of sandblasting and fibronectin immobilization enhanced the cell viability and fibronectin immobilization providing better arrangements of attached cells.

  13. Biodegradation of phenol by using free and immobilized cells of Acinetobacter sp. BS8Y.

    Science.gov (United States)

    Jiang, Lichun; Ruan, Qiping; Li, Rulan; Li, Tiandong

    2013-03-01

    Strain BS8Y with high biodegradation activity and high tolerance of phenol was isolated from activated sludge in an insulating material plant of China. This strain was capable of removing 99.2% of the initial 600 mg/l phenol in liquid minimal medium within 24 h and tolerating phenol at concentrations of up to 1,200 mg/ml. DNA sequencing and homologous analysis of the 16S rRNA gene identified that the strain BS8Y belonged to an Acinetobacter species. Polyvinyl alcohol was used as gel matrix to immobilize the strain BS8Y. The factors affecting the phenol degradation by immobilized cells and the phenol removal efficiency of free and immobilized cells were investigated; the stability of the immobilized cells is also reported. The results show that the immobilized cells could tolerate a higher phenol level and protected the bacteria much more effectively against changes in temperature and pH. The phenol degradation efficiency was high at up to 96% within 30 h, with an initial concentration of 800 mg/l phenol, and the immobilized cells showed better performance than the suspended cells. Reusability tests revealed that the immobilized cells were stable enough even after reuse for ten times or storing at 4°C for 35 d. These results demonstrate that immobilized Acinetobacter sp. BS8Y possesses a good application potential in the treatment of phenol-containing wastewater.

  14. Biodegradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1

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    Preeti N. Tallur

    2015-09-01

    Full Text Available Pyrethroid pesticide cypermethrin is a environmental pollutant because of its widespread use, toxicity and persistence. Biodegradation of such chemicals by microorganisms may provide an cost-effective method for their detoxification. We have investigated the degradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1 in various matrices such as, polyurethane foam (PUF, polyacrylamide, sodium alginate and agar. The optimum temperature and pH for the degradation of cypermethrin by immobilized cells of Micrococcus sp. were found to be 30 °C and 7.0, respectively. The rate of degradation of 10 and 20 mM of cypermethrin by freely suspended cells were compared with that of immobilized cells in batches and semi-continuous with shaken cultures. PUF-immobilized cells showed higher degradation of cypermethrin (10 mM and 20 mM than freely suspended cells and cells immobilized in other matrices. The PUF-immobilized cells of Micrococcus sp. strain CPN 1 were retain their degradation capacity. Thus, they can be reused for more than 32 cycles, without losing their degradation capacity. Hence, the PUF-immobilized cells of Micrococcus sp. could potentially be used in the bioremediation of cypermethrin contaminated water.

  15. Biodegradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1.

    Science.gov (United States)

    Tallur, Preeti N; Mulla, Sikandar I; Megadi, Veena B; Talwar, Manjunatha P; Ninnekar, Harichandra Z

    2015-01-01

    Pyrethroid pesticide cypermethrin is a environmental pollutant because of its widespread use, toxicity and persistence. Biodegradation of such chemicals by microorganisms may provide an cost-effective method for their detoxification. We have investigated the degradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1 in various matrices such as, polyurethane foam (PUF), polyacrylamide, sodium alginate and agar. The optimum temperature and pH for the degradation of cypermethrin by immobilized cells of Micrococcus sp. were found to be 30 °C and 7.0, respectively. The rate of degradation of 10 and 20 mM of cypermethrin by freely suspended cells were compared with that of immobilized cells in batches and semi-continuous with shaken cultures. PUF-immobilized cells showed higher degradation of cypermethrin (10 mM and 20 mM) than freely suspended cells and cells immobilized in other matrices. The PUF-immobilized cells of Micrococcus sp. strain CPN 1 were retain their degradation capacity. Thus, they can be reused for more than 32 cycles, without losing their degradation capacity. Hence, the PUF-immobilized cells of Micrococcus sp. could potentially be used in the bioremediation of cypermethrin contaminated water.

  16. Production of xanthan gum by free and immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii.

    Science.gov (United States)

    Niknezhad, Seyyed Vahid; Asadollahi, Mohammad Ali; Zamani, Akram; Biria, Davoud

    2016-01-01

    Production of xanthan gum using immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii grown on glucose or hydrolyzed starch as carbon sources was investigated. Calcium alginate (CA) and calcium alginate-polyvinyl alcohol-boric acid (CA-PVA) beads were used for the immobilization of cells. Xanthan titers of 8.2 and 9.2g/L were obtained for X. campestris cells immobilized in CA-PVA beads using glucose and hydrolyzed starch, respectively, whereas those for X. pelargonii were 8 and 7.9 g/L, respectively. Immobilized cells in CA-PVA beads were successfully employed in three consecutive cycles for xanthan production without any noticeable degradation of the beads whereas the CA beads were broken after the first cycle. The results of this study suggested that immobilized cells are advantageous over the free cells for xanthan production. Also it was shown that the cells immobilized in CA-PVA beads are more efficient than cells immobilized in CA beads for xanthan production. PMID:26526173

  17. Comparison of Di-n-methyl Phthalate Biodegradation by Free and Immobilized Microbial Cells

    Institute of Scientific and Technical Information of China (English)

    JIAN-LONG WANG; YU-CAI YE; WEI-ZHONG WU

    2003-01-01

    Objective To compare the biodegradation of di-n-methyl pathalate by free and immobilizedmicrobial cells. Methods The enrichment and isolation technique was used to isolate themicroorganism. The PAV-entrapment method was utilized to immobilize the microorganisms. Thescanning electron microscophy (SEM) was used to observe the growth and distribution of microbialcells immobilized inside the PVA bead gels. The GC/MS method was used to identify the mainintermediates of DMP degradation. Results The microbial cells could grow quite well in PVA gel.The metabolic pathway did not change before and after immobilization of the microbial cells. Thedegradation rate of immobilized cells was higher than that of free cells. Conclusion Theimmobilized microbial cells possess advantages than free cells when applied to the biodegradation oftoxic organic pollutants.

  18. Development of High-Productivity Continuous Ethanol Production using PVA-Immobilized Zymomonas mobilis in an Immobilized-Cells Fermenter

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    Nurhayati Nurhayati

    2015-07-01

    Full Text Available Ethanol as one of renewable energy was being considered an excellent alternative clean-burning fuel to replace gasoline. Continuous ethanol fermentation systems had offered important economic advantages compared to traditional systems. Fermentation rates were significantly improved, especially when continuous fermentation was integrated with cell immobilization techniques to enrich the cells concentration in fermentor. Growing cells of Zymomonas mobilis immobilized in polyvinyl alcohol (PVA gel beads were employed in an immobilized-cells fermentor for continuous ethanol fermentation from glucose. The glucose loading, dilution rate, and cells loading were varied in order to determine which best condition employed in obtaining both high ethanol production and low residual glucose with high dilution rate. In this study, 20 g/L, 100 g/L, 125 g/L and 150 g/L of glucose concentration and 20% (w/v, 40% (w/v and 50% (w/v of cells loading were employed with range of dilution rate at 0.25 to 1 h-1. The most stable production was obtained for 25 days by employing 100 g/L of glucose loading. Meanwhile, the results also exhibited that 125 g/L of glucose loading as well as 40% (w/v of cells loading yielded high ethanol concentration, high ethanol productivity, and acceptable residual glucose at 62.97 g/L, 15.74 g/L/h and 0.16 g/L, respectively. Furthermore, the dilution rate of 4 hour with 100 g/L and 40% (w/v of glucose and cells loading was considered as the optimum condition with ethanol production, ethanol productivity and residual glucose obtained were 49.89 g/L, 12.47 g/L/h, and 2.04 g/L, respectively. This recent study investigated ethanol inhibition as well. The present research had proved that high sugar concentration was successfully converted to ethanol. These achieved results were promising for further study.

  19. Phospholipid polymer-based antibody immobilization for cell rolling surfaces in stem cell purification system.

    Science.gov (United States)

    Mahara, Atsushi; Chen, Hao; Ishihara, Kazuhiko; Yamaoka, Tetsuji

    2014-01-01

    We previously developed an antibody-conjugated cell rolling column that successfully separates stem cell subpopulations depending on the cell surface marker density, but a large amount of the injected cells were retained in the column because of non-specific interactions. In this study, an amphiphilic copolymer, poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (nBMA)-co-N-vinyl formamide (NVf)], with phospholipid polar side groups was designed as a novel antibody-immobilizing modifier. The formamide groups in NVf units were converted to active maleimide groups. A plastic flow microfluidic chamber was coated with the copolymers, and a reduced anti-CD90 antibody was immobilized. The adipose tissue-derived stem cells isolated from the rat were injected into the flow chamber, and their rolling behavior was observed under a microscope with a high-speed camera. Non-specific cell adhesion was reduced strongly by means of this immobilization method because of the MPC unit, resulting in a high percentage of rolling cells. These results demonstrate that a surface coated with phospholipid polar groups can be used in an effective stem cell separation system based on the cell rolling process.

  20. Hydrophilic PCU scaffolds prepared by grafting PEGMA and immobilizing gelatin to enhance cell adhesion and proliferation

    International Nuclear Information System (INIS)

    Gelatin contains many functional motifs which can modulate cell specific adhesion, so we modified polycarbonate urethane (PCU) scaffold surface by immobilization of gelatin. PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatins onto the surface of aminated PCU scaffolds. To increase the immobilization amount of gelatin, poly(ethylene glycol) methacrylate (PEGMA) was grafted onto PCU scaffolds by surface initiated atom transfer radical polymerization. Then, following amination and immobilization, PCU-g-PEGMA-g-gelatin scaffolds were obtained. Both modified scaffolds were characterized by chemical and biological methods. After immobilization of gelatin, the microfiber surface became rough, but the original morphology of scaffolds was maintained successfully. PCU-g-PEGMA-g-gelatin scaffolds were more hydrophilic than PCU-g-gelatin scaffolds. Because hydrophilic PEGMA and gelatin were grafted and immobilized onto the surface, the PCU-g-PEGMA-g-gelatin scaffolds showed low platelet adhesion, perfect anti-hemolytic activity and excellent cell growth and proliferation capacity. It could be envisioned that PCU-g-PEGMA-g-gelatin scaffolds might have potential applications in tissue engineering artificial scaffolds. - Graphical abstract: PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatin onto the surface of aminated PCU scaffolds (method a). To increase the immobilization amount of gelatin, PEGMAs were grafted onto the scaffold surface by SI-ATRP. PCU-g-PEGMA-g-gelatin scaffolds were prepared by method b. The gelatin modified scaffolds exhibited high hydrophilicity, low platelet adhesion, perfect anti-hemolytic activity, and excellent cell adhesion and proliferation capacity. They might have potential applications as tissue engineering scaffolds for artificial blood vessels. - Highlights: • Hydrophilic scaffolds were prepared by grafting PEGMA and immobilization of gelatins. • Grafting PEGMA enhanced the immobilization amount of gelatin

  1. Hydrophilic PCU scaffolds prepared by grafting PEGMA and immobilizing gelatin to enhance cell adhesion and proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Changcan; Yuan, Wenjie; Khan, Musammir; Li, Qian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Feng, Yakai, E-mail: yakaifeng@tju.edu.cn [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin) Tianjin 300072 (China); Yao, Fanglian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Zhang, Wencheng, E-mail: wenchengzhang@yahoo.com [Department of Physiology and Pathophysiology, Logistics University of Chinese People' s Armed Police Force, Tianjin 300162 (China)

    2015-05-01

    Gelatin contains many functional motifs which can modulate cell specific adhesion, so we modified polycarbonate urethane (PCU) scaffold surface by immobilization of gelatin. PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatins onto the surface of aminated PCU scaffolds. To increase the immobilization amount of gelatin, poly(ethylene glycol) methacrylate (PEGMA) was grafted onto PCU scaffolds by surface initiated atom transfer radical polymerization. Then, following amination and immobilization, PCU-g-PEGMA-g-gelatin scaffolds were obtained. Both modified scaffolds were characterized by chemical and biological methods. After immobilization of gelatin, the microfiber surface became rough, but the original morphology of scaffolds was maintained successfully. PCU-g-PEGMA-g-gelatin scaffolds were more hydrophilic than PCU-g-gelatin scaffolds. Because hydrophilic PEGMA and gelatin were grafted and immobilized onto the surface, the PCU-g-PEGMA-g-gelatin scaffolds showed low platelet adhesion, perfect anti-hemolytic activity and excellent cell growth and proliferation capacity. It could be envisioned that PCU-g-PEGMA-g-gelatin scaffolds might have potential applications in tissue engineering artificial scaffolds. - Graphical abstract: PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatin onto the surface of aminated PCU scaffolds (method a). To increase the immobilization amount of gelatin, PEGMAs were grafted onto the scaffold surface by SI-ATRP. PCU-g-PEGMA-g-gelatin scaffolds were prepared by method b. The gelatin modified scaffolds exhibited high hydrophilicity, low platelet adhesion, perfect anti-hemolytic activity, and excellent cell adhesion and proliferation capacity. They might have potential applications as tissue engineering scaffolds for artificial blood vessels. - Highlights: • Hydrophilic scaffolds were prepared by grafting PEGMA and immobilization of gelatins. • Grafting PEGMA enhanced the immobilization amount of gelatin

  2. Combinational Effect of Cell Adhesion Biomolecules and Their Immobilized Polymer Property to Enhance Cell-Selective Adhesion

    Directory of Open Access Journals (Sweden)

    Rio Kurimoto

    2016-01-01

    Full Text Available Although surface immobilization of medical devices with bioactive molecules is one of the most widely used strategies to improve biocompatibility, the physicochemical properties of the biomaterials significantly impact the activity of the immobilized molecules. Herein we investigate the combinational effects of cell-selective biomolecules and the hydrophobicity/hydrophilicity of the polymeric substrate on selective adhesion of endothelial cells (ECs, fibroblasts (FBs, and smooth muscle cells (SMCs. To control the polymeric substrate, biomolecules are immobilized on thermoresponsive poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide (poly(NIPAAm-co-CIPAAm-grafted glass surfaces. By switching the molecular conformation of the biomolecule-immobilized polymers, the cell-selective adhesion performances are evaluated. In case of RGDS (Arg-Gly-Asp-Ser peptide-immobilized surfaces, all cell types adhere well regardless of the surface hydrophobicity. On the other hand, a tri-Arg-immobilized surface exhibits FB-selectivity when the surface is hydrophilic. Additionally, a tri-Ile-immobilized surface exhibits EC-selective cell adhesion when the surface is hydrophobic. We believe that the proposed concept, which is used to investigate the biomolecule-immobilized surface combination, is important to produce new biomaterials, which are highly demanded for medical implants and tissue engineering.

  3. Bioproduction of usnic acid from acetate by kaolinite immobilized cells of Cladonia substellata Vain.

    Directory of Open Access Journals (Sweden)

    Eugenia C. Pereira

    2014-02-01

    Full Text Available Cells of the lichen Cladonia substellata, immobilized in kaolinite and supplied with acetate, produce at room temperature large amounts of usnic acid which can be recovered from the washing solution.

  4. Comparative analysis of hydrogen-producing bacteria and its immobilized cells for characteristics of hydrogen production

    Institute of Scientific and Technical Information of China (English)

    王相晶; 任南琪; 向文胜; 王爱杰; 林明; 郭婉茜

    2003-01-01

    A strain of hydrogen producing bacteria was immobilized by polyvinyl alcohol-boric acid method,with the addition of a small amount of calcium alginate. The immobilized cells were insensitive to the presence of traces of O2. Moreover, the immobilized cells increased both the evolution rate and the yield of hydrogen production. Batch experiments with a medium containing 10 g/L glucose demonstrated the yields of hydrogen production by the immobilized and free cells were 2.14 mol/mol glucose and 1.69 mol/mol glucose, respectively.In continuous cultures atmedium retention time of 2. 0 h, the yield and the evolution rate of hydrogen producmedium retention time of 6. 0 h, the yield and the evolution rate of hydrogen production by free cells were only 1.75 mol/mol glucose and 362.9ml/(L·h),respectively.

  5. Analysis of secondary cells with lithium anodes and immobilized fused-salt electrolytes

    Science.gov (United States)

    Cairns, E. J.; Rogers, G. L.; Shimotake, H.

    1969-01-01

    Secondary cells with liquid lithium anodes, liquid bismuth or tellurium cathodes, and fused lithium halide electrolytes immobilized as rigid pastes operate between 380 and 485 degrees. Applications include power sources in space, military vehicle propulsion and special commercial vehicle propulsion.

  6. ENHANCED PRODUCTION OF PECTINOLYTIC ENZYMES FROM IMMOBILIZED CELLS OF MIXED ASPERGILLUS SPECIES

    Directory of Open Access Journals (Sweden)

    Shruti Singh

    2012-12-01

    Full Text Available The cells of isolated mixed culture of Aspergillus fumigatus and Aspergillus sydowii were immobilized in calcium alginate beads. Studies were carried out on different parameters like alginate concentration, incubation time and bead inoculum which affects the productivity and stability of the immobilized system. The best enzymatic activities were obtained with 3% alginate concentration, 48h of incubation time and 200 beads/flask of inoculum. Optimization of these factors causes an increase in enzymatic activities and the possibility of semicontinuous cultivation. Immobilized cells could be reused in five successive reaction cycles with a slight decrease in activities.

  7. Microwave-synthesized magnetic chitosan microparticles for the immobilization of yeast cells.

    Science.gov (United States)

    Safarik, Ivo; Pospiskova, Kristyna; Maderova, Zdenka; Baldikova, Eva; Horska, Katerina; Safarikova, Mirka

    2015-01-01

    An extremely simple procedure has been developed for the immobilization of Saccharomyces cerevisiae cells on magnetic chitosan microparticles. The magnetic carrier was prepared using an inexpensive, simple, rapid, one-pot process, based on the microwave irradiation of chitosan and ferrous sulphate at high pH. Immobilized yeast cells have been used for sucrose hydrolysis, hydrogen peroxide decomposition and the adsorption of selected dyes. PMID:24753015

  8. Recent insights into the cell immobilization technology applied for dark fermentative hydrogen production.

    Science.gov (United States)

    Kumar, Gopalakrishnan; Mudhoo, Ackmez; Sivagurunathan, Periyasamy; Nagarajan, Dillirani; Ghimire, Anish; Lay, Chyi-How; Lin, Chiu-Yue; Lee, Duu-Jong; Chang, Jo-Shu

    2016-11-01

    The contribution and insights of the immobilization technology in the recent years with regards to the generation of (bio)hydrogen via dark fermentation have been reviewed. The types of immobilization practices, such as entrapment, encapsulation and adsorption, are discussed. Materials and carriers used for cell immobilization are also comprehensively surveyed. New development of nano-based immobilization and nano-materials has been highlighted pertaining to the specific subject of this review. The microorganisms and the type of carbon sources applied in the dark hydrogen fermentation are also discussed and summarized. In addition, the essential components of process operation and reactor configuration using immobilized microbial cultures in the design of varieties of bioreactors (such as fixed bed reactor, CSTR and UASB) are spotlighted. Finally, suggestions and future directions of this field are provided to assist the development of efficient, economical and sustainable hydrogen production technologies.

  9. Engineering cholesterol-based fibers for antibody immobilization and cell capture

    Science.gov (United States)

    Cohn, Celine

    In 2015, the United States is expected to have nearly 600,000 deaths attributed to cancer. Of these 600,000 deaths, 90% will be a direct result of cancer metastasis, the spread of cancer throughout the body. During cancer metastasis, circulating tumor cells (CTCs) are shed from primary tumors and migrate through bodily fluids, establishing secondary cancer sites. As cancer metastasis is incredibly lethal, there is a growing emphasis on developing "liquid biopsies" that can screen peripheral blood, search for and identify CTCs. One popular method for capturing CTCs is the use of a detection platform with antibodies specifically suited to recognize and capture cancer cells. These antibodies are immobilized onto the platform and can then bind and capture cells of interest. However, current means to immobilize antibodies often leave them with drastically reduced function. The antibodies are left poorly suited for cell capture, resulting in low cell capture efficiencies. This body of work investigates the use of lipid-based fibers to immobilize proteins in a way that retains protein function, ultimately leading to increased cell capture efficiencies. The resulting increased efficiencies are thought to arise from the retained three-dimensional structure of the protein as well as having a complete coating of the material surface with antibodies that are capable of interacting with their antigens. It is possible to electrospin cholesterol-based fibers that are similar in design to the natural cell membrane, providing proteins a more natural setting during immobilization. Such fibers have been produced from cholesterol-based cholesteryl succinyl silane (CSS). These fibers have previously illustrated a keen aptitude for retaining protein function and increasing cell capture. Herein the work focuses on three key concepts. First, a model is developed to understand the immobilization mechanism used by electrospun CSS fibers. The antibody immobilization and cell capturing

  10. OPTIMIZATION OF XYLANASE PRODUCTION FROM FREE AND IMMOBILIZED CELLS OF FUSARIUM SOLANI F7

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    Vijai Kumar Gupta

    2009-08-01

    Full Text Available The aim of the present investigation was to characterize a xylanase-producing Fusarium solani isolate and to optimize cultural conditions for xylanase enzyme production from free and immobilized cells. Screening of Fusarium solani isolate was based on the diameter of the clear zone formation in oat spelt xylan agar plates. Fusarium solani isolate F7 was selected and optimized for xylanase enzyme production using cheaper substrates such as wheat straw, rice straw, rice bran, and wood husk. Maximum enzyme activity was observed in wheat straw (78.32 U ml-1 for free cells and 94.68 U ml-1 for immobilized cells. Optimum pH and temperature for xylanase activity were found to be 5.5 and 30°C at 3% substrate concentration for free cells and 5.0 and 30°C at 3% substrate concentration for immobilized cells. In the purification step, 75% ammonium sulphate saturation was found to be suitable, giving maximum xylanase activity. Production of xylanase was greater from immobilized cells than from free cells. Purified xylanase from free cells yielded a single band with a molecular weight of 89kDa, while it was 92.8kDa for immobilized cells. The use of wheat straw as a major carbon source is particularly valuable, because oat spelt xylan is very expensive. The Fusarium solani F7 isolate proved to be a promising microorganism for xylanase production.

  11. [Study on immobilized cells for producing alpha-amylase by using polyving alcohol as the carrier(II): The effect of fermentating conditions on the ability producing alpha-amylase of the cells immobilized with polyving alcohol as the corrier and continuous fermentation of the immobilized cells in CSTR].

    Science.gov (United States)

    Liu, Z; Wang, J; Li, Z

    1998-03-01

    The effects of fermentating conditions on the ability of immobilized cells with PVA as carrier for producing alpha-amylase were studied. The continuous fermentation with the immobilized cells were tested in continuous flow stirred tank reactor (CSTR). The results showed that the adaptability of the immobilized Bacillus substilis to pH increased after immobilization. In CSTR, the immobilized cells can be fermentated continuously for 360 hrs and the activity of alpha-amylase can be kept on the level of about 170 u/ml.

  12. Ethanol fermentation of molasses by Saccharomyces cerevisiae cells immobilized onto sugar beet pulp

    Directory of Open Access Journals (Sweden)

    Vučurović Vesna M.

    2012-01-01

    Full Text Available Natural adhesion of Saccharomyces cerevisiae onto sugar beet pulp (SBP is a very simple and cheap immobilization method for retaining high cells density in the ethanol fermentation system. In the present study, yeast cells were immobilized by adhesion onto SBP suspended in the synthetic culture media under different conditions such as: glucose concentration (100, 120 and 150 g/l, inoculum concentration (5, 10 and 15 g/l dry mass and temperature (25, 30, 35 and 40°C. In order to estimate the optimal immobilization conditions the yeast cells retention (R, after each immobilization experiment was analyzed. The highest R value of 0.486 g dry mass yeast /g dry mass SBP was obtained at 30°C, glucose concentration of 150 g/l, and inoculum concentration of 15 g/l. The yeast immobilized under these conditions was used for ethanol fermentation of sugar beet molasses containing 150.2 g/l of reducing sugar. Efficient ethanol fermentation (ethanol concentration of 70.57 g/l, fermentation efficiency 93.98% of sugar beet molasses was achieved using S. cerevisiae immobilized by natural adhesion on SBP. [Projekat Ministarstva nauke Republike Srbije, br. TR-31002

  13. Biodegradation of pesticide profenofos by the free and immobilized cells of Pseudoxanthomonas suwonensis strain HNM.

    Science.gov (United States)

    Talwar, Manjunatha P; Ninnekar, Harichandra Z

    2015-09-01

    Profenofos is an organophosphate pesticide used extensively in agriculture to control pests. A bacterium capable of degrading profenofos was isolated from pesticide-contaminated soil samples and identified as Pseudoxanthomonas suwonensis strain HNM based on its morphological and biochemical characteristics and phylogenetic analysis of 16S rRNA gene sequences. 4-Bromo-2-chlorophenol was identified as a metabolite of profenofos degradation by HPLC and GC-MS analysis. The organism degraded profenofos by hydrolysis to yield 4-bromo-2-chlorophenol which was further utilized as carbon source for growth. The organism utilized various organophosphate pesticides such as temephos, quinalphos, and chloropyrifos as carbon sources. The optimum conditions for degradation of profenofos by P. suwonensis strain HMN were found to be at pH 7 and 30 °C. We have investigated the rate of degradation of profenofos by the free and immobilized cells of P. suwonensis strain HNM in various matrices such as sodium alginate (SA), sodium alginate-polyvinyl alcohol (SA-PVA), and SA-bentonite clay. The rate of degradation of 3 and 6 mM profenofos by the freely suspended cells were compared with that by immobilized cells in batches and semi-continuous with shaken cultures. The SA-bentonite clay-immobilized cells showed higher rate of degradation of 3 and 6 mM profenofos then freely suspended cells and cells immobilized in SA and SA-PVA. The SA-bentonite clay-immobilized cells of P. suwonensis strain HNM could be reused for more than 32 cycles without losing their degradation capacity. Thus, the immobilized cells are more efficient than freely suspended cells for the degradation of organophosphate pesticide contaminated water. PMID:25832924

  14. Biodegradation of pesticide profenofos by the free and immobilized cells of Pseudoxanthomonas suwonensis strain HNM.

    Science.gov (United States)

    Talwar, Manjunatha P; Ninnekar, Harichandra Z

    2015-09-01

    Profenofos is an organophosphate pesticide used extensively in agriculture to control pests. A bacterium capable of degrading profenofos was isolated from pesticide-contaminated soil samples and identified as Pseudoxanthomonas suwonensis strain HNM based on its morphological and biochemical characteristics and phylogenetic analysis of 16S rRNA gene sequences. 4-Bromo-2-chlorophenol was identified as a metabolite of profenofos degradation by HPLC and GC-MS analysis. The organism degraded profenofos by hydrolysis to yield 4-bromo-2-chlorophenol which was further utilized as carbon source for growth. The organism utilized various organophosphate pesticides such as temephos, quinalphos, and chloropyrifos as carbon sources. The optimum conditions for degradation of profenofos by P. suwonensis strain HMN were found to be at pH 7 and 30 °C. We have investigated the rate of degradation of profenofos by the free and immobilized cells of P. suwonensis strain HNM in various matrices such as sodium alginate (SA), sodium alginate-polyvinyl alcohol (SA-PVA), and SA-bentonite clay. The rate of degradation of 3 and 6 mM profenofos by the freely suspended cells were compared with that by immobilized cells in batches and semi-continuous with shaken cultures. The SA-bentonite clay-immobilized cells showed higher rate of degradation of 3 and 6 mM profenofos then freely suspended cells and cells immobilized in SA and SA-PVA. The SA-bentonite clay-immobilized cells of P. suwonensis strain HNM could be reused for more than 32 cycles without losing their degradation capacity. Thus, the immobilized cells are more efficient than freely suspended cells for the degradation of organophosphate pesticide contaminated water.

  15. Simultaneous Removal of Thiophene and Dibenzothiophene by Immobilized Pseudomonas delafieldii R-8 cells

    Institute of Scientific and Technical Information of China (English)

    唐煌; 李强; 王泽龙; 闫道江; 邢建民

    2012-01-01

    Biodesulfurization (BDS) is a promising technology for deep desulfurization. In this work, Pseudomonas delafieldii R-8 cells are immobilized in calcium alginate beads and used for BDS of transportation fuels. It is found that thiophene and dibenzothiophene (DBT) can be simultaneously metabolized by immobilized R-8 cells. The initial sulfur content in the model oil is 300 mg·kg-1 (thiophene " DI3T= 1 " 1). After 10 h of treatment, the thiophene concentration is reduced by 40%, while DBT is reduced by 25%. The utilization rate of thiophene is faster than that of DBT. Moreover, the oil/water ratio of alginate immobilized cells is studied to reduce the water volume in desulfurization systems. Long-term recycling of BDS by alginate immobilized cells is carried out with oil/water ratio at 5 : 1. The immobilized cells are successfully reused over 15 batch cycles. In the last batch, the desulfurization activity remains at least 75% of the first batch.

  16. On chip single-cell separation and immobilization using optical tweezers and thermosensitive hydrogel.

    Science.gov (United States)

    Arai, Fumihito; Ng, Chinaik; Maruyama, Hisataka; Ichikawa, Akihiko; El-Shimy, Haitham; Fukuda, Toshio

    2005-12-01

    A novel approach appropriate for rapid separation and immobilization of a single cell by concomitantly utilizing laser manipulation and locally thermosensitive hydrogelation is proposed in this paper. We employed a single laser beam as optical tweezers for separating a target cell and locating it adjacent to a fabricated, transparent micro heater. Simultaneously, the target cell is immobilized or partially entrapped by heating the thermosensitive hydrogel with the micro heater. The state of the thermosensitive hydrogel can be switched from sol to gel and gel to sol by controlling the temperature through heating and cooling by the micro heater. After other unwanted cells are removed by the high-speed cleaning flow in the microchannel, the entrapped cell is successfully isolated. It is possible to collect the immobilized target cell for analysis or culture by switching off the micro heater and releasing the cell from the entrapment. We demonstrated that the proposed approach is feasible for rapid manipulation, immobilization, cleaning, isolation and extraction of a single cell. The experimental results are shown here. PMID:16286972

  17. Nitrilase-catalysed conversion of acrylonitrile by free and immobilized cells of Streptomyces sp.

    Indian Academy of Sciences (India)

    V K Nigam; A K Khandelwal; R K Gothwal; M K Mohan; B Choudhury; A S Vidyarthi; P Ghosh

    2009-03-01

    The biotransformation of acrylonitrile was investigated using thermophilic nitrilase produced from a new isolate Streptomyces sp. MTCC 7546 in both the free and immobilized state. Under optimal conditions, the enzyme converts nitriles to acids without the formation of amides. The whole cells of the isolate were immobilized in agar-agar and the beads so formed were evaluated for 25 cycles at 50°C. The enzyme showed a little loss of activity during reuse. Seventy-one per cent of 0.5 M acrylonitrile was converted to acid at 6 h of incubation at a very low density of immobilized cells, while 100% conversion was observed at 3 h by free cells.

  18. Production of fructooligosaccharides and b-fructofuranosidase by batch and repeated batch fermentation with immobilized cells of Penicillium expansum

    OpenAIRE

    Mussatto, Solange I.; Prata, Margarida Barros; Rodrigues, L. R.; Teixeira, J. A.

    2012-01-01

    The production of fructooligosaccharides (FOS) and b-fructofuranosidase (FFase) by immobilized cells of Penicillium expansum was evaluated. In an initial stage, different low-cost materials including synthetic fiber, polyurethane foam, stainless steel sponge, loofah sponge, and cork oak were tested as carrier for the fungus immobilization. Additionally, the influence of the inoculum age (1 or 3 weeks) on cells immobilization, FOS and FFase production was also verified....

  19. [Comparison of fibroblastic cell compatibility of type I collagen-immobilized titanium between electrodeposition and immersion].

    Science.gov (United States)

    Kyuragi, Takeru

    2014-03-01

    Titanium is widely used for medical implants. While many techniques for surface modification have been studied for optimizing its biocompatibility with hard tissues, little work has been undertaken to explore ways of maximizing its biocompatibility with soft tissues. We investigated cell attachment to titanium surfaces modified with bovine Type I collagen immobilized by either electrodeposition or a conventional immersion technique. The apparent thickness and durability of the immobilized collagen layer were evaluated prior to incubation of the collagen-immobilized titanium surfaces with NIH/3T3 mouse embryonic fibroblasts. The initial cell attachment and expression of actin and vinculin were evaluated. We determined that the immobilized collagen layer was much thicker and more durable when placed using the electrodeposition technique than the immersion technique. Both protocols produced materials that promoted better cell attachment, growth and structural protein expression than titanium alone. However, electrodeposition was ultimately superior to immersion because it is quicker to perform and produces a more durable collagen coating. We conclude that electrodeposition is an effective technique for immobilizing type I collagen on titanium surfaces, thus improving their cytocompatibility with fibroblasts. PMID:24812763

  20. Enzymatic production of atranorin: a component of the oak moss absolute by immobilized lichen cells.

    Science.gov (United States)

    Vicente, C; Fontaniella, B; Millanes, A M; Sebastián, B; Legaz, M E

    2003-04-01

    Cells of the lichen, Evernia prunastri, immobilized in calcium alginate were able to produce the depside atranorin from acetate. The synthesis of the depside was enhanced by molecular oxygen and NADH. This enhancement suggested the participation of an oxidase and an alcohol dehydrogenase to produce an aldehyde-substituted phenolic acid, hematommic acid, as the most probable precursor of atranorin. The participation of both enzymes was confirmed by loading immobilized cells with sodium azide, an inhibitor of several metallo-oxidases, and pyrazole, an inhibitor of alcohol dehydrogenase, which impeded atranorin production and accumulated beta-methyl orsellinate (after azide loading) or its alcohol derivative (after pirazole treatment). PMID:18494879

  1. Immobilization of cells with nitrilase activity from a thermophilic bacterial strain.

    Science.gov (United States)

    Kabaivanova, L; Dobreva, E; Dimitrov, P; Emanuilova, E

    2005-01-01

    Cells of the moderately thermophilic Bacillus sp. UG-5B strain, producing nitrilase (EC3.5.5.1), which converts nitriles directly to the corresponding acid and ammonia, were immobilized using different types of matrices and techniques. A variety of sol-gel silica hybrids were tested for entrapment and adsorption of bacterial cells as well as chemical binding on polysulphone membranes. Activation of the matrix surface with formaldehyde led to an increase in immobilization efficiency and operational stability of the biocatalysts. Among the supports screened, membranes gave the best results for enzyme activity and especially operational stability, with retention of 100% activity after eight reaction cycles.

  2. Immobilization of anode-attached microbes in a microbial fuel cell.

    KAUST Repository

    Wagner, Rachel C

    2012-01-03

    Current-generating (exoelectrogenic) bacteria in bioelectrochemical systems (BESs) may not be culturable using standard in vitro agar-plating techniques, making isolation of new microbes a challenge. More in vivo like conditions are needed where bacteria can be grown and directly isolated on an electrode. While colonies can be developed from single cells on an electrode, the cells must be immobilized after being placed on the surface. Here we present a proof-of-concept immobilization approach that allows exoelectrogenic activity of cells on an electrode based on applying a layer of latex to hold bacteria on surfaces. The effectiveness of this procedure to immobilize particles was first demonstrated using fluorescent microspheres as bacterial analogs. The latex coating was then shown to not substantially affect the exoelectrogenic activity of well-developed anode biofilms in two different systems. A single layer of airbrushed coating did not reduce the voltage produced by a biofilm in a microbial fuel cell (MFC), and more easily applied dip-and-blot coating reduced voltage by only 11% in a microbial electrolysis cell (MEC). This latex immobilization procedure will enable future testing of single cells for exoelectrogenic activity on electrodes in BESs.

  3. Hexavalent chromium reduction by immobilized cells of Bacillus sphaericus AND 303

    Directory of Open Access Journals (Sweden)

    Arundhati Pal

    2013-06-01

    Full Text Available Bacillus sphaericus AND 303, a Cr(VI-resistant and reducing bacterium reported from serpentine outcrops of Andaman was evaluated for Cr(VI reduction using immobilized cells under batch culture. Screening of inert matrices for entrapment of whole cells indicated that polyvinyl alchohol-alginate was the most effective one reducing 87.5% of 20 µM Cr(VI in 24 h. The rate of chromate reduction was dependent on initial Cr(VI and biomass concentrations. The PVA cell beads were recycled three times without cell leakage and disintegration. The reduction efficiency was improved in the presence of glucose and glycerol as electron donors leading to complete reduction. However, the presence of additional metal ions was inhibitory to Cr(VI reduction. It could be emphasized that PVA-alginate immobilized cells of B. sphaericus AND 303 could be used as a continuous bioprocess in treating Cr(VI contaminated effluents.

  4. Removal of Cadmium and Zinc from Soil using Immobilized Cell of Biosurfactant Producing Bacteria

    Directory of Open Access Journals (Sweden)

    Charoon Sarin

    2010-07-01

    Full Text Available Immobilized biosurfactant producing bacteria (Bacillus subtilis TP8 and Pseudomonas fluorescens G7 were assessed for survival in heavy metal contaminated soil and for their ability to remove cadmium and zinc from contaminated soil. P. fluorescens G7 was considered to be a good candidate for bioremediation of heavy metals because of its high minimum inhibitory concentrations (MIC for each heavy metal and because of the obviously increased numbers of cell surviving after incubation in the heavy metal contaminated soil up to 4 weeks. The results of soil remediation showed that approximately 19% of Zn and 16.7% of Cd could be removed by this immobilized biosurfactant producing bacteria after incubation for 2 weeks. The results confirm the potential applicability of the immobilized biosurfactant producing bacteria for heavy metal bioremediation.

  5. Chitin hydrolysis assisted by cell wall degrading enzymes immobilized of Thichoderma asperellum on totally cinnamoylated D-sorbitol beads

    International Nuclear Information System (INIS)

    In this study, cell wall degrading enzymes produced by Thrichoderma asperellum (TCWDE) were immobilized on totally cinnamoylated D-sorbitol (TCNSO) beads and used for chitin hydrolysis. In order to optimize immobilization efficiency, the reaction time was varied from 2 to 12 h and reactions were conducted in the presence or absence of Na2SO4. Immobilized enzymes were analysed concerning to thermal and operational stability. Immobilization in presence of Na2SO4 was 54% more efficient than immobilization in absence of salt. After optimization, 32% of the total enzyme offered was immobilized, with 100% of bounding efficiency, measured as the relation between protein and enzyme immobilized. Free and TCNSO–TCWDE presented very similar kinetics with maximum hydrolysis reached at 90 min of reaction. Thermal stability of both free and TCNSO–TCWDE was similar, with losses in activity after 55 °C. Moreover, free and TCNSO–TCWDE retained 100% activity after 3 h incubation at 55 °C. TCNSO–TCWDE were used in a bath-wise reactor during 14 cycles, producing 1825 μg of N-acetylglucosamine (NAG) maintaining 83% of initial activity. - Highlights: • TCWDE immobilized on TCNSO, a support with highly hydrophobic character • New immobilization strategy for immobilization on a hydrophobic support • TCNSO–TCWDE were retained during washes and during incubation at 55 °C for 3 h

  6. Chitin hydrolysis assisted by cell wall degrading enzymes immobilized of Thichoderma asperellum on totally cinnamoylated D-sorbitol beads

    Energy Technology Data Exchange (ETDEWEB)

    Fernandes, Kátia F., E-mail: katia@icb.ufg.br [Departamento de Bioquímica e Biologia Molecular, Instituo de Ciências Biológicas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970 Goiânia, GO (Brazil); Grupo de Química de Carbohidratos y Biotecnología de Alimentos (QCBA), Departamento de Química Orgánica, Facultad de Química, Universidad de Murcia, E-30100 Espinardo, Murcia (Spain); Cortijo-Triviño, David [Grupo de Química de Carbohidratos y Biotecnología de Alimentos (QCBA), Departamento de Química Orgánica, Facultad de Química, Universidad de Murcia, E-30100 Espinardo, Murcia (Spain); Batista, Karla A.; Ulhoa, Cirano J. [Departamento de Bioquímica e Biologia Molecular, Instituo de Ciências Biológicas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970 Goiânia, GO (Brazil); García-Ruiz, Pedro A. [Grupo de Química de Carbohidratos y Biotecnología de Alimentos (QCBA), Departamento de Química Orgánica, Facultad de Química, Universidad de Murcia, E-30100 Espinardo, Murcia (Spain)

    2013-07-01

    In this study, cell wall degrading enzymes produced by Thrichoderma asperellum (TCWDE) were immobilized on totally cinnamoylated D-sorbitol (TCNSO) beads and used for chitin hydrolysis. In order to optimize immobilization efficiency, the reaction time was varied from 2 to 12 h and reactions were conducted in the presence or absence of Na{sub 2}SO{sub 4}. Immobilized enzymes were analysed concerning to thermal and operational stability. Immobilization in presence of Na{sub 2}SO{sub 4} was 54% more efficient than immobilization in absence of salt. After optimization, 32% of the total enzyme offered was immobilized, with 100% of bounding efficiency, measured as the relation between protein and enzyme immobilized. Free and TCNSO–TCWDE presented very similar kinetics with maximum hydrolysis reached at 90 min of reaction. Thermal stability of both free and TCNSO–TCWDE was similar, with losses in activity after 55 °C. Moreover, free and TCNSO–TCWDE retained 100% activity after 3 h incubation at 55 °C. TCNSO–TCWDE were used in a bath-wise reactor during 14 cycles, producing 1825 μg of N-acetylglucosamine (NAG) maintaining 83% of initial activity. - Highlights: • TCWDE immobilized on TCNSO, a support with highly hydrophobic character • New immobilization strategy for immobilization on a hydrophobic support • TCNSO–TCWDE were retained during washes and during incubation at 55 °C for 3 h.

  7. Continuous ethanol production using immobilized yeast cells entrapped in loofa-reinforced alginate carriers

    Directory of Open Access Journals (Sweden)

    Phoowit Bangrak

    2011-06-01

    Full Text Available A culture of Saccharomyces cerevisiae M30 entrapped in loofa-reinforced alginate was used for continuous ethanol fermentation in a packed-bed reactor with initial sugar concentrations of 200-248 g/L. Maximum ethanol productivity of 11.5 g/(L·h was obtained at an ethanol concentration of 57.4 g/L, an initial sugar concentration of 220 g/L and a dilution rate (D of 0.2 h-1. However, a maximum ethanol concentration of 82.1 g/L (productivity of 9.0 g/(L·h was obtained at a D of 0.11 h-1. Ethanol productivity in the continuous culture was 6-8-fold higher than that in the batch culture. Due to the developed carrier's high biocompatibility, high porosity, and good mechanical strength, advantages such as cell regeneration, reusability, altered mechanical strength, and high capacity to trap active cells in the reactor were achieved in this study. The immobilized cell reactor was successfully operated for 30 days without any loss in ethanol productivity. The average conversion yield was 0.43-0.45 throughout the entire operation, with an immobilization yield of 47.5%. The final total cell concentration in the reactor was 37.3 g/L (17.7 g/L immobilized cells and 19.6 g/L suspended cells. The concentration of suspended cells in the effluent was 0.8 g/L.

  8. Immobilization of Escherichia coli Cells Containing Aspartase Activity with Polyurethane and Its Application for l-Aspartic Acid Production

    OpenAIRE

    Fusee, Murray C.; Swann, Wayne E.; Calton, Gary J.

    1981-01-01

    Whole cells of Escherichia coli containing aspartase activity were immobilized by mixing a cell suspension with a liquid isocyanate-capped polyurethane prepolymer (Hypol). The immobilized cell preparation was used to convert ammonium fumarate to l-aspartic acid. Properties of the immobilized E. coli cells containing aspartase were investigated with a batch reactor. A 1.67-fold increase in the l-aspartic acid production rate was observed at 37°C as compared to 25°C operating temperature. The p...

  9. Gibberellic acid production by free and immobilized cells in different culture systems.

    Science.gov (United States)

    Durán-Páramo, Enrique; Molina-Jiménez, Héctor; Brito-Arias, Marco A; Robles-Martínez, Fabián

    2004-01-01

    Gibberellic acid production was studied in different fermentation systems. Free and immobilized cells of Gibberella fujikuroi cultures in shake-flask, stirred and fixed-bed reactors were evaluated for the production of gibberellic acid (GA3). Gibberellic acid production with free cells cultured in a stirred reactor reached 0.206 g/L and a yield of 0.078 g of GA3/g biomass.

  10. Enhanced dibenzothiophene biodesulfurization by immobilized cells of Brevibacterium lutescens in n-octane-water biphasic system.

    Science.gov (United States)

    Dai, Yong; Shao, Rong; Qi, Gang; Ding, Bin-Bin

    2014-11-01

    In this study, it was the first report that the Brevibacterium lutescens CCZU12-1 was employed as a sulfur removing bacteria. Using dibenzothiophene (DBT) as the sole sulfur source, B. lutescens could selectively degrade DBT into 2-hydroxybiphenyl (2-HBP) via the "4S" pathway. In the basal salt medium (BSM) supplemented with 0.25 mM DBT and 0.5 g/L Tween-80, high desulfurization rate (100 %) was obtained by growth cells after 60 h. Furthermore, the n-octane-water (10:90, v/v) biphasic system was built for the biodesulfurization by resting cells. Moreover, a combination of magnetic nano Fe3O4 particles with calcium alginate immobilization was used for enhancing biodesulfurization. In this n-octane-water biphasic system, immobilized B. lutescens cells could be reused for not less than four times. Therefore, B. lutescens CCZU12-1 shows high potential in the biodesulfurization. PMID:25173674

  11. Bioleaching of Primary Nickel Ore Using Acidithiobacillus ferrooxidans LR Cells Immobilized in Glass Beads

    Directory of Open Access Journals (Sweden)

    Ellen Cristine Giese

    2015-06-01

    Full Text Available Sulphide minerals are one of the most important sources of value metals. For several years, a large number of hydrometallurgical and biotechnological processes have been developed to leach low-grade sulphide ores and the conditions are well established. However, the management of microorganisms in the bioleaching process is not easy to handle. In this paper, the use of immobilized cells of Acidithiobacillus ferrooxidans LR in glass beads in bioleaching of primary nickel ore was evaluated. The column experiments inoculated with immobilized cells of A. ferrooxidans LR showed the same efficiency than the conventional method using free cells and is promising for application on a larger scale as it ensuring integrity and activity of biomining microorganisms and reduce process costs. DOI: http://dx.doi.org/10.17807/orbital.v7i2.698 

  12. New method for selection of hydrogen peroxide adapted bifidobacteria cells using continuous culture and immobilized cell technology

    Directory of Open Access Journals (Sweden)

    Meile Leo

    2010-07-01

    Full Text Available Abstract Background Oxidative stress can severely compromise viability of bifidobacteria. Exposure of Bifidobacterium cells to oxygen causes accumulation of reactive oxygen species, mainly hydrogen peroxide, leading to cell death. In this study, we tested the suitability of continuous culture under increasing selective pressure combined with immobilized cell technology for the selection of hydrogen peroxide adapted Bifidobacterium cells. Cells of B. longum NCC2705 were immobilized in gellan-xanthan gum gel beads and used to continuously ferment MRS medium containing increasing concentration of H2O2 from 0 to 130 ppm. Results At the beginning of the culture, high cell density of 1013 CFU per litre of reactor was tested. The continuous culture gradually adapted to increasing H2O2 concentrations. However, after increasing the H2O2 concentration to 130 ppm the OD of the culture decreased to 0. Full wash out was prevented by the immobilization of the cells in gel matrix. Hence after stopping the stress, it was possible to re-grow the cells that survived the highest lethal dose of H2O2 and to select two adapted colonies (HPR1 and HPR2 after plating of the culture effluent. In contrast to HPR1, HPR2 showed stable characteristics over at least 70 generations and exhibited also higher tolerance to O2 than non adapted wild type cells. Preliminary characterization of HPR2 was carried out by global genome expression profile analysis. Two genes coding for a protein with unknown function and possessing trans-membrane domains and an ABC-type transporter protein were overexpressed in HPR2 cells compared to wild type cells. Conclusions Our study showed that continuous culture with cell immobilization is a valid approach for selecting cells adapted to hydrogen peroxide. Elucidation of H2O2 adaptation mechanisms in HPR2 could be helpful to develop oxygen resistant bifidobacteria.

  13. Cell culture plastics with immobilized interleukin-4 for monocyte differentiation

    DEFF Research Database (Denmark)

    Hansen, Morten; Hjortø, Gertrud Malene; Met, Ozcan;

    2011-01-01

    Standard cell culture plastic was surface modified by passive adsorption or covalent attachment of interleukin (IL)-4 and investigated for its ability to induce differentiation of human monocytes into mature dendritic cells, a process dose-dependently regulated by IL-4. Covalent attachment of IL-...

  14. Cell culture plastics with immobilized interleukin-4 for monocyte differentiation

    DEFF Research Database (Denmark)

    Hansen, Morten; Hjortø, Gertrud Malene; Met, Özcan;

    2011-01-01

    Standard cell culture plastic was surface modified by passive adsorption or covalent attachment of interleukin (IL)-4 and investigated for its ability to induce differentiation of human monocytes into mature dendritic cells, a process dose-dependently regulated by IL-4. Covalent attachment of IL-4...

  15. Internalization: acute apoptosis of breast cancer cells using herceptin-immobilized gold nanoparticles

    Directory of Open Access Journals (Sweden)

    Rathinaraj P

    2015-02-01

    Full Text Available Pierson Rathinaraj,1 Ahmed M Al-Jumaily,1 Do Sung Huh21Institute of Biomedical Technologies, Auckland University of Technology, Auckland, New Zealand; 2Department of Nano science and Engineering, Inje University, Gimhea, South KoreaAbstract: Herceptin, the monoclonal antibody, was successfully immobilized on gold nanoparticles (GNPs to improve their precise interactions with breast cancer cells (SK-BR3. The mean size of the GNPs (29 nm, as determined by dynamic light scattering, enlarged to 82 nm after herceptin immobilization. The in vitro cell culture experiment indicated that human skin cells (FB proliferated well in the presence of herceptin-conjugated GNP (GNP–Her, while most of the breast cancer cells (SK-BR3 had died. To elucidate the mechanism of cell death, the interaction of breast cancer cells with GNP–Her was tracked by confocal laser scanning microscopy. Consequently, GNP–Her was found to be bound precisely to the membrane of the breast cancer cell, which became almost saturated after 6 hours incubation. This shows that the progression signal of SK-BR3 cells is retarded completely by the precise binding of antibody to the human epidermal growth factor receptor 2 receptor of the breast cancer cell membrane, causing cell death.Keywords: herceptin, gold nanoparticles, SK-BR3 cells, intracellular uptake

  16. Optimizing Immobilized Enzyme Performance in Cell-Free Environments to Produce Liquid Fuels

    Energy Technology Data Exchange (ETDEWEB)

    Belfort, Georges [Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Chemical and Biological Engineering; Grimaldi, Joseph J. [Rensselaer Polytechnic Inst., Troy, NY (United States). Dept. of Chemical and Biological Engineering

    2015-01-27

    Limitations on biofuel production using cell culture (Escherichia coli, Clostridium, Saccharomyces cerevisiae, brown microalgae, blue-green algae and others) include low product (alcohol) concentrations (≤0.2 vol%) due to feedback inhibition, instability of cells, and lack of economical product recovery processes. To overcome these challenges, an alternate simplified biofuel production scheme was tested based on a cell-free immobilized enzyme system. Using this cell free system, we were able to obtain about 2.6 times higher concentrations of iso-butanol using our non-optimized system as compared with live cell systems. This process involved two steps: (i) converts acid to aldehyde using keto-acid decarboxylase (KdcA), and (ii) produces alcohol from aldehyde using alcohol dehydrogenase (ADH) with a cofactor (NADH) conversion from inexpensive formate using a third enzyme, formate dehydrogenase (FDH). To increase stability and conversion efficiency with easy separations, the first two enzymes were immobilized onto methacrylate resin. Fusion proteins of labile KdcA (fKdcA) were expressed to stabilize the covalently immobilized KdcA. Covalently immobilized ADH exhibited long-term stability and efficient conversion of aldehyde to alcohol over multiple batch cycles without fusions. High conversion rates and low protein leaching were achieved by covalent immobilization of enzymes on methacrylate resin. The complete reaction scheme was demonstrated by immobilizing both ADH and fKdcA and using FDH free in solution. The new system without in situ removal of isobutanol achieved a 55% conversion of ketoisovaleric acid to isobutanol at a concentration of 0.5 % (v/v). Further increases in titer will require continuous removal of the isobutanol using our novel brush membrane system that exhibits a 1.5 fold increase in the separation factor of isobutanol from water versus that obtained for commercial silicone rubber membranes. These bio-inspired brush membranes are based on the

  17. A novel cell weighing method based on the minimum immobilization pressure for biological applications

    International Nuclear Information System (INIS)

    A novel weighing method for cells with spherical and other regular shapes is proposed in this paper. In this method, the relationship between the cell mass and the minimum aspiration pressure to immobilize the cell (referred to as minimum immobilization pressure) is derived for the first time according to static theory. Based on this relationship, a robotic cell weighing process is established using a traditional micro-injection system. Experimental results on porcine oocytes demonstrate that the proposed method is able to weigh cells at an average speed of 16.3 s/cell and with a success rate of more than 90%. The derived cell mass and density are in accordance with those reported in other published results. The experimental results also demonstrated that this method is able to detect less than 1% variation of the porcine oocyte mass quantitatively. It can be conducted by a pair of traditional micropipettes and a commercial pneumatic micro-injection system, and is expected to perform robotic operation on batch cells. At present, the minimum resolution of the proposed method for measuring the cell mass can be 1.25 × 10−15 kg. Above advantages make it very appropriate for quantifying the amount of the materials injected into or moved out of the cells in the biological applications, such as nuclear enucleations and embryo microinjections

  18. A novel cell weighing method based on the minimum immobilization pressure for biological applications

    Science.gov (United States)

    Zhao, Qili; Shirinzadeh, Bijan; Cui, Maosheng; Sun, Mingzhu; Liu, Yaowei; Zhao, Xin

    2015-07-01

    A novel weighing method for cells with spherical and other regular shapes is proposed in this paper. In this method, the relationship between the cell mass and the minimum aspiration pressure to immobilize the cell (referred to as minimum immobilization pressure) is derived for the first time according to static theory. Based on this relationship, a robotic cell weighing process is established using a traditional micro-injection system. Experimental results on porcine oocytes demonstrate that the proposed method is able to weigh cells at an average speed of 16.3 s/cell and with a success rate of more than 90%. The derived cell mass and density are in accordance with those reported in other published results. The experimental results also demonstrated that this method is able to detect less than 1% variation of the porcine oocyte mass quantitatively. It can be conducted by a pair of traditional micropipettes and a commercial pneumatic micro-injection system, and is expected to perform robotic operation on batch cells. At present, the minimum resolution of the proposed method for measuring the cell mass can be 1.25 × 10-15 kg. Above advantages make it very appropriate for quantifying the amount of the materials injected into or moved out of the cells in the biological applications, such as nuclear enucleations and embryo microinjections.

  19. A novel cell weighing method based on the minimum immobilization pressure for biological applications

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Qili [Robotics and Mechatronics Research Laboratory, Department of Mechanical and Aerospace Engineering, Monash University, Clayton 3800 (Australia); Institute of Robotics and Automatic Information System, Nankai University, Tianjin 300071 (China); Shirinzadeh, Bijan [Robotics and Mechatronics Research Laboratory, Department of Mechanical and Aerospace Engineering, Monash University, Clayton 3800 (Australia); Cui, Maosheng [Biotechnology Lab of Animal Reproduction, Tianjin Animal Sciences, Tianjin 300112 (China); Sun, Mingzhu; Liu, Yaowei; Zhao, Xin, E-mail: zhaoxin@nankai.edu.cn [Institute of Robotics and Automatic Information System, Nankai University, Tianjin 300071 (China)

    2015-07-28

    A novel weighing method for cells with spherical and other regular shapes is proposed in this paper. In this method, the relationship between the cell mass and the minimum aspiration pressure to immobilize the cell (referred to as minimum immobilization pressure) is derived for the first time according to static theory. Based on this relationship, a robotic cell weighing process is established using a traditional micro-injection system. Experimental results on porcine oocytes demonstrate that the proposed method is able to weigh cells at an average speed of 16.3 s/cell and with a success rate of more than 90%. The derived cell mass and density are in accordance with those reported in other published results. The experimental results also demonstrated that this method is able to detect less than 1% variation of the porcine oocyte mass quantitatively. It can be conducted by a pair of traditional micropipettes and a commercial pneumatic micro-injection system, and is expected to perform robotic operation on batch cells. At present, the minimum resolution of the proposed method for measuring the cell mass can be 1.25 × 10{sup −15 }kg. Above advantages make it very appropriate for quantifying the amount of the materials injected into or moved out of the cells in the biological applications, such as nuclear enucleations and embryo microinjections.

  20. Immobilization of RGD Peptides onto Decellularized Valve Scaffolds to Promote Cell Adhesion

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Porcine aortic valves were decellularized with trypsinase/EDTA and Triton-100. With the help of a coupling reagent Sulfo-LC-SPDP, the biological valve scaffolds were immobilized with one of RGD(arginine-glycine-aspartic acid) containing peptides, called GRGDSPC peptide. Myofibroblasts harvested from rats were seeded onto them. Based on the spectra of X-ray photoelectron spectroscopy, we could find conjugation of GRGDSPC peptide and the scaffolds. Cell count by both microscopy and MTT assay showed that myofibroblasts were easier to adhere to the modified scaffolds. It is proved that it is feasible to immobilize RGD peptides onto decellularized valve scaffolds, and effective to promote cell adhesion, which is beneficial for constructing tissue engineering heart valves in vitro.

  1. Continuous Ethanol Production Using Immobilized-Cell/Enzyme Biocatalysts in Fluidized-Bed Bioreactor (FBR)

    Energy Technology Data Exchange (ETDEWEB)

    Nghiem, NP

    2003-11-16

    The immobilized-cell fluidized-bed bioreactor (FBR) was developed at Oak Ridge National Laboratory (ORNL). Previous studies at ORNL using immobilized Zymomonas mobilis in FBR at both laboratory and demonstration scale (4-in-ID by 20-ft-tall) have shown that the system was more than 50 times as productive as industrial benchmarks (batch and fed-batch free cell fermentations for ethanol production from glucose). Economic analysis showed that a continuous process employing the FBR technology to produce ethanol from corn-derived glucose would offer savings of three to six cents per gallon of ethanol compared to a typical batch process. The application of the FBR technology for ethanol production was extended to investigate more complex feedstocks, which included starch and lignocellulosic-derived mixed sugars. Economic analysis and mathematical modeling of the reactor were included in the investigation. This report summarizes the results of these extensive studies.

  2. Production of biofuels from pretreated microalgae biomass by anaerobic fermentation with immobilized Clostridium acetobutylicum cells.

    Science.gov (United States)

    Efremenko, E N; Nikolskaya, A B; Lyagin, I V; Senko, O V; Makhlis, T A; Stepanov, N A; Maslova, O V; Mamedova, F; Varfolomeev, S D

    2012-06-01

    The purpose of this work was to study the possible use of pretreated biomass of various microalgae and cyanobacteria as substrates for acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum cells immobilized into poly(vinyl alcohol) cryogel. To this end, the biochemical composition of photosynthetic microorganisms cultivated under various conditions was studied. The most efficient technique for pretreating microalgal biomass for its subsequent conversion into biofuels appeared to be thermal decomposition at 108 °C. For the first time the maximum productivity of the ABE fermentation in terms of hydrogen (8.5 mmol/L medium/day) was obtained using pretreated biomass of Nannochloropsis sp. Maximum yields of butanol and ethanol were observed with Arthrospira platensis biomass used as the substrate. Immobilized Clostridium cells were demonstrated to be suitable for multiple reuses (for a minimum of five cycles) in ABE fermentation for producing biofuels from pretreated microalgal biomass.

  3. OPTIMIZATION OF XYLANASE PRODUCTION FROM FREE AND IMMOBILIZED CELLS OF FUSARIUM SOLANI F7

    OpenAIRE

    Vijai Kumar Gupta; Rajeeva Gaur; Santosh Kumar Yadava; Nandan Singh Darmwal

    2009-01-01

    The aim of the present investigation was to characterize a xylanase-producing Fusarium solani isolate and to optimize cultural conditions for xylanase enzyme production from free and immobilized cells. Screening of Fusarium solani isolate was based on the diameter of the clear zone formation in oat spelt xylan agar plates. Fusarium solani isolate F7 was selected and optimized for xylanase enzyme production using cheaper substrates such as wheat straw, rice straw, rice bran, and wood husk. Max...

  4. Study of acetic acid production by immobilized acetobacter cells: oxygen transfer

    Energy Technology Data Exchange (ETDEWEB)

    Ghommidh, C.; Navarro, J.M.; Durand, G.

    1982-03-01

    The immobilization of living Acetobacter cells by adsorption onto a large-surface-area ceramic support was studied in a pulsed flow reactor. The high oxygen transfer capability of the reactor enabled acetic acid production rates up to 10.4 g/L/h to be achieved. Using a simple mathematical model incorporating both internal and external mass transfer coefficients, it was shown that oxygen transfer in the microbial film controls the reactor productivity. (Refs. 10).

  5. Bioleaching of Primary Nickel Ore Using Acidithiobacillus ferrooxidans LR Cells Immobilized in Glass Beads

    OpenAIRE

    Ellen Cristine Giese; Patrícia Morgado Vaz

    2015-01-01

    Sulphide minerals are one of the most important sources of value metals. For several years, a large number of hydrometallurgical and biotechnological processes have been developed to leach low-grade sulphide ores and the conditions are well established. However, the management of microorganisms in the bioleaching process is not easy to handle. In this paper, the use of immobilized cells of Acidithiobacillus ferrooxidans LR in glass beads in bioleaching of primary nickel ore was evaluated. The...

  6. The production of sorbitol by permeabilized and immobilized cells of Z. mobilis in sucrose

    Directory of Open Access Journals (Sweden)

    Josiane Alessandra Vignoli

    2006-07-01

    Full Text Available The production of sorbitol by permeabilized and immobilized cells of Zymomonas mobilis in Luffa cylindrica was investigated in sucrose medium. A full 2³ factorial design was used to verify the influence of each factor and its interactions. The cell permeabilization showed a significant and negative effect upon the production of sorbitol, while the time of cultivation and the immobilization process were significant and positive. The results demonstrated that the cell immobilization and the time of cultivation of 36 h presented higher production of sorbitol.Foi investigado a produção de sorbitol em meio de sacarose por células de Z. mobilis permeabilizadas e imobilizadas em Luffa cylindrica. Este trabalho avaliou o efeito da permeabilização de células de Z. mobilis tratadas com cetilmetilamoniobrometo e imobilizadas em Luffa cylindrinca. Um planejamento fatorial completo 2³ foi utilizado para verificar a influência dos fatores e suas interações. A permeabilização da célula mostrou um efeito significante e negativo sobre a produção de sorbitol, enquanto o tempo de cultivo e o processo de imobilização foram significantes e positivos. Os resultados mostraram que a imobilização das células e o tempo de cultivo de 36h forneceram concentração mais elevadas de sorbitol.

  7. Enzymatic Synthesis of Agmatine by Immobilized Escherichia coli Cells with Arginine Decarboxylase Activity

    Institute of Scientific and Technical Information of China (English)

    ZHANG Wei-guo; ZHAO Gen-hai; LIU Jun-zhong; LIU Qian; JIAO Qing-cai

    2011-01-01

    A new method for the enzymatic synthesis of agmatine by immobilized Escherichia coli cells with arginine decarboxylase(ADC)activity was established and a series of optimal reaction conditions was set down.The arginine decarboxylase showed the maximum activity when the pyridoxal phosphate(PLP)concentration was 50 mmol/L,pH=7 and 45 ℃.The arginine decarboxylase exhibited the maximum production efficiency when the substrate concentration was 100 mmol/L and the reaction time was 15 h.It was also observed that the appropriate concentration of Mg2+,especially at 0.5 mmol/L promoted the arginine decarboxylase activity; Mn2+ had little effect on the arginine decarboxylase activity.The inhibition of Cu2+ and Zn2+ to the arginine decarboxylase activity was significant.The immobilized cells were continuously used 6 times and the average conversion rate during the six-time usage was 55.6%.The immobilized cells exhibited favourable operational stability.After optimization,the maximally cumulative amount of agmatine could be up to 20 g/L.In addition,this method can also catalyze D,L-arginine to agmatine,leaving the pure optically D-arginine simultaneously.The method has a very important guiding significance to the enzymatic preparation of agmatine.

  8. Raspberry wine fermentation with suspended and immobilized yeast cells of two strains of Saccharomyces cerevisiae.

    Science.gov (United States)

    Djordjević, Radovan; Gibson, Brian; Sandell, Mari; de Billerbeck, Gustavo M; Bugarski, Branko; Leskošek-Čukalović, Ida; Vunduk, Jovana; Nikićević, Ninoslav; Nedović, Viktor

    2015-01-01

    The objectives of this study were to assess the differences in fermentative behaviour of two different strains of Saccharomyces cerevisiae (EC1118 and RC212) and to determine the differences in composition and sensory properties of raspberry wines fermented with immobilized and suspended yeast cells of both strains at 15 °C. Analyses of aroma compounds, glycerol, acetic acid and ethanol, as well as the kinetics of fermentation and a sensory evaluation of the wines, were performed. All fermentations with immobilized yeast cells had a shorter lag phase and faster utilization of sugars and ethanol production than those fermented with suspended cells. Slower fermentation kinetics were observed in all the samples that were fermented with strain RC212 (suspended and immobilized) than in samples fermented with strain EC1118. Significantly higher amounts of acetic acid were detected in all samples fermented with strain RC212 than in those fermented with strain EC1118 (0.282 and 0.602 g/l, respectively). Slightly higher amounts of glycerol were observed in samples fermented with strain EC1118 than in those fermented with strain RC212.

  9. Application in the Ethanol Fermentation of Immobilized Yeast Cells in Matrix of Alginate/Magnetic Nanoparticles, on Chitosan-Magnetite Microparticles and Cellulose-coated Magnetic Nanoparticles

    CERN Document Server

    Ivanova, Viara; Hristov, Jordan

    2011-01-01

    Saccharomyces cerevisiae cells were entrapped in matrix of alginate and magnetic nanoparticles and covalently immobilized on magnetite-containing chitosan and cellulose-coated magnetic nanoparticles. Cellulose-coated magnetic nanoparticles with covalently immobilized thermostable {\\alpha}-amylase and chitosan particles with immobilized glucoamylase were also prepared. The immobilized cells and enzymes were applied in column reactors - 1/for simultaneous corn starch saccharification with the immobilized glucoamylase and production of ethanol with the entrapped or covalently immobilized yeast cells, 2/ for separate ethanol fermentation of the starch hydrolysates with the fixed yeasts. Hydrolysis of corn starch with the immobilized {\\alpha}-amylase and glucoamylase, and separate hydrolysis with the immobilized {\\alpha}-amylase were also examined. In the first reactor the ethanol yield reached approx. 91% of the theoretical; the yield was approx. 86% in the second. The ethanol fermentation was affected by the typ...

  10. Oxidation reactions are required to produce atranorin from acetate by alginate-immobilized cells of Cladonia verticillaris.

    OpenAIRE

    Sebastián, B.; Fontaniella, B.; E. C. Pereira; Vicente, C

    2000-01-01

    Atranorin, a p-depside of the b-orcinol series, is produced by several Cladonia species. Immobilized cells of Cladonia verticillaris in calcium alginate are able to produce atranorin when they are supplied with 1.0 mM acetate as a precursor. Production of the depside is enhanced by adding an oxidant agent (NAD+ or FMN) to the incubation media and its secretion to these media is facilitated by permeabilizing the immobilized cells with 2 % iso-propanol.

  11. Bioethanol Production from Uncooked Raw Starch by Immobilized Surface-engineered Yeast Cells

    Science.gov (United States)

    Chen, Jyh-Ping; Wu, Kuo-Wei; Fukuda, Hideki

    Surface-engineered yeast Saccharomyces cerevisiae codisplaying Rhizopus oryzae glucoamylase and Streptococcus bovis α-amylase on the cell surface was used for direct production of ethanol from uncooked raw starch. By using 50 g/L cells during batch fermentation, ethanol concentration could reach 53 g/L in 7 days. During repeated batch fermentation, the production of ethanol could be maintained for seven consecutive cycles. For cells immobilized in loofa sponge, the concentration of ethanol could reach 42 g/L in 3 days in a circulating packed-bed bioreactor. However, the production of ethanol stopped thereafter because of limited contact between cells and starch. The bioreactor could be operated for repeated batch production of ethanol, but ethanol concentration dropped to 55% of its initial value after five cycles because of a decrease in cell mass and cell viability in the bioreactor. Adding cells to the bioreactor could partially restore ethanol production to 75% of its initial value.

  12. Ethanol production from molasses by immobilized cells of zymomonas mobilis EMCC 1546

    International Nuclear Information System (INIS)

    Ethanol production from beet molasses by zymomonas mobilis EMCC 1546 was studied using continuous processes in which immobilized bacterial cells of Z.mobilis EMCC 1546 was grown on both sodium alginate and polyvinyl alcohol(PVA). The fermentation was performed in a shaking incubation and 1-liter ferment or with final working 750 ml. The initial sugar concentration studied was 50, 100,150, 200 and 250 g/l. The results showed that optimum initial sugar for ethanol production was 200 g/l. In batch fermentation, the highest ethanol concentration was 28.50 g/. Also effect of gamma irradiation was studied to enhance ethanol production. The highest ethanol production at dose dose 0.25 kGy was 34.82 g/l. The results showed that continuous fermentation, at dilution rate 1.36 (I/h), helped to increase the ethanol production significantly and continuous fermentation with immobilized cells in alginate gave higher ethanol production, 35.8 (g/I), as compared with those immobilized in hydrogel (PVA)

  13. Cane molasses fermentation for continuous ethanol production in an immobilized cells reactor by Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Ghorbani, Farshid; Younesi, Habibollah; Esmaeili Sari, Abbas [Department of Environmental Science, Faculty of Natural Resources and Marine Sciences, Tarbiat Modares University, Noor, P.O. Box: 64414-356 (Iran); Najafpour, Ghasem [Department of Chemical Engineering, Faculty of Engineering, Noshirvani University of Technology, Babol (Iran)

    2011-02-15

    Sodium-alginate immobilized yeast was employed to produce ethanol continuously using cane molasses as a carbon source in an immobilized cell reactor (ICR). The immobilization of Saccharomyces cerevisiae was performed by entrapment of the cell cultured media harvested at exponential growth phase (16 h) with 3% sodium alginate. During the initial stage of operation, the ICR was loaded with fresh beads of mean diameter of 5.01 mm. The ethanol production was affected by the concentration of the cane molasses (50, 100 and 150 g/l), dilution rates (0.064, 0.096, 0.144 and 0.192 h{sup -1}) and hydraulic retention time (5.21, 6.94, 10.42 and 15.63 h) of the media. The pH of the feed medium was set at 4.5 and the fermentation was carried out at an ambient temperature. The maximum ethanol production, theoretical yield (Y{sub E/S}), volumetric ethanol productivity (Q{sub P}) and total sugar consumption was 19.15 g/l, 46.23%, 2.39 g l{sup -1} h{sup -1} and 96%, respectively. (author)

  14. Radioresistance of mice cells immobilized by adhesion in glass layer

    International Nuclear Information System (INIS)

    Phagocytic leukocytes are involved in bio compatibility and biodegradation processes at which materials utilized in different at which materials utilized in different types of implants are submitted. In this work round shape glass cover slips were implanted subcutaneously in 45-day-old C57B1J 6 mice and later irradiated with a 60 Co sublethal whole-body dose of 4.0 Gy. Cover slips were removed 1,3,7 and 14 days post-implant and dyed by the hematoxylin-eosin technique. Macrophage and giant cell estimations were done in a microscope by means of an integrator eyepiece. The modifications found permit to conclude that they to exist significant differences in macrophages as a function of time after implant but not as a consequence of irradiation. (author)

  15. Cell Proliferation on Macro/Nano Surface Structure and Collagen Immobilization of 3D Polycaprolactone Scaffolds.

    Science.gov (United States)

    Park, Young-Ouk; Myung, Sung-Woon; Kook, Min-Suk; Jung, Sang-Chul; Kim, Byung-Hoon

    2016-02-01

    In this study, 3D polycaprolactone (PCL) scaffolds were fabricated by 3D printing technique. The macro/nano morphology of, 3D PCL scaffolds surface was etched with oxygen plasma. Acrylic acid (AA) plasma-polymerization was performed to functionalize the macro/nano surface with carboxyl groups and then collagen was immobilized with plasma-polymerized 3D PCL scaffolds. After O2 plasma and AA plasma-polymerization, contact angles were decreased. The FE-SEM and AFM results showed that O2 plasma is increased the surface roughness. The MTT assay results showed that proliferation of the M3CT3-E1 cells increased on the oxygen plasma treated and collagen immobilized 3D PCL scaffolds. PMID:27433597

  16. BIOSORPTION OF TEXTILE DYE USING IMMOBILIZED BACTERIAL (PSEUDOMONAS AERUGINOSA AND FUNGAL (PHANEROCHATE CHRYSOSPORIUM CELLS

    Directory of Open Access Journals (Sweden)

    Natarajan Saravanan

    2013-01-01

    Full Text Available Wastewater containing dyes presents a serious problem due to its high toxicity which leads to creating enormous environmental pollution and ecological hazards. Therefore the removal of the high stable dyes from the textile effluents is of major importance. The purpose of this study is to remove the reactive dye Procion Blue 2G from textile dye solution by biosorption process using immobilized cells of Pseudomonas aeruginosa and Phanerochate chrysosporium. It was found that maximum dye uptake is 1.648 mg g-1 of bead for P. aeruginosa and it is 1.242 mg g-1 of bead for P. chrysosporium. Both the results are derived from higher initial dye concentration (100 mg L-1 and high cell concentration (in terms of volume of inoculum 20 mL and at low mass of biosorbent (5 g of bead. Comparatively better results are produced by the beads having the cells of P. aeruginosa than P. chrysosporium. Further, due to the cell immobilization, both the cell beads can be utilized repeatedly in continuous reactors by selecting suitable eluent in industrial scale with the advantage of avoiding wash out of cells.

  17. The use of covalently immobilized stem cell factor to selectively affect hematopoietic stem cell activity within a gelatin hydrogel

    Science.gov (United States)

    Mahadik, B.P.; Haba, S. Pedron; Skertich, L.J.; Harley, B.A.C.

    2015-01-01

    Hematopoietic stem cells (HSCs) are a rare stem cell population found primarily in the bone marrow and responsible for the production of the body’s full complement of blood and immune cells. Used clinically to treat a range of hematopoietic disorders, there is a significant need to identify approaches to selectively expand their numbers ex vivo. Here we describe a methacrylamide-functionalized gelatin (GelMA) hydrogel for in vitro culture of primary murine HSCs. Stem cell factor (SCF) is a critical biomolecular component of native HSC niches in vivo and is used in large dosages in cell culture media for HSC expansion in vitro. We report a photochemistry based approach to covalently immobilize SCF within GelMA hydrogels via acrylate-functionalized polyethylene glycol (PEG) tethers. PEG-functionalized SCF retains the native bioactivity of SCF but can be stably incorporated and retained within the GelMA hydrogel over 7 days. Freshly-isolated murine HSCs cultured in GelMA hydrogels containing covalently-immobilized SCF showed reduced proliferation and improved selectivity for maintaining primitive HSCs. Comparatively, soluble SCF within the GelMA hydrogel network induced increased proliferation of differentiating hematopoietic cells. We used a microfluidic templating approach to create GelMA hydrogels containing gradients of immobilized SCF that locally direct HSC response. Together, we report a biomaterial platform to examine the effect of the local presentation of soluble vs. matrix-immobilized biomolecular signals on HSC expansion and lineage specification. This approach may be a critical component of a biomaterial-based artificial bone marrow to provide the correct sequence of niche signals to grow HSCs in the laboratory. PMID:26232879

  18. Immobilization by Polyurethane of Pseudomonas dacunhae Cells Containing l-Aspartate β-Decarboxylase Activity and Application to l-Alanine Production

    OpenAIRE

    Fusee, Murray C.; Weber, Jennifer E.

    1984-01-01

    Whole cells of Pseudomonas dacunhae containing l-aspartate β-decarboxylase activity were immobilized by mixing a cell suspension with a liquid isocyanate-capped polyurethane prepolymer (Hypol; W. R. Grace & Co., Lexington, Mass.). The immobilized cell preparation was used to convert l-aspartic acid to l-alanine. Properties of the immobilized P. dacunhae cells containing aspartate β-decarboxylase activity were investigated with batch reactors. Retention of enzyme activity was observed to be as...

  19. Biochips for cell biology by combined dip-pen nanolithography and DNA-directed protein immobilization.

    Science.gov (United States)

    Arrabito, Giuseppe; Reisewitz, Stephanie; Dehmelt, Leif; Bastiaens, Philippe I; Pignataro, Bruno; Schroeder, Hendrik; Niemeyer, Christof M

    2013-12-20

    A general methodology for patterning of multiple protein ligands with lateral dimensions below those of single cells is described. It employs dip pen nanolithography (DPN) patterning of DNA oligonucleotides which are then used as capture strands for DNA-directed immobilization (DDI) of oligonucleotide-tagged proteins. This study reports the development and optimization of PEG-based liquid ink, used as carrier for the immobilization of alkylamino-labeled DNA oligomers on chemically activated glass surfaces. The resulting DNA arrays have typical spot sizes of 4-5 μm with a pitch of 12 μm micrometer. It is demonstrated that the arrays can be further functionalized with covalent DNA-streptavidin (DNA-STV) conjugates bearing ligands recognized by cells. To this end, biotinylated epidermal growth factor (EGF) is coupled to the DNA-STV conjugates, the resulting constructs are hybridized with the DNA arrays and the resulting surfaces used for the culturing of MCF-7 (human breast adenocarcinoma) cells. Owing to the lateral diffusion of transmembrane proteins in the cell's plasma membrane, specific recruitment and concentration of EGF receptor can be induced specifically at the sites where the ligands are bound on the solid substrate. This is a clear demonstration that this method is suitable for precise functional manipulations of subcellular areas within living cells.

  20. Nitrilase-catalyzed conversion of (R,S)-mandelonitrile by immobilized recombinant Escherichia coli cells harboring nitrilase.

    Science.gov (United States)

    Zhang, Xin-Hong; Liu, Zhi-Qiang; Xue, Ya-Ping; Xu, Ming; Zheng, Yu-Guo

    2016-07-01

    (R)-(-)-Mandelic acid (R-MA) is widely used both as a versatile intermediate for pharmaceuticals and a resolving agent in chiral resolution processes. In the current study, to improve the stability of operation, recombinant Escherichia coli cells expressing nitrilase from Alcaligenes faecalis were immobilized for the enantioselective hydrolysis of (R,S)-mandelonitrile to R-MA. Different immobilization methods including entrapment matrices, entrapment matrices cross-linked by cross-linking and polymerization agents, and direct cross-linking cells using glutaraldehyde (GA) or bionic silicon were investigated. To facilitate industrial solid-liquid separation, the direct cross-linking recombinant E. coli cells using diatomite/GA/polyethyleneimine with 135.95% relative activity compared with free cells was chosen using water as the reaction medium. The operational stability of the immobilized cells was obviously superior to that of free cells, without significant activity loss after 28 cycles of batch reaction and the successive production of R-MA could reach 1.88 M. Moreover, the immobilized cells showed good storage stability with about 52% relative activity after storing for 30 days at 4 °C. Therefore, the immobilized biocatalyst is very promising for upscale production of optically pure R-MA with high performance and low cost.

  1. An Evaluation of Kinetic Parameters of Cadmium and Copper Biosorption by Immobilized Cells

    Directory of Open Access Journals (Sweden)

    Nelly Georgieva

    2007-10-01

    Full Text Available Bioremediation is the use of living organisms to reduce or eliminate environmental hazards resulting from the accumulation of toxic chemicals and other hazardous wastes. This technology is based on the utilization of microorganisms to transform organic and inorganic compounds. The filamentous yeast Trichosporon cutaneum strain R57, immobilized and free cells was cultivated as batch culture on a liquid medium in the presence of various concentrations of cadmium and copper ions. The simultaneous uptake and accumulation of Cd2+ and Cu2+ ions by Tr. cutaneum cells depending on the initial concentration of Cd2+ and Cu2+ in the medium were studied. The potential use of the free and immobilized cells of Trichosporon cutaneum to remove cadmium and copper ions, from aqueous solutions was evaluated. Two important physicochemical aspects for the evaluation of the sorption process as a unit operation are the equilibrium of sorption and the kinetics. The Cd2+ and Cu2+ ions biosorption capacities of all tested adsorbent were presented as a function of the initial concentration of metal ions within the aqueous biosorption medium. The individual, as well as bicomponent sorption kinetics of copper and cadmium ions by immobilised cells of Trichosporon cutaneum R57 is presented. A second order kinetic model obtains kinetic parameters for the copper and cadmium ions.

  2. Chitin hydrolysis assisted by cell wall degrading enzymes immobilized of Thichoderma asperellum on totally cinnamoylated D-sorbitol beads.

    Science.gov (United States)

    Fernandes, Kátia F; Cortijo-Triviño, David; Batista, Karla A; Ulhoa, Cirano J; García-Ruiz, Pedro A

    2013-07-01

    In this study, cell wall degrading enzymes produced by Thrichoderma asperellum (TCWDE) were immobilized on totally cinnamoylated D-sorbitol (TCNSO) beads and used for chitin hydrolysis. In order to optimize immobilization efficiency, the reaction time was varied from 2 to 12 h and reactions were conducted in the presence or absence of Na2SO4. Immobilized enzymes were analysed concerning to thermal and operational stability. Immobilization in presence of Na2SO4 was 54% more efficient than immobilization in absence of salt. After optimization, 32% of the total enzyme offered was immobilized, with 100% of bounding efficiency, measured as the relation between protein and enzyme immobilized. Free and TCNSO-TCWDE presented very similar kinetics with maximum hydrolysis reached at 90 min of reaction. Thermal stability of both free and TCNSO-TCWDE was similar, with losses in activity after 55 °C. Moreover, free and TCNSO-TCWDE retained 100% activity after 3h incubation at 55 °C. TCNSO-TCWDE were used in a bath-wise reactor during 14 cycles, producing 1825 μg of N-acetylglucosamine (NAG) maintaining 83% of initial activity.

  3. Microchip-based Integration of Cell Immobilization, Electrophoresis, Post-column Derivatization, and Fluorescence Detection for Monitoring the Release of Dopamine from PC 12 Cells

    OpenAIRE

    Li, Michelle W.; Martin, R. Scott

    2008-01-01

    In this paper, we describe the fabrication and evaluation of a multilayer microchip device that can be used to quantitatively measure the amount of catecholamines released from PC 12 cells immobilized within the same device. This approach allows immobilized cells to be stimulated on-chip and, through rapid actuation of integrated microvalves, the products released from the cells are repeatedly injected into the electrophoresis portion of the microchip, where the analytes are separated based u...

  4. Phytoremediation of Benzophenone and Bisphenol A by Glycosylation with Immobilized Plant Cells

    Directory of Open Access Journals (Sweden)

    Kei Shimoda

    2009-01-01

    Full Text Available Benzophenone and bisphenol A are environmental pollutions, which have been listed among “chemicals suspected of having endocrine disrupting effects” by the World Wildlife Fund, the National Institute of Environmental Health Sciences in the USA and the Japanese Environment Agency. The cultured cells of Nicotiana tabacum glycosylated benzophenone to three glycosides, 4-O-β-D-glucopyranosylbenzophenone (9%, diphenylmethyl β-D-glucopyranoside (14%, and diphenylmethyl 6-O-(β-D-glucopyranosyl-β-D-glucopyranoside (12% after 48 h incubation. On the other hand, incubation of benzophenone with immobilized cells of N. tabacum in sodium alginate gel gave products in higher yields, i.e. the yields of 4-O-β-D-glucopyranosylbenzophenone, diphenylmethyl β-D-glucopyranoside, and diphenylmethyl 6-O-(β-D-glucopyranosyl-β-D-glucopyranoside were 15, 27, and 22%, respectively. Bisphenol A was converted into three glycosides, 2,2-bis(4-β-D-glucopyranosyloxyphenylpropane (16%, 2-(4-β-D-glucopyranosyloxy-3-hydroxyphenyl-2-(4-β-D-gluco- pyranosyloxyphenyl propane (8%, and 2-(3-β-D-glucopyranosyloxy-4-hydroxyphenyl-2-(4-β-D-glucopyranosyloxyphe nylpropane (5%. Also the use of immobilized N. tabacum cells improved the yield of products; the glycosylation of bisphenol A with immobilized N. tabacum gave 2,2-bis(4-β-D-glucopyranosyloxyphenylpropane (24%, 2-(4-β-D-gluco- pyranosyloxy-3-hydroxyphenyl-2-(4-β-D-glucopyranosyloxyphenyl propane (15%, and 2-(3-β-D-glucopyranosyloxy- 4-hydroxyphenyl-2-(4-β-D-glucopyranosyloxyphenylpropane (11%.

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

    Science.gov (United States)

    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

  6. Drying of immobilized yeast cells in a spouted bed dryer with a moving draft tube

    OpenAIRE

    Dragan Povrenović; Viktor Nedović

    2010-01-01

    Brewery yeast cells immobilized in Ca-alginate were dried in a laboratory scale spouted bed with a draft tube.The experiment was conducted under variable temperatures and air flow rates. The temperature and air velocity at the bottom of the column have been varied in the range from 30 to 60 °C and from 6 to 10 m/s in a duration of 60 min. The moisture of dryied particles was in the interval of 10.00 to 21.00 g/g, while the water activity was in the range of 0.40 to 0.45 what ensures the prese...

  7. Hydroxylation and biodegration of 6-methylquinoline by pseudomonads in aqueous and nonaqueous immobilized-cell bioreactors

    Energy Technology Data Exchange (ETDEWEB)

    Rothenburger, S.; Atlas, R.M. (Univ. of Louisville, KY (United States))

    1993-07-01

    Heterocyclic nitrogen compounds, including quinoline, methylquinolines, and isoquinoline, occur in creosote and various fossil fuels and are significant pollutants in soils and water. The removal of quinolines and other nitrogen and sulfur heterocyclic compounds from fuels could prevent the formation of air pollutants when fuels are burned. Although biodegradation of several quinolines has been reported, methylquinolines are particularly resistant to biodegradation. This paper describes the biotransformation and biodegradation of 6-methylquinoline by Pseudomona putida. The biotransformation of quinolines by immobilized cells in aqueous and nonaqueous systems, relevant for bioremediation systems, is also reported. 23 refs., 7 figs., 1 tab.

  8. Ferrous Sulphate Oxidation Using Acidithiobacillus Ferrooxidans Cells Immobilized in Ceramic Beads

    OpenAIRE

    Junfeng, Y.; Guoliang, L.; Wei, C.

    2007-01-01

    The immobilization of Acidithiobacillus ferrooxidans cells on ceramic beads as carrier is described. The effects of ferrous ion concentration and dilution on the kinetics of ferrous ion oxidation in a packed-bed bioreactor were studied. In a medium containing 13.91 g of ferrous ion per litre, the fastest oxidation rate was 4.21 g L–1 at a dilution rate of 0.8 h–1. The corresponding conversion was X = 70 %. At ferrous ion mass concentrations greater than = 8.34 g L–1 and dilution rates greate...

  9. Study on Immobilized Algal Cells for Treatment and Recycle of Refinery Wastewater

    Institute of Scientific and Technical Information of China (English)

    Yu Baocheng; Liu Deqi; Dong Lihua; Li Gang

    2005-01-01

    Compared to the algal oxidation pond, treatment of wastewater using the immobilized algal cell technology has excellent effect, which not only can effectively avoid the disadvantage of oxidation pond,but can also remarkably improve the efficiency of treating system and the effluent quality. When the treating system operates under an optimal control conditions, such as a 55% loading rate, an illumination intensity of 2500-3500 lux and a hydraulic residence time of 4 hours, the COD and ammonia nitrogen removal can reach 90%. Water after deep treatment can comply with the requirement of the refinery for the quality of recycled water.

  10. Oxygen supply for CHO cells immobilized on a packed-bed of Fibra-Cel disks.

    Science.gov (United States)

    Meuwly, F; Loviat, F; Ruffieux, P-A; Bernard, A R; Kadouri, A; von Stockar, U

    2006-03-01

    Packed-bed bioreactors (PBR) have proven to be efficient systems to culture mammalian cells at very high cell density in perfusion mode, thus leading to very high volumetric productivity. However, the immobilized cells must be continuously supplied with all nutrients in sufficient quantities to remain viable and productive over the full duration of the perfusion culture. Among all nutrients, oxygen is the most critical since it is present at very low concentration due to its low solubility in cell culture medium. This work presents the development of a model for oxygenation in a packed-bed bioreactor system. The experimental system used to develop the model was a packed-bed of Fibra-Cel disk carriers used to cultivate Chinese Hamster Ovary cells at high density ( approximately 6.1 x 10(7) cell/mL) in perfusion mode. With the help of this model, it was possible to identify if a PBR system is operated in optimal or sub-optimal conditions. Using the model, two options were proposed, which could improve the performance of the basal system by about twofold, that is, by increasing the density of immobilized cells per carrier volume from 6.1 x 10(7) to 1.2 x 10(8) cell/mL, or by increasing the packed-bed height from 0.2 to 0.4 m. Both strategies would be rather simple to test and implement in the packed-bed bioreactor system used for this study. As a result, it would be possible to achieve a substantial improvement of about twofold higher productivity as compared with the basal conditions. PMID:16358288

  11. Oxygen supply for CHO cells immobilized on a packed-bed of Fibra-Cel disks.

    Science.gov (United States)

    Meuwly, F; Loviat, F; Ruffieux, P-A; Bernard, A R; Kadouri, A; von Stockar, U

    2006-03-01

    Packed-bed bioreactors (PBR) have proven to be efficient systems to culture mammalian cells at very high cell density in perfusion mode, thus leading to very high volumetric productivity. However, the immobilized cells must be continuously supplied with all nutrients in sufficient quantities to remain viable and productive over the full duration of the perfusion culture. Among all nutrients, oxygen is the most critical since it is present at very low concentration due to its low solubility in cell culture medium. This work presents the development of a model for oxygenation in a packed-bed bioreactor system. The experimental system used to develop the model was a packed-bed of Fibra-Cel disk carriers used to cultivate Chinese Hamster Ovary cells at high density ( approximately 6.1 x 10(7) cell/mL) in perfusion mode. With the help of this model, it was possible to identify if a PBR system is operated in optimal or sub-optimal conditions. Using the model, two options were proposed, which could improve the performance of the basal system by about twofold, that is, by increasing the density of immobilized cells per carrier volume from 6.1 x 10(7) to 1.2 x 10(8) cell/mL, or by increasing the packed-bed height from 0.2 to 0.4 m. Both strategies would be rather simple to test and implement in the packed-bed bioreactor system used for this study. As a result, it would be possible to achieve a substantial improvement of about twofold higher productivity as compared with the basal conditions.

  12. The production of cellulase in a spouted bed fermentor using cells immobilized in biomass support particles.

    Science.gov (United States)

    Webb, C; Fukuda, H; Atkinson, B

    1986-01-01

    Continuous cellulase production by Trichoderma viride QM 9123, immobilized in 6 mm diameter, spherical, stainless steel biomass support particles, has been achieved using a medium containing glucose as the main carbon source. Experiments were carried out in a 10-L spouted bed fermentor. In this type of reactor-recycled broth is used to create a jet at the base of a bed of particles, causing the particles to spout and circulate. During the circulation, particles pass through a region of high shear near the jet inlet. This effectively prevents a buildup of excess biomass and thus enables steady-state conditions to be achieved during continuous operation. Continuous production of cellulase was achieved at significantly higher yield and productivity than in conventional systems. At a dilution rate of 0.15 h(-1) (nominal washout rate for freely suspended cells is 0.012 h(-1)), the yield of cellulase on glucose was 31% higher than that measured during batch operation, while the volumetric productivity (31.5 FPA U/L. h) was 53% greater than in the batch system. The specific cellulase productivity of the immobilized cells was more than 3 times that of freely suspended cells, showing that diffusional limitations can be beneficial. This offers significant opportunity for the further development of biomass support particles and associated bioreactors. PMID:18553840

  13. Immobilized Kluyveromyces marxianus cells in carboxymethyl cellulose for production of ethanol from cheese whey: experimental and kinetic studies.

    Science.gov (United States)

    Roohina, Fatemeh; Mohammadi, Maedeh; Najafpour, Ghasem D

    2016-09-01

    Cheese whey fermentation to ethanol using immobilized Kluyveromyces marxianus cells was investigated in batch and continuous operation. In batch fermentation, the yeast cells were immobilized in carboxymethyl cellulose (CMC) polymer and also synthesized graft copolymer of CMC with N-vinyl-2-pyrrolidone, denoted as CMC-g-PVP, and the efficiency of the two developed cell entrapped beads for lactose fermentation to ethanol was examined. The yeast cells immobilized in CMC-g-PVP performed slightly better than CMC with ethanol production yields of 0.52 and 0.49 g ethanol/g lactose, respectively. The effect of supplementation of cheese whey with lactose (42, 70, 100 and 150 g/l) on fermentative performance of K. marxianus immobilized in CMC beads was considered and the results were used for kinetic studies. The first order reaction model was suitable to describe the kinetics of substrate utilization and modified Gompertz model was quite successful to predict the ethanol production. For continuous ethanol fermentation, a packed-bed immobilized cell reactor (ICR) was operated at several hydraulic retention times; HRTs of 11, 15 and 30 h. At the HRT of 30 h, the ethanol production yield using CMC beads was 0.49 g/g which implies that 91.07 % of the theoretical yield was achieved. PMID:27126500

  14. Application of immobilized cell preparation obtained from biomass of Gluconacetobacter xylinus bacteria in biotransformation of glycerol to dihydroxyacetone

    Directory of Open Access Journals (Sweden)

    Lidia Stasiak-Różańska

    2011-03-01

    Full Text Available Introduction. Dihydroxyacetone (DHA, being a product of glycerol oxidation by acetic acid bacteria, is an important compound widely applied in the cosmetic, food, and pharmaceutical industry, as well as in medicine. Biotransformation of glycerol to DHA is catalyzed by glycerol dehydrogenase (GlyDH, EC 1.1.1.6 bound with the cytoplasmic membrane of bacteria. An attempt was undertaken in this study to conduct glycerol biotransformation with immobilized fractions of a cell preparation with GlyDH activity. The content of dihydroxyacetone obtained with the cell preparation was compared with its content achieved in the reaction with immobilized viable cells of G. xylinus. Material and methods. Cell walls of Gluconacetobacter xylinus bacteria were disintegrated enzymatically. The resultant preparation was immobilized on calcium alginate or first separated into two fractions (precipitate and supernatant by centrifugation and then immobilized. DHA content was determined colorimetrically after the reaction with 3,5-dinitrosalicilic acid. Glycerol content was assayed with the refractometric method. Results. After 20 days of the process, the concentration of DHA obtained with immobilized whole cells reached 25 g/l. In turn, the content of DHA obtained in the same period with immobilized fractions of the cell preparation accounted for 16.9 g/l and 8.95 g/l (depending on the fraction applied. Conclusions. DHA may be obtained in the process independent of G. xylinus metabolic activity using a preparation which displays the catalytic activity of glycerol dehydrogenase and obtained as a result of disintegration of live bacterial cells. The application of such a preparation may in the future eliminate technological problems posed by the presence of bacterial cells and their metabolites in the culture medium.

  15. Drying of immobilized yeast cells in a spouted bed dryer with a moving draft tube

    Directory of Open Access Journals (Sweden)

    Dragan Povrenović

    2010-07-01

    Full Text Available Brewery yeast cells immobilized in Ca-alginate were dried in a laboratory scale spouted bed with a draft tube.The experiment was conducted under variable temperatures and air flow rates. The temperature and air velocity at the bottom of the column have been varied in the range from 30 to 60 °C and from 6 to 10 m/s in a duration of 60 min. The moisture of dryied particles was in the interval of 10.00 to 21.00 g/g, while the water activity was in the range of 0.40 to 0.45 what ensures the preservation of immobilized yeast as a starter and provides the biological activity of dried particles. A rehidration process of dryied particles proved that dried particles could completely restore their original shape and starting volume, while the mechanical resistance is somewhat reduced. The cells preserved in this way completely restore their catalytical activity after the rehidration.

  16. Immobilization of pamidronic acids on the nanotube surface of titanium discs and their interaction with bone cells

    Science.gov (United States)

    Koo, Tae-Hyung; Borah, Jyoti S.; Xing, Zhi-Cai; Moon, Sung-Mo; Jeong, Yongsoo; Kang, Inn-Kyu

    2013-03-01

    Self-assembled layers of vertically aligned titanium nanotubes were fabricated on a Ti disc by anodization. Pamidronic acids (PDAs) were then immobilized on the nanotube surface to improve osseointegration. Wide-angle X-ray diffraction, X-ray photoelectron microscopy, and scanning electron microscopy were employed to characterize the structure and morphology of the PDA-immobilized TiO2 nanotubes. The in vitro behavior of osteoblast and osteoclast cells cultured on an unmodified and surface-modified Ti disc was examined in terms of cell adhesion, proliferation, and differentiation. Osteoblast adhesion, proliferation, and differentiation were improved substantially by the topography of the TiO2 nanotubes, producing an interlocked cell structure. PDA immobilized on the TiO2 nanotube surface suppressed the viability of the osteoclasts and reduced their bone resorption activity.

  17. Fermentation efficiency of cells immobilized on delignified brewers' spent grains after low- and high-temperature thin layer thermal drying.

    Science.gov (United States)

    Tsaousi, Konstantina; Koutinas, Athanasios A; Bekatorou, Argyro; Loukatos, Paul

    2010-09-01

    Low-cost dried yeasts immobilized on delignified brewers' spent grains for use in wine making and brewing were produced by simple thermal drying techniques. To optimize the thermal drying process, vacuum and air stream conditions were examined. Drying of thin layers of the biocatalysts was performed at low (30-38 degrees C) and high temperatures (40-70 degrees C). The fermentation efficiency of the thermally dried biocatalysts was acceptable, with immobilized cells showing a significantly higher thermotolerance compared with free cells. Immobilized cells dried at high temperatures presented slightly improved glucose fermentation efficiency compared with the low-temperature dried biocatalysts. Gas chromatography-mass spectrometry analysis of aroma volatiles of the fermented products revealed an increase of esters, lower higher alcohol formation, and significantly lower concentration of carbonylic compounds.

  18. VITALITY AND MORPHOLOGY OF TUMOR CELLS TREATED WITH 4-TIAZOLIDINONE DERIVATIVES IMMOBILIZED ON NANOSCALE POLYMER CARRIER

    Directory of Open Access Journals (Sweden)

    N. M. Boiko

    2015-02-01

    Full Text Available A nanoscale polymeric carrier was used for delivery of novel anticancer compounds – 4-tiazolidinone derivatives – to tumor cells of different lines. It was found that such way of delivery of the above mentioned compounds to target cells significantly (approximately 10 times decreased acting cytotoxic dose of some of these compounds with preservation of similar level of their antineoplastic effect in vitro towards various mammalian tumor cells. The microscopic investigation of these cells demonstrated that under the action of some immobilized 4-tiazolidonone derivatives, there was an increase (up to 40% of the part of apoptotic cells, as well as an appearance of 10% of cells with morphologically changed nucleus, and up to 35% of cells with an increased intensity of red fluorescence of acridine orange in the lysosomes, compared with such indicators observed under the action of free form of those compounds. Thus, the applied nanoscale carrier is a perspective polymer system for delivery of anticancer drugs to target cells.

  19. Power generation enhancement in novel microbial carbon capture cells with immobilized Chlorella vulgaris

    Science.gov (United States)

    Zhou, Minghua; He, Huanhuan; Jin, Tao; Wang, Hongyu

    2012-09-01

    With the increasing concerns for global climate change, a sustainable, efficient and renewable energy production from wastewater is imperative. In this study, a novel microbial carbon capture cell (MCC), is constructed for the first time by the introduction of immobilized microalgae (Chlorella vulgaris) into the cathode chamber of microbial fuel cells (MFCs) to fulfill the zero discharge of carbon dioxide. This process can achieve an 84.8% COD removal, and simultaneously the maximum power density can reach 2485.35 mW m-3 at a current density of 7.9 A m-3 and the Coulombic efficiency is 9.40%, which are 88% and 57.7% greater than that with suspended C. vulgaris, respectively. These enhancements in performance demonstrate the feasibility of an economical and effective approach for the simultaneous wastewater treatment, electricity generation and biodiesel production from microalgae.

  20. Glucosyltransferase production by Klebsiella sp. K18 and conversion of sucrose to palatinose using immobilized cells.

    Science.gov (United States)

    Orsi, Daniela C; Kawaguti, Haroldo Y; Sato, Hélia H

    2009-01-01

    The strain Klebsiella sp. K18 produces the enzyme glucosyltransferase and catalyses the conversion of sucrose to palatinose, an alternative sugar that presents low cariogenicity. Response Surface Methodology was successfully employed to determine the optimal concentration of culture medium components. Maximum glucosyltransferase production (21.78 U mL(-1)) was achieved using the optimized medium composed by sugar cane molasses (80 g L(-1)), bacteriological peptone (7 g L(-1)) and yeast extract (20 g L(-1)), after 8 hours of fermentation at 28°C. The conversion of sucrose to palatinose was studied utilizing immobilized cells in calcium alginate. The effects of the alginate concentration (2-4%), cell mass concentration (20-40%) and substrate concentration (25-45%) were evaluated and the yield of palatinose was approximately 62.5%. PMID:24031319

  1. Immobilized WNT Proteins Act as a Stem Cell Niche for Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Molly Lowndes

    2016-07-01

    Full Text Available The timing, location, and level of WNT signaling are highly regulated during embryonic development and for the maintenance of adult tissues. Consequently the ability to provide a defined and directed source of WNT proteins is crucial to fully understand its role in tissue development and to mimic its activity in vitro. Here we describe a one-step immobilization technique to covalently bind WNT3A proteins as a basal surface with easy storage and long-lasting activity. We show that this platform is able to maintain adult and embryonic stem cells while also being adaptable for 3D systems. Therefore, this platform could be used for recapitulating specific stem cell niches with the goal of improving tissue engineering.

  2. Production of phenolics by immobilized cells of the lichen Pseudevernia furfuracea: the role of epiphytic bacteria.

    Science.gov (United States)

    Blanch, M; Blanco, Y; Fontaniella, B; Legaz, M E; Vicente, C

    2001-06-01

    Immobilized lichen cells from the thalli of the lichen Pseudevernia furfuracea, supplied with acetate as the only source of carbon, continuously produced phenolic substances, atranorin and physodic acid, over 23 days. Epiphytic bacteria associated with the lichen thallus grew actively, probably using both acetate and reduced compounds supplied by lichen cells, since their active growth was avoided by including 10 microM 3,3'-dichlorophenyl-1,1'dimethylurea in the bath solution. Penicillin largely impeded the growth of epiphytic bacteria and decreased phenolic production, which was recovered only at the end of the experimental period, just when the bacteria started a slow, but active growth. We suggest the cooperation of epiphytic bacteria in the biosynthesis of both atranotrin and physodic acid. PMID:11770830

  3. Wet Chemistry and Peptide Immobilization on Polytetrafluoroethylene for Improved Cell-adhesion.

    Science.gov (United States)

    Gabriel, Matthias; Niederer, Kerstin; Frey, Holger

    2016-01-01

    Endowing materials surface with cell-adhesive properties is a common strategy in biomaterial research and tissue engineering. This is particularly interesting for already approved polymers that have a long standing use in medicine because these materials are well characterized and legal issues associated with the introduction of newly synthesized polymers may be avoided. Polytetrafluoroethylene (PTFE) is one of the most frequently employed materials for the manufacturing of vascular grafts but the polymer lacks cell adhesion promoting features. Endothelialization, i.e., complete coverage of the grafts inner surface with a confluent layer of endothelial cells is regarded key to optimal performance, mainly by reducing thrombogenicity of the artificial interface. This study investigates the growth of endothelial cells on peptide-modified PTFE and compares these results to those obtained on unmodified substrate. Coupling with the endothelial cell adhesive peptide Arg-Glu-Asp-Val (REDV) is performed via activation of the fluorin-containing polymer using the reagent sodium naphthalenide, followed by subsequent conjugation steps. Cell culture is accomplished using Human Umbilical Vein Endothelial Cells (HUVECs) and excellent cellular growth on peptide-immobilized material is demonstrated over a two-week period. PMID:27584937

  4. Optimization of Two-species Whole-cell Immobilization System Constructed with Marine-derived Fungi and Its Biological Degradation Ability

    Institute of Scientific and Technical Information of China (English)

    陈慧英; 王明霞; 沈煜斌; 姚善泾

    2014-01-01

    Mycelia pellet formed spontaneously in the process of cultivation was exploited as a biological carrier for whole-cell immobilization due to its unique structural characteristic. An innovative two-species whole-cell im-mobilization system was achieved by inoculating the marine-derived fungus Pestalotiopsis sp. J63 spores into cul-ture medium containing another fungus Penicillium janthinellum P1 pre-grown mycelia pellets for 2 days without any pretreatment. In order to evaluate the biological degradation capacity of this novel constructed immobilization system, the immobilized pellets were applied to treat paper mill effluent and decolorize dye Azure B. The use of the constructed immobilization system in the effluent resulted in successful and rapid biodegradation of numerous in-soluble fine fibers. The optimum conditions of immobilized procedure for maximum biodegradation capacity were determined using orthogonal design with biomass of P1 pellets 10 g (wet mass), concentration of J63 spore 2×109 ml-1, and immobilization time 2 d. The results demonstrate that immobilized pellets have more than 99%biodegra-dation capacity in a ten-hour treatment process. The kinetics of biodegradation fits the Michaelis-Menten equation well. Besides, the decolorization capability of immobilized pellets is more superior than that of P1 mycelia pellets. Overall, the present study offers a simple and reproducible way to construct a two-species whole-cell immobiliza-tion system for sewage treatment.

  5. Immobilization of Escherichia coli Cells Containing Aspartase Activity with Polyurethane and Its Application for l-Aspartic Acid Production

    Science.gov (United States)

    Fusee, Murray C.; Swann, Wayne E.; Calton, Gary J.

    1981-01-01

    Whole cells of Escherichia coli containing aspartase activity were immobilized by mixing a cell suspension with a liquid isocyanate-capped polyurethane prepolymer (Hypol). The immobilized cell preparation was used to convert ammonium fumarate to l-aspartic acid. Properties of the immobilized E. coli cells containing aspartase were investigated with a batch reactor. A 1.67-fold increase in the l-aspartic acid production rate was observed at 37°C as compared to 25°C operating temperature. The pH optimum was broad, ranging from 8.5 to 9.2. Increasing the concentration of ammonium fumarate to 1.5 M from 1.0 M negatively affected the reaction rate. l-Aspartic acid was produced at an average rate of 2.18 × 10−4 mol/min per g (wet weight) of immobilized E. coli cells with a 37°C substrate solution consisting of 1.0 M ammonium fumarate with 1 mM Mg2+ (pH 9.0). PMID:16345865

  6. Preparation and immobilization of noble metal nanoparticles for plasmonic solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ruoli; Pitzer, Martin; Hu, DongZhi; Schaadt, Daniel M. [Institut fuer Angewandte Physik, Karlsruher Institut fuer Technologie (KIT), Karlsruhe (Germany); DFG Centrum fuer Funktionelle Nanostrukturen (CFN), KIT (Germany); Fruk, Ljiljana [DFG Centrum fuer Funktionelle Nanostrukturen (CFN), KIT (Germany)

    2011-07-01

    Thin-film solar cells are of high interest due to good electrical properties and low material consumption. Traditional thin-film cells, however, have considerable transmission losses because of the reduced absorption volume. A promising way to enhance absorption in the active layer is the light-trapping by plasmonic nanostructures. Metallic nanoparticles have in particular shown large enhancement of the photocurrent in thin-film devices. In this poster, we present preparation of Au,Ag and Pt nanoparticles by polyol method and seed mediated methods for use in plasmonic solar cells. Polyol method typically uses ethylene glycol as the solvent and reducing agent,and in seed-mediated synthesis small nanoparticle seeds are first prepared and then used to promote the growth of different shapes of nanoparticles. We particularly focus on the use of nanocubes and nanospheres for solar cell design. Following the nanoparticle preparation, a new method to immobilize particles on GaAs surfaces via covalent chemical bonds has been developed which prevents agglomerations and allows control of the surface density. Photocurrent spectra of GaAs pin solar cells with and without particles have been recorded. These measurements show the dependence of the photocurrent enhancement on particle material, shape and density.

  7. Biodiesel Production: Utilization of Loofah Sponge to Immobilize Rhizopus chinensis CGMCC #3.0232 Cells as a Whole-Cell Biocatalyst.

    Science.gov (United States)

    He, Qiyang; Xia, Qianjun; Wang, Yuejiao; Li, Xun; Zhang, Yu; Hu, Bo; Wang, Fei

    2016-07-28

    Rhizopus chinensis cells immobilized on loofah (Luffa cylindrica) sponges were used to produce biodiesel via the transesterification of soybean oil. In whole-cell immobilization, loofah sponge is considered to be a superior alternative to conventional biomass carriers because of its biodegradable and renewable properties. During cell cultivation, Rhizopus chinensis mycelia can spontaneously and firmly adhere to the surface of loofah sponge particles. The optimal conditions for processing 9.65 g soybean oil at 40°C and 180 rpm using a 3:1 methanol-to-oil molar ratio were found to be 8% cell addition and 3-10% water content (depending on the oil's weight). Under optimal conditions, an over 90% methyl ester yield was achieved after the first reaction batch. The operational stability of immobilized Rhizopus chinensis cells was assayed utilizing a 1:1 methanol-to-oil molar ratio, thus resulting in a 16.5-fold increase in half-life when compared with immobilized cells of the widely studied Rhizopus oryzae. These results suggest that transesterification of vegetable oil using Rhizopus chinensis whole cells immobilized onto loofah sponge is an effective approach for biodiesel production. PMID:27090185

  8. SURVIVAL OF LIVER CELLS, IMMOBILIZED ON 3D-MATRIXES, IN LIVER FAILURE MODEL

    Directory of Open Access Journals (Sweden)

    M. Y. Shagidulin

    2011-01-01

    Full Text Available It was examined a new method for correction of hepatic failure by transplantation of liver support biounit (liver cells, immobilized on biocompatible and biodegradable 3D-matrixes ElastoPOB® into small intestine mesentery. It was determined that after modeling of acute hepatic failure on dogs by 65–70% liver resection and transplantation liver support biounit the restoration of disturbed biochemical indecies (such as total protein, lactate, cytolytic ensymes-ALT, AST, ALP, LDH, fibrinogen, protrombine index and others took place more rapidly on 9–14th day instead of 18th day in control. It was made a preposition about efficiency of the suggested method for correction both acute hepatic failure because even 90 days after transplantation of liver support biounit alive hepatocytes and neogenic plethoric vessels, growing through matrix were revealed. 

  9. Performance of a magnetically stabilized bed reactor with immobilized yeast cells.

    Science.gov (United States)

    Ivanova, V; Hristov, J; Dobreva, E; al-Hassan, Z; Penchev, I

    1996-05-01

    This paper is focused on the possibility to apply the magnetic stabilization technique in bioprocessing. The feasibility of a continuous ethanol fermentation process with immobilized Saccharomyces cerevisiae cells in a magnetically stabilized bed (MSB) was demonstrated. The fermentation processes were carried out in an external magnetic field, transverse to the fluid flow. The flexibility to change the bed expansion owing to the independent change of the fluid flow and the field intensity (the "magnetization FIRST" mode) permitted the creation of fixed beds with different particle arrangements, which affected the bed porosity, the effective fluid-particle contact area, and the mass transfer processes on the particle-fluid interface. As a result, higher ethanol concentration, ethanol production, and glucose uptake rates than in conventional packed bed reactor were reached.

  10. Flavor formation and cell physiology during the production of alcohol-free beer with immobilized Saccharomyces cerevisiae

    NARCIS (Netherlands)

    Iersel, van M.F.M.; Dieren, van B.; Rombouts, F.M.; Abee, T.

    1999-01-01

    Production of alcohol-free beer by limited fermentation is optimally performed in a packed-bed reactor operating in downflow. This ensures a highly controllable system with optimal reactor design. In the present study, we report on changes in the physiology of immobilized yeast cells in the reactor.

  11. Optimization of ethanol production from carob pod extract using immobilized Saccharomyces cerevisiae cells in a stirred tank bioreactor.

    Science.gov (United States)

    Ercan, Yatmaz; Irfan, Turhan; Mustafa, Karhan

    2013-05-01

    In this study, optimization of ethanol production from carob pod extract was carried out by immobilized Saccharomyces cerevisiae. Results showed that Ca-alginate concentration and the amount of immobilized cells had significant effects on yield. Optimum conditions for ethanol fermentation were determined to be 2% Ca-alginate concentration, 150 rpm agitation rate, 5% yeast cells entrapped in beads and pH 5.5. After validation experiments; ethanol concentration, yield, production rate and sugar utilization rate were respectively 40.10 g/L, 46.32%, 3.19 g/L/h and 90.66%; and the fermentation time was decreased to 24 h. In addition, the immobilized cells were shown to be reusable for five cycles, though a decrease in yield was observed. Finally, carob pod extract was used for ethanol fermentation by controlled and uncontrolled pH without any enrichment, and the results suggest that carob extract can be utilized effectively by immobilized-cell fermentation without the use of enrichments to facilitate yeast growth.

  12. Hydrogenases as catalysts for fuel cells: Strategies for efficient immobilization at electrode interfaces

    International Nuclear Information System (INIS)

    Highlights: ► This review examines hydrogenases as suitable biocatalysts for H2/O2 biofuel cells. ► It focuses on a O2, CO and temperature-resistant hydrogenase from Aquifex aeolicus. ► Electrically connected hydrogenase amount increases on carbon nanotube network. ► Hydrogenase orientation at the interface controls the electron transfer process. ► Hydrogenase insertion into liposomes enhances its stability. - Abstract: Hydrogenases are the key enzymes for hydrogen metabolism in many microorganisms. Due to the high efficiency they develop for H2 oxidation, research in the last five years has aimed towards their use as biocatalysts for H2/O2 biofuel cells to replace platinum-based chemical catalysts. We report in this review the major issues that have been addressed in view of the future development of such a novel biotechnological device. This includes enhancing the stability of either the enzyme itself or its immobilization onto conductive supports, increasing the amount of electrically connected enzymes and, finally, controlling hydrogenase orientation at the electrode surface, and hence the electron transfer process. We specifically focus on a particular [NiFe] membrane-bound hydrogenase purified from the hyperthermophilic and microaerophilic bacterium Aquifex aeolicus. This enzyme resists to O2, CO, and high temperatures making it potentially efficient as a biocatalyst. Recent progress in these domains strengthens the credibility of a viable H2/O2 biofuel cell and opens new avenues for biofuel cell design.

  13. Decolorization of industrial synthetic dyes using engineered Pseudomonas putida cells with surface-immobilized bacterial laccase

    Directory of Open Access Journals (Sweden)

    Wang Wei

    2012-06-01

    Full Text Available Abstract Background Microbial laccases are highly useful in textile effluent dye biodegradation. However, the bioavailability of cellularly expressed or purified laccases in continuous operations is usually limited by mass transfer impediment or enzyme regeneration difficulty. Therefore, this study develops a regenerable bacterial surface-displaying system for industrial synthetic dye decolorization, and evaluates its effects on independent and continuous operations. Results A bacterial laccase (WlacD was engineered onto the cell surface of the solvent-tolerant bacterium Pseudomonas putida to construct a whole-cell biocatalyst. Ice nucleation protein (InaQ anchor was employed, and the ability of 1 to 3 tandemly aligned N-terminal repeats to direct WlacD display were compared. Immobilized WlacD was determined to be surface-displayed in functional form using Western blot analysis, immunofluorescence microscopy, flow cytometry, and whole-cell enzymatic activity assay. Engineered P. putida cells were then applied to decolorize the anthraquinone dye Acid Green (AG 25 and diazo-dye Acid Red (AR 18. The results showed that decolorization of both dyes is Cu2+- and mediator-independent, with an optimum temperature of 35°C and pH of 3.0, and can be stably performed across a temperature range of 15°C to 45°C. A high activity toward AG25 (1 g/l with relative decolorization values of 91.2% (3 h and 97.1% (18 h, as well as high activity to AR18 (1 g/l by 80.5% (3 h and 89.0% (18 h, was recorded. The engineered system exhibited a comparably high activity compared with those of separate dyes in a continuous three-round shake-flask decolorization of AG25/AR18 mixed dye (each 1 g/l. No significant decline in decolorization efficacy was noted during first two-rounds but reaction equilibriums were elongated, and the residual laccase activity eventually decreased to low levels. However, the decolorizing capacity of the system was easily retrieved

  14. Immobilization free electrochemical biosensor for folate receptor in cancer cells based on terminal protection.

    Science.gov (United States)

    Ni, Jiancong; Wang, Qingxiang; Yang, Weiqiang; Zhao, Mengmeng; Zhang, Ying; Guo, Longhua; Qiu, Bin; Lin, Zhenyu; Yang, Huang-Hao

    2016-12-15

    The determination of folate receptor (FR) that over expressed in vast quantity of cancerous cells frequently is significant for the clinical diagnosis and treatment of cancers. Many DNA-based electrochemical biosensors have been developed for FR detection with high selectivity and sensitivity, but most of them need complicated immobilization of DNA on the electrode surface firstly, which is tedious and therefore results in the poor reproducibility. In this study, a simple, sensitive, and selective electrochemical FR biosensor in cancer cells has been proposed, which combines the advantages of the convenient immobilization-free homogeneous indium tin oxide (ITO)-based electrochemical detection strategy and the high selectivity of the terminal protection of small molecule linked DNA. The small molecule of folic acid (FA) and an electroactive molecule of ferrocence (Fc) were tethered to 3'- and 5'-end of an arbitrary single-stranded DNA (ssDNA), respectively, forming the FA-ssDNA-Fc complex. In the absence of the target FR, the FA-ssDNA-Fc was degraded by exonuclease I (Exo I) from 3'-end and produced a free Fc, diffusing freely to the ITO electrode surface and resulting in strong electrochemical signal. When the target FR was present, the FA-ssDNA-Fc was bound to FR through specific interaction with FA anchored at the 3'-end, effectively protecting the ssDNA strand from hydrolysis by Exo I. The FR-FA-ssDNA-Fc could not diffuse easily to the negatively charged ITO electrode surface due to the electrostatic repulsion between the DNA strand and the negatively charged ITO electrode, so electrochemical signal reduced. The decreased electrochemical signal has a linear relationship with the logarithm of FR concentration in range of 10fM to 10nM with a detection limit of 3.8fM (S/N=3). The proposed biosensor has been applied to detect FR in HeLa cancer cells, and the decreased electrochemical signal has a linear relationship with the logarithm of cell concentration ranging

  15. Magnetic immobilization of Bacillus subtilis natto cells for menaquinone-7 fermentation.

    Science.gov (United States)

    Ebrahiminezhad, Alireza; Varma, Vikas; Yang, Shuyi; Berenjian, Aydin

    2016-01-01

    Production of menaquinone-7 (MK-7) by Bacillus subtilis natto is associated with major drawbacks. To address the current challenges in MK-7 fermentation, studying the effect of magnetic nanoparticles on the bacterial cells can open up a new domain for intensified bioprocesses. This article introduces the new concept of application of iron oxide nanoparticles (IONs) as a pioneer tool for MK-7 process intensification. In this order, IONs with the average size of 11 nm were successfully fabricated and characterized for possible in situ removal of target substances from the fermentation media. The prepared particles were used for decoration and immobilization of B. subtilis natto cells. Presence of iron oxide nanoparticles significantly enhanced the MK-7 specific yield (15 %) as compared to the control samples. In addition, fabricated IONs showed a promising ability for in situ recovery of bacterial cells from the fermentation media with more than 95 % capture efficiency. Based on the results, IONs can be implemented successfully as a novel tool for MK-7 production. This study provides a considerable interest for industrial application of magnetic nanoparticles and their future role in designing an intensified biological process.

  16. Immobilization of Microbial Cells for Alcoholic and Malolactic Fermentation of Wine and Cider

    Science.gov (United States)

    Kourkoutas, Yiannis; Manojlović, Verica; Nedović, Viktor A.

    Wine- or cider-making is highly associated with biotechnology owing to the traditional nature of must fermentation.. Nowadays, there have been considerable developments in wine- or cider-making techniques affecting all phases of wine or cider production, but more importantly, the fermentation process. It is well-known that the transformation of grape must by microbial activity results in the production of wine, and the fermentation of apples (or sometimes pears) in the production of cider. In this process, a variety of compounds affecting the organoleptic profile of wine or cider are synthesized. It is also common sense that in wine- or cider-making, the main objective is to achieve an adequate quality of the product. The technological progress and the improved quality of the wines or ciders have been associated with the control of technical parameters. Herein, cell immobilization offers numerous advantages, such as enhanced fermentation productivity, ability for cell recycling, application of continuous configurations, enhanced cell stability and viability, and improvement of quality (Margaritis and Merchant 1984; Stewart and Russel 1986; Kourkoutas et al. 2004a).

  17. Modulation of Protein Adsorption and Cell Proliferation on Polyethylene Immobilized Graphene Oxide Reinforced HDPE Bionanocomposites.

    Science.gov (United States)

    Upadhyay, Rahul; Naskar, Sharmistha; Bhaskar, Nitu; Bose, Suryasarathi; Basu, Bikramjit

    2016-05-18

    The uniform dispersion of nanoparticles in a polymer matrix, together with an enhancement of interfacial adhesion is indispensable toward achieving better mechanical properties in the nanocomposites. In the context to biomedical applications, the type and amount of nanoparticles can potentially influence the biocompatibility. To address these issues, we prepared high-density polyethylene (HDPE) based composites reinforced with graphene oxide (GO) by melt mixing followed by compression molding. In an attempt to tailor the dispersion and to improve the interfacial adhesion, we immobilized polyethylene (PE) onto GO sheets by nucleophilic addition-elimination reaction. A good combination of yield strength (ca. 20 MPa), elastic modulus (ca. 600 MPa), and an outstanding elongation at failure (ca. 70%) were recorded with 3 wt % polyethylene grafted graphene oxide (PE-g-GO) reinforced HDPE composites. Considering the relevance of protein adsorption as a biophysical precursor to cell adhesion, the protein adsorption isotherms of bovine serum albumin (BSA) were determined to realize three times higher equilibrium constant (Keq) for PE-g-GO-reinforced HDPE composites as compared to GO-reinforced composites. To assess the cytocompatibility, we grew osteoblast cell line (MC3T3) and human mesenchymal stem cells (hMSCs) on HDPE/GO and HDPE/PE-g-GO composites, in vitro. The statistically significant increase in metabolically active cell over different time periods in culture for up to 6 days in MC3T3 and 7 days for hMSCs was observed, irrespective of the substrate composition. Such observation indicated that HDPE with GO or PE-g-GO addition (up to 3 wt %) can be used as cell growth substrate. The extensive proliferation of cells with oriented growth pattern also supported the fact that tailored GO addition can support cellular functionality in vitro. Taken together, the experimental results suggest that the PE-g-GO in HDPE can effectively be utilized to enhance both mechanical and

  18. Evaluation of Osteoblast-Like Cell Viability and Differentiation on the Gly-Arg-Gly-Asp-Ser Peptide Immobilized Titanium Dioxide Nanotube via Chemical Grafting.

    Science.gov (United States)

    Kim, Ga-Hyun; Kim, Il-Shin; Park, Sang-Won; Lee, Kwangmin; Yun, Kwi-Dug; Kim, Hyun-Seung; Oh, Gye-Jeong; Ji, Min-Kyung; Lim, Hyun-Pil

    2016-02-01

    This study examined the effect of the immobilization of the Gly-Arg-Gly-Asp-Ser (GRGDS) peptide on titanium dioxide (TiO2) nanotube via chemical grafting on osteoblast-like cell (MG-63) viability and differentiation. The specimens were divided into two groups; TiO2 nanotubes and GRGDS-immobilized TiO2 nanotubes. The surface characteristics of GRGDS-immobilized TiO2 nanotubes were observed by using X-ray photoelectron spectroscopy (XPS) and a field emission scanning electron microscope (FE-SEM). The morphology of cells on specimens was observed by FE-SEM after 2 hr and 24 hr. The level of cell viability was investigated via a tetrazolium (XTT) assay after 2 and 4 days. Alkaline phosphatase (ALP) activity was evaluated to measure the cell differentiation after 4 and 7 days. The presence of nitrogen up-regulation or C==O carbons con- firmed that TiO2 nanotubes were immobilized with GRGDS peptides. Cell adhesion was enhanced on the GRGDS-immobilized TiO2 nanotubes compared to TiO2 nanotubes. Furthermore, significantly increased cell spreading and proliferation were observed with the cells grown on GRGDS-immobilized TiO2 nanotubes (P nanotubes and TiO2 nanotubes. These results suggest that the GRGDS-immobilized TiO2 nanotubes might be effective in improving the osseointegration of dental implants.

  19. Development of thrombus-resistant and cell compatible crimped polyethylene terephthalate cardiovascular grafts using surface co-immobilized heparin and collagen

    Energy Technology Data Exchange (ETDEWEB)

    Al Meslmani, Bassam, E-mail: almeslmanib@yahoo.com [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Mahmoud, Gihan, E-mail: mahmoudg@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Department of Pharmaceutics and Industrial Pharmacy, Helwan University, Ain Helwan, 11795 Cairo (Egypt); Strehlow, Boris, E-mail: strehlo4@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Mohr, Eva, E-mail: mohr@staff.uni-marburg.de [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany); Leichtweiß, Thomas, E-mail: Thomas.leichtweiss@phys.chemie.uni-giessen.de [Institute of Physical Chemistry, Justus-Liebig-University, Heinrich-Buff-Ring 58, 35392 Giessen (Germany); Bakowsky, Udo, E-mail: ubakowsky@aol.com [Department of Pharmaceutical Technology and Biopharmaceutics, Marburg University, Ketzerbach 63, 35037 Marburg (Germany)

    2014-10-01

    Short-term patency of polyethylene terephthalate (PET) cardiovascular grafts is determined mainly by the inherent thrombogenicity and improper endothelialization following grafts implantation. The aim of the present study was to immobilize heparin to develop thrombus resistant grafts. Additionally, collagen was co-immobilized to enhance the host cell compatibility. The synthetic woven and knitted forms of crimped PET grafts were surface modified by Denier reduction to produce functional carboxyl groups. The produced groups were used as anchor sites for covalent immobilization of heparin or co-immobilization of heparin/collagen by the end-point method. The modified surface was characterized using Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The biological activity of immobilized molecules was investigated in vitro using direct blood coagulation test, and “platelet deposition under flow condition. Furthermore, the biocompatibility of modified grafts with host cells was assessed using L929 cell as model. All modified grafts showed significant resistance against fibrin and clot formation. The number of deposited platelets on heparin-immobilized woven and knitted grafts obviously decreased by 3 fold and 2.8 fold per unit surface area respectively, while the heparin/collagen co-immobilized grafts showed only a decrease by 1.7 and 1.8 fold compared to unmodified PET. Heparin-immobilized grafts reported no significant effect on L929 cells adhesion and growth (P > 0.05), conversely, collagen co-immobilization considerably increased cell adhesion almost ∼ 1.3 fold and 2 fold per unit surface area for woven and knitted grafts respectively. Our results emphasize that immobilization of heparin minimized the inherent thrombogenicity of the PET grafts. The simultaneous co-immobilization of collagen supported host cell adhesion and growth required for the grafts biocompatibility. - Highlight: • Heparin and collagen were co-immobilized on

  20. Properties of catechol 1,2-dioxygenase in the cell free extract and immobilized extract of Mycobacterium fortuitum

    Directory of Open Access Journals (Sweden)

    A.S. Silva

    2013-01-01

    Full Text Available Polycyclic aromatic hydrocarbons (PAH are carcinogenic compounds which contaminate water and soil, and the enzymes can be used for bioremediation of these environments. This study aimed to evaluate some environmental conditions that affect the production and activity of the catechol 1,2-dioxygenase (C12O by Mycobacterium fortuitum in the cell free and immobilized extract in sodium alginate. The bacterium was grown in mineral medium and LB broth containing 250 mg L-1 of anthracene (PAH. The optimum conditions of pH (4.0-9.0, temperature (5-70 ºC, reaction time (10-90 min and the effect of ions in the enzyme activity were determined. The Mycobacterium cultivated in LB shown higher growth and the C12O activity was two-fold higher to that in the mineral medium. To both extracts the highest enzyme activity was at pH 8.0, however, the immobilized extract promoted the increase in the C12O activity in a pH range between 4.0 and 8.5. The immobilized extract increased the enzymatic activity time and showed the highest C12O activity at 45 ºC, 20 ºC higher than the greatest temperature in the cell free extract. The enzyme activity in both extracts was stimulated by Fe3+, Hg2+ and Mn2+ and inhibited by NH4+ and Cu2+, but the immobilization protected the enzyme against the deleterious effects of K+ and Mg2+ in tested concentrations. The catechol 1,2-dioxygenase of Mycobacterium fortuitum in the immobilized extract has greater stability to the variations of pH, temperature and reaction time, and show higher activity in presence of ions, comparing to the cell free extract.

  1. Interaction force measurement between E. coli cells and nanoparticles immobilized surfaces by using AFM.

    Science.gov (United States)

    Zhang, Wen; Stack, Andrew G; Chen, Yongsheng

    2011-02-01

    To better understand environmental behaviors of nanoparticles (NPs), we used the atomic force microscopy (AFM) to measure interaction forces between E. coli cells and NPs immobilized on surfaces in an aqueous environment. The results showed that adhesion force strength was significantly influenced by particle size for both hematite (α-Fe(2)O(3)) and corundum (α-Al(2)O(3)) NPs whereas the effect on the repulsive force was not observed. The adhesion force decreased from 6.3±0.7nN to 0.8±0.4nN as hematite NPs increased from 26nm to 98nm in diameter. Corundum NPs exhibited a similar dependence of adhesion force on particle size. The Johnson-Kendall-Roberts (JKR) model was employed to estimate the contact area between E. coli cells and NPs, and based on the JKR model a new model that considers local effective contact area was developed. The prediction of the new model matched the size dependence of adhesion force in experimental results. Size effects on adhesion forces may originate from the difference in local effective contact areas as supported by our model. These findings provide fundamental information for interpreting the environmental behaviors and biological interactions of NPs, which barely have been addressed. PMID:20932723

  2. Interaction force measurement between E. coli cells and nanoparticles immobilized surfaces by using AFM

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Wen [Georgia Institute of Technology; Chen, Yongsheng [Georgia Institute of Technology

    2011-01-01

    To better understand environmental behaviors of nanoparticles (NPs), we used the atomic force microscopy (AFM) to measure interaction forces between E. coli cells and NPs immobilized on surfaces in an aqueous environment. The results showed that adhesion force strength was significantly influenced by particle size for both hematite ( -Fe2 O3 ) and corundum ( -Al2 O3 ) NPs whereas the effect on the repulsive force was not observed. The adhesion force decreased from 6.3 0.7 nN to 0.8 0.4 nN as hematite NPs increased from 26 nm to 98 nm in diameter. Corundum NPs exhibited a similar dependence of adhesion force on particle size. The Johnson Kendall Roberts (JKR) model was employed to estimate the contact area between E. coli cells and NPs, and based on the JKR model a new model that considers local effective contact area was developed. The prediction of the new model matched the size dependence of adhesion force in experimental results. Size effects on adhesion forces may originate from the difference in local effective contact areas as supported by our model. These findings provide fundamental information for interpreting the environmental behaviors and biological interactions of NPs, which barely have been addressed.

  3. Ethanol production from mahula (Madhuca latifolia L.) flowers with immobilized cells of Saccharomyces cerevisiae in Luffa cylindrica L. sponge discs

    Energy Technology Data Exchange (ETDEWEB)

    Behera, Shuvashish; Mohanty, Rama Chandra [Department of Botany, Utkal University, Vanivihar, Bhubaneswar 751 004, Orissa (India); Ray, Ramesh Chandra [Microbiology Laboratory, Central Tuber Crops Research Institute (Regional Centre), Bhubaneswar 751 019, Orissa (India)

    2011-01-15

    The dried spongy fruit of luffa (Luffa cylindrica L.), a cucurbitaceous crop available in abundance in tropical and sub-tropical countries has been found to be a promising material for immobilizing microbial cells. The aim of the present study was to examine the ethanol production from mahula flowers in submerged fermentation using whole cells of Saccharomyces cerevisiae immobilized in luffa sponge discs. The cells not only survived but also were physiologically active in three more cycles of fermentation without significant reduction (<5%) in ethanol production. After 96 h, there was 91.1% sugar conversion producing 223.2 g ethanol/kg flowers (1st cycle) which was 0.99%, 2.3% and 3.2% more than 2nd (221 g ethanol/kg flowers), 3rd (218 g ethanol/kg flowers) and 4th (216 g ethanol/kg flowers) cycle of fermentation, respectively. Furthermore, ethanol production by immobilized cells was 8.96% higher than the free cells. (author)

  4. The Experimental Study of the Performance of Nano-Thin Polyelectrolyte Shell for Dental Pulp Stem Cells Immobilization.

    Science.gov (United States)

    Grzeczkowicz, A; Granicka, L H; Maciejewska, I; Strawski, M; Szklarczyk, M; Borkowska, M

    2015-12-01

    Carious is the most frequent disease of mineralized dental tissues which might result in dental pulp inflammation and mortality. In such cases an endodontic treatment is the only option to prolong tooth functioning in the oral cavity; however, in the cases of severe pulpitis, especially when complicated with periodontal tissue inflammation, the endodontic treatment might not be enough to protect against tooth loss. Thus, keeping the dental pulp viable and/or possibility of the reconstruction of a viable dental pulp complex, appears to become a critical factor for carious and/or pulp inflammation treatment. The nowadays technologies, which allow handling dental pulp stem cells (DPSC), seem to bring us closer to the usage of dental stem cells for tooth tissues reconstruction. Thus, DPSC immobilized within nano-thin polymeric shells, allowing for a diffusion of produced factors and separation from bacteria, may be considered as a cover system supporting technology of dental pulp reconstruction. The DPSC were immobilized using a layer-by-layer technique within nano-thin polymeric shells constructed and modified by nanostructure involvement to ensure the layers stability and integrity as well as separation from bacterial cells. The cytotoxity of the material used for membrane production was assessed on the model of adherent cells. The performance of DPSC nano-coating was assessed in vitro. Membrane coatings showed no cytotoxicity on the immobilized cells. The presence of coating shell was confirmed with flow cytometry, atomic force microscopy and visualized with fluorescent microscopy. The transfer of immobilized DPSC within the membrane system ensuring cells integrity, viability and protection from bacteria should be considered as an alternative method for dental tissues transportation and regeneration. PMID:26682375

  5. Immobilization of Electroporated Cells for Fabrication of Cellular Biosensors: Physiological Effects of the Shape of Calcium Alginate Matrices and Foetal Calf Serum

    OpenAIRE

    Nikos Katsanakis; Andreas Katsivelis; Spiridon Kintzios

    2009-01-01

    In order to investigate the physiological effect of transfected cell immobilization in calcium alginate gels, we immobilized electroporated Vero cells in gels shaped either as spherical beads or as thin membrane layers. In addition, we investigated whether serum addition had a positive effect on cell proliferation and viability in either gel configuration. The gels were stored for four weeks in a medium supplemented or not with 20% (v/v) foetal calf serum. Throughout a culture period of four ...

  6. Removal of lead in wastewater by immobilized inactivated cells of Rhizopus oligosporus

    Institute of Scientific and Technical Information of China (English)

    于霞; 柴立元; 闵小波

    2003-01-01

    A novel technology for lead removal with nonliving Rhizopus oligosporus immobilized in calcium alginate was studied. The results show that the main influencing factors include pH value and interfering cations. pH value has different effects on biosorption of various heavy metals and lead adsorption can be proceeded by controlling pH value in a range of 2-5; interfering cations especially Cu( Ⅱ ) can make the adsorption amount of Pb( Ⅱ ) decrease by immobilized Rhizopus oligosporus. Desorption efficiency of different eluants and kinetics were investigated. Citrate the reaction equilibrium reaches 3 h. Immobilized biomass keeps high lead biosorption capacity after five cycles of regeneration.

  7. An ethanol biosensor based on a bacterial cell-immobilized eggshell membrane

    Institute of Scientific and Technical Information of China (English)

    Guang Ming Wen; Shao Min Shuang; Chuan Dong; Martin M.F. Choi

    2012-01-01

    An ethanol biosensor was fabricated based on a Methylobacterium organophilium-immobilized eggshell membrane and an oxygen (O2) electrode.A linear response for ethanol was obtained in the range of 0.050-7.5 mmol/L with a detection limit of 0.025 mmol/L (S/N =3) and a R.S.D.of 2.1%.The response time was less than 100 s at room temperature and ambient pressure.The optimal loading of bacterial cells on the biosensor membrane is 40 mg (wet weight).The optimal working conditions for the microbial biosensor are pH 7.0 phosphate buffer (50 mmol/L) at 20-25 ℃.The interference test,operational and storage stability of the biosensor are studied in detail.Finally,the biosensor is applied to determine the ethanol contents in various alcohol samples and the results are comparable to that obtained by gas chromatographic method and the results are satisfactory.Our proposed biosensor provides a convenient,simple and reliable method to determine ethanol content in alcoholic drinks.

  8. New biosensor for detection of copper ions in water based on immobilized genetically modified yeast cells.

    Science.gov (United States)

    Vopálenská, Irena; Váchová, Libuše; Palková, Zdena

    2015-10-15

    Contamination of water by heavy metals represents a potential risk for both aquatic and terrestrial organisms, including humans. Heavy metals in water resources can come from various industrial activities, and drinking water can be ex-post contaminated by heavy metals such as Cu(2+) from house fittings (e.g., water reservoirs) and pipes. Here, we present a new copper biosensor capable of detecting copper ions at concentrations of 1-100 μM. This biosensor is based on cells of a specifically modified Saccharomyces cerevisiae strain immobilized in alginate beads. Depending on the concentration of copper, the biosensor beads change color from white, when copper is present in concentrations below the detection limit, to pink or red based on the increase in copper concentration. The biosensor was successfully tested in the determination of copper concentrations in real samples of water contaminated with copper ions. In contrast to analytical methods or other biosensors based on fluorescent proteins, the newly designed biosensor does not require specific equipment and allows the quick detection of copper in many parallel samples.

  9. New biosensor for detection of copper ions in water based on immobilized genetically modified yeast cells.

    Science.gov (United States)

    Vopálenská, Irena; Váchová, Libuše; Palková, Zdena

    2015-10-15

    Contamination of water by heavy metals represents a potential risk for both aquatic and terrestrial organisms, including humans. Heavy metals in water resources can come from various industrial activities, and drinking water can be ex-post contaminated by heavy metals such as Cu(2+) from house fittings (e.g., water reservoirs) and pipes. Here, we present a new copper biosensor capable of detecting copper ions at concentrations of 1-100 μM. This biosensor is based on cells of a specifically modified Saccharomyces cerevisiae strain immobilized in alginate beads. Depending on the concentration of copper, the biosensor beads change color from white, when copper is present in concentrations below the detection limit, to pink or red based on the increase in copper concentration. The biosensor was successfully tested in the determination of copper concentrations in real samples of water contaminated with copper ions. In contrast to analytical methods or other biosensors based on fluorescent proteins, the newly designed biosensor does not require specific equipment and allows the quick detection of copper in many parallel samples. PMID:25982723

  10. A new technological approach proposed for distillate production using immobilized cells.

    Science.gov (United States)

    Loukatos, Paul; Kanellaki, Maria; Komaitis, Michael; Athanasiadis, Ilias; Koutinas, Athanasios A

    2003-01-01

    A new technological approach to distillate production using immobilized cells was investigated. The effect of temperature on the main volatile by-products in distillates was determined. Wines produced by delignified cellulose-, gluten- and kissiris-supported biocatalysis were used as starting materials. The produced distillates were analyzed for ethanol, methanol, acetaldehyde, ethyl acetate, propanol-1, isobutanol and amyl alcohol content. The results showed that distillates from delignified cellulosic material (DCM) at 16 degrees C contained smaller amounts of amyl alcohols, 57% of that produced by gluten and 32% of that produced by kissiris. The ethyl acetate content of distillates from DCM improved the aroma of distillates. These results agree with those of sensory evaluation. Subsequently, the scale-up for low-temperature distillate production at 16 degrees C using DCM was further investigated. A new version of an industrial multi-stage fixed bed tower (MFBT) bioreactor with a capacity of 11,000 l proved to be suitable for continuous fermentation by DCM-supported biocatalysis. Economic analysis showed a reduction in the cost of almost 30% for distillate production and 78% for wine production.

  11. Immobilization of Electroporated Cells for Fabrication of Cellular Biosensors: Physiological Effects of the Shape of Calcium Alginate Matrices and Foetal Calf Serum

    Directory of Open Access Journals (Sweden)

    Nikos Katsanakis

    2009-01-01

    Full Text Available In order to investigate the physiological effect of transfected cell immobilization in calcium alginate gels, we immobilized electroporated Vero cells in gels shaped either as spherical beads or as thin membrane layers. In addition, we investigated whether serum addition had a positive effect on cell proliferation and viability in either gel configuration. The gels were stored for four weeks in a medium supplemented or not with 20% (v/v foetal calf serum. Throughout a culture period of four weeks, cell proliferation and cell viability were assayed by optical microscopy after provision of Trypan Blue. Non-elaborate culture conditions (room temperature, non-CO2 enriched culture atmosphere were applied throughout the experimental period in order to evaluate cell viability under less than optimal storage conditions. Immobilization of electroporated cells was associated with an initially reduced cell viability, which was gradually increased. Immobilization was associated with maintenance of cell growth for the duration of the experimental period, whereas electroporated cells essentially died after a week in suspension culture. Considerable proliferation of immobilized cells was observed in spherical alginate beads. In both gel configurations, addition of serum was associated with increased cell proliferation. The results of the present study could contribute to an improvement of the storability of biosensors based on electroporated, genetically or membrane-engineered cells.

  12. Time-Course Changes of Steroidogenic Gene Expression and Steroidogenesis of Rat Leydig Cells after Acute Immobilization Stress

    Directory of Open Access Journals (Sweden)

    Han Lin

    2014-11-01

    Full Text Available Leydig cells secrete testosterone, which is essential for male fertility and reproductive health. Stress increases the secretion of glucocorticoid (corticosterone, CORT; in rats, which decreases circulating testosterone levels in part through a direct action by binding to the glucocorticoid receptors (NR3C1 in Leydig cells. The intratesticular CORT level is dependent on oxidative inactivation of glucocorticoid by 11β-hydroxysteroid dehydrogenase 1 (HSD11B1 in Leydig cells. In the present study, we investigated the time-course changes of steroidogenic gene expression levels after acute immobilization stress in rats. The plasma CORT levels were significantly increased 0.5, 1, 3 and 6 h after immobilization stress, while plasma testosterone levels were significantly reduced 3 and 6 h, after stress and luteinizing hormone (LH did not change. Immobilization stress caused the down-regulation of Scarb1, Star and Cyp17a1 expression levels in the rat testis starting at the first hour of stress, ahead of the significant decreases of plasma testosterone levels. Other mRNA levels, including Cyp11a1, Hsd3b1 and Hsd17b3, began to decline after 3 h. Hsd11b1 and Nos2 mRNA levels did not change during the course of stress. Administration of glucocorticoid antagonist RU486 significantly restored plasma testosterone levels. In conclusion, Scarb1, Star and Cyp17a1 expression levels are more sensitive to acute stress, and acute immobilization stress causes the decline of the steroidogenic pathway via elevating the levels of glucocorticoid, which binds to NR3C1 in Leydig cells to inhibit steroidogenic gene expression.

  13. Use of an Immobilized Monoclonal Antibody to Examine Integrin &agr;5&bgr;1 Signaling Independent of Cell Spreading

    Directory of Open Access Journals (Sweden)

    Bao Wenjie

    2002-01-01

    Full Text Available Cell attachment to the extracellular matrix (ECM engages integrin signaling into the cell, but part of the signaling response also stem from cell spreading (3. To analyze specific integrin signaling-mediated responses independent of cell spreading, we developed a method engaging integrin signaling by use of an immobilized anti-integrin monoclonal antibody (mab directed against the fibronectin (FN receptor integrin &agr;5&bgr;1. ECV 304 cells were plated onto FN or immobilized mab JBS5 (anti-integrin &agr;5&bgr;1 or onto poly-L-lysin (P-L-L, which mediates integrin-independent attachment. Cells attached and spread on FN, while cells on JBS5 or P-L-L attached but did not spread. Importantly, plating onto FN or mab JBS5 gave rise to identical integrin-induced responses, including a down-regulation of the cyclin-dependent kinase (Cdk2 inhibitors p21CIP1 and p27KIP1, while attachment to P-L-L did not. We conclude that engagement of the FN-receptor integrin &agr;5&bgr;1 induces integrin signaling regulating the Cdk2-inhibitors independent of cell spreading and present a method for how integrin signaling can be analyzed separate from the effects of cell spreading.

  14. Interaction between immobilized polyelectrolyte complex nanoparticles and human mesenchymal stromal cells

    Directory of Open Access Journals (Sweden)

    Woltmann B

    2014-05-01

    Full Text Available Beatrice Woltmann,1 Bernhard Torger,2,3 Martin Müller,2,3,* Ute Hempel1,*1Dresden University of Technology, Faculty of Medicine Carl Gustav Carus, Institute of Physiological Chemistry, Dresden, Germany; 2Leibniz Institute of Polymer Research Dresden, Department of Polyelectrolytes and Dispersions, Dresden, Germany; 3Dresden University of Technology, Department of Chemistry and Food Chemistry, Dresden, Germany*These authors contributed equally to this workBackground: Implant loosening or deficient osseointegration is a major problem in patients with systemic bone diseases (eg, osteoporosis. For this reason, the stimulation of the regional cell population by local and sustained drug delivery at the bone/implant interface to induce the formation of a mechanical stable bone is promising. The purpose of this study was to investigate the interaction of polymer-based nanoparticles with human bone marrow-derived cells, considering nanoparticles’ composition and surface net charge.Materials and methods: Polyelectrolyte complex nanoparticles (PECNPs composed of the polycations poly(ethyleneimine (PEI, poly(L-lysine (PLL, or (N,N-diethylaminoethyldextran (DEAE in combination with the polyanions dextran sulfate (DS or cellulose sulfate (CS were prepared. PECNPs’ physicochemical properties (size, net charge were characterized by dynamic light scattering and particle charge detector measurements. Biocompatibility was investigated using human mesenchymal stromal cells (hMSCs cultured on immobilized PECNP films (5–50 nmol·cm-2 by analysis for metabolic activity of hMSCs in dependence of PECNP surface concentration by MTS (3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium, inner salt assay, as well as cell morphology (phase contrast microscopy.Results: PECNPs ranging between ~50 nm and 150 nm were prepared. By varying the ratio of polycations and polyanions, PECNPs with a slightly positive (PEC+NP or negative (PEC

  15. Effect of concentration and substrate flow rate on isomaltulose production from sucrose by Erwinia sp. cells immobilized in calcium-alginate using packed bed reactor.

    Science.gov (United States)

    Kawaguti, Haroldo Yukio; Harumi Sato, Hélia

    2010-09-01

    Isomaltulose was obtained from sucrose solution by immobilized cells of Erwinia sp. D12 using a batch and a continuous process. Parameters for sucrose conversion into isomaltulose were evaluated using both experimental design and response surface methodology. Erwinia sp. D12 cells were immobilized in different alginates, and the influence of substrate flow rate and concentration parameters to produce isomaltulose from sucrose were observed. Response surface methodology demonstrated that packed bed columns containing cells immobilized in low-viscosity sodium alginate (250 cP) presented a mean isomaltulose conversion rate of 47%. In a continuous process, both sucrose substrate concentration and substrate flow rate parameters had a significant effect (p < 0.05) and influenced the conversion of sucrose into isomaltulose. Higher conversion rates of sucrose into isomaltulose, from 53-75% were obtained using 75 g of immobilized cells at a substrate flow rate of 0.6 mL/min.

  16. Biosorption of copper (II) onto immobilized cells of Pycnoporus sanguineus from aqueous solution: Equilibrium and kinetic studies

    International Nuclear Information System (INIS)

    The ability of white-rot fungus, Pycnoporus sanguineus to adsorb copper (II) ions from aqueous solution is investigated in a batch system. The live fungus cells were immobilized into Ca-alginate gel to study the influence of pH, initial metal ions concentration, biomass loading and temperature on the biosorption capacity. The optimum uptake of Cu (II) ions was observed at pH 5 with a value of 2.76 mg/g. Biosorption equilibrium data were best described by Langmuir isotherm model followed by Redlich-Peterson and Freundlich models, respectively. The biosorption kinetics followed the pseudo-second order and intraparticle diffusion equations. The thermodynamic parameters enthalpy change (10.16 kJ/mol) and entropy change (33.78 J/mol K) were determined from the biosorption equilibrium data. The FTIR analysis showed that -OH, -NH, C-H, C=O, -COOH and C-N groups were involved in the biosorption of Cu (II) ions onto immobilized cells of P. sanguineus. The immobilized cells of P. sanguineus were capable of removing Cu (II) ions from aqueous solution

  17. Immobilization of Lactobacillus rhamnosus in mesoporous silica-based material: An efficiency continuous cell-recycle fermentation system for lactic acid production.

    Science.gov (United States)

    Zhao, Zijian; Xie, Xiaona; Wang, Zhi; Tao, Yanchun; Niu, Xuedun; Huang, Xuri; Liu, Li; Li, Zhengqiang

    2016-06-01

    Lactic acid bacteria immobilization methods have been widely used for lactic acid production. Until now, the most common immobilization matrix used is calcium alginate. However, Ca-alginate gel disintegrated during lactic acid fermentation. To overcome this deficiency, we developed an immobilization method in which Lactobacillus rhamnosus cells were successfully encapsulated into an ordered mesoporous silica-based material under mild conditions with a high immobilization efficiency of 78.77% by using elemental analysis. We also optimized the cultivation conditions of the immobilized L. rhamnosus and obtained a high glucose conversion yield of 92.4%. Furthermore, L. rhamnosus encapsulated in mesoporous silica-based material exhibited operational stability during repeated fermentation processes and no decrease in lactic acid production up to 8 repeated batches.

  18. Immobilization of Lactobacillus rhamnosus in mesoporous silica-based material: An efficiency continuous cell-recycle fermentation system for lactic acid production.

    Science.gov (United States)

    Zhao, Zijian; Xie, Xiaona; Wang, Zhi; Tao, Yanchun; Niu, Xuedun; Huang, Xuri; Liu, Li; Li, Zhengqiang

    2016-06-01

    Lactic acid bacteria immobilization methods have been widely used for lactic acid production. Until now, the most common immobilization matrix used is calcium alginate. However, Ca-alginate gel disintegrated during lactic acid fermentation. To overcome this deficiency, we developed an immobilization method in which Lactobacillus rhamnosus cells were successfully encapsulated into an ordered mesoporous silica-based material under mild conditions with a high immobilization efficiency of 78.77% by using elemental analysis. We also optimized the cultivation conditions of the immobilized L. rhamnosus and obtained a high glucose conversion yield of 92.4%. Furthermore, L. rhamnosus encapsulated in mesoporous silica-based material exhibited operational stability during repeated fermentation processes and no decrease in lactic acid production up to 8 repeated batches. PMID:26803707

  19. Tris-sucrose buffer system: a new specially designed medium for extracellular invertase production by immobilized cells of isolated yeast Cryptococcus laurentii MT-61.

    Science.gov (United States)

    Aydogan, Mehmet Nuri; Taskin, Mesut; Canli, Ozden; Arslan, Nazli Pinar; Ortucu, Serkan

    2014-01-01

    The aims of the present study were to isolate new yeasts with high extracellular (exo) invertase activity and to investigate the usability of buffer systems as invertase production media by immobilized yeast cells. Among 70 yeast isolates, Cryptococcus laurentii MT-61 had the highest exo-invertase activity. Immobilization of yeast cells was performed using sodium alginate. Higher exo-invertase activity for immobilized cells was achieved in tris-sucrose buffer system (TSBS) compared to sodium acetate buffer system and potassium phosphate buffer system. TSBS was prepared by dissolving 30 g of sucrose in 1 L of tris buffer solution. The optimum pH, temperature, and incubation time for invertase production with immobilized cells were determined as 8.0, 35 °C and 36 h in TSBS, respectively. Under optimized conditions, maximum exo-invertase activity was found to be 28.4 U/mL in sterile and nonsterile TSBS. Immobilized cells could be reused in 14 and 12 successive cycles in sterile and nonsterile TSBS without any loss in the maximum invertase activity, respectively. This is the first report which showed that immobilized microbial cells could be used as a biocatalyst for exo-invertase production in buffer system. As an additional contribution, a new yeast strain with high invertase activity was isolated.

  20. Electrochemical characterization of methanol/O2 biofuel cell: Use of laccase biocathode immobilized with polypyrrole film and PAMAM dendrimers

    International Nuclear Information System (INIS)

    This paper describes the performance of a mediated electron transfer (MET) biocathode for a methanol/O2 biofuel cell. To this end, we employed PAMAM (polyamidoamine) dendrimers for the immobilization of laccase using 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) diammonium salt (ABTS) in solution or entrapped into polypyrrole films. We used the enzyme immobilized onto the carbon platform obtained either in the presence or in the absence of the electropolymerized film to determine kinetic parameters. The results point to a very similar kinetic rate conversion in both situations; however, substrate affinity seems to increase in the bioelectrode containing the entrapped substrate molecules. The electrochemical characterization tests confirmed that the electropolymerized polypyrrole film was able to retain entrapped ABTS molecules. Additionally, laccase provides enhanced catalytic oxidation current for the mediator compared with the control sample containing PAMAM dendrimer only. Compared to the control sample, which gave power density values around 0.7 μW cm−2, tests employing ABTS as mediator furnished 6 μW cm−2 when the mediator was added in solution and around 25 μW cm−2 when it was entrapped into the biocathode layers. Overall, the developed biocathode is environmentally friendly for immobilization of the enzyme laccase, being satisfactorily stable in the kinetic tests and affording good power data in the biofuel cell tests

  1. Optimizing immobilized enzyme performance in cell-free environments to produce liquid fuels.

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Sanat

    2015-02-05

    The overall goal of this project was to optimize enzyme performance for the production of bio-diesel fuel. Enzyme immobilization has attracted much attention as a means to increase productivity. Mesorporous silica materials have been known to be best suited for immobilizing enzymes. A major challenge is to ensure that the enzymatic activity is retained after immobilization. Two major factors which drive enzymatic deactivation are protein-surface and inter-protein interactions. Previously, we studied protein stability inside pores and how to optimize protein-surface interactions to minimize protein denaturation. In this work we studied eh effect of surface curvature and chemistry on inter-protein interactions. Our goal was to find suitable immobilization supports which minimize these inter-protein interactions. Our studies carried out in the frame work of Hydrophobic-Polar (HP) model showed that enzymes immobilized inside hydrophobic pores of optimal sizes are best suited to minimize these inter-protein interactions. Besides, this study is also of biological importance to understand the role of chaperonins in protein disaggregation. Both of these aspects profited immensely with collaborations with our experimental colleague, Prof. Georges Belfort (RPI), who performed the experimental analog of our theoretical works.

  2. Immobilization by Polyurethane of Pseudomonas dacunhae Cells Containing l-Aspartate β-Decarboxylase Activity and Application to l-Alanine Production

    Science.gov (United States)

    Fusee, Murray C.; Weber, Jennifer E.

    1984-01-01

    Whole cells of Pseudomonas dacunhae containing l-aspartate β-decarboxylase activity were immobilized by mixing a cell suspension with a liquid isocyanate-capped polyurethane prepolymer (Hypol; W. R. Grace & Co., Lexington, Mass.). The immobilized cell preparation was used to convert l-aspartic acid to l-alanine. Properties of the immobilized P. dacunhae cells containing aspartate β-decarboxylase activity were investigated with batch reactors. Retention of enzyme activity was observed to be as much as 100% when cell lysis was allowed to occur before immobilization. The pH and temperature optima were determined to be 5.5 and 45°C, respectively. Immobilized P. dacunhael-aspartate β-decarboxylase activity was stabilized by the addition of 0.1 mM pyridoxal-5-phosphate and 0.1 mM α-ketoglutaric acid to a 1.7 M ammonium aspartate (pH 5.5) substrate solution. Under conditions of semicontinuous use in a batch reactor, a 2.5% loss in immobilized l-aspartate β-decarboxylase activity was observed over a 31-day period. PMID:16346636

  3. Ethanol production by repeated batch and continuous fermentations of blackstrap molasses using immobilized yeast cells on thin-shell silk cocoons

    International Nuclear Information System (INIS)

    Highlights: → Thin-shell silk cocoons for immobilization of Saccharomycescerevisiae. → Advantages: high mechanical strength, light weight, biocompatibility and high surface area. → Enhanced cell stability and ethanol productivity by the immobilization system. -- Abstract: A thin-shell silk cocoon (TSC), a residual from the silk industry, is used as a support material for the immobilization of Saccharomyces cerevisiae M30 in ethanol fermentation because of its properties such as high mechanical strength, light weight, biocompatibility and high surface area. In batch fermentation with blackstrap molasses as the main fermentation substrate, an optimal ethanol concentration of 98.6 g/L was obtained using a TSC-immobilized cell system at an initial reducing sugar concentration of 240 g/L. The ethanol concentration produced by the immobilized cells was 11.5% higher than that produced by the free cells. Ethanol production in five-cycle repeated batch fermentation demonstrated the enhanced stability of the immobilized yeast cells. Under continuous fermentation in a packed-bed reactor, a maximum ethanol productivity of 19.0 g/(L h) with an ethanol concentration of 52.8 g/L was observed at a 0.36 h-1 dilution rate.

  4. Quantitation of residual trypsin in cell-based therapeutics using immobilized α-1-antitrypsin or SBTI in an ELISA format.

    Science.gov (United States)

    Braatz, James A; Elias, Christopher; Finny, Joseph G; Tran, Huan; McCaman, Michael

    2015-02-01

    An Enzyme-Linked Immunosorbent Assay (ELISA) has been developed for the quantitation of porcine trypsin as a process residual in cell therapy products based on its capture by either of two immobilized anti-trypsins, α-1-antitrypsin (α1AT) or soybean trypsin inhibitor (SBTI) followed by detection with a polyclonal goat anti-porcine trypsin-IgG conjugated with peroxidase. It was demonstrated that an extended range of antigen quantitation could be achieved that covered nearly three orders of magnitude of trypsin concentration. The utility of the assay was demonstrated by its application to samples generated in a cell-based therapeutic manufacturing setting.

  5. Hydrogen Photoproduction by Nutrient-Deprived Chalamydomonas reinhardtii Cells Immobilized Within Thin Alginate Films Under Aerobic and Anaerobic Conditions

    Energy Technology Data Exchange (ETDEWEB)

    Kosourov, S. N.; Seibert, M.

    2009-01-01

    A new technique for immobilizing H{sub 2}-photoproducing green algae within a thin (<400 {micro}m) alginate film has been developed. Alginate films with entrapped sulfur/phosphorus-deprived Chlamydomonas reinhardtii, strain cc124, cells demonstrate (a) higher cell density (up to 2,000 {micro}g Chl mL{sup -1} of matrix), (b) kinetics of H{sub 2} photoproduction similar to sulfur-deprived suspension cultures, (c) higher specific rates (up to 12.5 {micro}mol mg{sup -1} Chl h{sup -1}) of H{sub 2} evolution, (d) light conversion efficiencies to H{sub 2} of over 1% and (e) unexpectedly high resistance of the H{sub 2}-photoproducing system to inactivation by atmospheric O{sub 2}. The algal cells, entrapped in alginate and then placed in vials containing 21% O{sub 2} in the headspace, evolved up to 67% of the H{sub 2} gas produced under anaerobic conditions. The results indicate that the lower susceptibility of the immobilized algal H{sub 2}-producing system to inactivation by O{sub 2} depends on two factors: (a) the presence of acetate in the medium, which supports higher rates of respiration and (b) the capability of the alginate polymer itself to effectively separate the entrapped cells from O{sub 2} in the liquid and headspace and restrict O{sub 2} diffusion into the matrix. The strategy presented for immobilizing algal cells within thin polymeric matrices shows the potential for scale-up and possible future applications.

  6. Hemicellulosic Ethanol Production by Immobilized Wild Brazilian Yeast Scheffersomyces shehatae UFMG-HM 52.2: Effects of Cell Concentration and Stirring Rate.

    Science.gov (United States)

    Antunes, F A F; Santos, J C; Chandel, A K; Milessi, T S S; Peres, G F D; da Silva, S S

    2016-02-01

    The use of sugarcane bagasse hemicellulosic hydrolysates presents an interesting alternative to second generation (2G) ethanol production. Techniques to enhance the fermentation process, e.g., the use of immobilized cells, is one of the key factors for efficient production. Here, the effect of two important parameters (cell concentration in immobilized system and stirring rate) on the 2G ethanol production using the wild Brazilian yeast S. shehatae UFMG-HM 52.2 immobilized in calcium alginate matrix are presented. A 2(2) full factorial design of experiments was carried out to evaluate the effect of cell concentrations in sodium alginate solution for immobilized bead production (3.0, 6.0, and 9.0 g/L) and stirring rate (150, 200, and 250 rpm) for 2G ethanol production. Statistical analysis showed that the use of both variables at low levels enhanced ethanol yield (YP/S). Under these process conditions, YP/S of 0.31 g/g and ethanol productivity (Qp) of 0.12 g/L h were achieved. Results showed the potential of this immobilized yeast in 2G ethanol production from C5 sugars and demonstrate the importance of adequate cell concentration in immobilized systems, a finding that stands to increase bioprocesses yields and productivity.

  7. KINETIC STUDIES ON BIODEGRADATION OF LIPIDS FROM OLIVE OIL MILL WASTEWATERS WITH FREE AND IMMOBILIZED Bacillus sp. CELLS

    Directory of Open Access Journals (Sweden)

    Anca-Irina Galaction

    2012-03-01

    Full Text Available The studies on the biodegradation of lipids from olive oil mill wastewater with free and immobilized Bacillus sp. cells indicated that the maximum specific rate of the process is reached at pH = 8. The use of immobilized cells allows to increasing the number of biodegradation process cycles, but reduces the rate of the process. In this case, the process rate depends on the biocatalysts size and cells concentration inside them. Thus, at bacterial cells concentration of 9 g d.w./100 mL biocatalyst, the apparent specific rate varied from 4.65 to 1.46×10-2 h-1 by increasing the biocatalyst particles diameter from 3 to 4.2 mm.The cumulated influences of the particles size and cells concentration have been included in a mathematical model for the apparent specific rate of lipids biodegradation. The model offers a good concordance with the experimental data, the average deviation being of +/- 7.38%.

  8. Enhanced production of alkaline thermostable keratinolytic protease from calcium alginate immobilized cells of thermoalkalophilic Bacillus halodurans JB 99 exhibiting dehairing activity.

    Science.gov (United States)

    Shrinivas, Dengeti; Kumar, Raghwendra; Naik, G R

    2012-01-01

    The thermoalkalophilic Bacillus halodurans JB 99 cells known for production of novel thermostable alkaline keratinolytic protease were immobilized in calcium alginate matrix. Batch and repeated batch cultivation using calcium alginate immobilized cells were studied for alkaline protease production in submerged fermentation. Immobilized cells with 2.5% alginate and 350 beads/flask of initial cell loading showed enhanced production of alkaline protease by 23.2% (5,275 ± 39.4 U/ml) as compared to free cells (4,280 ± 35.4 U/ml) after 24 h. In the semicontinuous mode of cultivation, immobilized cells under optimized conditions produced an appreciable level of alkaline protease in up to nine cycles and reached a maximal value of 5,975 U/ml after the seventh cycle. The enzyme produced from immobilized cells efficiently degraded chicken feathers in the presence of a reducing agent which can help the poultry industry in the management of keratin-rich waste and obtaining value-added products.

  9. Bioactivity of immobilized hyaluronic acid derivatives regarding protein adsorption and cell adhesion

    DEFF Research Database (Denmark)

    Köwitsch, Alexander; Yang, Yuan; Ma, Ning;

    2011-01-01

    Hyaluronic acid (HA) was chemically modified either by oxidation to obtain aldehyde-HA (aHA) or 3,3'-dithiobis(propanoic hydrazide) to obtain thiol-HA (tHA) that was covalently immobilized on model substrata such as amino-terminated surfaces or gold. Knowledge about the effect of modification with...

  10. 海澡酸钙胶囊化重组E.coli BL21(DE3)生产靛蓝%Production of Indigo by Immobilization of E.coli BL21 (DE3) Cells in Calcium- Alginate Gel Capsules

    Institute of Scientific and Technical Information of China (English)

    陆燕; 梅乐和

    2007-01-01

    The ability of catalyzing indole into indigo of gene engineering strain expressing P450 BM3 immobilized by entrapment in calcium-alginate gel capsules was examined, and various characteristics of immobilized cells were assessed.Optimum conditions for cells activity were not affected after immobilization, and pH and temperature for both free and immobilized cells were found to be pH 7.5 and 35℃, respectively.The immobilized cells exhibited a markedly improved thermal stability than free cells.After five repeated experiments, the yield of indigo with the immobilized cells retained over 94% of their original activity, which indicated that the operational stability for recycling in batch processes was improved.

  11. Aerobic decolorization and degradation of Acid Orange G (AOG) by suspended growing cells and immobilized cells of a yeast strain Candida tropicalis TL-F1.

    Science.gov (United States)

    Tan, Liang; Li, Hua; Ning, Shuxiang; Hao, Jia

    2014-10-01

    In this study, aerobic decolorization and degradation of azo dye Acid Orange G (AOG) by both suspended growing cells and immobilized cells of a yeast strain Candida tropicalis TL-F1 were studied. The effects of different parameters on decolorization of AOG by both growing suspended and immobilized strain TL-F1 were investigated. Furthermore, a possible decolorization mechanism of AOG was proposed through analyzing metabolic intermediates using UV-vis and high-performance liquid chromatography-mass spectrometry (HPLC-MS) methods. Strain TL-F1 could decolorize AOG in both liquid and solid mediums through degradation. The optimal conditions for decolorization with suspended growing cells of strain TL-F1 were as follows: 6-10 g/L sucrose, 5-7 g/L urea, ≥6 % (v/v) inoculation size, ≥160 rpm, 35-40 °C, and pH 5.0-6.0; and those for immobilized cells, the conditions were as follows: 4-6 g/L glucose, 0.2-0.4 g/L urea, 6-10 g/L (wet cell pellets) inoculation size, ≥160 rpm, 35-40 °C, and pH 5.0-7.0. Results of UV-vis scanning spectra suggested that AOG was decolorized through biodegradation, and the possible pathway was proposed through the results of HPLC-MS analysis and related literature. This is a systematic research on aerobic decolorization and degradation of AOG by both suspended and immobilized cells of a C. tropicalis strain.

  12. Biodegradation of benzidine based azodyes Direct red and Direct blue by the immobilized cells of Pseudomonas fluorescens D41.

    Science.gov (United States)

    Puvaneswari, N; Muthukrishnan, J; Gunasekaran, P

    2002-10-01

    Benzidine based azodyes are proven carcinogens, mutagens and have been linked to bladder cancer of human beings and laboratory animals. The textile and dyestuff manufacturing industry are the two major sources that released azodyes in their effluents. The dye, Direct blue contains two carcinogenic compounds namely benzidine (BZ), 4-amino biphenyl (4-ABP), while the dye Direct red has benzidine (BZ). Among 40 isolates of Pseudomonas fluorescens screened, one isolate designated as D41 was found to be capable of extensively degrading the dyes Direct blue and Direct red. Immobilized cells of P. fluorescens D41 efficiently degraded Direct red (82%) and Direct blue (71%) in the presence of glucose.

  13. Production of isomaltulose obtained by Erwinia sp. cells submitted to different treatments and immobilized in calcium alginate

    Directory of Open Access Journals (Sweden)

    Haroldo Yukio Kawaguti

    2011-03-01

    Full Text Available In recent decades, there has been an increase in the studies of isomaltulose obtainment, due to its physicochemical properties and physiological health benefits. These properties, which include low cariogenicity, low glycemic index and greater stability, allow the use of this sweetener as a substitute for sucrose in foods; besides the fact that it can be converted to isomalt, a dietary non-cariogenic sugar alcohol used in pharmaceuticals as well as in the food industry. Isomaltulose (6-O-α-D-glucopyronosyl-1-6-D-fructofuranose is a disaccharide reducer obtained by the enzymatic conversion of sucrose - the α-glucosyltransferase enzyme. Different treatments were performed for the preparation of whole cells; lysed cells; and crude enzyme extract of Erwinia sp. D12 strain immobilized in calcium alginate. The packed bed column of granules, containing Erwinia sp. cells sonicated and immobilized in calcium alginate (CSI, reached a maximum conversion of 53-59% sucrose into isomaltulose and it presented activity for 480 hours. The converted syrup was purified and the isomaltulose crystallization was performed through the lowering of temperature. The isomaltulose crystals presented purity of 96.5%.

  14. Osteoinductive Effects of Free and Immobilized Bone Forming Peptide-1 on Human Adipose-Derived Stem Cells.

    Directory of Open Access Journals (Sweden)

    Wenyue Li

    Full Text Available Most synthetic polymeric materials currently used for bone tissue engineering lack specific signals through which cells can identify and interact with the surface, resulting in incompatibility and compromised osteogenic activity. Soluble inductive factors also have issues including a short half-live in vivo. Bone forming peptide-1 is a truncated peptide from the immature form of bone morphogenetic protein-7 (BMP-7 that displays higher osteogenic activity than full-length, mature BMP-7. In this study, we used a mussel-inspired immobilization strategy mediated by polymerization of dopamine to introduce recently discovered stimulators of bone forming peptide-1 (BFP-1 onto the surface of poly-lactic-co-glycolic acid (PLGA substrate to form a biomaterial that overcomes these challenges. Human adipose-derived stem cells (hASCs, being abundant and easy accessible, were used to test the osteogenic activity of BFP-1 and the novel biomaterial. Under osteoinductive conditions, cells treated with both BFP-1 alone and BFP-1-coated biomaterials displayed elevated expression of the osteogenic markers alkaline phosphatase (ALP, osteocalcin (OC, and RUNX2. Furthermore, hASCs associated with poly-dopamine-assisted BFP-1-immobilized PLGA (pDA-BFP-1-PLGA scaffolds promoted in vivo bone formation in nude mice. Our novel materials may hold great promise for future bone tissue engineering applications.

  15. Preservation of Bacillus firmus Strain 37 and Optimization of Cyclodextrin Biosynthesis by Cells Immobilized on Loofa Sponge

    Directory of Open Access Journals (Sweden)

    Cristiane Moriwaki

    2012-08-01

    Full Text Available The preservation of Bacillus firmus strain 37 cells by lyophilization was evaluated and response surface methodology (RSM was used to optimize the β-cyclodextrin (β-CD production by cells immobilized on loofa sponge. Interactions were studied with the variables temperature, pH and dextrin concentration using a central composite design (CCD. Immobilization time influence on β-CD production was also investigated. B. firmus strain 37 cells remained viable after one year of storage, showing that the lyophilization is a suitable method for preservation of the microorganism. From the three-dimensional diagrams and contour plots, the best conditions for β-CD production were determined: temperature 60 °C, pH 8, and 18% dextrin. Considering that the amount of dextrin was high, a new assay was carried out, in which dextrin concentrations of 10, 15, and 18% were tested and the temperature of 60 °C and pH 8 were maintained. The results achieved showed very small differences and therefore, for economic reasons, the use of 10% dextrin is suggested. Increasing the immobilization time of cells immobilized on synthetic sponge the β-CD production decreased and did not change for cells immobilized on loofa sponge. The results of this research are important for microorganism preservation and essential in the optimization of the biosynthesis of CD.

  16. 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.

    Science.gov (United States)

    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.

  17. Elongation of lifetime of photosynthetic biofuel-cells containing immobilized algae; Koteika aiso wo mochiita kogosei biseibutsu denchi no chojumyoka

    Energy Technology Data Exchange (ETDEWEB)

    Yagishita, T.; Sawayama, S.; Inoue, S.; Ogi, T. [National Institute for Resources and Environment, Tsukuba (Japan)

    1994-12-08

    An experimental study is performed for elongation of lifetime of photosynthetic biofuel-cells using the living blue-green algae and a mediator. In the experiment, correlation between a current generated from cultured Anabaena and the life of the cells is investigated. Anabaena is recovered from the cells after the cells are operated for 10 hours in the dark and is cultured for 10 hours under irradiation with a Xe lamp and ventilation of 3 % CO2. Thereafter, immobilized Anabaena is returned into the cells and the cells are again actuated in repetition. Three load resistances 1 K ohm, 700 ohm, 400 ohm are employed and operation time of the current is lengthened under any conditions compared with the case where the cells are continuously operated. Further, provided a generated current is limited to 0.6 mA or lower, the current is not lowered even if the cells are operated for 90 hours. It is concluded that provided Anabaena is cultured after the electricity of 6.4 mA/h per the amount of chlorophyl in Anabaena is taken out, an output of the cells is kept unchanged for a long time. 7 refs., 4 figs., 1 tab.

  18. Manipulation of culture strategies to enhance capsaicin biosynthesis in suspension and immobilized cell cultures of Capsicum chinense Jacq. cv. Naga King Chili.

    Science.gov (United States)

    Kehie, Mechuselie; Kumaria, Suman; Tandon, Pramod

    2014-06-01

    Manipulation of culture strategies was adopted to study the influence of nutrient stress, pH stress and precursor feeding on the biosynthesis of capsaicin in suspension and immobilized cell cultures of C. chinense. Cells cultured in the absence of one of the four nutrients (ammonium and potassium nitrate for nitrate and potassium stress, potassium dihydrogen orthophosphate for phosphorus stress, and sucrose for sugar stress) influenced the accumulation of capsaicin. Among the stress factors studied, nitrate stress showed maximal capsaicin production on day 20 (505.9 ± 2.8 μg g(-1) f.wt) in immobilized cell, whereas in suspension cultures the maximum accumulation (345.5 ± 2.9 μg g(-1) f.wt) was obtained on day 10. Different pH affected capsaicin accumulation; enhanced accumulation of capsaicin (261.6 ± 3.4 μg g(-1) f.wt) was observed in suspension cultures at pH 6 on day 15, whereas in case of immobilized cultures the highest capsaicin content (433.3 ± 3.3 μg g(-1) f.wt) was obtained at pH 5 on day 10. Addition of capsaicin precursors and intermediates significantly enhanced the biosynthesis of capsaicin, incorporation of vanillin at 100 μM in both suspension and immobilized cell cultures resulted in maximum capsaicin content with 499.1 ± 5.5 μg g(-1) f.wt on day 20 and 1,315.3 ± 10 μg g(-1) f.wt on day 10, respectively. Among the different culture strategies adopted to enhance capsaicin biosynthesis in cell cultures of C. chinense, cells fed with vanillin resulted in the maximum capsaicin accumulation. The rate of capsaicin production was significantly higher in immobilized cells as compared to freely suspended cells. PMID:24141419

  19. Manipulation of culture strategies to enhance capsaicin biosynthesis in suspension and immobilized cell cultures of Capsicum chinense Jacq. cv. Naga King Chili.

    Science.gov (United States)

    Kehie, Mechuselie; Kumaria, Suman; Tandon, Pramod

    2014-06-01

    Manipulation of culture strategies was adopted to study the influence of nutrient stress, pH stress and precursor feeding on the biosynthesis of capsaicin in suspension and immobilized cell cultures of C. chinense. Cells cultured in the absence of one of the four nutrients (ammonium and potassium nitrate for nitrate and potassium stress, potassium dihydrogen orthophosphate for phosphorus stress, and sucrose for sugar stress) influenced the accumulation of capsaicin. Among the stress factors studied, nitrate stress showed maximal capsaicin production on day 20 (505.9 ± 2.8 μg g(-1) f.wt) in immobilized cell, whereas in suspension cultures the maximum accumulation (345.5 ± 2.9 μg g(-1) f.wt) was obtained on day 10. Different pH affected capsaicin accumulation; enhanced accumulation of capsaicin (261.6 ± 3.4 μg g(-1) f.wt) was observed in suspension cultures at pH 6 on day 15, whereas in case of immobilized cultures the highest capsaicin content (433.3 ± 3.3 μg g(-1) f.wt) was obtained at pH 5 on day 10. Addition of capsaicin precursors and intermediates significantly enhanced the biosynthesis of capsaicin, incorporation of vanillin at 100 μM in both suspension and immobilized cell cultures resulted in maximum capsaicin content with 499.1 ± 5.5 μg g(-1) f.wt on day 20 and 1,315.3 ± 10 μg g(-1) f.wt on day 10, respectively. Among the different culture strategies adopted to enhance capsaicin biosynthesis in cell cultures of C. chinense, cells fed with vanillin resulted in the maximum capsaicin accumulation. The rate of capsaicin production was significantly higher in immobilized cells as compared to freely suspended cells.

  20. Drying of Micro-Encapsulated Lactic Acid Bacteria-Effects of Trehalose and Immobilization on Cell Survival and Release Properties

    Institute of Scientific and Technical Information of China (English)

    LI Xiaoyan; CHEN Xiguang

    2009-01-01

    Lactic acid bacteria (LAB) were encapsulated with alginate, gelatin and trehalose additives by the extrusion method and dried at 4℃. The microcapsules were generally spherical and had a wrinkled surface with a size of 1.7mm±0.2mm. Trehalose as a carbohydrate source in the culture medium could reduce acid production and performed no function in the positive proliferation of LAB. Using trehalose as a carbohydrate source and protective medium simultaneously had a benefit in the protection of LAB cells during the storage at 4℃. The density of hve LAB cells could be 10- CFU g-1 after 8 weeks of storage. Cells of LAB could be con-tinuously released from the capsules from the acidic (pH 1.2) to neutral conditions (plt 6.8). The release amounts and proliferation speeds of LAB cells in neutral medium were much larger and faster than those m acidic conditions. Additionally, immobilization of LAB could improve the survival of cells when they, were exposed to acidic medium (pH 1.2) with a survival rate of 76 %.

  1. Embryonic Stem Cells Maintain an Undifferentiated State on Dendrimer-Immobilized Surface with d-Glucose Display

    Directory of Open Access Journals (Sweden)

    Masahito Taya

    2011-12-01

    Full Text Available In serial passaging cultures of mouse embryonic stem (ES cells, we employed a dendrimer-immobilized substrate that displayed d-glucose as a terminal ligand. The d-glucose-displaying dendrimer (GLU/D surface caused the ES cells to form loosely attached spherical colonies, while those on a gelatin-coated surface formed flatter colonies that were firmly attached to the surface. Despite the morphological similarities between the colonies on the GLU/D surface and aggregates on a conventional bacteriological dish, immunostaining and RT-PCR analyses revealed the maintenance of cells within the spherical colonies on the GLU/D surface in an undifferentiated state with very low expressions of primitive endoderm markers. On the bacteriological dish, however, the cells within the aggregates showed a different cellular state with partial differentiation into the primitive endoderm lineage, and the expression level increased gradually along with the number of passages. These results indicate that the GLU/D surface can be a potential tool for controlling the ES cell morphology and then govern their self-renewal and fate.

  2. Drying of micro-encapsulated lactic acid bacteria — Effects of trehalose and immobilization on cell survival and release properties

    Science.gov (United States)

    Li, Xiaoyan; Chen, Xiguang

    2009-03-01

    Lactic acid bacteria (LAB) were encapsulated with alginate, gelatin and trehalose additives by the extrusion method and dried at 4 °C. The microcapsules were generally spherical and had a wrinkled surface with a size of 1.7 mm ± 0.2 mm. Trehalose as a carbohydrate source in the culture medium could reduce acid production and performed no function in the positive proliferation of LAB. Using trehalose as a carbohydrate source and protective medium simultaneously had a benefit in the protection of LAB cells during the storage at 4 °C. The density of live LAB cells could be 107 CFU g-1 after 8 weeks of storage. Cells of LAB could be continuously released from the capsules from the acidic (pH 1.2) to neutral conditions (pH 6.8). The release amounts and proliferation speeds of LAB cells in neutral medium were much larger and faster than those in acidic conditions. Additionally, immobilization of LAB could improve the survival of cells when they were exposed to acidic medium (pH 1.2) with a survival rate of 76 %.

  3. Kinetic evaluation of nitrification performance in an immobilized cell membrane bioreactor.

    Science.gov (United States)

    Güven, D; Ubay Çokgör, E; Sözen, S; Orhon, D

    2016-01-01

    High rate membrane bioreactor (MBR) systems operated at extremely low sludge ages (superfast membrane bioreactors (SFMBRs)) are inefficient to achieve nitrogen removal, due to insufficient retention time for nitrifiers. Moreover, frequent chemical cleaning is required due to high biomass flux. This study aims to satisfy the nitrification in SFMBRs by using sponge as carriers, leading to the extension of the residence time of microorganisms. In order to test the limits of nitrification, bioreactor was run under 52, 5 and 2 days of carrier residence time (CRT), with a hydraulic retention time of 6 h. Different degrees of nitrification were obtained for different CRTs. Sponge immobilized SFMBR operation with short CRT resulted in partial nitrification indicating selective dominancy of ammonia oxidizers. At higher CRT, simultaneous nitrification-denitrification was achieved when accompanying with oxygen limitation. Process kinetics was determined through evaluation of the results by a modeling study. Nitrifier partition in the reactor was also identified by model calibration.

  4. Immobilization of a Metal-Nitrogen-Carbon Catalyst on Activated Carbon with Enhanced Cathode Performance in Microbial Fuel Cells.

    Science.gov (United States)

    Yang, Wulin; Logan, Bruce E

    2016-08-23

    Applications of microbial fuel cells (MFCs) are limited in part by low power densities mainly due to cathode performance. Successful immobilization of an Fe-N-C co-catalyst on activated carbon (Fe-N-C/AC) improved the oxygen reduction reaction to nearly a four-electron transfer, compared to a twoelectron transfer achieved using AC. With acetate as the fuel, the maximum power density was 4.7±0.2 W m(-2) , which is higher than any previous report for an air-cathode MFC. With domestic wastewater as a fuel, MFCs with the Fe-N-C/AC cathode produced up to 0.8±0.03 W m(-2) , which was twice that obtained with a Pt-catalyzed cathode. The use of this Fe-N-C/AC catalyst can therefore substantially increase power production, and enable broader applications of MFCs for renewable electricity generation using waste materials.

  5. Determination of ethanol in acetic acid-containing samples by a biosensor based on immobilized Gluconobacter cells

    Directory of Open Access Journals (Sweden)

    VALENTINA A. KRATASYUK

    2012-11-01

    Full Text Available Reshetilov AN, Kitova AE, Arkhipova AV, Kratasyuk VA, Rai MK. 2012. Determination of ethanol in acetic acid containing samples by a biosensor based on immobilized Gluconobacter cells. Nusantara Bioscience 4: 97-100. A biosensor based on Gluconobacter oxydans VKM B-1280 bacteria was used for detection of ethanol in the presence of acetic acid. It was assumed that this assay could be useful for controlling acetic acid production from ethanol and determining the final stage of the fermentation process. Measurements were made using a Clark electrode-based amperometric biosensor. The effect of pH of the medium on the sensor signal and the analytical parameters of the sensor (detection range, sensitivity were investigated. The residual content of ethanol in acetic acid samples was analyzed. The results of the study are important for monitoring the acetic acid production process, as they represent a method of tracking its stages

  6. The microalga Chlamydomonas reinhardtii CW-15 as a solar cell for hydrogen peroxide photoproduction. Comparison between free and immobilized cells and thylakoids for energy conversion efficiency

    Energy Technology Data Exchange (ETDEWEB)

    Scholz, W.; Galvan, F.; Rosa, F.F. de la [Instituto de Bioquimica Vegetal y Fotosintesis, Universidad de Sevilla y CSIC, Sevilla (Spain)

    1995-11-28

    Immobilized cells and thylakoid vesicles of the microalga Chlamydomonas reinhardtii CW-15 have been developed as a solar cell because of their capabilities of producing hydrogen peroxide. This compound is an efficient and clean fuel used for rocket propulsion, motors and for heating. Hydrogen peroxide is produced by the photosystem in a catalyst cycle in which a redox mediator (methyl viologen) is reduced by electrons obtained from water by the photosynthetic apparatus of the microalga and it is re-oxidized by the oxygen dissolved in the solution. The photoproduction has been investigated using a discontinuous system with whole cells, or thylakoid vesicles, free or immobilized on alginate. The stimulation by azide as an inhibitor of catalase has also been analyzed. Under determined optimum conditions, the photoproduction by Ca-alginate entrapped cells, with a rate of 33 {mu}mol H{sub 2}O{sub 2}/mg Chl.h, was maintained for several hours with an energy conversion efficiency of 0.25%

  7. Mapping and identification of interferon gamma-regulated HeLa cell proteins separated by immobilized pH gradient two-dimensional gel electrophoresis

    DEFF Research Database (Denmark)

    Shaw, AC; Rossel Larsen, M; Roepstorff, P;

    1999-01-01

    magnitude of IFN-gamma responsive genes has been reported previously. Our goal is to identify and map IFN-gamma-regulated HeLa cell proteins to the two-dimensional polyacrylamide gel electrophoresis with the immobilized pH gradient (IPG) two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) system...

  8. Actin Immobilization on Chitin for Purifying Myosin II: A Laboratory Exercise That Integrates Concepts of Molecular Cell Biology and Protein Chemistry

    Science.gov (United States)

    de Souza, Marcelle Gomes; Grossi, Andre Luiz; Pereira, Elisangela Lima Bastos; da Cruz, Carolina Oliveira; Mendes, Fernanda Machado; Cameron, Luiz Claudio; Paiva, Carmen Lucia Antao

    2008-01-01

    This article presents our experience on teaching biochemical sciences through an innovative approach that integrates concepts of molecular cell biology and protein chemistry. This original laboratory exercise is based on the preparation of an affinity chromatography column containing F-actin molecules immobilized on chitin particles for purifying…

  9. Effects of initial pH value of the medium on the alcoholic fermentation performance of Saccharomyces cerevisiae cells immobilized on nipa leaf sheath pieces

    OpenAIRE

    Hoang Duc Toan Le; Van Viet Man Le

    2014-01-01

    Immobilized yeast on nipa leaf sheath pieces was applied to ethanol fermentation using the medium with different initial pH values (5.1, 4.5, 4.0, and 3.5). Control samples with the free yeast were also carried out under the same conditions. Low pH value of 4.0 or 3.5 significantly reduced yeast growth and increased the residual sugar level in the fermentation broths for both the immobilized and free cells. In all cases, the ethanol content produced and ethanol formation rate of the ...

  10. Effects of initial pH value of the medium on the alcoholic fermentation performance of Saccharomyces cerevisiae cells immobilized on nipa leaf sheath pieces

    Directory of Open Access Journals (Sweden)

    Hoang Duc Toan Le

    2014-12-01

    Full Text Available Immobilized yeast on nipa leaf sheath pieces was applied to ethanol fermentation using the medium with different initial pH values (5.1, 4.5, 4.0, and 3.5. Control samples with the free yeast were also carried out under the same conditions. Low pH value of 4.0 or 3.5 significantly reduced yeast growth and increased the residual sugar level in the fermentation broths for both the immobilized and free cells. In all cases, the ethanol content produced and ethanol formation rate of the immobilized yeast were 13-33% and 35-69%, respectively, higher than those of the free yeast. In addition, the residual sugar content in the immobilized yeast cultures was 2.1-20.5 times lower than that in the free yeast cultures. The yeast immobilized on nipa leaf stem pieces exhibited higher alcoholic fermentation performance than the free yeast in medium with low pH value. This support was potential for further research for application in ethanol industry.

  11. Immobilization of cross linked Col-I–OPN bone matrix protein on aminolysed PCL surfaces enhances initial biocompatibility of human adipogenic mesenchymal stem cells (hADMSC)

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Young-Hee; Jyoti, Md. Anirban; Song, Ho-Yeon, E-mail: songmic@sch.ac.kr

    2014-06-01

    In bone tissue engineering surface modification is considered as one of the important ways of fabricating successful biocompatible material. Addition of biologically active functionality on the surfaces has been tried for improving the overall biocompatibility of the system. In this study poly-ε-caprolactone film surfaces have been modified through aminolysis and immobilization process. Collagen type I (COL-I) and osteopontin (OPN), which play an important role in osteogenesis, was immobilized onto PCL films followed by aminolysis treatment using 1,6-hexanediamine. Characterization of animolysed and immobilized surfaces were done by a number techniques using scanning electron microscopy (SEM), FT-IR, XPS, ninhydrin staining, SDS-PAGE and confocal microscopy and compared between the modified and un-modified surfaces. Results of the successive experiments showed that aminolysis treatment was homogeneously achieved which helped to entrap or immobilize Col-I–OPN proteins on surfaces of PCL film. In vitro studies with human adipogenic mesenchymal stem cells (hADMSC) also confirmed the attachment and proliferation of cells was better in modified PCL surfaces than the unmodified surfaces. SEM, confocal microscopy and MTT assay showed a significant increase in cell spreading, attachment and proliferations on the biofunctionalized surfaces compared to the unmodified PCL surfaces at all-time points indicating the success of surface biofunctionalization.

  12. Optimization of date syrup for enhancement of the production of citric acid using immobilized cells of Aspergillus niger.

    Science.gov (United States)

    Mostafa, Yasser S; Alamri, Saad A

    2012-04-01

    Date syrup as an economical source of carbohydrates and immobilized Aspergillus niger J4, which was entrapped in calcium alginate pellets, were employed for enhancing the production of citric acid. Maximum production was achieved by pre-treating date syrup with 1.5% tricalcium phosphate to remove heavy metals. The production of citric acid using a pretreated medium was 38.87% higher than an untreated one that consumed sugar. The appropriate presence of nitrogen, phosphate and magnesium appeared to be important in order for citric acid to accumulate. The production of citric acid and the consumed sugar was higher when using 0.1% ammonium nitrate as the best source of nitrogen. The production of citric acid increased significantly when 0.1 g/l of KH2PO4 was added to the medium of date syrup. The addition of magnesium sulfate at the rate of 0.20 g/l had a stimulating effect on the production of citric acid. Maximum production of citric acid was obtained when calcium chloride was absent. One of the most important benefits of immobilized cells is their ability and stability to produce citric acid under a repeated batch culture. Over four repeated batches, the production of citric acid production was maintained for 24 days when each cycle continued for 144 h. The results obtained in the repeated batch cultivation using date syrup confirmed that date syrup could be used as a medium for the industrial production of citric acid.

  13. Alkali-Soluble Pectin Is the Primary Target of Aluminum Immobilization in Root Border Cells of Pea (Pisum sativum)

    Science.gov (United States)

    Yang, Jin; Qu, Mei; Fang, Jing; Shen, Ren Fang; Feng, Ying Ming; Liu, Jia You; Bian, Jian Feng; Wu, Li Shu; He, Yong Ming; Yu, Min

    2016-01-01

    We investigated the hypothesis that a discrepancy of Al binding in cell wall constituents determines Al mobility in root border cells (RBCs) of pea (Pisum sativum), which provides protection for RBCs and root apices under Al toxicity. Plants of pea (P. sativum L. ‘Zhongwan no. 6’) were subjected to Al treatments under mist culture. The concentration of Al in RBCs was much higher than that in the root apex. The Al content in RBCs surrounding one root apex (104 RBCs) was approximately 24.5% of the total Al in the root apex (0–2.5 mm), indicating a shielding role of RBCs for the root apex under Al toxicity. Cell wall analysis showed that Al accumulated predominantly in alkali-soluble pectin (pectin 2) of RBCs. This could be attributed to a significant increase of uronic acids under Al toxicity, higher capacity of Al adsorption in pectin 2 [5.3-fold higher than that of chelate-soluble pectin (pectin 1)], and lower ratio of Al desorption from pectin 2 (8.5%) compared with pectin 1 (68.5%). These results indicate that pectin 2 is the primary target of Al immobilization in RBCs of pea, which impairs Al access to the intracellular space of RBCs and mobility to root apices, and therefore protects root apices and RBCs from Al toxicity. PMID:27679639

  14. Alkali-Soluble Pectin Is the Primary Target of Aluminum Immobilization in Root Border Cells of Pea (Pisum sativum).

    Science.gov (United States)

    Yang, Jin; Qu, Mei; Fang, Jing; Shen, Ren Fang; Feng, Ying Ming; Liu, Jia You; Bian, Jian Feng; Wu, Li Shu; He, Yong Ming; Yu, Min

    2016-01-01

    We investigated the hypothesis that a discrepancy of Al binding in cell wall constituents determines Al mobility in root border cells (RBCs) of pea (Pisum sativum), which provides protection for RBCs and root apices under Al toxicity. Plants of pea (P. sativum L. 'Zhongwan no. 6') were subjected to Al treatments under mist culture. The concentration of Al in RBCs was much higher than that in the root apex. The Al content in RBCs surrounding one root apex (10(4) RBCs) was approximately 24.5% of the total Al in the root apex (0-2.5 mm), indicating a shielding role of RBCs for the root apex under Al toxicity. Cell wall analysis showed that Al accumulated predominantly in alkali-soluble pectin (pectin 2) of RBCs. This could be attributed to a significant increase of uronic acids under Al toxicity, higher capacity of Al adsorption in pectin 2 [5.3-fold higher than that of chelate-soluble pectin (pectin 1)], and lower ratio of Al desorption from pectin 2 (8.5%) compared with pectin 1 (68.5%). These results indicate that pectin 2 is the primary target of Al immobilization in RBCs of pea, which impairs Al access to the intracellular space of RBCs and mobility to root apices, and therefore protects root apices and RBCs from Al toxicity.

  15. Co-immobilization of glucoamylase and glucose oxidase for electrochemical sequential enzyme electrode for starch biosensor and biofuel cell.

    Science.gov (United States)

    Lang, Qiaolin; Yin, Long; Shi, Jianguo; Li, Liang; Xia, Lin; Liu, Aihua

    2014-01-15

    A novel electrochemical sequential biosensor was constructed by co-immobilizing glucoamylase (GA) and glucose oxidase (GOD) on the multi-walled carbon nanotubes (MWNTs)-modified glassy carbon electrode (GCE) by chemical crosslinking method, where glutaraldehyde and bovine serum albumin was used as crosslinking and blocking agent, respectively. The proposed biosensor (GA/GOD/MWNTs/GCE) is capable of determining starch without using extra sensors such as Clark-type oxygen sensor or H2O2 sensor. The current linearly decreased with the increasing concentration of starch ranging from 0.005% to 0.7% (w/w) with the limit of detection of 0.003% (w/w) starch. The as-fabricated sequential biosensor can be applicable to the detection of the content of starch in real samples, which are in good accordance with traditional Fehling's titration. Finally, a stable starch/O2 biofuel cell was assembled using the GA/GOD/MWNTs/GCE as bioanode and laccase/MWNTs/GCE as biocathode, which exhibited open circuit voltage of ca. 0.53 V and the maximum power density of 8.15 μW cm(-2) at 0.31 V, comparable with the other glucose/O2 based biofuel cells reported recently. Therefore, the proposed biosensor exhibited attractive features such as good stability in weak acidic buffer, good operational stability, wide linear range and capable of determination of starch in real samples as well as optimal bioanode for the biofuel cell. PMID:23954673

  16. Comparative study of bio-ethanol production from mahula (Madhuca latifolia L.) flowers by Saccharomyces cerevisiae cells immobilized in agar agar and Ca-alginate matrices

    Energy Technology Data Exchange (ETDEWEB)

    Behera, Shuvashish; Mohanty, Rama Chandra [Department of Botany, Utkal University, Vani Vihar, Bhubaneswar 751004, Orissa (India); Kar, Shaktimay; Ray, Ramesh Chandra [Microbiology Laboratory, Central Tuber Crops Research Institute (Regional Centre), Bhubaneswar 751019, Orissa (India)

    2010-01-15

    Batch fermentation of mahula (Madhuca latifolia L., a tree commonly found in tropical rain forest) flowers was carried out using immobilized cells (in agar agar and calcium alginate) and free cells of Saccharomyces cerevisiae. The ethanol yields were 151.2, 154.5 and 149.1 g kg{sup -1} flowers using immobilized (in agar agar and calcium alginate) and free cells, respectively. Cell entrapment in calcium alginate was found to be marginally superior to those in agar agar (2.2% more) as well as over free cell (3.5% more) as regard to ethanol yield from mahula flowers is concerned. Further, the immobilized cells were physiologically active at least for three cycles [150.6, 148.5 and 146.5 g kg{sup -1} (agar agar) and 152.8, 151.5 and 149.5 g kg{sup -1} flowers (calcium alginate) for first, second and third cycle, respectively] of ethanol fermentation without apparently lowering the productivity. Mahula flowers, a renewable, non-food-grade cheap carbohydrate substrate from non-agricultural environment such as forest can serve as an alternative to food grade sugar/starchy crops such as maize, sugarcane for bio-ethanol production. (author)

  17. CHANGES IN LIPID CONTENT OF WINE YEASTS DURING FERMENTATION BY IMMOBILIZED CELLS

    Directory of Open Access Journals (Sweden)

    Fedor Malik

    2010-05-01

    Full Text Available Comparison of the lipid composition of immobilised and non-immobilised cells of the wine cell strain Saccharomyces cerevisiae 6C subjected to ethanol stress indicates that the whole impact of the ethanol stress on the fatty acids composition is less influenced with immobilised cells as with non- immobilised ones. The ethanol stress raised in immobilised and free cells occurrence of palmitoleic acid to the detriment of palmitic acid. The character of changes in lipid composition during immobilisation probably has an impact upon slightly increased stress resistance. The immobilised cells are as well resistive against passive membrane fluidisation by ethanol. doi:10.5219/56

  18. Immobilization of Trichosporon cutaneum R 57 Cells onto Methylcellulose/SiO2 Hybrids and Biosorption of Cadmium and Copper Ions

    Directory of Open Access Journals (Sweden)

    Georgieva N.

    2009-12-01

    Full Text Available Methylcellulose/Silica (MC/SiO2 hybrids were synthesized via poly step sol-gel method. SiO2 was included into the hybrids from two silica precursors - methyltriethoxysilane (MTES and ethyltrimethoxysilane (ETMS with different quantity of organic part-5, 20 and 50 wt.%. The filamentous yeasts Trichosporon cutaneum strain R 57 was immobilized onto the synthesized MC/SiO2 hybrids. After immobilization the hybrid materials were used in the processes of sorption of cadmium and copper ions. The obtained results of protein content analysis indicated that the amount of protein increased with increasing of MC in the hybrids. It was established that the maximal efficiency of copper and cadmium removal were observed for hybrid materials containing MTES and 50 wt.% MC - 66% and 26% respectively. For ETMS and 50 wt.% MC a high value of copper removal was 56% and for cadmium - 45% removal, respectively. FTIR analysis of free and immobilized cells with metal ions was conducted. SEM images showed successful immobilization of the yeasts cells. Second order model was employed in order to investigate the kinetics of copper and cadmium biosorption.

  19. AN INTEGRATIVE WAY OF TEACHING MOLECULAR CELL BIOLOGY AND PROTEIN CHEMISTRY USING ACTIN IMMOBILIZATION ON CHITIN FOR PURIFYING MYOSIN II.

    Directory of Open Access Journals (Sweden)

    M.G. Souza

    2007-05-01

    Full Text Available Our intent is to present our experience on teaching Molecular Cell Biology andProtein Chemistry at UNIRIO through an innovative approach that includes myosin IIextraction and purification. We took advantage of the properties of muscle contractionand propose a simple method for purifying myosin II by affinity chromatography. Thisoriginal method is based on the preparation of an affinity column containing actinmolecules covalently bound to chitin particles. We propose a three-week syllabus thatincludes lectures and bench experimental work. The syllabus favors the activelearning of protein extraction and purification, as well as, of scientific concepts suchas muscle contraction, cytoskeleton structure and its importance for the living cell. Italso promotes the learning of the biotechnological applications of chitin and theapplications of protein immobilization in different industrial fields. Furthermore, theactivities also target the development of laboratorial technical abilities, thedevelopment of problem solving skills and the ability to write up a scientific reportfollowing the model of a scientific article. It is very important to mention that thissyllabus can be used even in places where a facility such as ultra-centrifugation islacking.

  20. Fabrication of Aligned Carbon Nanotube/Polycaprolactone/Gelatin Nanofibrous Matrices for Schwann Cell Immobilization

    OpenAIRE

    Shiao-Wen Tsai; Chun-Chiang Huang; Lih-Rou Rau; Fu-Yin Hsu

    2014-01-01

    In this study, we utilized a mandrel rotating collector consisting of two parallel, electrically conductive pieces of tape to fabricate aligned electrospun polycaprolactone/gelatin (PG) and carbon nanotube/polycaprolactone/gelatin (PGC) nanofibrous matrices. Furthermore, we examined the biological performance of the PGC nanofibrous and film matrices using an in vitro culture of RT4-D6P2T rat Schwann cells. Using cell adhesion tests, we found that carbon nanotube inhibited Schwann cell attach...

  1. A process for the treatment of olive mill waste waters by immobilized cells.

    Directory of Open Access Journals (Sweden)

    ElYachioui, M.

    2005-06-01

    Full Text Available Mould strains were immobilized on sawdust from woods as a solid material for the treatment of Olive Mill Waste (OMW waters. Assays were carried out in flasks. The treatment process was monitored by physico-chemical determinations including pH, polyphenols and COD, which were followed up during the incubation time. In parallel the chemical inhibitory activity of OMW was confirmed biologically by the determination of some microorganisms in the medium including the plate count, yeasts and lactic acid bacteria. Results indicated that the polyphenol degradation level was 87 %. The COD was also reduced by 60 %. The pH of the effluent increased from 4.5 to 6.6. The microbial profiles showed their best growth during the treatment period indicating a removal of the inhibitory activities from the OMW waters. The growth patterns of all microorganism groups were similar and could reach high levels in the effluent.Cepas de moho fueron inmovilizadas sobre serrín de madera como material sólido para el tratamiento de aguas residuales de un molino de aceituna (OMW. Los ensayos se realizaron en matraces. El proceso de tratamiento se monitorizó mediante determinaciones físico-químicas incluyendo pH, polifenoles y DQO, que también se analizaron durante el tiempo de incubación. En paralelo, la actividad inhibidora química de las OMW se confirma biológicamente mediante su efecto sobre algunos microorganismos incluyendo levaduras y bactérias ácido lácticas. Los resultados indicaron que los polifenoles se degradan hasta un nivel del 87 %. La DQO se redujo también al 60 %. El pH del efluente aumentó de 4.5 a 6.6. Los perfiles microbiológicos mostraron un mejor crecimiento a medida que avanzaba el tratamiento indicando una supresión de las actividades inhibidoras de las aguas (OMW. El comportamiento del crecimiento de todos los grupos de microorganismos fue similar y puede alcanzar altos niveles en el efluente

  2. A Ca-alginate particle co-immobilized with Phanerochaete chrysosporium cells and the combined cross-linked enzyme aggregates from Trametes versicolor.

    Science.gov (United States)

    Li, Yanchun; Wang, Zhi; Xu, Xudong; Jin, Liqiang

    2015-12-01

    For improving stability of immobilized white-rot fungus to treat various effluents, Phanerochaete chrysosporium cells and the combined cross-link enzyme aggregates (combi-CLEAs) prepared from Trametes versicolor were co-immobilized into the Ca-alginate gel particles in this paper. The activity yields of obtained combi-CLEAs were 42.7% for lignin peroxidases (LiPs), 31.4% for manganese peroxidases (MnPs) and 40.4% for laccase (Lac), respectively. And their specific activities were 30.2U/g as combi-CLEAs-LiPs, 9.5 U/g as combi-CLEAs-MnPs and 28.4 U/g as combi-CLEAs-Lac. Further, the present of the combi-CLEAs in the particles extremely improved their ability to degrade the dyes. Compared to the immobilized Ph. chrysosporium without the combi-CLEAs, the co-immobilized particles enhanced the decolorized rate of Acid Violet 7 (from 45.2% to 93.4%) and Basic Fuchsin (from 12.1% to 67.9%). In addition, the addition of the combi-CLEAs improved the adaptability of the white-rot fungal particles to adverse environmental conditions. PMID:26413897

  3. Analysis of cell performance and thermal regeneration of a lithium-tin cell having an immobilized fused-salt electrolyte

    Science.gov (United States)

    Cairns, E. J.; Shimotake, H.

    1969-01-01

    Cell performance and thermal regeneration of a thermally regenerative cell uses lithium and tin and a fused-salt electrolyte. The emf of the Li-Sn cell, as a function of cathode-alloy composition, is shown to resemble that of the Na-Bi cell.

  4. Immobilization of Enzymes by Electrochemical and Chemical Oxidative Polymerization of L-DOPA to Fabricate Amperometric Biosensors and Biofuel Cells.

    Science.gov (United States)

    Dai, Mengzhen; Sun, Lingen; Chao, Long; Tan, Yueming; Fu, Yingchun; Chen, Chao; Xie, Qingji

    2015-05-27

    Electrochemical/chemical oxidative synthesis and biosensing/biofuel cell applications of poly(L-DOPA) (PD) are studied versus polydopamine (PDA) as a recent hotspot biomaterial. The enzyme electrode developed by coelectrodeposition of PD and glucose oxidase (GOx), uricase, or tyrosinase shows biosensing performance superior to that of the corresponding PDA-based enzyme electrode. The chemical oxidative polymerization of L-DOPA (PDC) by NaAuCl4 in GOx-containing neutral aqueous solution is used to immobilize GOx and gold nanoparticles (AuNPs). The thus-prepared chitosan (CS)/GOx-PDC-AuNPs/Au(plate)/Au electrode working in the first-generation biosensing mode responds linearly to glucose concentration with a sensitivity of 152 μA mM(-1) cm(-2), which is larger than those of the CS/GOx-PDAC-AuNPs/Au(plate)/Au electrode, the CS/GOx-poly(3-anilineboronic acid) (PABA)-AuNPs/Au(plate)/Au electrode, and the most reported GOx-based enzyme electrodes. This PDC-based enzyme electrode also works well in the second-generation biosensing mode and as an excellent bioanode in biofuel cell construction, probably because PD as an amino acid polymer has the higher biocompatibility and the more favorable affinity to the enzyme than PDA. The PD material of great convenience in synthesis, outstanding biocompatibility for preparing high-performance bionanocomposites, and strong capability of multifunctional coatings on many surfaces may find wide applications in diversified fields including biotechnology and surface-coating. PMID:25938891

  5. Potential of Immobilized Whole-Cell Methylocella tundrae as a Biocatalyst for Methanol Production from Methane.

    Science.gov (United States)

    Mardina, Primata; Li, Jinglin; Patel, Sanjay K S; Kim, In-Won; Lee, Jung-Kul; Selvaraj, Chandrabose

    2016-07-28

    Methanol is a versatile compound that can be biologically synthesized from methane (CH4) by methanotrophs using a low energy-consuming and environment-friendly process. Methylocella tundrae is a type II methanotroph that can utilize CH4 as a carbon and energy source. Methanol is produced in the first step of the metabolic pathway of methanotrophs and is further oxidized into formaldehyde. Several parameters must be optimized to achieve high methanol production. In this study, we optimized the production conditions and process parameters for methanol production. The optimum incubation time, substrate, pH, agitation rate, temperature, phosphate buffer and sodium formate concentration, and cell concentration were determined to be 24 h, 50% CH4, pH 7, 150 rpm, 30°C, 100 mM and 50 mM, and 18 mg/ml, respectively. The optimization of these parameters significantly improved methanol production from 0.66 to 5.18 mM. The use of alginate-encapsulated cells resulted in enhanced methanol production stability and reusability of cells after five cycles of reuse under batch culture conditions. PMID:27012239

  6. Laminin Peptide-Immobilized Hydrogels Modulate Valve Endothelial Cell Hemostatic Regulation.

    Directory of Open Access Journals (Sweden)

    Liezl Rae Balaoing

    Full Text Available Valve endothelial cells (VEC have unique phenotypic responses relative to other types of vascular endothelial cells and have highly sensitive hemostatic functions affected by changes in valve tissues. Furthermore, effects of environmental factors on VEC hemostatic function has not been characterized. This work used a poly(ethylene glycol diacrylate (PEGDA hydrogel platform to evaluate the effects of substrate stiffness and cell adhesive ligands on VEC phenotype and expression of hemostatic genes. Hydrogels of molecular weights (MWs 3.4, 8, and 20 kDa were polymerized into platforms of different rigidities and thiol-modified cell adhesive peptides were covalently bound to acrylate groups on the hydrogel surfaces. The peptide RKRLQVQLSIRT (RKR is a syndecan-1 binding ligand derived from laminin, a trimeric protein and a basement membrane matrix component. Conversely, RGDS is an integrin binding peptide found in many extracellular matrix (ECM proteins including fibronectin, fibrinogen, and von Willebrand factor (VWF. VECs adhered to and formed a stable monolayer on all RKR-coated hydrogel-MW combinations. RGDS-coated platforms supported VEC adhesion and growth on RGDS-3.4 kDa and RGDS-8 kDa hydrogels. VECs cultured on the softer RKR-8 kDa and RKR-20 kDa hydrogel platforms had significantly higher gene expression for all anti-thrombotic (ADAMTS-13, tissue factor pathway inhibitor, and tissue plasminogen activator and thrombotic (VWF, tissue factor, and P-selectin proteins than VECs cultured on RGDS-coated hydrogels and tissue culture polystyrene controls. Stimulated VECs promoted greater platelet adhesion than non-stimulated VECs on their respective culture condition; yet stimulated VECs on RGDS-3.4 kDa gels were not as responsive to stimulation relative to the RKR-gel groups. Thus, the syndecan binding, laminin-derived peptide promoted stable VEC adhesion on the softer hydrogels and maintained VEC phenotype and natural hemostatic function. In

  7. Laminin Peptide-Immobilized Hydrogels Modulate Valve Endothelial Cell Hemostatic Regulation.

    Science.gov (United States)

    Balaoing, Liezl Rae; Post, Allison Davis; Lin, Adam Yuh; Tseng, Hubert; Moake, Joel L; Grande-Allen, K Jane

    2015-01-01

    Valve endothelial cells (VEC) have unique phenotypic responses relative to other types of vascular endothelial cells and have highly sensitive hemostatic functions affected by changes in valve tissues. Furthermore, effects of environmental factors on VEC hemostatic function has not been characterized. This work used a poly(ethylene glycol) diacrylate (PEGDA) hydrogel platform to evaluate the effects of substrate stiffness and cell adhesive ligands on VEC phenotype and expression of hemostatic genes. Hydrogels of molecular weights (MWs) 3.4, 8, and 20 kDa were polymerized into platforms of different rigidities and thiol-modified cell adhesive peptides were covalently bound to acrylate groups on the hydrogel surfaces. The peptide RKRLQVQLSIRT (RKR) is a syndecan-1 binding ligand derived from laminin, a trimeric protein and a basement membrane matrix component. Conversely, RGDS is an integrin binding peptide found in many extracellular matrix (ECM) proteins including fibronectin, fibrinogen, and von Willebrand factor (VWF). VECs adhered to and formed a stable monolayer on all RKR-coated hydrogel-MW combinations. RGDS-coated platforms supported VEC adhesion and growth on RGDS-3.4 kDa and RGDS-8 kDa hydrogels. VECs cultured on the softer RKR-8 kDa and RKR-20 kDa hydrogel platforms had significantly higher gene expression for all anti-thrombotic (ADAMTS-13, tissue factor pathway inhibitor, and tissue plasminogen activator) and thrombotic (VWF, tissue factor, and P-selectin) proteins than VECs cultured on RGDS-coated hydrogels and tissue culture polystyrene controls. Stimulated VECs promoted greater platelet adhesion than non-stimulated VECs on their respective culture condition; yet stimulated VECs on RGDS-3.4 kDa gels were not as responsive to stimulation relative to the RKR-gel groups. Thus, the syndecan binding, laminin-derived peptide promoted stable VEC adhesion on the softer hydrogels and maintained VEC phenotype and natural hemostatic function. In conclusion

  8. Efficient biosynthesis of γ-decalactone in ionic liquids by immobilized whole cells of Yarrowia lipolytica G3-3.21 on attapulgite.

    Science.gov (United States)

    Zhao, Yuping; Xu, Yan; Jiang, Changxing

    2015-10-01

    In this study, the biosynthesis of γ-decalactone (GDL) was successfully conducted in an ionic liquid (IL)-containing cosolvent system using immobilized cells of Yarrowia lipolytica G3-3.21 on attapulgite (ATG). We found the immobilized Y. lipolytica G3-3.21 cells in N-butyl-pyridinium tetrafluoroborate ([BPy]BF4) solution gave the highest activity of C16-Acyl-CoA oxidase and the maximum yield of GDL. The optimum immobilization conditions for the highest yield of GDL were 20 g/L of ATG, 1.5 % of CaCl2 and 2 % of sodium alginate (NaAlg). The optimal [BPy]BF4 content, buffer pH, reaction temperature, shaking speed, castor oil and glucose contents were 7.5 %, 26 °C, 150 rpm, 100 g/L and 10 %, respectively. Under the optimized conditions, the GDL yield was up to 8.05 g/L. After ten times of reuse, the GDL yield was 7.51 g/L, corresponding to 93.3 % of that obtained in the first batch, suggesting a good reusability and potential for industrial applications. PMID:26091898

  9. Silk fibroin immobilization on poly(ethylene terephthalate) films: Comparison of two surface modification methods and their effect on mesenchymal stem cells culture

    International Nuclear Information System (INIS)

    Silk fibroin (SF) has played a curial role for the surface modification of conventional materials to improve the biocompatibility, and SF modified poly(ethylene terephthalate) (PET) materials have potential applications on tissue engineering such as artificial ligament, artificial vessel, artificial heart valve sewing cuffs dacron and surgical mesh engineering. In this work, SF was immobilized onto PET film via two different methods: 1) plasma pretreatment followed by SF dip coating (PET-SF) and 2) plasma-induce acrylic acid graft polymerization and subsequent covalent immobilization of SF on PET film (PET-PAA-SF). It could be found that plasma treatment provided higher surface roughness which was suitable for further SF dip coating, while grafted poly(acrylic acid) (PAA) promised the covalent bonding between SF and PAA. ATR-FTIR adsorption band at 3284 cm−1, 1623 cm−1 and 1520 cm−1 suggested the successful introduction of SF onto PET surface, while the amount of immobilized SF of PET-SF was higher than PET-PAA-SF according to XPS investigation (0.29 vs 0.23 for N/C ratio). Surface modified PET film was used as substrate for mesenchymal stem cells (MSCs) culture, the cells on PET-SF surface exhibited optimum density compared to PET-PAA-SF according to CCK-8 assays, which indicated that plasma pretreatment followed by SF dip coating was a simple and effective way to prepare biocompatible PET surface. Highlights: ► Silk fibroins were immobilized onto PET films with or without the linker of PAA. ► Various techniques were performed to characterize the modified surfaces ► Plasma treatment followed by SF dip coating introduced more SF onto PET films. ► Compare to PET-PAA-SF, PET-SF has better biocompatibility base on MSCs culture

  10. Complete biodegradation of chlorpyrifos by engineered Pseudomonas putida cells expressing surface-immobilized laccases.

    Science.gov (United States)

    Liu, Jin; Tan, Luming; Wang, Jing; Wang, Zhiyong; Ni, Hong; Li, Lin

    2016-08-01

    The long-term abuse use of chlorpyrifos-like pesticides in agriculture and horticulture has resulted in significant soil or water contamination and a worldwide ecosystem threat. In this study, the ability of a solvent-tolerant bacterium, Pseudomonas putida MB285, with surface-displayed bacterial laccase, to biodegrade chlorpyrifos was investigated. The results of compositional analyses of the degraded products demonstrate that the engineered MB285 was capable of completely eliminating chlorpyrifos via direct biodegradation, as determined by high-performance liquid chromatography and gas chromatography-mass spectrometry assays. Two intermediate metabolites, namely 3,5,6-trichloro-2-pyridinol (TCP) and diethyl phosphate, were temporarily detectable, verifying the joint and stepwise degradation of chlorpyrifos by surface laccases and certain cellular enzymes, whereas the purified free laccase incompletely degraded chlorpyrifos into TCP. The degradation reaction can be conducted over a wide range of pH values (2-7) and temperatures (5-55 °C) without the need for Cu(2+). Bioassays using Caenorhabditis elegans as an indicator organism demonstrated that the medium was completely detoxified of chlorpyrifos by degradation. Moreover, the engineered cells exhibited a high capacity of repeated degradation and good performance in continuous degradation cycles, as well as a high capacity to degrade real effluents containing chlorpyrifos. Therefore, the developed system exhibited a high degradation capacity and performance and constitutes an improved approach to address chlorpyrifos contamination in chlorpyrifos-remediation practice. PMID:27231878

  11. Efficient anti-Prelog enantioselective reduction of acetyltrimethylsilane to (R-1-trimethylsilylethanol by immobilized Candida parapsilosis CCTCC M203011 cells in ionic liquid-based biphasic systems

    Directory of Open Access Journals (Sweden)

    Zhang Bo-Bo

    2012-08-01

    Full Text Available Abstract Background Biocatalytic asymmetric reductions with whole cells can offer high enantioselectivity, environmentally benign processes and energy-effective operations and thus are of great interest. The application of whole cell-mediated bioreduction is often restricted if substrate and product have low water solubility and/or high toxicity to the biocatalyst. Many studies have shown that a biphasic system is often useful in this instance. Hence, we developed efficient biphasic reaction systems with biocompatible water-immiscible ionic liquids (ILs, to improve the biocatalytic anti-Prelog enantioselective reduction of acetyltrimethylsilane (ATMS to (R-1-trimethylsilylethanol {(R-1-TMSE}, which is key synthon for a large number of silicon-containing drugs, using immobilized Candida parapsilosis CCTCC M203011 cells as the biocatalyst. Results It was found that the substrate ATMS and the product 1-TMSE exerted pronounced toxicity to immobilized Candida parapsilosis CCTCC M203011 cells. The biocompatible water-immiscible ILs can be applied as a substrate reservoir and in situ extractant for the product, thus greatly enhancing the efficiency of the biocatalytic process and the operational stability of the cells as compared to the IL-free aqueous system. Various ILs exerted significant but different effects on the bioreduction and the performances of biocatalysts were closely related to the kinds and combination of cation and anion of ILs. Among all the water-immiscible ILs investigated, the best results were observed in 1-butyl-3-methylimidazolium hexafluorophosphate (C4mim·PF6/buffer biphasic system. Furthermore, it was shown that the optimum substrate concentration, volume ratio of buffer to IL, buffer pH, reaction temperature and shaking rate for the bioreduction were 120 mM, 8/1 (v/v, 6.0, 30°C and 180 r/min, respectively. Under these optimized conditions, the initial reaction rate, the maximum yield and the product e.e. were 8.1

  12. Use of Saccharum spontaneum (wild sugarcane) as biomaterial for cell immobilization and modulated ethanol production by thermotolerant Saccharomyces cerevisiae VS3.

    Science.gov (United States)

    Chandel, Anuj K; Narasu, M Lakshmi; Chandrasekhar, G; Manikyam, A; Rao, L Venkateswar

    2009-04-01

    Saccharum spontaneum is a wasteland weed consists of 45.10+/-0.35% cellulose and 22.75+/-0.28% of hemicellulose on dry solid (DS) basis. Aqueous ammonia delignified S. spontaneum yielded total reducing sugars, 53.91+/-0.44 g/L (539.10+/-0.55 mg/g of substrate) with a hydrolytic efficiency of 77.85+/-0.45%. The enzymes required for hydrolysis were prepared from culture supernatants of Aspergillus oryzae MTCC 1846. A maximum of 0.85+/-0.07 IU/mL of filter paperase (FPase), 1.25+/-0.04 IU/mL of carboxy methyl cellulase (CMCase) and 55.56+/-0.52 IU/mL of xylanase activity was obtained after 7 days of incubation at 28+/-0.5 degrees C using delignified S. spontaneum as carbon source under submerged fermentation conditions. Enzymatic hydrolysate of S. spontaneum was then tested for ethanol production under batch and repeated batch production system using "in-situ" entrapped Saccharomyces cerevisiae VS3 cells in S. spontaneum stalks (1 cm x 1 cm) size. Immobilization was confirmed by the scanning electron microscopy (SEM). Batch fermentation of VS3 free cells and immobilized cells showed ethanol production, 19.45+/-0.55 g/L (yield, 0.410+/-0.010 g/g) and 21.66+/-0.62 g/L (yield, 0.434+/-0.021 g/g), respectively. Immobilized VS3 cells showed maximum ethanol production (22.85+/-0.44 g/L, yield, 0.45+/-0.04 g/g) up to 8th cycle during repeated batch fermentation followed by a gradual reduction in subsequent cycles of fermentation. PMID:19114303

  13. The combined effects of matrix stiffness and growth factor immobilization on the bioactivity and differentiation capabilities of adipose-derived stem cells.

    Science.gov (United States)

    Banks, Jessica M; Mozdzen, Laura C; Harley, Brendan A C; Bailey, Ryan C

    2014-10-01

    Biomaterial designs are increasingly incorporating multiple instructive signals to induce a desired cell response. However, many approaches do not allow orthogonal manipulation of immobilized growth factor signals and matrix stiffness. Further, few methods support patterning of biomolecular signals across a biomaterial in a spatially-selective manner. Here, we report a sequential approach employing carbodiimide crosslinking and benzophenone photoimmobilization chemistries to orthogonally modify the stiffness and immobilized growth factor content of a model collagen-GAG (CG) biomaterial. We subsequently examined the singular and combined effects of bone morphogenetic protein (BMP-2), platelet derived growth factor (PDGF-BB), and CG membrane stiffness on the bioactivity and osteogenic/adipogenic lineage-specific gene expression of adipose derived stem cells, an increasingly popular cell source for regenerative medicine studies. We found that the stiffest substrates direct osteogenic lineage commitment of ASCs regardless of the presence or absence of growth factors, while softer substrates require biochemical cues to direct cell fate. We subsequently describe the use of this approach to create overlapping patterns of growth factors across a single substrate. These results highlight the need for versatile approaches to selectively manipulate the biomaterial microenvironment to identify synergies between biochemical and mechanical cues for a range of regenerative medicine applications.

  14. 固定化细胞滤床和生物膜滤床净化二甲苯的比较%Comparison of air-borne xylene biodegradation between immobilized-cell biofilter and biofilm attached biofilter

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The Bacillus firmus was immobilized into Ca- alginate beads according to the different initial biomass concentration, calcification time and activation time. Three types of immobilized Bacillus firmus beads were packed respectively in trickling biofilter to purify xylene contained waste gases, and the performance of immobilized-cell biofilter was compared with traditional biofilm attached biofilter packed with two types of ceramic pellets. The results showed that three types of immobilized beads had different capabilities for removing xylene and life-spans. Higher initial biomass in immobilized beads resulted in better performance but shorter life-span. Activation process can remarkably enhance the activity of bacteria, and the removal efficiency of xylene can substantially be improved. Calcification time had influence on life-span of immobilized beads. Without acclimation, the cell-entrapped biofilter can obtain the maximum elimination capacity of 92.4 g/(m3·h). However, compared with biofilm attached biofilter, it has a poorer intrinsic drawback in volatile organic compounds (VOCs) removal due to the existence of excess mass transfer resistance.

  15. Physiological and Morphological Modifications in Immobilized Gibberella fujikuroi Mycelia

    OpenAIRE

    Saucedo, José Edmundo Nava; Barbotin, Jean-Noël; Thomas, Daniel

    1989-01-01

    Constraints created by immobilization conditions modified the physiological behavior and morphological characteristics of Gibberella fujikuroi mycelia in comparison with their development in free-cell conditions. G. fujikuroi mycelia were immobilized in different support matrices (polyurethane, carrageenan, and alginate) and showed a variety of reactions in response to the different microenvironmental factors encountered during and after immobilization. The best support with respect to gibber...

  16. Matrix-immobilized BMP-2 on microcontact printed fibronectin as in vitro tool to study BMP-mediated signaling and cell migration

    Directory of Open Access Journals (Sweden)

    Kristin eHauff

    2015-05-01

    Full Text Available During development, bone morphogenetic proteins (BMPs exert important functions in several tissues by regulating signaling for cell differentiation and migration. In vivo the extracellular matrix (ECM not only provides a support for adherent cells, but also presents a reservoir of growth factors (GFs. Several constituents of the ECM provide adhesive cues, which serve as binding sites for cell transmembrane receptors, such as integrins, which convey adhesion-mediated signaling to the intracellular compartment. Integrins do not function alone but rather crosstalk and cooperate with other receptors, such as GF receptors, in regulating cell responses to extracellular signals. To this, we present here the immobilization of BMP-2 onto cellular fibronectin (cFN, a key protein of the ECM, to investigate their impact on GF-mediated signaling and migration.Following biotinylation, BMP-2 was linked to biotinylated cFN using NeutrAvidin (NA as cross-linker. Characterization with QCM-D and ELISA confirmed the efficient immobilization of BMP-2 on cFN over a period of 24 h.To validate the bioactivity of matrix-immobilized BMP-2 (iBMP-2 we investigated short- and long-term responses of C2C12 myoblasts in comparison to soluble BMP-2 (sBMP-2 or in absence of GFs. Similarly to sBMP-2, iBMP-2 triggered Smad 1/5 phosphorylation and translocation into the nucleus corresponding to the activation of BMP-mediated Smad-dependent pathway. Additionally, successful suppression of myotube formation was observed after six days.We next implemented this approach to fabricate cFN micro patterned stripes by soft lithography. These stripes only allowed cell-surface interaction on the pattern due to passivation of the surface in between, thus serving as platform for studies on directed cell migration. During a 10 h-period, cells showed an increased migratory activity upon BMP-2 exposure.Thus, this versatile tool retains the GF's bioactivity and allows the presentation of ECM

  17. Electron tomography of cryo-immobilized plant tissue: a novel approach to studying 3D macromolecular architecture of mature plant cell walls in situ.

    Directory of Open Access Journals (Sweden)

    Purbasha Sarkar

    Full Text Available Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (∼ 2 nm, and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF, cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we

  18. Immobilization of nitrite oxidizing bacteria using biopolymeric chitosan media

    Institute of Scientific and Technical Information of China (English)

    Pranee Lertsutthiwong; Duangcheewan Boonpuak; Wiboonluk Pungrasmi; Sorawit Powtongsook

    2013-01-01

    The effects of chitosan characteristics including the degree of deacetylation,molecular weight,particle size,pH pretreatment and immobilization time on the immobilization of nitrite-oxidizing bacteria (NOB) on biopolymeric chitosan were investigated.Nitrite removal efficiency of immobilized NOB depended on the degree of deacetylation,particle size,pH pretreatment on the surface of chitosan and immobilization time.Scanning electron microscope characterization illustrated that the number of NOB cells attached to the surface of chitosan increased with an increment of immobilization time.The optimal condition for NOB immobilization on chitosan was achieved during a 24-hr immobilization period using chitosan with the degree of deacetylation larger than 80% and various particle size ranges between 1-5 mm at pH 6.5.In general,the NOB immobilized on chitosan flakes has a high potential to remove excess nitrite from wastewater and aquaculture systems.

  19. A High-Throughput Oxidative Stress Biosensor Based on Escherichia coli roGFP2 Cells Immobilized in a k-Carrageenan Matrix

    Directory of Open Access Journals (Sweden)

    Lia Ooi

    2015-01-01

    Full Text Available Biosensors fabricated with whole-cell bacteria appear to be suitable for detecting bioavailability and toxicity effects of the chemical(s of concern, but they are usually reported to have drawbacks like long response times (ranging from hours to days, narrow dynamic range and instability during long term storage. Our aim is to fabricate a sensitive whole-cell oxidative stress biosensor which has improved properties that address the mentioned weaknesses. In this paper, we report a novel high-throughput whole-cell biosensor fabricated by immobilizing roGFP2 expressing Escherichia coli cells in a k-carrageenan matrix, for the detection of oxidative stress challenged by metalloid compounds. The E. coli roGFP2 oxidative stress biosensor shows high sensitivity towards arsenite and selenite, with wide linear range and low detection limit (arsenite: 1.0 × 10−3–1.0 × 101 mg·L−1, LOD: 2.0 × 10−4 mg·L−1; selenite: 1.0 × 10−5–1.0 × 102 mg·L−1, LOD: 5.8 × 10−6 mg·L−1, short response times (0–9 min, high stability and reproducibility. This research is expected to provide a new direction in performing high-throughput environmental toxicity screening with living bacterial cells which is capable of measuring the bioavailability and toxicity of environmental stressors in a friction of a second.

  20. Immobilization of biomacromolecules on poly-L-lactide surface via a layer-by-layer method for the improving of its cytocompatibility to bone marrow stromal cells

    Institute of Scientific and Technical Information of China (English)

    L(U) Delong; MENG Sheng; ZHONG Wei; DU Qiangguo; GONG Li; LIU Jinfen; Dusan Bakos

    2005-01-01

    Hyaluronic acid (HA) and chitosan (CS) were immobilized on the surface of poly-L-lactide (PLLA) by the following procedure: Firstly, PLLA was aminolyzed with 1, 6-hexanediamine, and part of the PLLA surface ester groups were converted to free amino groups. Then negatively charged hyaluronic acid and positively charged chitosan were deposited onto the surface of aminolyzed PLLA film in a layer-by-layer assembly manner. The effect of the layer-by- layer deposition was evaluated by ATR-FTIR spectroscopy, Raman spectroscopy and static contact angle measurements. The cytocompatibility of PLLA sample to bone marrow stromal cells (BMSCs) was improved after modification with chitosan and HA. The cell attachment, activity, and proliferation on CS/HA modified PLLA films were enhanced comparing with the control. The cells cultured on the modified PLLA samples excreted abundant cytoplasm and can differentiate to vascular smooth muscle (SM)-like (SM-like) cells. A macroporous three-dimensional PLLA scaffold was prepared by integrating both the technique of freeze-drying and particle leaching. Layer-by-layer modification by HA/CS and cell culture was also applied on this scaffold. The scaffold cultured with BMSCs for 2 weeks has been tested successfully in vivo as a patch for repairing the artificial incision on canine pulmonary artery.

  1. Biological hydrogen production by immobilized cells of Clostridium tyrobutyricum JM1 isolated from a food waste treatment process.

    Science.gov (United States)

    Jo, Ji Hye; Lee, Dae Sung; Park, Donghee; Park, Jong Moon

    2008-09-01

    A fermentative hydrogen-producing bacterium, Clostridium tyrobutyricum JM1, was isolated from a food waste treating process using 16S rRNA gene sequencing and amplified ribosomal DNA restriction analysis (ARDRA). A fixed-bed bioreactor packed with polyurethane foam as support matrix for the growth of the isolate was operated at different hydraulic retention time (HRT) to evaluate its performance for hydrogen production. The reactor achieved the maximal hydrogen production rate of 7.2 l H(2)l(-1)d(-1) at 2h HRT, where hydrogen content in biogas was 50.0%, and substrate conversion efficiency was 97.4%. The maximum hydrogen yield was 223 ml (g-hexose)(-1) with an influent glucose concentration of 5 g l(-1). Therefore, the immobilized reactor using C. tyrobutyricum JM1 was an effective and stable system for continuous hydrogen production.

  2. Immobilization Technologies in Probiotic Food Production

    OpenAIRE

    Gregoria Mitropoulou; Viktor Nedovic; Arun Goyal; Yiannis Kourkoutas

    2013-01-01

    Various supports and immobilization/encapsulation techniques have been proposed and tested for application in functional food production. In the present review, the use of probiotic microorganisms for the production of novel foods is discussed, while the benefits and criteria of using probiotic cultures are analyzed. Subsequently, immobilization/encapsulation applications in the food industry aiming at the prolongation of cell viability are described together with an evaluation of their poten...

  3. Immobilization Technologies in Probiotic Food Production

    Directory of Open Access Journals (Sweden)

    Gregoria Mitropoulou

    2013-01-01

    Full Text Available Various supports and immobilization/encapsulation techniques have been proposed and tested for application in functional food production. In the present review, the use of probiotic microorganisms for the production of novel foods is discussed, while the benefits and criteria of using probiotic cultures are analyzed. Subsequently, immobilization/encapsulation applications in the food industry aiming at the prolongation of cell viability are described together with an evaluation of their potential future impact, which is also highlighted and assessed.

  4. 3-Chloro-1,2-propanediol biodegradation by Ca-alginate immobilized Pseudomonas putida DSM 437 cells applying different processes: mass transfer effects.

    Science.gov (United States)

    Konti, Aikaterini; Mamma, Diomi; Hatzinikolaou, Dimitios G; Kekos, Dimitris

    2016-10-01

    3-Chloro-1,2-propanediol (3-CPD) biodegradation by Ca-alginate immobilized Pseudomonas putida cells was performed in batch system, continuous stirred tank reactor (CSTR), and packed-bed reactor (PBR). Batch system exhibited higher biodegradation rates and 3-CPD uptakes compared to CSTR and PBR. The two continuous systems (CSTR and PBR) when compared at 200 mg/L 3-CPD in the inlet exhibited the same removal of 3-CPD at steady state. External mass-transfer limitations are found negligible at all systems examined, since the observable modulus for external mass transfer Ω ≪ 1 and the Biot number Bi > 1. Intra-particle diffusion resistance had a significant effect on 3-CPD biodegradation in all systems studied, but to a different extent. Thiele modulus was in the range of 2.5 in batch system, but it was increased at 11 when increasing cell loading in the beads, thus lowering significantly the respective effectiveness factor. Comparing the systems at the same cell loading in the beads PBR was less affected by internal diffusional limitations compared to CSTR and batch system, and, as a result, exhibited the highest overall effectiveness factor. PMID:27262716

  5. 3-Chloro-1,2-propanediol biodegradation by Ca-alginate immobilized Pseudomonas putida DSM 437 cells applying different processes: mass transfer effects.

    Science.gov (United States)

    Konti, Aikaterini; Mamma, Diomi; Hatzinikolaou, Dimitios G; Kekos, Dimitris

    2016-10-01

    3-Chloro-1,2-propanediol (3-CPD) biodegradation by Ca-alginate immobilized Pseudomonas putida cells was performed in batch system, continuous stirred tank reactor (CSTR), and packed-bed reactor (PBR). Batch system exhibited higher biodegradation rates and 3-CPD uptakes compared to CSTR and PBR. The two continuous systems (CSTR and PBR) when compared at 200 mg/L 3-CPD in the inlet exhibited the same removal of 3-CPD at steady state. External mass-transfer limitations are found negligible at all systems examined, since the observable modulus for external mass transfer Ω ≪ 1 and the Biot number Bi > 1. Intra-particle diffusion resistance had a significant effect on 3-CPD biodegradation in all systems studied, but to a different extent. Thiele modulus was in the range of 2.5 in batch system, but it was increased at 11 when increasing cell loading in the beads, thus lowering significantly the respective effectiveness factor. Comparing the systems at the same cell loading in the beads PBR was less affected by internal diffusional limitations compared to CSTR and batch system, and, as a result, exhibited the highest overall effectiveness factor.

  6. Immobilized waste leaching

    International Nuclear Information System (INIS)

    The main mechanism by which the immobilized radioactive materials can return to biosphere is the leaching due to the intrusion of water into the repositories. Some mathematical models and experiments utilized to evaluate the leaching rates in different immobilization matrices are described. (author)

  7. Studies on enzyme production technology of immobilized cell from F. yellowsea YS-9412-130%黄海黄杆菌YS-9412-130固定化细胞产酶技术

    Institute of Scientific and Technical Information of China (English)

    张云波; 王跃军; 洪义国; 孙谧; 刘惠; 王春波; 刘晓萍

    2000-01-01

    3种不同的包埋材料固定化YS-9412-130菌体细胞,进行半连续发酵。结果表明,用2.5%卡拉胶固定化细胞,产酶效率最高;添加明胶对产酶不利而添加豆饼粉与玉米粉后,酶活力有显著提高;卡拉胶固定化细胞发酵液中游离菌体浓度随发酵时间变化略有上升,总体水平较低。发酵液酶活随菌体浓度的增大而呈上升趋势;固定化细胞半连续发酵效率远高于游离细胞分批发酵的效率。%The cells of F. Yellowsea YS-9412-130 immobilized with three different imbedding materials are fermented semi-continuously. The result shows that the enzyme-producing rate reaches the highest value when cells are immobilized with 2.5% carrageenan, adding soy bean cake meal and cornsteep meal can improve the enzyme activity greatly, while adding gelalin is bad to enzyme activity; the concentration of free cell in the fermented broth of carrageenan-immobilized cells arises slightly with fermentation time going, however the level in general is low; the enzyme activity of fermented broth shows a rising trend with the increase of the cells concentration; the efficiency of semi-continuous fermentation of immobilized cells is much higher than that of batch-fermentation with free cells.

  8. Modulation of Selectin-Mediated Adhesion of Flowing Lymphoma and Bone Marrow Cells by Immobilized SDF-1

    Directory of Open Access Journals (Sweden)

    Elizabeth A. Hedges

    2014-08-01

    Full Text Available The α-chemokine, stromal-derived factor-1 (SDF-1, has been linked to the homing of circulating tumor cells to bone. SDF-1 is expressed by bone microvascular cells and osteoblasts and normally functions to attract blood-borne hematopoietic stem and progenitor cells to marrow. It has been shown that treatment of cancer cells with soluble SDF-1 results in a more aggressive phenotype; however, the relevance of the administration of the soluble protein is unclear. As such, a flow device was functionalized with P-selectin and SDF-1 to mimic the bone marrow microvasculature and the initial steps of cell adhesion. The introduction of SDF-1 onto the adhesive surface was found to significantly enhance the adhesion of lymphoma cells, as well as low-density bone marrow cells (LDBMC, both in terms of the number of adherent cells and the strength of cell adhesion. Thus, SDF-1 has a synergistic effect with P-selectin on cancer cell adhesion and may be sufficient to promote preferential metastasis to bone.

  9. A bioanode based on MWCNT/protein-assisted co-immobilization of glucose oxidase and 2,5-dihydroxybenzaldehyde for glucose fuel cells.

    Science.gov (United States)

    Yu, Chung-Mu; Yen, Miao-Ju; Chen, Lin-Chi

    2010-07-15

    This paper describes an easy-to-prepare, robust bioanode constructed on a polyester-supported screen-printed carbon paste electrode (SPCE) for glucose biofuel cells. To prepare the bioanode, carboxylated multi-walled carbon nanotubes (MWCNTs) were drop-coated on the SPCE first, and then a crosslinked matrix composed of glucose oxidase (GOx), 2,5-dihydroxybenzaldehyde (DHB), bovine serum albumin (BSA) and glutaraldehyde was coated atop the MWCNTs. It was found that the MWCNTs assisted the immobilization of the crosslinked matrix, enhanced the electron-shuttling process, and showed electrocatalytic effect to gluconic acid, which allowed squeeze more electrons out of a glucose molecule. Inside the matrix, DHB mediators could couple to GOx and BSA via the Schiff base reaction, and GOx and BSA could crosslink to each other with glutaraldehyde. From cyclic voltammetry, it was estimated that 3.63 nmol cm(-2) of DHB was anchored on the bioanode, and no mediator leaching was observed. The bioanode also attained reproducible flow-injection analysis (FIA) signals for glucose sensing (RSD=4.99%) and retained 84% of the initial response after keeping in a buffer at 4 degrees C for a week. In addition, the bioanode obeyed the Michaelis-Menten kinetics. Finally, we demonstrated that a glucose biofuel cell assembled with an optimal bioanode and a laccase/ABTS cathode generated an electric power of 45 microW cm(-2) from 1M glucose at 37 degrees C. PMID:20472420

  10. Hydrogen Production with High Evolution Rate and High Yield by Immobilized Cells of Hydrogen-producing Bacteria Strain B49 in a Column Reactor

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    To improve the hydrogen evolution rate in continuous hydrogen production of a novel fermentative hydrogen-producing bacteria strain B49 (AF481148 in EMBL), 4 % immobilized cells by polyvinyl alcohol-boric acid method, with the addition of a small amount of calcium alginate in a column reactor obtain hydrogen yield of 2.31 mol H2/mol glucose and hydrogen evolution rate of 1435.4 ml/L culture*h respectively at medium retention time of 2.0 h with a medium containing 10g glucose/L. Moreover, as the cell density in gel beads is increased to 8%, hydrogen yield and hydrogen evolution rate for 10g glucose/L are 2.34 mol H2/mol glucose and 2912.4 ml/L culture*h respectively at medium retention time of 1.0 h, and for molasses wastewater COD of 7505.9 mg/L hydrogen production potential of 205.6 ml/g COD and hydrogen evolution rate of 2057.7 ml/L culture*h at hydraulic retention time of 0.75 h are observed. In the continuous culture pH value keeps around 3.9 by self-regulating.

  11. Using a water-immiscible ionic liquid to improve asymmetric reduction of 4-(trimethylsilyl-3-butyn-2-one catalyzed by immobilized Candida parapsilosis CCTCC M203011 cells

    Directory of Open Access Journals (Sweden)

    Smith Thomas J

    2009-10-01

    Full Text Available Abstract Background Whole cells are usually employed for biocatalytic reduction reactions to ensure efficient coenzyme regeneration and to avoid problems with enzyme purification and stability. The efficiency of whole cell-catalyzed bioreduction is frequently restricted by pronounced toxicity of substrate and/or product to the microbial cells and in many instances the use of two-phase reaction systems can solve such problems. Therefore, we developed new, biphasic reaction systems with biocompatible water-immiscible ionic liquids (ILs as alternatives to conventional organic solvents, in order to improve the asymmetric reduction of 4-(trimethylsilyl-3-butyn-2-one (TMSB to (S-4-(trimethylsilyl-3-butyn-2-ol {(S-TMSBOL}, a key intermediate for synthesis of 5-lipoxygenase inhibitors, using immobilized Candida parapsilosis CCTCC M203011 cells as the biocatalyst. Results Various ILs exerted significant but different effects on the bioreduction. Of all the tested water-immiscible ILs, the best results were observed with 1-butyl-3-methylimidazolium hexafluorophosphate (C4MIM·PF6, which exhibited not only good biocompatibility with the cells but also excellent solvent properties for the toxic substrate and product, thus markedly improving the efficiency of the bioreduction and the operational stability of the cells as compared to the IL-free aqueous system. 2-Propanol was shown to be the most suitable co-substrate for coenzyme regeneration, and it was found that the optimum volume ratio of buffer to C4MIM·PF6, substrate concentration, buffer pH, 2-propanol concentration and reaction temperature were 4/1 (v/v, 24 mM, 5.5, 130 mM and 30°C, respectively. Under these optimized conditions, the maximum yield and the product e.e. wer 97.7% and >99%, respectively, which are much higher than the corresponding values previously reported. The efficient whole-cell biocatalytic process was shown to be feasible on a 250-mL scale. Conclusion The whole cell

  12. The Application of Dielectric Spectroscopy and Biocalorimetry for the Monitoring of Biomass in Immobilized Mammalian Cell Cultures

    Directory of Open Access Journals (Sweden)

    Harriet E. Cole

    2015-05-01

    Full Text Available The purpose of this study was to introduce dielectric spectroscopy and biocalorimetry as monitoring methods to follow immobilised Chinese Hamster Ovary (CHO cell culture development. The theory behind both monitoring techniques is explained and perfusion cultures are performed in a Reaction Calorimeter (eRC1 from Mettler Toledo as an application example. The findings of this work show that dielectric spectroscopy gives highly reliable information upon the viable cell density throughout the entire culture. On the other hand, the RC1 could only provide accurate data from day 5, when the cell density exceeded 4 × 106 vcells∙mL−1 (viable cell per mL working volume (WV. The method validation showed the limit of detection (LOD for 1.4 L cultures to be 8.86 × 106 vcells∙mL−1, a viable cell density commonly achieved in fed-batch and the early stages of a perfusion culture. This work suggests that biocalorimetry should be possible to implement at industrial scale to monitor CHO cell cultures.

  13. Radiation technology for immobilization of bioactive materials

    International Nuclear Information System (INIS)

    Within the framework of the Agency's coordinated research programme on ''Application of Radiation Technology in Immobilization of Bioactive Materials'', the third and final research coordination meeting was held at Beijing University, Beijing, People's Republic of China, 15-18 June 1987. The present publication compiles all presentations made at the meeting. Fundamental processes for the immobilization of enzymes, antibodies, cells and drugs were developed and established using gamma radiation, electron beams and plasma discharge. Applications of various biofunctional components, immobilized by radiation techniques in different processes, were studied. A range of backbone polymers has been examined together with various monomers. Coupling procedures have been developed which are relevant to our particular requirements. Enzymes of various types and characteristics have been immobilized with considerable efficiency. The immobilized biocatalysts have been shown to possess significant activity and retention of activity on storage. There appears to be a high degree of specificity associated with the properties of the immobilised biocatalysts, their activity and the ease of their preparation. Novel additives which lower the total radiation dose in grafting have been discovered and their value in immobilization processes assessed. Potential applications include: medical (diagnostic, therapeutic), and industrial processes (fermentation, bioseparation, etc.). Refs, figs and tabs

  14. Asymmetric Synthesis of (—)—1—Trimethylsiyl—ethanol with Immobilized Saccharomyces Cerevisiae Cells in Water/Organic Solvent Biphasic System

    Institute of Scientific and Technical Information of China (English)

    娄文勇; 宗敏华; 范晓丹

    2003-01-01

    Asymmetric synthesis of (-)-1-trimethylsilyl-ethanol with immobilized Saccharomyces cerevisiae cells in water/organic solvent biphasic system was studied,The effects of shake speed,hydrophobictiy of organic solvent ,volume ratio of water phase to organic phase,pH value of aqueous phase and reaction temperature on the initial reaction rate,maximum yield and enantiomeric excess(ee) of the product were systematically explored,All the above-mentioned factors had significant infuence on the reaction.n-Hexane was found to be the best organic solvent for the reaction.The optimum shake speed,volume ratio of water phase to organic phase,pH value and reaction temperature were 150 r.min-1,1/2,8 and 30℃ respectively,under which the maximum yield and enantiomeric excess of the product were as high as 96.8% and 95.7%,which are 15% and 16% higher than those of the corresponding reaction performed in aqueous phase ,To our best knowledge,this is the most satisfactory result obtained.

  15. Back propagation neural network model for predicting the performance of immobilized cell biofilters handling gas-phase hydrogen sulphide and ammonia.

    Science.gov (United States)

    Rene, Eldon R; López, M Estefanía; Kim, Jung Hoon; Park, Hung Suck

    2013-01-01

    Lab scale studies were conducted to evaluate the performance of two simultaneously operated immobilized cell biofilters (ICBs) for removing hydrogen sulphide (H2S) and ammonia (NH3) from gas phase. The removal efficiencies (REs) of the biofilter treating H2S varied from 50 to 100% at inlet loading rates (ILRs) varying up to 13 g H2S/m(3) ·h, while the NH3 biofilter showed REs ranging from 60 to 100% at ILRs varying between 0.5 and 5.5 g NH3/m(3) ·h. An application of the back propagation neural network (BPNN) to predict the performance parameter, namely, RE (%) using this experimental data is presented in this paper. The input parameters to the network were unit flow (per min) and inlet concentrations (ppmv), respectively. The accuracy of BPNN-based model predictions were evaluated by providing the trained network topology with a test dataset and also by calculating the regression coefficient (R (2)) values. The results from this predictive modeling work showed that BPNNs were able to predict the RE of both the ICBs efficiently.

  16. Back Propagation Neural Network Model for Predicting the Performance of Immobilized Cell Biofilters Handling Gas-Phase Hydrogen Sulphide and Ammonia

    Directory of Open Access Journals (Sweden)

    Eldon R. Rene

    2013-01-01

    Full Text Available Lab scale studies were conducted to evaluate the performance of two simultaneously operated immobilized cell biofilters (ICBs for removing hydrogen sulphide (H2S and ammonia (NH3 from gas phase. The removal efficiencies (REs of the biofilter treating H2S varied from 50 to 100% at inlet loading rates (ILRs varying up to 13 g H2S/m3·h, while the NH3 biofilter showed REs ranging from 60 to 100% at ILRs varying between 0.5 and 5.5 g NH3/m3·h. An application of the back propagation neural network (BPNN to predict the performance parameter, namely, RE (% using this experimental data is presented in this paper. The input parameters to the network were unit flow (per min and inlet concentrations (ppmv, respectively. The accuracy of BPNN-based model predictions were evaluated by providing the trained network topology with a test dataset and also by calculating the regression coefficient (R2 values. The results from this predictive modeling work showed that BPNNs were able to predict the RE of both the ICBs efficiently.

  17. Immobilization of aluminum with mucilage secreted by root cap and root border cells is related to aluminum resistance in Glycine max L.

    Science.gov (United States)

    Cai, Miaozhen; Wang, Ning; Xing, Chenghua; Wang, Fangmei; Wu, Kun; Du, Xing

    2013-12-01

    The root cap and root border cells (RBCs) of most plant species produced pectinaceous mucilage, which can bind metal cations. In order to evaluate the potential role of root mucilage on aluminum (Al) resistance, two soybean cultivars differing in Al resistance were aeroponic cultured, the effects of Al on root mucilage secretion, root growth, contents of mucilage-bound Al and root tip Al, and the capability of mucilage to bind Al were investigated. Increasing Al concentration and exposure time significantly enhanced mucilage excretion from both root caps and RBCs, decreased RBCs viability and relative root elongation except roots exposed to 400 μM Al for 48 h in Al-resistant cultivar. Removal of root mucilage from root tips resulted in a more severe inhibition of root elongation. Of the total Al accumulated in root, mucilage accounted 48-72 and 12-27 %, while root tip accounted 22-52 and 73-88 % in Al-resistant and Al-sensitive cultivars, respectively. A (27)Al nuclear magnetic resonance spectrum of the Al-adsorbed mucilage showed Al tightly bound to mucilage. Higher capacity to exclude Al in Al-resistant soybean cultivar is related to the immobilization and detoxification of Al by the mucilage secreted from root cap and RBCs.

  18. Dynamics of yeast immobilized-cell fluidized-bed bioreactors systems in ethanol fermentation from lactose-hydrolyzed whey and whey permeate.

    Science.gov (United States)

    Gabardo, Sabrina; Pereira, Gabriela Feix; Klein, Manuela P; Rech, Rosane; Hertz, Plinho F; Ayub, Marco Antônio Záchia

    2016-01-01

    We studied the dynamics of ethanol production on lactose-hydrolyzed whey (LHW) and lactose-hydrolyzed whey permeate (LHWP) in batch fluidized-bed bioreactors using single and co-cultures of immobilized cells of industrial strains of Saccharomyces cerevisiae and non-industrial strains of Kluyveromyces marxianus. Although the co-culture of S. cerevisiae CAT-1 and K. marxianus CCT 4086 produced two- to fourfold the ethanol productivity of single cultures of S. cerevisiae, the single cultures of the K. marxianus CCT 4086 produced the best results in both media (Y EtOH/S = 0.47-0.49 g g(-1) and Q P = 1.39-1.68 g L(-1) h(-1), in LHW and LHWP, respectively). Ethanol production on concentrated LHWP (180 g L(-1)) reached 79.1 g L(-1), with yields of 0.46 g g(-1) for K. marxianus CCT 4086 cultures. Repeated batches of fluidized-bed bioreactor on concentrated LHWP led to increased ethanol productivity, reaching 2.8 g L(-1) h(-1).

  19. Anhydride-functional silane immobilized onto titanium surfaces induces osteoblast cell differentiation and reduces bacterial adhesion and biofilm formation.

    Science.gov (United States)

    Godoy-Gallardo, Maria; Guillem-Marti, Jordi; Sevilla, Pablo; Manero, José M; Gil, Francisco J; Rodriguez, Daniel

    2016-02-01

    Bacterial infection in dental implants along with osseointegration failure usually leads to loss of the device. Bioactive molecules with antibacterial properties can be attached to titanium surfaces with anchoring molecules such as silanes, preventing biofilm formation and improving osseointegration. Properties of silanes as molecular binders have been thoroughly studied, but research on the biological effects of these coatings is scarce. The aim of the present study was to determine the in vitro cell response and antibacterial effects of triethoxysilypropyl succinic anhydride (TESPSA) silane anchored on titanium surfaces. X-ray photoelectron spectroscopy confirmed a successful silanization. The silanized surfaces showed no cytotoxic effects. Gene expression analyses of Sarcoma Osteogenic (SaOS-2) osteoblast-like cells cultured on TESPSA silanized surfaces reported a remarkable increase of biochemical markers related to induction of osteoblastic cell differentiation. A manifest decrease of bacterial adhesion and biofilm formation at early stages was observed on treated substrates, while favoring cell adhesion and spreading in bacteria-cell co-cultures. Surfaces treated with TESPSA could enhance a biological sealing on implant surfaces against bacteria colonization of underlying tissues. Furthermore, it can be an effective anchoring platform of biomolecules on titanium surfaces with improved osteoblastic differentiation and antibacterial properties.

  20. Dynamic Presentation of Immobilized Ligands Regulated through Biomolecular Recognition

    OpenAIRE

    Liu, Bo; Liu, Yang; Riesberg, Jeremiah J.; Shen, Wei

    2010-01-01

    To mimic the dynamic regulation of signaling ligands immobilized on extracellular matrices or on the surfaces of neighboring cells for guidance of cell behavior and fate selection, we have harnessed biomolecular recognition in combination with polymer engineering to create dynamic surfaces on which the accessibility of immobilized ligands to cell surface receptors can be reversibly interconverted under physiological conditions. The cell-adhesive RGD peptide is chosen as a model ligand. RGD is...

  1. Hyaluronic Acid Immobilized Polyacrylamide Nanoparticle Sensors for CD44 Receptor Targeting and pH Measurement in Cells

    DEFF Research Database (Denmark)

    Sun, Honghao; Benjaminsen, Rikke Vicki; Almdal, Kristoffer;

    2012-01-01

    Our ability to design receptor-targeted nanocarriers aimed at drug release after endocytosis is limited by the current knowledge of intracellular nanoparticle (NP) trafficking. It is not clear if NP size, surface chemistry, and/or targeting of cell surface receptors changes the intracellular fate...

  2. Escherichia coli NemA is an efficient chromate reductase that can be biologically immobilized to provide a cell free system for remediation of hexavalent chromium.

    Directory of Open Access Journals (Sweden)

    Katherine J Robins

    Full Text Available Hexavalent chromium is a serious and widespread environmental pollutant. Although many bacteria have been identified that can transform highly water-soluble and toxic Cr(VI to insoluble and relatively non-toxic Cr(III, bacterial bioremediation of Cr(VI pollution is limited by a number of issues, in particular chromium toxicity to the remediating cells. To address this we sought to develop an immobilized enzymatic system for Cr(VI remediation. To identify novel Cr(VI reductase enzymes we first screened cell extracts from an Escherichia coli library of soluble oxidoreductases derived from a range of bacteria, but found that a number of these enzymes can reduce Cr(VI indirectly, via redox intermediates present in the crude extracts. Instead, activity assays for 15 candidate enzymes purified as His6-tagged proteins identified E. coli NemA as a highly efficient Cr(VI reductase (k(cat/K(M= 1.1×10(5 M(-1 s(-1 with NADH as cofactor. Fusion of nemA to the polyhydroxyalkanoate synthase gene phaC from Ralstonia eutropha enabled high-level biosynthesis of functionalized polyhydroxyalkanoate granules displaying stable and active NemA on their surface. When these granules were combined with either Bacillus subtilis glucose dehydrogenase or Candida boidinii formate dehydrogenase as a cofactor regenerating partner, high levels of chromate transformation were observed with only low initial concentrations of expensive NADH cofactor being required, the overall reaction being powered by consumption of the cheap sacrificial substrates glucose or formic acid, respectively. This system therefore offers promise as an economic solution for ex situ Cr(VI remediation.

  3. Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column

    Institute of Scientific and Technical Information of China (English)

    SHEN Shaochuan; WANG Liangyan; SUN Zongtao; LI Mingfeng; LIU Chengzhi; TIAN Bing; YUN Junxian

    2013-01-01

    Geranylgeranyl diphosphate synthase (GGPPS) plays a key role in the biosynthesis of antioxidative carotenoid from the extremely radioresistant bacterium Deinococcus radiodurans.In this work,the recombinant GGPPS expressed in Escherichia coli by cloning and transforming the gene dr1395 of D.radiodurans was isolated rapidly by an immobilized metal affinity supermacroporous cryogel,i.e.,Cu2+-iminodiacetic acid (IDA)-cryogel.The properties of the Cu2+-IDA-cryogel were characterized using capillary-based mathematical model and experimental measurements.The obtained protein samples were analyzed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).The porosity of the present Cu2+-IDA-cryogel is 90.4% and the water permeability is 5.04×10-12 m2.From the capillary-based model,this cryogel presents a slightly wide normal pore (capillary)size distribution with the mean diameter of 55.2 μm,the standard deviation of 28.0 μm and the half of skeleton wall thickness of 2.8 μm.The pore size distribute from about 10 to 141 μm and the effective tortuosity of these capillary pores increases from 2.60 to 9.05.The isolation of the GGPPS from cell homogenate can be achieved at the flow velocity of 3.40× 10-4 m·s-1 by the Cu2+-IDA-cryogel bed.High-purity GGPPS (about 91.4%) is obtained according to the SDS-PAGE analysis of the elution samples,indicating that the present method is a promising,simple and effective approach to isolate GGPPS from cell homogenate of engineering strains.

  4. Cell surface polypeptide CshA mediates binding of Streptococcus gordonii to other oral bacteria and to immobilized fibronectin.

    OpenAIRE

    McNab, R; Holmes, A.R.; Clarke, J M; Tannock, G W; Jenkinson, H F

    1996-01-01

    Isogenic mutants of Streptococcus gordonii DL1 (Challis) in which the genes encoding high-molecular-mass cell surface polypeptides CshA and/or CshB were inactivated were deficient in binding to four strains of Actinomyces naeslundii and two strains of Streptococcus oralis. Lactose-sensitive interactions of S. gordonii with A. naeslundii ATCC 12104 and PK606 were associated with expression of cshA but not of cshB. Lactose-insensitive interactions of S. gordonii with A. naeslundii T14V and WVU6...

  5. An automated method for determining the cytoadhesion of Plasmodium falciparum-infected erythrocytes to immobilized cells

    DEFF Research Database (Denmark)

    Hempel, Casper; Boisen, Ida M; Efunshile, Akinwale;

    2015-01-01

    BACKGROUND: Plasmodium falciparum exports antigens to the surface of infected erythrocytes causing cytoadhesion to the host vasculature. This is central in malaria pathogenesis but in vitro studies of cytoadhesion rely mainly on manual counting methods. The current study aimed at developing an...... automated high-throughput method for this purpose utilizing the pseudoperoxidase activity of intra-erythrocytic haemoglobin. METHODS: Chinese hamster ovary (CHO) cells were grown to confluence in chamber slides and microtiter plates. Cytoadhesion of co-cultured P. falciparum, selected for binding to CHO...

  6. THERMAL ACTIVATION OF IMMOBILIZED PAPAIN

    Institute of Scientific and Technical Information of China (English)

    1998-01-01

    Papain (Papainase, EC 3.4.22.2) was immobilized on porous silica beads by cross linking with glutaraldehyde. The thermal activation of this immobilized papain in aqueous system was found at a temperature range from 50 to 90℃. The higher the temperature, the more active the immobilized papain will possess. At the same time,the durability of the immobilized papain on heating was greatly improved. The effect of additives and salts on the activity of the immobilized papain were also studied. The results showed that the additives and some of the salts studied could markedly enhance the activity of the immobilized papain at elevated temperature.

  7. Performance of three pilot-scale immobilized-cell biotrickling filters for removal of hydrogen sulfide from a contaminated air steam.

    Science.gov (United States)

    Chen, Yiqing; Fan, Zhidong; Ma, Lixia; Yin, Juan; Luo, Man; Cai, Wangfeng

    2014-11-01

    Hydrogen sulfide (H2S) is a major malodorous compound emitted from wastewater treatment plants. In this study, the performance of three pilot-scale immobilized-cell biotrickling filters (BTFs) spacked with combinations of bamboo charcoal and ceramsite in different ratios was investigated in terms of H2S removal. Extensive tests were performed to determine the removal characteristics, pressure drops, metabolic products, and removal kinetics of the BTFs. The BTFs were operated in continuous mode at low loading rates varying from 0.59 to 5.00 g H2S m(-3) h(-1) with an empty bed retention time (EBRT) of 25 s. The removal efficiency (RE) for each BTF was >99% in the steady-state period, and high standards were met for the exhaust gas. It was found that a multilayer BTF had a slight advantage over a perfectly mixed BTF for the removal of H2S. Furthermore, an impressive amount >97% of the H2S was eliminated by 10% of packing materials near the inlet of the BTF. The modified Michaelis-Menten equation was adopted to describe the characteristics of the BTF, and K s and V m values for the BTF with pure bamboo charcoal packing material were 3.68 ppmv and 4.26 g H2S m(-3) h(-1), respectively. Both bamboo charcoal and ceramsite demonstrated good performance as packing materials in BTFs for the removal of H2S, and the results of this study could serve as a guide for further design and operation of industrial-scale systems. PMID:25313280

  8. Effects of multiple polyaniline layers immobilized on carbon nanotube and glutaraldehyde on performance and stability of biofuel cell

    Science.gov (United States)

    Christwardana, Marcelinus; Kwon, Yongchai

    2015-12-01

    Enzymatic biofuel cell (EBC) employing new catalyst for anode electrode is fabricated. The new catalyst consists of glucose oxidase (GOx), polyaniline (PANI) and carbon nanotube (CNT) that are multiply stacked together and finally the stack layer is surrounded by glutaraldehyde (GA) (GA/[GOx/PANI/CNT]n). To evaluate how the GA/[GOx/PANI/CNT]n layer affects EBC performance and stability, electrochemical characterizations are implemented. Regarding optimization, GA/[GOx/PANI/CNT]3 is determined. For elucidating reaction mechanism between glucose and flavin adenine dinucleotide (FAD) of GA/[GOx/PANI/CNT]3, associated investigations are performed. In the evaluations, drop in reduction current peak of FAD is observed with provisions of glucose and O2, while glucose does not influence FAD reaction without O2, confirming O2 makes mediator role. When the GA/[GOx/PANI/CNT]3 layer is adopted, superior catalytic activity and EBC performance are gained (electron transfer rate constant of 5.1 s-1, glucose sensitivity of 150 ìA mM-1 cm-2, and EBC maximum power density (MPD) of 0.29 mW cm-2). Regarding EBC stability, MPD of EBC adopting GA/[GOx/PANI/CNT]3 maintains up to 93% of their initial value even after four weeks. Although GA is little effective for improving EBC performance, EBC stability is helped by GA due to its adhesion promotion capability with [GOx/PANI/CNT]n layer.

  9. The 13C/12C fractionation by microbial cells immobilized on a solid-phase carrier during the growth on glucose

    Science.gov (United States)

    Zyakun, Anatoly; Kochetkov, Vladimir

    2010-05-01

    Problem. In microbiological ecology, the level of basal СО2 respiration and the potential of microbial activity defined as substrate-induced respiration (SIR) are used as criteria of the metabolic state of soil microbiota. The peculiar feature of glucose metabolism in soil is its utilization by microbial cells immobilized on soil particles as a solid-phase carrier. The efficiency of substrate utilization and СО2 production in such cases depend on the rate of microorganisms' growth and colonization of the solid-phase carrier surface, where the substrate is located. The products of microbial metabolism are supposed to inherit the substrate isotope composition correct to the isotopic effects accompanying substrate utilization and metabolic transformations. However, all experiments in carbon isotope fractionation during microbial utilization of glucose as a substrate have been carried out with microorganisms growing in liquid media. Objective: Study of the kinetics of glucose utilization as a test substrate during the growth of soil microorganisms immobilized on a solid-phase carrier and ascertainment of peculiarities of the formation of carbon isotope composition of produced metabolic СО2. The objects of research were Pseudomonas aureofaciens BS1393(pBS216) (culture A) and Rhodococcus sp. 3-30 (culture B) as representatives of pseudomonades and rhodococci, which occur in the soils of different genesis and are of defining value in development and implementation of biotechnological schemes for degradation of toxic organic pollutants in the environment. Results and discussion. The cultures under study had different rates of growth on glucose. Specific rates of СО2 production during the growth of cultures A and B on glucose were 0.34 (± 0.05) and 0.078 (± 0.01) μg С-СО2 h-1, respectively. The lag periods of culture (A and B) growth were about 4.3 and 26 h, respectively. Comparison of the lag periods of these representatives of pseudomonades and rhodococci

  10. Effect of cell immobilization on the treatment of olive mill wastewater by a total phenols, acetic acid and formic acid degrading bacterium strain

    Directory of Open Access Journals (Sweden)

    Errami, Mohamed

    2005-06-01

    Full Text Available Olive mill wastewater (OMW is a pure vegetative by-product, containing a high organic and polyphenol content and is resistant to biodegradation. Its disposal lead to major environmental pollution problems in the Mediterranean basin. An aerobic bacterium was isolated from OMW. During three consecutive diluted and supplemented OMW treatment cycles, significant abatement of its phytotoxic substances was observed. In fact, total phenols, acetic and formic acids were reduced between 33 and 64 % when cells of the isolated bacterium were grown free; and between 62 and 78 % when cells of the same isolated bacterium were grown immobilized in a polyurethane sponge. These results suggest that the bacterium culture of the new isolate would decrease the OMW phytotoxicity. Phylogenetic analysis of 16S ribosomal DNA showed that all the related sequences are members of the Enterobacteriaceae family and revealed that the isolated bacterium was characterized as a Klebsiella oxytoca strain.El alpechín (OMW es un residuo puro de la extracción del aceite de oliva, que contiene una elevada carga orgánica y de polifenoles por lo que es resistente a la degradación. Su descarga produce graves problemas de contaminación medioambiental en toda el área mediterránea. Se ha aislado una bacteria anaerobia del OMW, que , durante tres ciclos consecutivos de tratamiento del OMW diluido y suplementado, produjo una disminución significativa de las sustancias fitotóxicas del residuo. De hecho, la concentración en fenoles totales, ácido acético y ácido fórmico se redujeron entre 33 y 64 % cuando las células no estaban inmovilizadas y entre el 62 y 78 % cuando las células bacterianas se inmovilizaron en una esponja de poliuretano. Estos resultados indican que el cultivo de la nueva bacteria aislada puede disminuir la fototoxicidad del alpechín. Análisis filogenético del ribosoma 16S de DNA demostró que todas las secuencias eran miembros de la familia

  11. Hierarchically Nanoporous Bioactive Glasses for High Efficiency Immobilization of Enzymes

    DEFF Research Database (Denmark)

    He, W.; Min, D.D.; Zhang, X.D.;

    2014-01-01

    Bioactive glasses with hierarchical nanoporosity and structures have been heavily involved in immobilization of enzymes. Because of meticulous design and ingenious hierarchical nanostructuration of porosities from yeast cell biotemplates, hierarchically nanostructured porous bioactive glasses can...

  12. Biodegradation of oil wastewater by free and immobilized Yarrowia lipolytica W29

    Institute of Scientific and Technical Information of China (English)

    WU Lan; GE Gang; WAN Jinbao

    2009-01-01

    The ability of Yarrowia lipolytica W29 immobilized by calcium alginate to degrade oil and chemical oxygen demand (COD) was examined. The degradation rules of oil and COD by immobilized cells with the cell density of 6.65 × 106 CFU/mL degraded 2 000 mg/L oil and 2 000 mg/L COD within 50 h at 30℃ ( pH 7.0, 150 r/min), similarly to those of free cells, and the degradation efficiencies of oil and COD by immobilized cells were above 80%, respectively. At the same time, temperature and initial oil, COD concentration affecting oil and COD degradation of immobilized cells were investigated, the results showed that immobilized cells had high thermostability compared to that of free cells, and substrate concentration significantly affected degrading ability of immobilized cells. Storage stability and reusability tests revealed that the oil degradation ability of immobilized cells was stable after storing at 4℃ for 30 d and reuse for 12 times, respectively, the COD degradation rate of immobilized cells was also maintained 82% at the sixth cycle. These results suggested that immobilized Yarrowia lipolytica might be applicable to a wastewater treatment system for the removal of oil and COD.

  13. Down-regulation of triose phosphate isomerase in Vineristine-resistant gastric cancer SGC7901 cell line identified by immobilized pH gradient two-dimensional gel electrophoresis and mierosequencing

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Objective:To exkplore new multidrug-resistance-related proteins in gastric SC7901 cells and clarify their mechanisms.Methods:Two-dimensional(2-D) polyacrylamide gel electrophoresis with immobilized pH gradients(IPG) was applied to compare the differential expression of multidrug-resistance-related proteins in gastric cancer SGC7901 cells and Vineristine-resistant SGC7901 cells (SGC7901/VCR) induced by vincristine sulfate.The 2-D gels were silver-stained.Then,preparative 2-D PAGE was performed.The differential proteins of PVDF membranes were cxcised and identified by N-terminal microsequencing.The mRNA expressions of differential proteins were detected in SGC 7901 cells and SGC7901/VCR cells by RT-PCR.Results:Approximatedly 680 protein sports were resolved on each 2-D gel by silver staining.Most protein spots showed no difference in composition,shape or density.25 proteins differed in abundance (6 higher in SGC7901/VCR cells;19 higher in 7901 cells);5 proteins were unique to one kind of cell or the othe(3 in SGC7901/VRC cells,2 in 7901 cells).One drug-resistance-related protein,which was down-regulated in SGC7901/VCR cells,was identified as trisephosphate isomerase(TPI),a glycolytic pathway enzyme.Conclusions:the results suggest that these differential proteins including TPI may be related to the Vincristine-resistant mechanism in human gastric cancer SGC7901/VCR cell line.

  14. Immobilization of Yarrowia lipolytica for aroma production from castor oil.

    Science.gov (United States)

    Braga, Adelaide; Belo, Isabel

    2013-04-01

    The main aim of this study was to compare different materials for Y. lipolytica immobilization that could be used in the production of γ-decalactone (a peach-like aroma) in order to prevent the toxic effect both of the substrate and the aroma upon the cells. Therefore, cells adsorption onto pieces of methyl polymethacrylate and of DupUM(®) was studied and further used in the biotransformation of castor oil into γ-decalactone. The highest aroma concentration was obtained with immobilized cells in DupUM(®), where reconsumption of the aroma by the cells was prevented, contrarily to what happens with free cells. This is a very promising result for γ-decalactone production, with potential to be used at an industrial level since the use of immobilized cells system will facilitate the conversion of a batch process into a continuous mode keeping high cell density and allowing easier recovery of metabolic products.

  15. Enzyme immobilization by means of ultrafiltration techniques.

    Science.gov (United States)

    Scardi, V; Cantarella, M; Gianfreda, L; Palescandolo, R; Alfani, F; Greco, G

    1980-01-01

    Unstirred, plane membrane, ultrafiltration cells have been used as enzymatic reactor units. Because of the concentration polarization phenomena which take place in the system, at steady-state the enzyme is confined (dynamically immobilized) within an extremely narrow region upstream the ultrafiltration membrane. Correspondingly its concentration attains fairly high values. Kinetic studies have been therefore performed under quite unusual experimental conditions in order to better approximate local enzyme concentration levels in immobilized enzyme systems. Studies have been also carried out on the kinetics of enzyme deactivation in the continuous presence of substrate and reaction products. Once the enzyme concentration profile is completely developed, further injection into the system of suitable amounts of an inert proteic macromolecule (albumin polymers) gives rise to the formation of a gel layer onto the ultrafiltration membrane within which the enzyme is entrapped (statically immobilized). The effect of this immobilization technique has been studied as far as the kinetics of the main reaction, the substrate mass transfer resistances and the enzyme stability are concerned. The rejective properties of such gel layers towards enzymatic molecules have been exploited in producing multilayer, multi-enzymatic reactors. PMID:7417597

  16. 新型固定化细胞膜反应器脱氮研究%Study on nitrogen removal from wastewater in a new co-immobilized cells membrane bioreactor

    Institute of Scientific and Technical Information of China (English)

    曹国民; 赵庆祥; 龚剑丽; 张彤

    2001-01-01

    研究了一种新型的废水生物脱氮反应器,即利用固定化细胞膜将反应器一隔为二,膜的一侧与好氧的氨氮废水接触,另一侧与缺氧的乙醇水溶液(反硝化碳源)接触.固定于膜中的硝化细菌将氨氮氧化成亚硝氮和硝氮,随即被同一膜中的反硝化细菌还原成氮气.硝化细菌和反硝化细菌混合固定于膜内时的氨氧化速率约为硝化细菌单独固定时的2倍.未发现碳源重复利用对脱氮过程产生不利影响.此新型反应器可以稳定运行50天以上.%A new bioreactor (double-chambered bioreactor) for the nitrogen removal from wastewater was described which consisted of a plate membrane containing nitrifying organisms and denitrifying organisms. The one side of the co-immobilized cells membrane was in aerobic contact with wastewater containing ammonia, while the other side of the membrane was in anoxic contact with ethanol solution (carbon sources for denitrification). Nitrifying bacteria oxidized ammonia to nitrite and nitrate in the membrane, and then denitrifying bacteria reduced nitrite and nitrate to nitrogen gas in the same membrane. The co-immobilized nitrifying organisms and denitrifying organisms accelerated the nitrification rate two times faster than the rate of the immobilized nitrifying organisms alone, although the initial densities of nitrifying organisms in the two membranes were the same. The effect of carbon sources reuse on biological nitrogen removal process wasn't found in the experiment. The co-immobilized cells membrane remained stable for a period of more then 50 days.

  17. Enhanced xylitol production using immobilized Candida tropicalis with non-detoxified corn cob hemicellulosic hydrolysate

    OpenAIRE

    Yewale, Tatyaso; Panchwagh, Shruti; Rajagopalan, Srinivasan; Dhamole, Pradip B.; Jain, Rishi

    2016-01-01

    This study reports an industrially applicable non-sterile xylitol fermentation process to produce xylitol from a low-cost feedstock like corn cob hydrolysate as pentose source without any detoxification. Different immobilization matrices/mediums (alginate, polyvinyl alcohol, agarose gel, polyacrylamide, gelatin, and κ-carrageenan) were studied to immobilize Candida tropicalis NCIM 3123 cells for xylitol production. Amongst this calcium alginate, immobilized cells produced maximum amount of xy...

  18. Enhanced Uranium Immobilization and Reduction by Geobacter sulfurreducens Biofilms

    OpenAIRE

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

    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 concentra...

  19. Kinetics of Phenol Biodegradation by an Immobilized Methanogenic Consortium †

    OpenAIRE

    Dwyer, Daryl F.; Krumme, Mary Lou; Boyd, Stephen A.; Tiedje, James M

    1986-01-01

    A phenol-degrading methanogenic enrichment was successfully immobilized in agar as shown by the stoichiometric conversion of phenol to CH4 and CO2. The enrichment contained members of three physiological groups necessary for the syntrophic mineralization of phenol: a phenol-oxidizing bacterium, a Methanothrix-like bacterium, and an H2-utilizing methanogen. The immobilization technique resulted in the cells being embedded in a long, thin agar strand (1 mm in diameter by 2 to 50 cm in length) t...

  20. POTENTIAL APPLICATIONS OF CHITOSAN NANOPARTICLES AS NOVEL SUPPORT IN ENZYME IMMOBILIZATION

    Directory of Open Access Journals (Sweden)

    Hoda Jafarizadeh Malmiri

    2012-01-01

    Full Text Available Chitosan is an attractive natural biopolymer from renewable resources with the presence of reactive amino and hydroxyl functional groups in its structure. Due to the good biocompatibility of chitosan, it can be used in magnetic-field assisted drug delivery, enzyme or cell immobilization and many other industrial applications. In the past decade, nanotechnology has been a considerable research interest in the area of preparation of immobilized enzyme carriers. This study looks at characteristics and applications of chitosan and chitosan nanoparticles and their potentials as suitable supports for enzyme immobilization. Results indicated that activity of immobilized enzymes and performance of enzyme immobilization onto chitosan nanoparticles are higher than chitosan macro and microparticles. As compared to other biopolymers nanoparticles, application of chitosan nanoparticles to immobilize enzymes strongly increases stability of immobilized enzymes and their easy separability from the reaction mixture at the end of the biochemical process.

  1. Tissue stiffness induced by prolonged immobilization of the rat knee joint and relevance of AGEs (pentosidine).

    Science.gov (United States)

    Lee, Sachiko; Sakurai, Takashi; Ohsako, Masafumi; Saura, Ryuichi; Hatta, Hideo; Atomi, Yoriko

    2010-12-01

    Joints, connective tissues consisting of extracellular matrix (ECM) with few blood vessels, transfer tension to the skeleton in response to environmental demand. Therefore, joint immobilization decreases active and passive mechanical stress, resulting in increased joint stiffness and tissue degeneration; however, the cause of joint stiffness is obscure. Using a rat knee immobilization model, we examined the relationship between range of motion (ROM) and cell numbers and ECM cross-links by accumulation of advanced glycation end products, pentosidine, in the posterior joint capsule of immobilized joints during 16 weeks of immobilization. The left knee joint was immobilized by internal fixation and compared with the non-immobilized right leg. As early as 2 weeks of immobilization, joint ROM and torque significantly decreased and in parallel, disordered alignment of collagen fiber bundles significantly increased, compared with non-immobilized joints. Those changes continued until 16 weeks of immobilization. Significant increases in pentosidine-positive areas after 8 weeks and significantly decreased cell numbers after 16 weeks of immobilization were also observed compared to the contralateral side. A significant negative correlation between tissue stiffness measured by restriction of ROM and accumulation of pentosidine was observed. This study is the first to show that immobilization of knee joints induces articular contracture associated with sequential changes of ECM alignment, influencing ROM and later pentosidine accumulation and decreased cell numbers during the 16-week immobilization period. Pentosidine appears to be an indicator toward a chronic tissue stiffness leading to decreased cell number rather than a cause of ROM restriction induced by joint immobilization.

  2. Bioreporter pseudomonas fluorescens HK44 immobilized in a silica matrix

    Directory of Open Access Journals (Sweden)

    Trogl J.

    2003-01-01

    Full Text Available The bioluminescent bioreporter Pseudomonas fluorescens HK44, the whole cell bacterial biosensor that responds to naphthalene and its metabolites via the production of visible light, was immobilized into a silica matrix by the sol-gel technique. The bioluminescence intensities were measured in the maximum of the bioluminescence band at X = 500 nm. The immobilized cells (>105 cells per g silica matrix produced light after induction by salicylate (cone. > 10 g/l, naphthalene and aminobenzoic acid. The bioluminescence intensities induced by 2,3-dihydroxynaphthalene 3-hydroxybenzoic acid and 4-hydroxybenzoic acid were comparable to a negative control. The cells in the silica layers on glass slides produced light in response to the presence of an inductor at least 8 months after immobilization, and >50 induction cycles. The results showed that these test slides could be used as assays for the multiple determination of water pollution.

  3. Immobilizing U from solution by immobilized sulfate-reducing bacteria of desulfovibrio desulfuricans

    Science.gov (United States)

    Xu, Hulfang; Barton, Larry L.

    2000-07-01

    As determined by transmission electron microscopy, the reduction of uranyl accetate by immobilized cells of Desulfovibrio desulfuricans results in the production of black uraninite nanocrystals precipitated outside the cell. Some nanocrystals are associated with outer membranes of the cell as revealed from cross sections of these metabolically active sulfate-reducing bacteria. The nanocrystals have an average diameter of 5 nm and have anhedral shape. It is proposed that cytochrome in these cells has an important role in the reduction of uranyl through transferring electron from molecular hydrogen or lactic acid to uranyl ions.

  4. Effect of oleic acid on the production of ethanol and fructose from glucose/fructose mixtures in an immobilized cell reactor

    Energy Technology Data Exchange (ETDEWEB)

    Guenette, M.E. [Ottawa Univ., ON (Canada). Dept. of Chemical Engineering]|[IOGEN Corp., Ottawa, ON (Canada); Duvnjak, Z. [Ottawa Univ., ON (Canada). Dept. of Chemical Engineering]|[IOGEN Corp., Ottawa, ON (Canada)

    1995-12-31

    Saccharomyces cerevisiae ATCC 39859 was immobilized onto small cubes of wood to produce ethanol and very enriched fructose syrup from glucose/fructose mixtures through the selective fermentation of glucose. A maximum ethanol productivity of 21.9 g/l.h was attained from a feed containing 9.7% (w/v) glucose and 9.9% (w/v) fructose. An ethanol concentration, glucose conversion and fructose yield of 29.6 g/l, 62% and 99% were obtained, respectively. This resulted in a final fructose/glucose ratio of 2.7. At lower ethanol productivity levels the fructose/glucose ratio increases, as does the ethanol concentration in the effluent. The addition of 30 mg/l oleic acid to the medium increased the ethanol productivity and its concentration by 13% at a dilution rate of 0.74 h{sup -1}. (orig.)

  5. Uptake of plutonium by immobilized bacteria

    International Nuclear Information System (INIS)

    The use of plastic-immobilized bacteria as a system for the concentration of plutonium from aqueous media is investigated. Previous research is reviewed quantifying free cell bacterial concentration of plutonium from solution or suspension. Our research indicates that the species Pseudomonas aeruginosa can be induced to attach firmly to a polymer substrate, while retaining its ability to concentrate plutonium. Melt-blown, filamentous polypropylene is shown to foster cell embedment and uptake capabilities surpassing various other substrates. Oxygen plasma treatment, used to enhance polypropylene wettability, is found to increase the rate of cell embedment significantly. Both embedment and uptake phenomena are found to be dependent upon cell viability. Potential applications for the cell/polymer system are discussed

  6. Poly(Dopamine-Assisted Immobilization of Xu Duan on 3D Printed Poly(Lactic Acid Scaffolds to Up-Regulate Osteogenic and Angiogenic Markers of Bone Marrow Stem Cells

    Directory of Open Access Journals (Sweden)

    Chia-Hung Yeh

    2015-07-01

    Full Text Available Three-dimensional printing is a versatile technique to generate large quantities of a wide variety of shapes and sizes of polymer. The aim of this study is to develop functionalized 3D printed poly(lactic acid (PLA scaffolds and use a mussel-inspired surface coating and Xu Duan (XD immobilization to regulate cell adhesion, proliferation and differentiation of human bone-marrow mesenchymal stem cells (hBMSCs. We prepared PLA scaffolds and coated with polydopamine (PDA. The chemical composition and surface properties of PLA/PDA/XD were characterized by XPS. PLA/PDA/XD controlled hBMSCs’ responses in several ways. Firstly, adhesion and proliferation of hBMSCs cultured on PLA/PDA/XD were significantly enhanced relative to those on PLA. In addition, the focal adhesion kinase (FAK expression of cells was increased and promoted cell attachment depended on the XD content. In osteogenesis assay, the osteogenesis markers of hBMSCs cultured on PLA/PDA/XD were significantly higher than seen in those cultured on a pure PLA/PDA scaffolds. Moreover, hBMSCs cultured on PLA/PDA/XD showed up-regulation of the ang-1 and vWF proteins associated with angiogenic differentiation. Our results demonstrate that the bio-inspired coating synthetic PLA polymer can be used as a simple technique to render the surfaces of synthetic scaffolds active, thus enabling them to direct the specific responses of hBMSCs.

  7. Treating Wastewater With Immobilized Enzymes

    Science.gov (United States)

    Jolly, Clifford D.

    1991-01-01

    Experiments show enzymes are immobilized on supporting materials to make biocatalyst beds for treatment of wastewater. With suitable combination of enzymes, concentrations of various inorganic and organic contaminants, including ammonia and urea, reduced significantly.

  8. Photochemistry of immobilized photoactive compounds

    NARCIS (Netherlands)

    Browne, Wesley R.

    2008-01-01

    The development of responsive molecular based materials and surfaces requires the incorporation of functional molecular components. In this regard thermo-, electro- and photochromic systems are of considerable interest. In this review, the immobilization of photoactive inorganic complexes is focused

  9. [Recovery of platinum with immobilized Citrobacter freudii XP05 biomass].

    Science.gov (United States)

    Hu, Hong-Bo; Liu, Yue-Ying; Fu, Jin-Kun; Xue, Ru; Gu, Ping-Ying

    2003-07-01

    The objective of this work was to develop a valuable adsorbent for recovery of platinum by studying the properties of Pt4+ -adsorption with immobilized Citrobacter freudii XP05 biomass. Five methods for immobilization of Citrobacter freudii XP05 biomass were compared. The method with gelatin-alginate sodium as entrapment matrix was considered to be the optimal. Spherical and uniform beads were produced and the SEM micrograph indicated that the cell of strain XP08 were uniformly dispersed within the matrix. The adsorption of Pt4+ by immobilized XP05 biomass was affected with adsorptive time, pH value of the solution, immobilized biomass concentration, Pt4+ initial concentration The adsorption was a rapid process. The optimal pH value for Pt4+ adsorption was 1.5, and its adsorptive capacity increased linearly with increasing Pt4+ initial concentrations in the range of 50 - 250 mg/L. The experimental data could be fitted to Langmuir and Freundlich models of adsorption isotherm. The adsorptive capacity reached 35.2 mg/g under the conditions of 250 Pt4+ mg/L, 2.0 g/L immobilized biomass, pH 1.5 and 30 degrees C for 60 min. 98.7% of Pt4+ adsorbed on immobilized biomass could be desorbed with 0.5 mol HC1/L. The characteristics of dynamic adsorption and desorption of immobilized XP05 biomass in packed-bed reactor were investigated. The saturation uptake was 24.66 mg Pt4+ /g under the conditions of flow rate 1.2 mL/min, pH 1.5, 50 mg Pt4+/L and 1.85 g biomass(dry weight) . Adsorptive efficiency of Pt4 + by the immobilized XP05 biomass was above 78% for 4 cycles of adsorption and desorption. The recovery of platinum from waste platinum catalyst was studied. The adsorptive capacity was 20.94 mg Pt4+/g immobilized biomass under the conditions of 4.0 g/L immobilized XP05 biomass, 117.76 mg Pt4+/L and pH 1.5 for 60 min. The immobilized XP05 biomass is potentially applicable to the recovery of platinum from waste and wastewater containing platinum. PMID:15969064

  10. Covalent co-immobilization of heparin/laminin complex that with different concentration ratio on titanium surface for selectively direction of platelets and vascular cells behavior

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jian; Chen, Yuan; Liu, Tao; Wang, Xue; Liu, Yang; Wang, Yuan; Chen, Junying, E-mail: chenjy@263.net; Huang, Nan

    2014-10-30

    Highlights: • Extracellular matrix inspired surface modification with fibronectin, heparin and VEGF to construct a favorable microenvironment for selectively anticoagulant and promote endothelialization. • Take the advantage of specific intermolecular interaction, the bioactivity of above biomolecules was more efficiently maintained in compared with the common used covalent immobilization method. • Poly-l-lysine was used as a novel interlayer for surface amination, and in comparison, PLL coating was more feasible and the degradation product had no harm to human body. - Abstract: Surface biofunctional modification of coronary artery stent to improve the hemocompatibility and selectively accelerate endothelium regeneration but prevent restenosis have been become a new hotspot. For this, a novel method was developed in this work by co-immobilization of Ln and heparin complex on poly-L-lysine modified Ti surface. Take the advantage of the specific interaction between Ln and heparin, Ln and heparin complexes with different concentration ratios were set up for creating different exposure density of these two types of biomolecules. According to biocompatibility evaluation results, the Hep/Ln complexes modified surface displayed less platelet adhesion and activation. Especially, on L(150)H and L(200)H surface, the AT III binding quantity, APTT value and anti-coagulation property of modified surface were significantly promoted. Furthermore, the adherent density and proliferation activity of ECs and EPCs were positively correlated with Ln concentration. Notably, the proliferation of both ECs and EPCs on L(100)H, L(150)H and L(200)H surface were greatly promoted. Another hand, the proliferation activity of SMCs was significantly inhibited on Hep/Ln modified surfaces, which was considered mainly due to the inhibitory effect of heparin to SMCs. According to the existing results, this study demonstrated that in a certain range of heparin and laminin concentration ratio

  11. Covalent co-immobilization of heparin/laminin complex that with different concentration ratio on titanium surface for selectively direction of platelets and vascular cells behavior

    International Nuclear Information System (INIS)

    Highlights: • Extracellular matrix inspired surface modification with fibronectin, heparin and VEGF to construct a favorable microenvironment for selectively anticoagulant and promote endothelialization. • Take the advantage of specific intermolecular interaction, the bioactivity of above biomolecules was more efficiently maintained in compared with the common used covalent immobilization method. • Poly-l-lysine was used as a novel interlayer for surface amination, and in comparison, PLL coating was more feasible and the degradation product had no harm to human body. - Abstract: Surface biofunctional modification of coronary artery stent to improve the hemocompatibility and selectively accelerate endothelium regeneration but prevent restenosis have been become a new hotspot. For this, a novel method was developed in this work by co-immobilization of Ln and heparin complex on poly-L-lysine modified Ti surface. Take the advantage of the specific interaction between Ln and heparin, Ln and heparin complexes with different concentration ratios were set up for creating different exposure density of these two types of biomolecules. According to biocompatibility evaluation results, the Hep/Ln complexes modified surface displayed less platelet adhesion and activation. Especially, on L(150)H and L(200)H surface, the AT III binding quantity, APTT value and anti-coagulation property of modified surface were significantly promoted. Furthermore, the adherent density and proliferation activity of ECs and EPCs were positively correlated with Ln concentration. Notably, the proliferation of both ECs and EPCs on L(100)H, L(150)H and L(200)H surface were greatly promoted. Another hand, the proliferation activity of SMCs was significantly inhibited on Hep/Ln modified surfaces, which was considered mainly due to the inhibitory effect of heparin to SMCs. According to the existing results, this study demonstrated that in a certain range of heparin and laminin concentration ratio

  12. Bioremediation of Bisphenol A and Benzophenone by Glycosylation with Immobilized Marine Microalga Pavlova sp.

    OpenAIRE

    Kei Shimoda; Hiroki Hamada

    2009-01-01

    Cultured cells of Pavlova sp. glycosylated bisphenol A to its mono-glucoside, 2-(4-β-D-glucopyranosyloxyphenyl)-2- hydroxyphenylpropane (9%). Use of immobilized Pavlova cells in sodium alginate gel improved yield of the product (17%). On the other hand, Pavlova cell cultures converted benzophenone into diphenylmethanol (49%) and diphenylmethyl β-D-glucopyranoside (6%). Incubation of benzophenone with immobilized Pavlova cells gave products in higher yields; the yields of diphenylmethanol and ...

  13. Immobilization of Acetobacter sp. CCTCC M209061 for efficient asymmetric reduction of ketones and biocatalyst recycling

    Directory of Open Access Journals (Sweden)

    Chen Xiao-Hong

    2012-09-01

    Full Text Available Abstract Background The bacterium Acetobacter sp. CCTCC M209061 is a promising whole-cell biocatalyst with exclusive anti-Prelog stereoselectivity for the reduction of prochiral ketones that can be used to make valuable chiral alcohols such as (R-4-(trimethylsilyl-3-butyn-2-ol. Although it has promising catalytic properties, its stability and reusability are relatively poor compared to other biocatalysts. Hence, we explored various materials for immobilizing the active cells, in order to improve the operational stability of biocatalyst. Results It was found that Ca-alginate give the best immobilized biocatalyst, which was then coated with chitosan to further improve its mechanical strength and swelling-resistance properties. Conditions were optimized for formation of reusable immobilized beads which can be used for repeated batch asymmetric reduction of 4′-chloroacetophenone. The optimized immobilized biocatalyst was very promising, with a specific activity of 85% that of the free-cell biocatalyst (34.66 μmol/min/g dw of cells for immobilized catalyst vs 40.54 μmol/min/g for free cells in the asymmetric reduction of 4′-chloroacetophenone. The immobilized cells showed better thermal stability, pH stability, solvent tolerance and storability compared with free cells. After 25 cycles reaction, the immobilized beads still retained >50% catalytic activity, which was 3.5 times higher than degree of retention of activity by free cells reused in a similar way. The cells could be recultured in the beads to regain full activity and perform a further 25 cycles of the reduction reaction. The external mass transfer resistances were negligible as deduced from Damkohler modulus Da η ∅ Conclusions Ca-alginate coated with chitosan is a highly effective material for immobilization of Acetobacter sp. CCTCC M209061 cells for repeated use in the asymmetric reduction of ketones. Only a small cost in terms of the slightly lower catalytic activity compared to

  14. Cultivation characteristics of immobilized Aspergillus oryzae for kojic acid production.

    Science.gov (United States)

    Kwak, M Y; Rhee, J S

    1992-04-15

    Aspergillus oryzae in situ grown from spores entrapped in calcium alginate gel beads was used for the production of kojic acid. The immobilized cells in flask cultures produced kojic acid in a linear proportion while maintaining the stable metabolic activity for a prolonged production period. Kojic acid was accumulated up to a high concentration of 83 g/L, at which the kojic acid began to crystallize, and, thus, the culture had to be replaced with fresh media for the next batch culture. The overall productivities of two consecutive cultivations were higher than that of free mycelial fermentation. However, the production rate of kojic acid by the immobilized cells was suddenly decreased with the appearance of central cavernae inside the immobilized gel beads after 12 days of the third batch cultivation. PMID:18601027

  15. Status of plutonium ceramic immobilization processes and immobilization forms

    Energy Technology Data Exchange (ETDEWEB)

    Ebbinghaus, B.B.; Van Konynenburg, R.A. [Lawrence Livermore National Lab., CA (United States); Vance, E.R.; Jostsons, A. [Australian Nuclear Science and Technology Organization, Menai (Australia)] [and others

    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 geologic 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.

  16. Cometabolic Degradation of Trichloroethene by Rhodococcus sp. Strain L4 Immobilized on Plant Materials Rich in Essential Oils▿ †

    OpenAIRE

    Suttinun, Oramas; Müller, Rudolf; Luepromchai, Ekawan

    2010-01-01

    The cometabolic degradation of trichloroethene (TCE) by Rhodococcus sp. L4 was limited by the loss of enzyme activity during TCE transformation. This problem was overcome by repeated addition of inducing substrates, such as cumene, limonene, or cumin aldehyde, to the cells. Alternatively, Rhodococcus sp. L4 was immobilized on plant materials which contain those inducers in their essential oils. Cumin seeds were the most suitable immobilizing material, and the immobilized cells tolerated up to...

  17. Covalent co-immobilization of heparin/laminin complex that with different concentration ratio on titanium surface for selectively direction of platelets and vascular cells behavior

    Science.gov (United States)

    Wang, Jian; Chen, Yuan; Liu, Tao; Wang, Xue; Liu, Yang; Wang, Yuan; Chen, Junying; Huang, Nan

    2014-10-01

    Surface biofunctional modification of coronary artery stent to improve the hemocompatibility and selectively accelerate endothelium regeneration but prevent restenosis have been become a new hotspot. For this, a novel method was developed in this work by co-immobilization of Ln and heparin complex on poly-L-lysine modified Ti surface. Take the advantage of the specific interaction between Ln and heparin, Ln and heparin complexes with different concentration ratios were set up for creating different exposure density of these two types of biomolecules. According to biocompatibility evaluation results, the Hep/Ln complexes modified surface displayed less platelet adhesion and activation. Especially, on L(150)H and L(200)H surface, the AT III binding quantity, APTT value and anti-coagulation property of modified surface were significantly promoted. Furthermore, the adherent density and proliferation activity of ECs and EPCs were positively correlated with Ln concentration. Notably, the proliferation of both ECs and EPCs on L(100)H, L(150)H and L(200)H surface were greatly promoted. Another hand, the proliferation activity of SMCs was significantly inhibited on Hep/Ln modified surfaces, which was considered mainly due to the inhibitory effect of heparin to SMCs. According to the existing results, this study demonstrated that in a certain range of heparin and laminin concentration ratio, the biological behavior of platelets, ECs, EPCs and SMCs could be selectively directed. We suggested that this article provided a potential method to construct an adequate platform on a stent surface for accelerate endothelialization with low side effects.

  18. Brain plasticity of rats exposed to prenatal immobilization stress

    OpenAIRE

    Badalyan B. Yu.; Tumasyan N. V.; Meliksetyan I. B.; Sahakyan I. K.; Abrahamyan S. S.; Galoyan A. A.

    2011-01-01

    Aim. This histochemical and immunohistochemical study was aimed at examining the brain cellular structures of newborn rats exposed to prenatal immobilization (IMO) stress. Methods. Histochemical method on detection of Ca2+-dependent acid phosphatase activity and ABC immunohistochemical technique. Results. Cell structures with radial astrocytes marker GFAP, neuroepithelial stem cell marker gene nestin, stem-cells marker and the hypothalamic neuroprotective proline-rich polypeptide PRP-1 (Galar...

  19. Immobilized surfactant-nanotube complexes support selectin-mediated capture of viable circulating tumor cells in the absence of capture antibodies.

    Science.gov (United States)

    Mitchell, Michael J; Castellanos, Carlos A; King, Michael R

    2015-10-01

    The metastatic spread of tumor cells from the primary site to anatomically distant organs leads to a poor patient prognosis. Increasing evidence has linked adhesive interactions between circulating tumor cells (CTCs) and endothelial cells to metastatic dissemination. Microscale biomimetic flow devices hold promise as a diagnostic tool to isolate CTCs and develop metastatic therapies, utilizing E-selectin (ES) to trigger the initial rolling adhesion of tumor cells under flow. To trigger firm adhesion and capture under flow, such devices also typically require antibodies against biomarkers thought to be expressed on CTCs. This approach is challenged by the fact that CTCs are now known to exhibit heterogeneous expression of conventional biomarkers. Here, we describe surfactant-nanotube complexes to enhance ES-mediated capture and isolation of tumor cells without the use of capture antibodies. While the majority of tumor cells exhibited weaker rolling adhesion on halloysite nanotubes (HNT) coated with ES, HNT functionalization with the sodium dodecanoate (NaL) surfactant induced a switch to firm cellular adhesion under flow. Conversely, surfactant-nanotube complexes significantly reduced the number of primary human leukocytes captured via ES-mediated adhesion under flow. The switch in tumor cell adhesion was exploited to capture and isolate tumor cells in the absence of EpCAM antibodies, commonly utilized as the gold standard for CTC isolation. Additionally, HNT-NaL complexes were shown to capture tumor cells with low to negligible EpCAM expression, that are not efficiently captured using conventional approaches.

  20. Skeletal fluorosis in immobilized extremities.

    Science.gov (United States)

    Rosenquist, J B

    1975-11-01

    The effect of immobilization on skeletal fluorosis was studied in growing rabbits. One hind leg was immobilized by an external fixation device extending below the wrist joint and above the knee joint, the extremity being in a straight position after severance of the sciatic nerve. The animals, aged 7 weeks at the beginning of the experiment, were given 10 mg of fluoride per kg body weight and day during 12 weeks. In the tibiae, development of the skeletal fluorosis was more irregular than that observed in previous studies of normally active animals, being most excessive in the mobile bone. The immobilization effect was most profound in the femora as the cortical thickness and the femur score were significantly higher than those in the mobile femora. It was suggested that an altered muscular activity was the reason for the observed changes. PMID:1189918

  1. Pathophysiology of immobilization osteoporosis

    Science.gov (United States)

    Doty, S. B.; DiCarlo, E. F.

    1995-01-01

    The reduction of gravity-related forces on the skeleton creates a type of osteoporosis that is unique because its severity is dependent on the mechanical stress bearing function of the skeleton as well as the length of time that the forces are absent or reduced. Bones that bear weight under normal conditions are more affected than bones that normally do not bear weight. The cytokine environment and the cells in the affected bones are altered in time so that stem cells produce fewer new cells and the differentiated cells tend to be less active. These alterations in the local environment of the affected parts appear to resemble those of age- and disease-associated systemic forms of osteoporosis. The osteoporosis produced as a result of the loss of normal activity however, appears to be at least partially reversible through remobilization, strenuous exercise, and--possibly in the future--cytokine therapy.

  2. Non-invasive screening for Alzheimer's disease by sensing salivary sugar using Drosophila cells expressing gustatory receptor (Gr5a immobilized on an extended gate ion-sensitive field-effect transistor (EG-ISFET biosensor.

    Directory of Open Access Journals (Sweden)

    Hui-Chong Lau

    Full Text Available Body fluids are often used as specimens for medical diagnosis. With the advent of advanced analytical techniques in biotechnology, the diagnostic potential of saliva has been the focus of many studies. We recently reported the presence of excess salivary sugars, in patients with Alzheimer's disease (AD. In the present study, we developed a highly sensitive, cell-based biosensor to detect trehalose levels in patient saliva. The developed biosensor relies on the overexpression of sugar sensitive gustatory receptors (Gr5a in Drosophila cells to detect the salivary trehalose. The cell-based biosensor was built on the foundation of an improved extended gate ion-sensitive field-effect transistor (EG-ISFET. Using an EG-ISFET, instead of a traditional ion-sensitive field-effect transistor (ISFET, resulted in an increase in the sensitivity and reliability of detection. The biosensor was designed with the gate terminals segregated from the conventional ISFET device. This design allows the construction of an independent reference and sensing region for simultaneous and accurate measurements of samples from controls and patients respectively. To investigate the efficacy of the cell-based biosensor for AD screening, we collected 20 saliva samples from each of the following groups: participants diagnosed with AD, participants diagnosed with Parkinson's disease (PD, and a control group composed of healthy individuals. We then studied the response generated from the interaction of the salivary trehalose of the saliva samples and the Gr5a in the immobilized cells on an EG-ISFET sensor. The cell-based biosensor significantly distinguished salivary sugar, trehalose of the AD group from the PD and control groups. Based on these findings, we propose that salivary trehalose, might be a potential biomarker for AD and could be detected using our cell-based EG-ISFET biosensor. The cell-based EG-ISFET biosensor provides a sensitive and direct approach for salivary sugar

  3. Hyaluronan Immobilized Polyurethane as a Blood Contacting Material

    International Nuclear Information System (INIS)

    Hyaluronan (hyaluronic acid, HA) was immobilized onto the surface of amino-functionalized polyurethane films with the goal of obtaining a novel kind of bio material which had the potential in blood-contacting applications. The amino-functionalized polyurethane was prepared by synthesized acidic polyurethane whose pendant carboxyl groups were treated with an excess amount of 1,3-diaminopropane in the presence of N,N-carbonyldiimidazole (CDI). Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), Raman spectroscopy (RS), scanning electron microscopy (SEM), and water contact angle measurement were used to confirm the surface changes at each step of treatment, both in morphologies and chemical compositions. APTT and PT results showed that HA immobilization could prolong the blood coagulation time, thus HA-immobilized polyurethane (PU-HA) exhibited improved blood compatibility. Cytotoxicity analysis showed that the PU-HA films synthesized in this study were cytocompatible and could support human vein endothelial cells (HUVECs) adhesion and proliferation.

  4. Ferrous ion oxidation by Thiobacillus ferrooxidans immobilized on activated carbon

    Institute of Scientific and Technical Information of China (English)

    ZHOU Ji-kui; QIN Wen-qing; NIU Yin-jian; LI Hua-xia

    2006-01-01

    The immobilization of Thiobacillus ferrooxidans on the activated carbon particles as support matrix was investigated. Cycling batch operation results in the complete oxidation of ferrous iron in 8 d when the modified 9 K medium is set to flow through the mini-bioreactor at a rate of 0.104 L/h at 25 ℃. The oxidation rate of ferrous iron with immobilized T. ferrooxidans is 9.38 g/(L·h). The results show that the immobilization of T. ferrooxidans on activated carbon can improve the rate of oxidation of ferrous iron. The SEM images show that a build-up of cells of T. ferrooxidans and iron precipitates is formed on the surface of activated carbon particles.

  5. Immobilization of Heparin: Approaches and Applications

    OpenAIRE

    Murugesan, Saravanababu; Xie, Jin; Linhardt, Robert J.

    2008-01-01

    Heparin, an anticoagulant, has been used in many forms to treat various diseases. These forms include soluble heparin and heparin immobilized to supporting matrices by physical adsorption, by covalent chemical methods and by photochemical attachment. These immobilization methods often require the use of spacers or linkers. This review examines and compares various techniques that have been used for the immobilization of heparin as well as applications of these immobilized heparins. In the app...

  6. Production of cyclodextrin glycosyltransferase by immobilized Bacillus sp. on chitosan matrix.

    Science.gov (United States)

    Eş, Ismail; Ribeiro, Maycon Carvalho; Dos Santos Júnior, Samuel Rodrigues; Khaneghah, Amin Mousavi; Rodriguez, Armando Garcia; Amaral, André Corrêa

    2016-10-01

    The whole-cell immobilization on chitosan matrix was evaluated. Bacillus sp., as producer of CGTase, was grown in solid-state and batch cultivation using three types of starches (cassava, potato and cornstarch). Biomass growth and substrate consumption were assessed by flow cytometry and modified phenol-sulfuric acid assays, respectively. Qualitative analysis of CGTase production was determined by colorless area formation on solid culture containing phenolphthalein. Scanning electron microscopy (SEM) analysis demonstrated that bacterial cells were immobilized on chitosan matrix efficiently. Free cells reached very high numbers during batch culture while immobilized cells maintained initial inoculum concentration. The maximum enzyme activity achieved by free cells was 58.15 U ml(-1) (36 h), 47.50 U ml(-1) (36 h) and 68.36 U ml(-1) (36 h) on cassava, potato and cornstarch, respectively. CGTase activities for immobilized cells were 82.15 U ml(-1) (18 h) on cassava, 79.17 U ml(-1) (12 h) on potato and 55.37 U ml(-1) (in 6 h and max 77.75 U ml(-1) in 36 h) on cornstarch. Application of immobilization technique increased CGTase activity significantly. The immobilized cells produced CGTase with higher activity in a shorter fermentation time comparing to free cells. PMID:27194141

  7. Papain immobilized polyurethane as an ureteral stent material.

    Science.gov (United States)

    Maria Manohar, Cynthya; Doble, Mukesh

    2016-05-01

    Long term use of polyurethane-based ureteral stent is hampered by the development of infection due to the formation of bacterial biofilm and salt deposition. Here papain, is covalently immobilized to polyurethane using glutarldehyde and is investigated as a possible anti-infective ureteral stent material. Fourier transform infrared spectrum confirmed its immobilization. Immobilized enzyme retained 85% of the activity of the free enzyme and about 12% loss of enzyme was observed from the polymer surface in one month. The modified polyurethane showed 8 log reduction in Staphylococcus aureus and 7 log reduction in Escherichia coli live colonies and 3-4 times decrease in the protein and carbohydrate in the biofilms than bare polymer. The amount of calcium and magnesium salts deposited on the polymer surface reduced by 40% after enzyme immobilization. 80% of L6 myoblast cells were viable on this material which indicated that it was noncytotoxic. A linear regression equation with hydrophilicity of the polymer surface and the cell surface hydrophobicity as the two independent variables was able to predict the number of live cells attached on the modified PU. This study indicated the possibility of using such an approach to overcome the problems of ureteral stent associated biofilm and salt encrustation. PMID:26853541

  8. Studies on the surface coat of Paramecium aurelia. II. Relationship to the immobilization antigen.

    Science.gov (United States)

    Wyroba, E

    1977-07-11

    Correlations between the presence of surface coat and immobilization antigen of Paramecium tetraurelia were studied. Supravital, partial removal of the surface coat resulted in accelerated response of monobacterially and axenically grown cells to the homologous antiserum. Ciliates pretreated with trypsin or pronase (0.5 mg/ml for 45 min at 0-4 degrees C) were immobilized approximately twice as fast as untreated control cells. The probable localization of at least part, of the immobilization antigen within the surface coat of P. tetraurelia is discussed.

  9. Histomorphometric analysis of the response of rat skeletal muscle to swimming, immobilization and rehabilitation

    Directory of Open Access Journals (Sweden)

    C.C.F. Nascimento

    2008-09-01

    Full Text Available The objective of the present study was to determine to what extent, if any, swimming training applied before immobilization in a cast interferes with the rehabilitation process in rat muscles. Female Wistar rats, mean weight 260.52 ± 16.26 g, were divided into 4 groups of 6 rats each: control, 6 weeks under baseline conditions; trained, swimming training for 6 weeks; trained-immobilized, swimming training for 6 weeks and then immobilized for 1 week; trained-immobilized-rehabilitated, swimming training for 6 weeks, immobilized for 1 week and then remobilized with swimming for 2 weeks. The animals were then sacrificed and the soleus and tibialis anterior muscles were dissected, frozen in liquid nitrogen and processed histochemically (H&E and mATPase. Data were analyzed statistically by the mixed effects linear model (P < 0.05. Cytoarchitectural changes such as degenerative characteristics in the immobilized group and regenerative characteristics such as centralized nucleus, fiber size variation and cell fragmentation in the groups submitted to swimming were more significant in the soleus muscle. The diameters of the lesser soleus type 1 and type 2A fibers were significantly reduced in the trained-immobilized group compared to the trained group (P < 0.001. In the tibialis anterior, there was an increase in the number of type 2B fibers and a reduction in type 2A fibers when trained-immobilized rats were compared to trained rats (P < 0.001. In trained-immobilized-rehabilitated rats, there was a reduction in type 2B fibers and an increase in type 2A fibers compared to trained-immobilized rats (P < 0.009. We concluded that swimming training did not minimize the deleterious effects of immobilization on the muscles studied and that remobilization did not favor tissue re-adaptation.

  10. Biodiesel production with immobilized lipase: A review.

    Science.gov (United States)

    Tan, Tianwei; Lu, Jike; Nie, Kaili; Deng, Li; Wang, Fang

    2010-01-01

    Fatty acid alkyl esters, also called biodiesel, are environmentally friendly and show great potential as an alternative liquid fuel. Biodiesel is produced by transesterification of oils or fats with chemical catalysts or lipase. Immobilized lipase as the biocatalyst draws high attention because that process is "greener". This article reviews the current status of biodiesel production with immobilized lipase, including various lipases, immobilization methods, various feedstocks, lipase inactivation caused by short chain alcohols and large scale industrialization. Adsorption is still the most widely employed method for lipase immobilization. There are two kinds of lipase used most frequently especially for large scale industrialization. One is Candida antartica lipase immobilized on acrylic resin, and the other is Candida sp. 99-125 lipase immobilized on inexpensive textile membranes. However, to further reduce the cost of biodiesel production, new immobilization techniques with higher activity and stability still need to be explored.

  11. [Modification of drug mutagenicity by their immobilization. Effect of prostatilen immobilized in polyvinyl alcohol in mice].

    Science.gov (United States)

    Mikheev, V S; Bolonina, V P; Gorbachev, A G

    1992-08-01

    Mutagenic drug effect of prostatilen and the possibility of modification were analysed in the sperm head anomalies (SHA) and the bone marrow cell aberrations (CA) tests in Mus musculus. It was found that intraperitoneal injection of 2.5 micrograms of prostatilen induced no significant increase in SHA and CA frequencies, the dose of 5 micrograms inducing both SHA and CA. Ultrafiltration of prostatilen led to decrease in its mutagenicity in the SHA test. Immobilization of the drug (5 and 10 micrograms) in polyvinyl alcohol reduced SHA and CA frequencies, the former decreasing to the control level.

  12. Kefir immobilized on corn grains as biocatalyst for lactic acid fermentation and sourdough bread making.

    Science.gov (United States)

    Plessas, Stavros; Alexopoulos, Athanasios; Bekatorou, Argyro; Bezirtzoglou, Eugenia

    2012-12-01

    The natural mixed culture kefir was immobilized on boiled corn grains to produce an efficient biocatalyst for lactic acid fermentation with direct applications in food production, such as sourdough bread making. The immobilized biocatalyst was initially evaluated for its efficiency for lactic acid production by fermentation of cheese whey at various temperatures. The immobilized cells increased the fermentation rate and enhanced lactic acid production compared to free kefir cells. Maximum lactic acid yield (68.8 g/100 g) and lactic acid productivity (12.6 g/L per day) were obtained during fermentation by immobilized cells at 37 °C. The immobilized biocatalyst was then assessed as culture for sourdough bread making. The produced sourdough breads had satisfactory specific loaf volumes and good sensory characteristics. Specifically, bread made by addition of 60% w/w sourdough containing kefir immobilized on corn was more resistant regarding mould spoilage (appearance during the 11(th) day), probably due to higher lactic acid produced (2.86 g/Kg of bread) compared to the control samples. The sourdough breads made with the immobilized biocatalyst had aroma profiles similar to that of the control samples as shown by headspace SPME GC-MS analysis.

  13. Production of gluconic acid by Aspergillus niger immobilized on polyurethane foam.

    Science.gov (United States)

    Vassilev, N B; Vassileva, M C; Spassova, D I

    1993-06-01

    Production of gluconic acid by cells of Aspergillus niger immobilized on polyurethane foam was studied in repeated-batch shake-flask and bubble-column fermentations. For passive immobilization, various amounts of polyurethane foam and spore suspension were tested in order to obtain a suitable combination for optimal concentration of immobilized biomass. Immobilized cells were successfully reused with higher levels of product formation being maintained for longer period (65-70 h) than free cells. The highest gluconic acid concentration of about 143 g l-1 was reached on hydrol-based production medium with 0.3-cm3 foam cubes in the bubble column, where the effect of more suitable aeration and particle volume: medium volume ratio scheme was also investigated.

  14. Nuclear waste immobilization. Progress report

    International Nuclear Information System (INIS)

    United States defense nuclear wastes are presently in tank storage, largely as sludges comprising Fe, Mn, Ni, U and Na oxides and hydroxides, together with 0.5 to 5 percent of fission products and actinides (exclusive of uranium). The relative proportions of Al, Fe, Mn, Ni, U and Na in the sludges from different tanks vary considerably, except that (Fe + Al + Mn) are by far the major components and Fe is more abundant than Mn. Typical compositions of some calcined sludges from Savannah River are given. This paper briefly describes how the SYNROC process, utilizing straightforward technology, can be readily adapted to the problem of defense waste immobilization, yielding a dense, inert, ceramic waste-form, SYNROC-D. Two classes of processes are discussed - one designed to immobilize sludges containing normal amounts of sodium and the other designed for otherwise similar sludges which are, however, strongly depleted in sodium as a result of more efficient washing procedures

  15. Immobile Complex Verbs in Germanic

    DEFF Research Database (Denmark)

    Vikner, Sten

    2005-01-01

    Certain complex verbs in Dutch, German, and Swiss German do not undergo verb movement. The suggestion to be made in this article is that these ‘‘immobile'' verbs have to fulfill both the requirements imposed on complex verbs of the V° type (=verbs with non-separable prefixes) and the requirements...... imposed on complex verbs of the V* type (=verbs with separable prefixes). This results in such verbs being morphologically unexceptional, i.e., having a full set of forms but syntactically peculiar (‘‘immobile''), i.e., they can only occur in their base position. Any movement is incompatible with either...... the V° requirements or the V* requirements. Haider (1993, p. 62) and Koopman (1995), who also discuss such immobile verbs, only account for verbs with two prefix-like parts (e.g., German uraufführen ‘to perform (a play) for the first time' or Dutch herinvoeren ‘to reintroduce'), not for the more...

  16. Influence of immobilization on the stability of pTG201 recombinant plasmid in some strains of Escherichia coli.

    OpenAIRE

    Nasri, M; Sayadi, S.; Barbotin, J N; Dhulster, P; Thomas, D.

    1987-01-01

    The stability of pTG201 plasmid was examined by continuous culture in three genetically different Escherichia coli hosts. Two types of experiment were carried out, one with free cells and one with immobilized cells. When cells were cultivated in free continuous culture in the absence of antibiotic selection, the plasmid was maintained with various degrees of stability in the three host organisms. By contrast, in continuous culture with immobilized cells, plasmid pTG201 was stably maintained i...

  17. Modifying enzyme activity and selectivity by immobilization

    OpenAIRE

    Rodrigues, Rafael C.; Ortiz, Claudia; Berenguer Murcia, Ángel; Torres, Rodrigo; Fernández Lafuente, Roberto

    2013-01-01

    Immobilization of enzymes may produce alterations in their observed activity, specificity or selectivity. Although in many cases an impoverishment of the enzyme properties is observed upon immobilization (caused by the distortion of the enzyme due to the interaction with the support) in some instances such properties may be enhanced by this immobilization. These alterations in enzyme properties are sometimes associated with changes in the enzyme structure. Occasionally, these variations will ...

  18. Inactivated Sendai Virus (HVJ-E Immobilized Electrospun Nanofiber for Cancer Therapy

    Directory of Open Access Journals (Sweden)

    Takaharu Okada

    2015-12-01

    Full Text Available Inactivated Hemagglutinating Virus of Japan Envelope (HVJ-E was immobilized on electrospun nanofibers of poly(ε-caprolactone by layer-by-layer (LbL assembly technique. The precursor LbL film was first constructed with poly-L-lysine and alginic acid via electrostatic interaction. Then the HVJ-E particles were immobilized on the cationic PLL outermost surface. The HVJ-E adsorption was confirmed by surface wettability test, scanning laser microscopy, scanning electron microscopy, and confocal laser microscopy. The immobilized HVJ-E particles were released from the nanofibers under physiological condition. In vitro cytotoxic assay demonstrated that the released HVJ-E from nanofibers induced cancer cell deaths. This surface immobilization technique is possible to perform on anti-cancer drug incorporated nanofibers that enables the fibers to show chemotherapy and immunotherapy simultaneously for an effective eradication of tumor cells in vivo.

  19. 钛表面固定特异性识别内皮祖细胞的多肽适配子%Immobilization of Peptide Aptamer of Specific Indentification of Endothelial Progenitor Cell on Titanium Surface

    Institute of Scientific and Technical Information of China (English)

    陈卓玥; 李全利; 赵元聪; 陈佳龙; 游天雪; 熊开琴; 黄楠

    2011-01-01

    在钛表面固定可与循环血液中的内皮祖细胞(EPC)特异性结合的多肽适配子,构建内皮祖细胞的特异性识别表面,用于心血管材料的表面改性.首先,采用固相合成法合成可与EPC特异性结合的多肽适配子,其序列为TPSLEQRTVYAK,并在羧基端进行生物素修饰;然后,采用磷酸处理钛表面,在钛表面获得化学键合的羟基,该羟基化表面与3-氨丙基三乙氧基硅烷反应,在钛表面获得游离的氨基,进一步通过碳二亚胺(EDC)介导,在钛表面接枝上生物素;最后,通过生物素-亲和素识别体系,实现EPC特异性多肽适配子在钛表面的固定.采用场发射扫描电子显微镜(SEM)、漫反射红外光谱(DR-FTIR)和免疫荧光分析等手段对样品进行了表征.本研究为多肽适配子在材料表面的固定提供了一种有效的方法,为进一步的生物医学应用研究提供了基础.%In vivo spontaneous endothelialization of cardiovascular materials is thought to be a promising approach to prevent the formation of thrombus and restenosis. Capturing endothelial progenitor cells (EPC)from blood and inducing EPC to grow on the surface of stents is a new strategy for this purpose. In this study,we developed a facile and effective approach to construct a surface that possessed a high affinity and specificity to EPCs by binding peptide aptamer. In order to introduce primary amine groups to covalently immobilize biotin, the titanium surface was treated by phosphoric acid solution to obtain the hydroxyl groups which were used to covalently immobilize aminopropyltriethoxysilane. Furthermore, the biotin was grafted onto the amine functionalized titanium surface by carbodiimide (EDC)-mediated. Finally, using layer-by-layer self-assembly method, biotinylated peptide aptamer was fixed on the titanium surface by the biofin-avidin recognition system.The results of fourier transform infrared spectroscopy ( FTIR), fluorescence labeling method and scanning

  20. Nitric Acid-Treated Carbon Fibers with Enhanced Hydrophilicity for Candida tropicalis Immobilization in Xylitol Fermentation

    OpenAIRE

    Le Wang; Na Liu; Zheng Guo; Dapeng Wu; Weiwei Chen; Zheng Chang; Qipeng Yuan; Ming Hui; Jinshui Wang

    2016-01-01

    Nitric acid (HNO3)-treated carbon fiber (CF) rich in hydrophilic groups was applied as a cell-immobilized carrier for xylitol fermentation. Using scanning electron microscopy, we characterized the morphology of the HNO3-treated CF. Additionally, we evaluated the immobilized efficiency (IE) of Candida tropicalis and xylitol fermentation yield by investigating the surface properties of nitric acid treated CF, specifically, the acidic group content, zero charge point, degree of moisture and cont...

  1. Immobilized epidermal growth factor stimulates persistent, directed keratinocyte migration via activation of PLCγ1.

    Science.gov (United States)

    Kim, Chloe S; Mitchell, Isaiah P; Desotell, Anthony W; Kreeger, Pamela K; Masters, Kristyn S

    2016-07-01

    Epidermal growth factor (EGF) is a critical element in dermal repair, but EGF-containing wound dressings have not been successful clinically. However, these dressings have delivered only soluble EGF, and the native environment provides both soluble and matrix-bound EGF. To address our hypothesis that tethered EGF can stimulate cell behaviors not achievable with soluble EGF, we examined single-cell movement and signaling in human immortalized HaCaT keratinocytes treated with soluble or immobilized EGF. Although both EGF treatments increased collective sheet displacement and individual cell speed, only cells treated with immobilized EGF exhibited directed migration, as well as 2-fold greater persistence compared with soluble EGF. Immunofluorescence showed altered EGF receptor (EGFR) trafficking, where EGFR remained membrane-localized in the immobilized EGF condition. Cells treated with soluble EGF demonstrated higher phosphorylated ERK1/2, and cells on immobilized EGF exhibited higher pPLCγ1, which was localized at the leading edge. Treatment with U0126 inhibited migration in both conditions, demonstrating that ERK1/2 activity was necessary but not responsible for the observed differences. In contrast, PLCγ1 inhibition with U73122 significantly decreased persistence on immobilized EGF. Combined, these results suggest that immobilized EGF increases collective keratinocyte displacement via an increase in single-cell migration persistence resulting from altered EGFR trafficking and PLCγ1 activation.-Kim, C. S., Mitchell, I. P., Desotell, A. W., Kreeger, P. K., Masters, K. S. Immobilized epidermal growth factor stimulates persistent, directed keratinocyte migration via activation of PLCγ1. PMID:27025961

  2. Production of Biodiesel Using Immobilized Lipase and the Characterization of Different Co-Immobilizing Agents and Immobilization Methods

    Directory of Open Access Journals (Sweden)

    Kang Zhao

    2016-08-01

    Full Text Available Lipase from Candida sp. 99–125 is widely employed to catalyzed transesterification and can be used for biodiesel production. In this study, the lipase was immobilized by combined adsorption and entrapment to catalyze biodiesel production from waste cooking oil (WCO via transesterification, and investigating co-immobilizing agents as additives according to the enzyme activity. The addition of the mixed co-immobilizing agents has positive effects on the activities of the immobilized lipase. Three different immobilizing methods were compared by the conversion ratio of biodiesel and structured by Atom Force Microscopy (AFM and Scanning Electron Microscopy (SEM, respectively. It was found that entrapment followed by adsorption was the best method. The effect of the co-immobilizing agent amount, lipase dosage, water content, and reuse ability of the immobilized lipase was investigated. By comparison with previous research, this immobilized lipase showed good reuse ability: the conversion ratio excesses 70% after 10 subsequent reactions, in particular, was better than Novozym435 and TLIM on waste cooking oil for one unit of lipase.

  3. Technetium Immobilization Forms Literature Survey

    Energy Technology Data Exchange (ETDEWEB)

    Westsik, Joseph H.; Cantrell, Kirk J.; Serne, R. Jeffrey; Qafoku, Nikolla

    2014-05-01

    Of the many radionuclides and contaminants in the tank wastes stored at the Hanford site, technetium-99 (99Tc) is one of the most challenging to effectively immobilize in a waste form for ultimate disposal. Within the Hanford Tank Waste Treatment and Immobilization Plant (WTP), the Tc will partition between both the high-level waste (HLW) and low-activity waste (LAW) fractions of the tank waste. The HLW fraction will be converted to a glass waste form in the HLW vitrification facility and the LAW fraction will be converted to another glass waste form in the LAW vitrification facility. In both vitrification facilities, the Tc is incorporated into the glass waste form but a significant fraction of the Tc volatilizes at the high glass-melting temperatures and is captured in the off-gas treatment systems at both facilities. The aqueous off-gas condensate solution containing the volatilized Tc is recycled and is added to the LAW glass melter feed. This recycle process is effective in increasing the loading of Tc in the LAW glass but it also disproportionally increases the sulfur and halides in the LAW melter feed which increases both the amount of LAW glass and either the duration of the LAW vitrification mission or the required supplemental LAW treatment capacity.

  4. Enhancement of nuclease P1 production by Penicillium citrinum YL104 immobilized on activated carbon filter sponge.

    Science.gov (United States)

    Zhao, Nan; Ren, Hengfei; Li, Zhenjian; Zhao, Ting; Shi, Xinchi; Cheng, Hao; Zhuang, Wei; Chen, Yong; Ying, Hanjie

    2015-02-01

    The efficiency of current methods for industrial production of the enzyme nuclease P1 is limited. In this study, we sought to improve fermentation methods for the production of nuclease P1. An immobilized fermentation system using an activated carbon filter sponge as a carrier was used for the production of nuclease P1. In an airlift internal loop reactor (ALR), the fermentation performance of three different fermentation modes, including free-cell fermentation, repeated-batch fermentation, and semi-continuous immobilized fermentation, were compared. The fermentation kinetics in the fermentation broth of the three fermentation modes, including dissolved oxygen (DO), pH value, cell concentration, residual sugar concentration, and enzyme activity, were tested. The productivity of semi-continuous immobilized fermentation reached 8.76 U/mL/h, which was 33.3 and 80.2% higher than that of repeated-batch fermentation and free-cell fermentation, respectively. The sugar consumption of free-cell, repeated-batch, and semi-continuous immobilized fermentations was 41.2, 30.8, and 25.9 g/L, respectively. These results showed that immobilized-cell fermentation by using Penicillium citrinum with activated carbon filter sponge in an ALR was advantageous for nuclease P1 production, especially in the semi-continuous immobilized fermentation mode. In spite of the significant improvement in nuclease P1 production in semi-continuous immobilized fermentation mode, the specific activity of nuclease P1 was almost equal among the three fermentation modes.

  5. Bioremediation of Petrochemical Wastewater Containing BTEX Compounds by a New Immobilized Bacterium Comamonas sp. JB in Magnetic Gellan Gum.

    Science.gov (United States)

    Jiang, Bei; Zhou, Zunchun; Dong, Ying; Wang, Bai; Jiang, Jingwei; Guan, Xiaoyan; Gao, Shan; Yang, Aifu; Chen, Zhong; Sun, Hongjuan

    2015-05-01

    In this study, we investigated the bioremediation of petrochemical wastewater containing BTEX compounds by immobilized Comamonas sp. JB cells. Three kinds of magnetic nanoparticles were evaluated as immobilization supports for strain JB. After comparison with Fe3O4 and a-Fe2O3 nanoparticles, r-Fe2O3 nanoparticle was selected as the optimal immobilization support. The highest biodegradation activity of r-Fe2O3-magnetically immobilized cells was obtained when the concentration of r-Fe2O3 nanoparticle was 120 mg L(-1). Additionally, the recycling experiments demonstrated that the degradation activity of r-Fe2O3-magnetically immobilized cells was still high and led to less toxicity than untreated wastewater during the eight recycles. qPCR suggested the concentration of strain JB in r-Fe2O3-magnetically immobilized cells was evidently increased after eight cycles of degradation experiments. These results supported developing efficient biocatalysts using r-Fe2O3-magnetically immobilized cells and provided a promising technique for improving biocatalysts used in the bioremediation of not only petrochemical wastewater but also other hazardous wastewater.

  6. Optimization of Adsorptive Immobilization of Alcohol Dehydrogenases

    NARCIS (Netherlands)

    Trivedi, Archana; Heinemann, Matthias; Spiess, Antje C.; Daussmann, Thomas; Büchs, Jochen

    2005-01-01

    In this work, a systematic examination of various parameters of adsorptive immobilization of alcohol dehydrogenases (ADHs) on solid support is performed and the impact of these parameters on immobilization efficiency is studied. Depending on the source of the enzymes, these parameters differently in

  7. Immobilized Lactase in the Biochemistry Laboratory

    Science.gov (United States)

    Allison, Matthew J.; Bering, C. Larry

    1998-10-01

    Immobilized enzymes have many practical applications. They may be used in clinical, industrial, and biotechnological laboratories and in many clinical diagnostic kits. For educational purposes, use of immobilized enzymes can easily be taught at the undergraduate or even secondary level. We have developed an immobilized enzyme experiment that combines many practical techniques used in the biochemistry laboratory and fits within a three-hour time frame. In this experiment, lactase from over-the-counter tablets for patients with lactose intolerance is immobilized in polyacrylamide, which is then milled into small beads and placed into a chromatography column. A lactose solution is added to the column and the eluant is assayed using the glucose oxidase assay, available as a kit. We have determined the optimal conditions to give the greatest turnover of lactose while allowing the immobilized enzymes to be active for long periods at room temperature.

  8. Immobilization mediated enhancement of phyllanthin and hypophyllanthin from Phyllanthus amarus

    Institute of Scientific and Technical Information of China (English)

    J.S.Thakur; R.K.Agarwal; M.D.Kharya

    2012-01-01

    Phyllanthus amarus plant is used in the traditional system of medicine as a hepatoprotective drug for which the major lignans phyllanthin and hypophyllanthin are responsible.So far,no significant work has been done on the culture of this plant.Realizing the hepatoprotective potential,the present investigation was undertaken.A cost effective process was developed for enhancing phyllanthin and hypophyllanthin utilizing the immobilization technique.HPTLC was used to compare the phyllanthin and hypophyllanthin contents in calcium alginate immobilized cells obtained from fresh grown plants and MS medium was supplemented with different abiotic elicitors,under aseptic conditions for the treatment with chitosan,copper sulphate,phenylalanine and silver nitrate solution to make the whole process commercially viable.It was revealed that silver nitrate and phenylalanine at low concentration enhances phyllanthin and hypophyllanthin yield as compared to control immobilized cell culture.The study revealed that an increase in the content of phyllanthin and hypophyllanthin was elicitor concentration dependent and silver nitrate treatment gave a maximum yield of hepatoprotective bioactives as compared to the other abiotic elicitors used.

  9. Enhanced accumulation of starch and total carbohydrates in alginate-immobilized Chlorella spp. induced by Azospirillum brasilense: II. Heterotrophic conditions.

    Science.gov (United States)

    Choix, Francisco J; de-Bashan, Luz E; Bashan, Yoav

    2012-10-10

    The effect of the bacterium Azospirillum brasilense jointly immobilized with Chlorella vulgaris or C. sorokiniana in alginate beads on total carbohydrates and starch was studied under dark and heterotrophic conditions for 144 h in synthetic growth medium supplemented with either d-glucose or Na-acetate as carbon sources. In all treatments, enhanced total carbohydrates and starch content per culture and per cell was obtained after 24h; only jointly immobilized C. vulgaris growing on d-glucose significantly increased total carbohydrates and starch content after 96 h. Enhanced accumulation of carbohydrate and starch under jointly immobilized conditions was variable with time of sampling and substrate used. Similar results occurred when the microalgae was immobilized alone. In both microalgae growing on either carbon sources, the bacterium promoted accumulation of carbohydrates and starch; when the microalgae were immobilized alone, they used the carbon sources for cell multiplication. In jointly immobilized conditions with Chlorella spp., affinity to carbon source and volumetric productivity and yield were higher than when Chlorella spp. were immobilized alone; however, the growth rate was higher in microalgae immobilized alone. This study demonstrates that under heterotrophic conditions, A. brasilense promotes the accumulation of carbohydrates in two strains Chlorella spp. under certain time-substrate combinations, producing mainly starch. As such, this bacterium is a biological factor that can change the composition of compounds in microalgae in dark, heterotrophic conditions.

  10. Luminescent Bacterial Sensors Made from Immobilized Films of Photobacterium Phosphoreum

    Institute of Scientific and Technical Information of China (English)

    YIN Ji-qiu; LI Xiao-zhou; ZHOU Chi; ZHANG Yi-hua

    2005-01-01

    A kind of luminous bacterial sensors that can quickly detect the acute toxicity of environmental pollutants were developed. The method is based on the detection of the cellular light of bright luminous bacillus by means of fixing cells so as to detect acute toxicity of luminous bacillus. The bacterial sensor is composed of immobilized film of photobacterium phosphoreum. These bacterial films are sensitive to detecting the toxicoids, which are difficult or even impossible to be measured by traditional analytical chemistry methods. The films should be stored at 4 ℃ and the stability of the sensors exceeds 1 month with no measurable deterioration of the signal. These results demonstrate that the immobilized film of P.phosphreum can be used to develop the on-line environmental contamination monitor.

  11. A study on the performance of hyaluronic acid immobilized chitosan film

    Energy Technology Data Exchange (ETDEWEB)

    Wang Yingjun; Guo Li; Ren Li; Yin Shiheng [Biomaterial Research Institute, College of Material Science and Engineering, South China University of Technology, Guangzhou, 510640 (China); Ge Jian; Gao Qianying [State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, 510060 (China); Luxbacher, Thomas; Luo Shijing, E-mail: imwangyj@scut.edu.c, E-mail: psliren@scut.edu.c [Anton Paar GmbH, Anton-Paar-Strasse 20, A-8054 Graz (Austria)

    2009-06-15

    In order to improve hydrophilicity and biocompatibility of chitosan, hyaluronic acid was immobilized onto the surface of chitosan film. The structure of films was characterized by Fourier transformed infrared spectroscopy with attenuated total reflectance (ATR-FTIR), x-ray photoelectron spectroscopy (XPS) and zeta potential. Results confirmed that hyaluronic acid was successfully immobilized on chitosan film. Transparency, water absorption percentage and contact angle of films were characterized. Results showed that there was no significant variation in transparency (p < 0.05) before and after immobilization, the maximum was up to 99% which was enough for corneal regeneration in clinical applications. After the immobilization, the time-dependent contact angle declined sharply (from 91.8 deg. to 67.7 deg. at 100 s). The hydrophilicity was significantly improved. The methylthiazol tetrazolium (MTT) (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay was used to assess cell viability and proliferation. Results showed that human cornea epithelial cells (HCEC) grew better on hyaluronic acid immobilized chitosan films than on chitosan films. The hyaluronic acid immobilized chitosan film could be a promising candidate material for corneal regeneration.

  12. Plutonium Immobilization Project Baseline Formulation

    Energy Technology Data Exchange (ETDEWEB)

    Ebbinghaus, B.

    1999-02-01

    A key milestone for the Immobilization Project (AOP Milestone 3.2a) in Fiscal Year 1998 (FY98) is the definition of the baseline composition or formulation for the plutonium ceramic form. The baseline formulation for the plutonium ceramic product must be finalized before the repository- and plant-related process specifications can be determined. The baseline formulation that is currently specified is given in Table 1.1. In addition to the baseline formulation specification, this report provides specifications for two alternative formulations, related compositional specifications (e.g., precursor compositions and mixing recipes), and other preliminary form and process specifications that are linked to the baseline formulation. The preliminary specifications, when finalized, are not expected to vary tremendously from the preliminary values given.

  13. Uranium Immobilization in Wetland Soils

    Science.gov (United States)

    Jaffe, Peter R.; Koster van Groos, Paul G.; Li, Dien; Chang, Hyun-Shik; Seaman, John C.; Kaplan, Daniel I.; Peacock, Aaron D.; Scheckel, Kirk

    2014-05-01

    stronger for the mesocosms with the higher Fe(II) load. Analysis via XANES showed that a fraction (up to ~1/3) of uranium was reduced to U(IV), for mesocosms operated under low iron loading, indicating that iron cycling in the rhizosphere also results in uranium reduction and immobilization. For mesocosms operating under the higher iron loading, the fraction of uranium immobilized as U(IV) was much lower, indicating that uranium co-precipitation with iron might have been the dominant immobilization process. In parallel to these mesocosm experiments, dialysis samplers have been deployed at the Savannah River National Laboratory near a creek with uranium contamination, to determine dissolved species, including Fe(II) and U(VI) in these wetland soils and their seasonal variability. The results show that there is a strong seasonal variability in dissolved iron and uranium, indicating a strong immobilization during the growing season, which is consistent with the mesocosm experimental results that the rhizosphere iron and uranium cycling are closely linked.

  14. Degradation of pyrene by immobilized microorganisms in saline-alkaline soil

    Institute of Scientific and Technical Information of China (English)

    Shanxian Wang; Xiaojun Li; Wan Liu; Peijun Li; Lingxue Kong; Wenjie Ren; Haiyan Wu; Ying TU

    2012-01-01

    Biodegradation of polycyclic aromatic hydrocarbons (PAHs) is very difficult in saline-alkaline soil due to the inhibition of microbial growth under saline-alkaline stress.The microorganisms that can most effectively degrade PAHs were screened by introducing microorganisms immobilized on farm byproducts and assessing the validity of the immobilizing technique for PAHs degradation in pyrene-contaminated saline-alkaline soil.Among the microorganisms examined,it was found that Mycobacterium sp.B2 is the best,and can degrade 82.2% and 83.2% of pyrene for free and immobilized cells after 30 days of incubation.The immobilization technique could increase the degradation of pyrene significantly,especially for fungi.The degradation of pyrene by the immobilized microorganisms Mucor sp.F2,fungal consortium MF and co-cultures of MB+MF was increased by 161.7% (P < 0.05),60.1% (P <0.05) and 59.6% (P < 0.05) after 30 days,respectively,when compared with free F2,MF and MB+MF.Scanning electron micrographs of the immobilized microstructure proved the positive effects of the immobilized microbial technique on pyrene remediation in salinealkaline soil,as the interspace of the carrier material structure was relatively large,providing enough space for cell growth.Co-cultures of different bacterial and fungal species showed different abilities to degrade PAHs.The present study suggests that Mycobacterium sp.B2 can be employed for in situ bioremediation of PAHs in saline-alkaline soil,and immobilization of fungi on farm byproducts and nutrients as carriers will enhance fungus PAH-degradation ability in saline-alkaline soil.

  15. In vivo biotinylation of recombinant beta-glucosidase enables simultaneous purification and immobilization on streptavidin coated magnetic particles

    DEFF Research Database (Denmark)

    Alftrén, Johan; Ottow, Kim Ekelund; Hobley, Timothy John

    2013-01-01

    Beta-glucosidase from Bacillus licheniformis was in vivo biotinylated in Escherichia coli and subsequently immobilized directly from cell lysate on streptavidin coated magnetic particles. In vivo biotinylation was mediated by fusing the Biotin Acceptor Peptide to the C-terminal of beta......-glucosidase and co-expressing the BirA biotin ligase. The approach enabled simultaneous purification and immobilization of the enzyme from crude cell lysate on magnetic particles because of the high affinity and strong interaction between biotin and streptavidin. After immobilization of the biotinylated beta...

  16. Treatment of landfill leachate by immobilized microorganisms

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    This paper focuses on the outcome and the main performance of the immobilized microbial that treats landfill leachate. Based on the analysis of COD and ammonia-nitrogen of the influent and effluent, research was done on the high removal efficiency of COD and ammonium nitrogen by immobilized microbial. The leachate composition was analyzed qualitatively using GC-MS before and after being treated. Biological loading of efficient microbial flora on the carrier was measured by Kjeldahl’s method. Finally, the patterns of immobilized microbe were observed through scanning electron microscopy (SEM). The results showed that in immobilized microorganisms system, the efficiencies of COD and nitrogen were 98.3% and 99.9%, respectively. There was a great reduction of organic components in effluent. When the immobilized biomass on the carrier was 38 g·L?1 (H2O), the filamentous microorganism was highly developed. There was no inhibitory effect on the nitrobacteria and nitrococcus, when ammonia was over 200 mg·L?1 and NH3 over 150 mg·L?1. At a high organic loading, it still had good nitrification. This paper also compares the performance of immobilized microbial with free microbial under the same condition. The immobilized microbial technology demonstrated better than the latter in all aspects.

  17. Treatment of landfill leachate by immobilized microorganisms

    Institute of Scientific and Technical Information of China (English)

    YE ZhengFang; YU HongYan; WEN LiLi; NI JinRen

    2008-01-01

    This paper focuses on the outcome and the main performance of the immobilized microbial that treats landfill leachate. Based on the analysis of COD and ammonia-nitrogen of the influent and effluent, research was done on the high removal efficiency of COD and ammonium nitrogen by immobilized microbial. The leachate composition was analyzed qualitatively using GC-MS before and after being treated. Biological loading of efficient microbial flora on the carrier was measured by Kjeldahl's method. Finally, the patterns of immobilized microbe were observed through scanning electron microscopy (SEM). The results showed that in immobilized microorganisms system, the efficiencies of COD and nitrogen were 98.3% and 99.9%, respectively. There was a great reduction of organic components in effluent. When the immobilized biomass on the carrier was 38 g·L-1 (H2O), the filamentous microorganism was highly developed. There was no inhibitory effect on the nitrobacteria and nitrococcus, when ammonia was over 200 mg·L-1 and NH3 over 150 mg·L-1, At a high organic loading, it still had good nitrification. This paper also compares the performance of immobilized microbial with free microbial under the same condition. The immobilized microbial technology demonstrated better than the latter in all aspects.

  18. IMMOBILIZATION OF Saccharomyces Cerevisiae USING POLY(ACRYLAMIDE) GEL FOR ASYMMETRIC SYNTHESIS OF R(-)-MANDELIC ACID

    Institute of Scientific and Technical Information of China (English)

    LI Zhongqin; GUO Daiping; HUANG Xinghua; YANG Kai; XU Xiaoping

    2006-01-01

    In this paper, the poly(acrylamide) hydrogel used to immobilize saccharomyces cerevisiae for asymmetric synthesis of R(-)-mandelic acid was prepared with free radical ploymerization in deionized water at room temperature under nitrogen atmosphere. The influence of the composition of hydrogel, loading amount of cells and culture conditions on the asymmetric synthesis was investigated. Results show that PAAm hydrogel is a feasible carrier for immobilization of cells which is a potential alternative method to prepare enantiomerically pure R(-)-mandelic acid.

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

    Energy Technology Data Exchange (ETDEWEB)

    Cao, Bin; Ahmed, B.; Kennedy, David W.; Wang, Zheming; Shi, Liang; Marshall, Matthew J.; Fredrickson, Jim K.; Isern, Nancy G.; Majors, Paul D.; Beyenal, Haluk

    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 upon 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.

  20. Improved Production of Cyclodextrins by Alkalophilic Bacilli Immobilized on Synthetic or Loofa Sponges

    Directory of Open Access Journals (Sweden)

    Graciette Matioli

    2012-10-01

    Full Text Available This study aimed to improve the production of β-cyclodextrin (β-CD by microbial cells immobilized on synthetic or loofa sponges both with and without the use of alginate or chitosan. The most suitable matrix for the immobilization of Bacillus firmus strain 7B was synthetic sponge and for Bacillus sphaericus strain 41 was loofa sponge. After 330 days of storage, the β-CD production by Bacillus firmus and Bacillus sphaericus remained at around 41% and 49%, respectively, of initial levels. After 24 days of immobilization on loofa sponge, Bacillus sphaericus strain 41 achieved an improved operational stability, reaching 86.6 mM β-CD after 20 days of production, compared to only 32.8 mM of β-CD produced by free Bacillus sphaericus strain 41 cells. The expected increase in β-CD production by immobilized cells of Bacillus firmus strain 7B on synthetic sponge for 4 days was not statistically different to that for cells immobilized for 24 days. The application of this process on an industrial scale using loofa sponge, an inexpensive and renewable matrix, will allow the stable production of β-CD.

  1. Immobilization of Peroxidase onto Magnetite Modified Polyaniline

    Directory of Open Access Journals (Sweden)

    Eduardo Fernandes Barbosa

    2012-01-01

    Full Text Available The present study describes the immobilization of horseradish peroxidase (HRP on magnetite-modified polyaniline (PANImG activated with glutaraldehyde. After the optimization of the methodology, the immobilization of HRP on PANImG produced the same yield (25% obtained for PANIG with an efficiency of 100% (active protein. The optimum pH for immobilization was displaced by the effect of the partition of protons produced in the microenvironment by the magnetite. The tests of repeated use have shown that PANImG-HRP can be used for 13 cycles with maintenance of 50% of the initial activity.

  2. Ion-implanted polytetrafluoroethylene enhances Saccharomyces cerevisiae biofilm formation for improved immobilization.

    Science.gov (United States)

    Tran, Clara T H; Kondyurin, Alexey; Hirsh, Stacey L; McKenzie, David R; Bilek, Marcela M M

    2012-11-01

    The surface of polytetrafluoroethylene (PTFE) was modified using plasma immersion ion implantation (PIII) with the aim of improving its ability to immobilize yeast. The density of immobilized cells on PIII-treated and -untreated PTFE was compared as a function of incubation time over 24 h. Rehydrated yeast cells attached to the PIII-treated PTFE surface more rapidly, with higher density, and greater attachment strength than on the untreated surface. The immobilized yeast cells were removed mechanically or chemically with sodium hydroxide and the residues left on the surfaces were analysed with Fourier transform infrared spectroscopy-attenuated total reflection (FTIR-ATR) and X-ray photoelectron spectroscopy (XPS). The results revealed that the mechanism of cell attachment on both surfaces differs and a model is presented for each. Rapid attachment on the PIII-treated surface occurs through covalent bonds of cell wall proteins and the radicals on the treated surface. In contrast, on the untreated surface, only physisorbed molecules were found in the residue and lipids were more highly concentrated than proteins. The presence of lipids in the residue was found to be a consequence of damage to the plasma membrane during the rehydration process and the increased cell stress was also apparent by the amount of Hsp12 in the protein residue. The immobilized yeast cells on PIII-treated PTFE were found to be as active as yeast cells in suspension.

  3. Hyaluronan Immobilized Polyurethane as a Blood Contacting Material

    Directory of Open Access Journals (Sweden)

    Feirong Gong

    2010-01-01

    Full Text Available Hyaluronan (hyaluronic acid, HA was immobilized onto the surface of amino-functionalized polyurethane films with the goal of obtaining a novel kind of biomaterial which had the potential in blood-contacting applications. The amino-functionalized polyurethane was prepared by synthesized acidic polyurethane whose pendant carboxyl groups were treated with an excess amount of 1,3-diaminopropane in the presence of N,N-carbonyldiimidazole (CDI. Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR, Raman spectroscopy (RS, scanning electron microscopy (SEM, and water contact angle measurement were used to confirm the surface changes at each step of treatment, both in morphologies and chemical compositions. APTT and PT results showed that HA immobilization could prolong the blood coagulation time, thus HA-immobilized polyurethane (PU-HA exhibited improved blood compatibility. Cytotoxicity analysis showed that the PU-HA films synthesized in this study were cytocompatible and could support human vein endothelial cells (HUVECs adhesion and proliferation.

  4. Immobilization of antibacterial chlorhexidine on stainless steel using crosslinking polydopamine film: Towards infection resistant medical devices.

    Science.gov (United States)

    Mohd Daud, Nurizzati; Saeful Bahri, Ihda Fithriyana; Nik Malek, Nik Ahmad Nizam; Hermawan, Hendra; Saidin, Syafiqah

    2016-09-01

    Chlorhexidine (CHX) is known for its high antibacterial substantivity and is suitable for use to bio-inert medical devices due to its long-term antibacterial efficacy. However, CHX molecules require a crosslinking film to be stably immobilized on bio-inert metal surfaces. Therefore, polydopamine (PDA) was utilized in this study to immobilize CHX on the surface of 316L type stainless steel (SS316L). The SS316L disks were pre-treated, modified with PDA film and immobilized with different concentrations of CHX (10mM-50mM). The disks were then subjected to various surface characterization analyses (ATR-FTIR, XPS, ToF-SIMS, SEM and contact angle measurement) and tested for their cytocompatibility with human skin fibroblast (HSF) cells and antibacterial activity against Escherichia coli and Staphylococcus aureus. The results demonstrated the formation of a thin PDA film on the SS316L surface, which acted as a crosslinking medium between the metal and CHX. CHX was immobilized via a reduction process that covalently linked the CHX molecules with the functional group of PDA. The immobilization of CHX increased the hydrophobicity of the disk surfaces. Despite this property, a low concentration of CHX optimized the viability of HSF cells without disrupting the morphology of adherent cells. The immobilized disks also demonstrated high antibacterial efficacy against both bacteria, even at a low concentration of CHX. This study demonstrates a strong beneficial effect of the crosslinked PDA film in immobilizing CHX on bio-inert metal, and these materials are applicable in medical devices. Specifically, the coating will restrain bacterial proliferation without suffocating nearby tissues. PMID:27153117

  5. Optimization of adsorptive immobilization of alcohol dehydrogenases.

    Science.gov (United States)

    Trivedi, Archana; Heinemann, Matthias; Spiess, Antje C; Daussmann, Thomas; Büchs, Jochen

    2005-04-01

    In this work, a systematic examination of various parameters of adsorptive immobilization of alcohol dehydrogenases (ADHs) on solid support is performed and the impact of these parameters on immobilization efficiency is studied. Depending on the source of the enzymes, these parameters differently influence the immobilization efficiency, expressed in terms of residual activity and protein loading. Residual activity of 79% was achieved with ADH from bakers' yeast (YADH) after optimizing the immobilization parameters. A step-wise drying process has been found to be more effective than one-step drying. A hypothesis of deactivation through bubble nucleation during drying of the enzyme/glass bead suspension at low drying pressure (300% residual activity was found after drying. Hyperactivation of the enzyme is probably caused by structural changes in the enzyme molecule during the drying process. ADH from Thermoanaerobacter species (ADH T) is found to be stable under drying conditions (>15 kPa) in contrast to LBADH and YADH.

  6. Immobilization of Rocky Flats Graphite Fines Residue

    International Nuclear Information System (INIS)

    The development of the immobilization process for graphite fines has proceeded through a series of experimental programs. The experimental procedures and results from each series of experiments are discussed in this report

  7. Immobilization of Rocky Flats Graphite Fines Residue

    Energy Technology Data Exchange (ETDEWEB)

    Rudisill, T.S.

    1999-04-06

    The development of the immobilization process for graphite fines has proceeded through a series of experimental programs. The experimental procedures and results from each series of experiments are discussed in this report.

  8. Immobilization of Bacillus sp. in mesoporous activated carbon for degradation of sulphonated phenolic compound in wastewater

    Energy Technology Data Exchange (ETDEWEB)

    Sekaran, G., E-mail: ganesansekaran@gmail.com [Environmental Technology Division, Council of Scientific and Industrial Research (CSIR), Central Leather Research Institute (CLRI), Adyar, Chennai-600 020 (India); Karthikeyan, S. [Environmental Technology Division, Council of Scientific and Industrial Research (CSIR), Central Leather Research Institute (CLRI), Adyar, Chennai-600 020 (India); Gupta, V.K. [Department of Chemistry, Indian Institute of Technology Roorkee, Roorkee-247 667 (India); Department of Chemistry, King Fahd University of Petroleum and Minerals, Dhahran 31261 (Saudi Arabia); Boopathy, R.; Maharaja, P. [Environmental Technology Division, Council of Scientific and Industrial Research (CSIR), Central Leather Research Institute (CLRI), Adyar, Chennai-600 020 (India)

    2013-03-01

    Xenobiotic compounds are used in considerable quantities in leather industries besides natural organic and inorganic compounds. These compounds resist biological degradation and thus they remain in the treated wastewater in the unaltered molecular configurations. Immobilization of organisms in carrier matrices protects them from shock load application and from the toxicity of chemicals in bulk liquid phase. Mesoporous activated carbon (MAC) has been considered in the present study as the carrier matrix for the immobilization of Bacillus sp. isolated from Effluent Treatment Plant (ETP) employed for the treatment of wastewater containing sulphonated phenolic (SP) compounds. Temperature, pH, concentration, particle size and mass of MAC were observed to influence the immobilization behavior of Bacillus sp. The percentage immobilization of Bacillus sp. was the maximum at pH 7.0, temperature 20 Degree-Sign C and at particle size 300 {mu}m. Enthalpy, free energy and entropy of immobilization were - 46.9 kJ mol{sup -1}, - 1.19 kJ mol{sup -1} and - 161.36 J K{sup -1} mol{sup -1} respectively at pH 7.0, temperature 20 Degree-Sign C and particle size 300 {mu}m. Higher values of {Delta}H{sup 0} indicate the firm bonding of the Bacillus sp. in MAC. Degradation of aqueous sulphonated phenolic compound by Bacillus sp. immobilized in MAC followed pseudo first order rate kinetics with rate constant 1.12 Multiplication-Sign 10{sup -2} min{sup -1}. Highlights: Black-Right-Pointing-Pointer Degradation on phenolic syntan using immobilized activated carbon as catalyst. Black-Right-Pointing-Pointer Bacillus sp. immobilized cell reactor removed all refractory organic loads. Black-Right-Pointing-Pointer The removal mechanism is due to co-metabolism between carbon and organisms. Black-Right-Pointing-Pointer The organics are completely metabolized rather than adsorption.

  9. Biodegradation of phenol by free and immobilized Acinetobacter sp.strain PD12

    Institute of Scientific and Technical Information of China (English)

    WANG Ying; TIAN Ye; HAN Bin; ZHAO Hua-bing; BI Jian-nan; CAI Bao-li

    2007-01-01

    A new phenol-degrading bacterium with high biodegradation activity and high tolerance of phenol, strain PD 12, was isolated from the activated sludge of Tianjin Jizhuangzi Wastewater Treatment Facility in China. This strain was capable of removing 500 mg phenol/L in liquid minimal medium by 99.6% within 9 h and metabolizing phenol at concentrations up to 1100 mg/L. DNA sequencing and homologous analysis of 16S rRNA gene identified PD12 to be an Acinetobacter sp. Polyvinyl alcohol (PVA) was used as a gel matrix to immobilize Acinetobacter sp. strain PD12 by repeated freezing and thawing. The factors affecting phenol degradation of immobilized cells were investigated, and the results showed that the immobilized cells could tolerate a high phenol level and protected the bacteria against changes in temperature and pH. Storage stability and reusability tests revealed that the phenol degradation functions of immobilized cells were stable after reuse for 50 times or storing at 4℃ for 50 d. These results indicate that immobilized Acinetobacter sp. strain PD 12 possesses a good application potential in the treatment of phenol-containing wastewater.

  10. Ceramification: A plutonium immobilization process

    Energy Technology Data Exchange (ETDEWEB)

    Rask, W.C. [Dept. of Energy, Golden, CO (United States); Phillips, A.G. [Rocky Flats Environmental Technology Site, Golden, CO (United States)

    1996-05-01

    This paper describes a low temperature technique for stabilizing and immobilizing actinide compounds using a combination process/storage vessel of stainless steel, in which measured amounts of actinide nitrate solutions and actinide oxides (and/or residues) are systematically treated to yield a solid article. The chemical ceramic process is based on a coating technology that produces rare earth oxide coatings for defense applications involving plutonium. The final product of this application is a solid, coherent actinide oxide with process-generated encapsulation that has long-term environmental stability. Actinide compounds can be stabilized as pure materials for ease of re-use or as intimate mixtures with additives such as rare earth oxides to increase their degree of proliferation resistance. Starting materials for the process can include nitrate solutions, powders, aggregates, sludges, incinerator ashes, and others. Agents such as cerium oxide or zirconium oxide may be added as powders or precursors to enhance the properties of the resulting solid product. Additives may be included to produce a final product suitable for use in nuclear fuel pellet production. The process is simple and reduces the time and expense for stabilizing plutonium compounds. It requires a very low equipment expenditure and can be readily implemented into existing gloveboxes. The process is easily conducted with less associated risk than proposed alternative technologies.

  11. Immobilization of Fast Reactor First Cycle Raffinate

    Energy Technology Data Exchange (ETDEWEB)

    Langley, K. F.; Partridge, B. A.; Wise, M.

    2003-02-26

    This paper describes the results of work to bring forward the timing for the immobilization of first cycle raffinate from reprocessing fuel from the Dounreay Prototype Fast Reactor (PFR). First cycle raffinate is the liquor which contains > 99% of the fission products separated from spent fuel during reprocessing. Approximately 203 m3 of raffinate from the reprocessing of PFR fuel is held in four tanks at the UKAEA's site at Dounreay, Scotland. Two methods of immobilization of this high level waste (HLW) have been considered: vitrification and cementation. Vitrification is the standard industry practice for the immobilization of first cycle raffinate, and many papers have been presented on this technique elsewhere. However, cementation is potentially feasible for immobilizing first cycle raffinate because the heat output is an order of magnitude lower than typical HLW from commercial reprocessing operations such as that at the Sellafield site in Cumbria, England. In fact, it falls within the upper end of the UK definition of intermediate level waste (ILW). Although the decision on which immobilization technique will be employed has yet to be made, initial development work has been undertaken to identify a suitable cementation formulation using inactive simulant of the raffinate. An approach has been made to the waste disposal company Nirex to consider the disposability of the cemented product material. The paper concentrates on the process development work that is being undertaken on cementation to inform the decision making process for selection of the immobilization method.

  12. Brain plasticity of rats exposed to prenatal immobilization stress

    Directory of Open Access Journals (Sweden)

    Badalyan B. Yu.

    2011-10-01

    Full Text Available Aim. This histochemical and immunohistochemical study was aimed at examining the brain cellular structures of newborn rats exposed to prenatal immobilization (IMO stress. Methods. Histochemical method on detection of Ca2+-dependent acid phosphatase activity and ABC immunohistochemical technique. Results. Cell structures with radial astrocytes marker GFAP, neuroepithelial stem cell marker gene nestin, stem-cells marker and the hypothalamic neuroprotective proline-rich polypeptide PRP-1 (Galarmin, a natural cytokine of a common precursor to neurophysin vasopressin associated glycoprotein have been revealed in several brain regions. Conclusions. Our findings indicate the process of generation of new neurons in response to IMO and PRP-1 involvement in this recovery mechanism, as PRP-1-Ir was detected in the above mentioned cell structures, as well as in the neurons and nerve fibers.

  13. Bioethanol production by reusable Saccharomyces cerevisiae immobilized in a macroporous monolithic hydrogel matrices.

    Science.gov (United States)

    Mulko, Lucinda; Rivarola, Claudia R; Barbero, Cesar A; Acevedo, Diego F

    2016-09-10

    Performance of yeasts on industrial processes can be dramatically improved by immobilization of the biocatalyst. The immobilization of Saccharomyces cerevisiae inside monolithic macroporous hydrogels were produced by in-situ polymerization of acrylamide around a live yeast suspension under cryogelation conditions. Preculture of the yeasts was not necessary and this innovative and simple procedure is amenable to scaling-up to industrial production. The yeasts were efficiently retained in monolithic hydrogels, presenting excellent mechanical properties and high cell viability. Macroporous hydrogels showed a fast mass transport allowing the hydrogel-yeast complexes achieved similar ethanol yield and productivity than free yeasts, which is larger than those reached with yeasts immobilized in compact hydrogels. Moreover, the same yeasts were able to maintain its activity by up to five reaction cycles with a cell single batch during fermentation reactions. PMID:27396938

  14. Bioethanol production by reusable Saccharomyces cerevisiae immobilized in a macroporous monolithic hydrogel matrices.

    Science.gov (United States)

    Mulko, Lucinda; Rivarola, Claudia R; Barbero, Cesar A; Acevedo, Diego F

    2016-09-10

    Performance of yeasts on industrial processes can be dramatically improved by immobilization of the biocatalyst. The immobilization of Saccharomyces cerevisiae inside monolithic macroporous hydrogels were produced by in-situ polymerization of acrylamide around a live yeast suspension under cryogelation conditions. Preculture of the yeasts was not necessary and this innovative and simple procedure is amenable to scaling-up to industrial production. The yeasts were efficiently retained in monolithic hydrogels, presenting excellent mechanical properties and high cell viability. Macroporous hydrogels showed a fast mass transport allowing the hydrogel-yeast complexes achieved similar ethanol yield and productivity than free yeasts, which is larger than those reached with yeasts immobilized in compact hydrogels. Moreover, the same yeasts were able to maintain its activity by up to five reaction cycles with a cell single batch during fermentation reactions.

  15. Improved enzyme immobilization for enhanced bioelectrocatalytic activity of porous electrodes

    Energy Technology Data Exchange (ETDEWEB)

    Szamocki, Rafael [University Bordeaux 1, CNRS, ISM, Ecole Nationale Superieur de Chimie et Physique de Bordeaux, 16 Avenue Pey Berland, 33607 Pessac (France); Department of Physical Chemistry, Universitaet des Saarlandes, 66123 Saarbruecken (Germany); Velichko, Alexandra; Muecklich, Frank [Department of Material Science, Universitaet des Saarlandes, 66123 Saarbruecken (Germany); Reculusa, Stephane; Ravaine, Serge [Centre de Recherche Paul Pascal-CRPP, 115 Avenue du Dr. Schweitzer, 33600 Pessac (France); Neugebauer, Sebastian; Schuhmann, Wolfgang [Department of Analytical Chemistry, Ruhr-Universitaet Bochum, Universitaetsstr. 150, 44780 Bochum (Germany); Hempelmann, Rolf [Department of Physical Chemistry, Universitaet des Saarlandes, 66123 Saarbruecken (Germany); Kuhn, Alexander [University Bordeaux 1, CNRS, ISM, Ecole Nationale Superieur de Chimie et Physique de Bordeaux, 16 Avenue Pey Berland, 33607 Pessac (France)

    2007-08-15

    Porous electrodes with increased surface area have been prepared using a template route via colloidal crystals. The ordered porous structure and the interconnections between the pores have been quantitatively characterized by Focused Ion Beam Tomography. The internal surfaces of the electrodes have been biofunctionalized with two enzymatic systems for glucose oxidation. In order to increase significantly the stability, the biocatalysts have been immobilized either by crosslinking or by incorporation in an electrodeposition paint. The modified porous electrodes show an increased overall signal and therefore a better detection limit and a higher sensitivity when used as sensors, and a potentially higher power output when employed in biofuel cells. (author)

  16. Phase composition of murataite ceramics for excess weapons plutonium immobilization

    Science.gov (United States)

    Sobolev, I. A.; Stefanovsky, S. V.; Myasoedov, B. F.; Kullako, Y. M.; Yudintsev, S. V.

    2000-07-01

    Among the host phases for actinides immobilization, murataite (cubic, space group Fm3m) with the general formula A4B2C7O22-x (A=Ca, Mn, Na, Ln, An; B=Mn, Ti, Zr, AnIV; C=Ti, Al, Fe; 0ceramics in detail has shown occurrence of several murataite varieties with three-, five-, and eight-fold fluorite unit cells. [1-3] The goal of the present step of work is to study an effect of waste elements on phase composition of murataite ceramic and isomorphic capacity of waste elements.

  17. Uranium sorption by Pseudomonas biomass immobilized in radiation polymerized polyacrylamide bio-beads.

    Science.gov (United States)

    D'Souza, S F; Sar, Pinaki; Kazy, Sufia K; Kubal, B S

    2006-01-01

    A Pseudomonas strain identified as a potent biosorbent of uranium (U) and thorium was immobilized in radiation-induced polyacrylamide matrix for its application in radionuclide containing wastewater treatment. The immobilized biomass exhibited a high U sorption of 202 mg g(-1) dry wt. with its optimum at pH 5.0. A good fit of experimental data to the Freundlich model suggested multilayered uranium binding with an affinity distribution among biomass metal binding sites. Scanning electron microscopy revealed a highly porous nature of the radiation-polymerized beads with bacterial cells mostly entrapped on pore walls. Energy dispersive X-ray analysis (EDXA) coupled with SEM ascertained the accumulation of uranium by the immobilized biomass without any physical damage to the cells. A significant (90%) part of biosorbed uranium was recovered using sodium bicarbonate with the immobilized biomass maintaining their U resorption capacity for multiple sorption-desorption cycles. Uranium loading and elution behavior of immobilized biomass evaluated within a continuous up-flow packed bed columnar reactor showed its effectiveness in removing uranium from low concentration (50 mg U L(-1)) followed by its recovery resulting in a 4-5-fold waste volume reduction. The data suggested the suitability of radiation polymerization in obtaining bacterial beads for metal removal and also the potential of Pseudomonas biomass in treatment of radionuclide containing waste streams. PMID:16484078

  18. Polishing of POME by Chlorella sp. in suspended and immobilized system

    Science.gov (United States)

    Lahin, F. A.; Sarbatly, R.; Suali, E.

    2016-06-01

    The effect of using suspended and immobilized growth of Chlorella sp. to treat POME was studied. Cotton and nylon ropes were used as the immobilization material in a rotating microalgae biofilm reactor. The result showed that POME treated in suspended growth system was able to remove 81.9% and 55.5% of the total nitrogen (TN) and total phosphorus (TP) respectively. Whereas the immobilized system showed lower removal of 77.22% and 53.02% for TN and TP. Lower performance of immobilized microalgae is due to the limited light penetration and supply of CO2 inside the immobilization materials. The rotating microalgae biofilm reactor was able to reduce the biochemical oxygen demand (BOD) to 90 mg/L and chemical oxygen demand (COD) to 720 mg/L. Higher BOD and COD reading were obtained in suspended growth due to the presence of small number of microalgae cell in the samples. This study shows that suspended growth system is able to remove higher percentages of nitrogen and phosphorus. However, an efficient separation method such as membrane filtration is required to harvest the cultivated microalgae cell to avoid organic matter release into water bodies.

  19. Biodegradation of Crude Oil in Contaminated Soils by Free and Immobilized Microorganisms

    Institute of Scientific and Technical Information of China (English)

    WANG Zhen-Yu; XU Ying; WANG Hao-Yun; ZHAO Jian; GAO Dong-Mei; LI Feng-Min; B. XING

    2012-01-01

    The efficiencies of free and immobilized bacterial cultures of petroleum hydrocarbon degraders were evaluated and compared in this study.Hydrocarbon-degrading microbial communities with high tolerance to and high degrading ability of crude oil were obtained from the soil contaminated with crude oil in the Yellow River Delta.Then,the microbial ceils were immobilized in sodium alginate (SA) beads and sodium alginate-diatomite (SAD) beads.The biodegradation of crude oil in soil by immobilized cells was compared with that by free cells at three inocuIation concentrations,1 × 104 colony forming units (cfu) kg-1 (low concentration,L),5 × 104 cfu kg-1 (medium concentration,M),and 1 × 105 cfu kg -1 (high concentration,H).At 20 d after inoculation,the maximum degradation rate in the immobilized systems reached 29.8% (SAD-M),significantly higher (P < 0.05) than that of the free cells (21.1%),and the SAD beads showed greater degradation than the SA beads.Moreover,both microbial populations and total microbial activity reached significantly higher level (P < 0.05) in the immobilized systems than free cell systems at a same initial inoculation amount.The scanning electronic microscope (SEM) images also confirmed the advantages of the immobilized microstructure of SAD beads.The enhanced degradation and bacterial growth in the SAD beads indicated the high potential of SAD beads as an effective option for bioremediation of crude oil-contaminated soils in the Yellow River Delta.

  20. Characteristics of Immobilized Urease on Grafted Alginate Bead Systems

    Directory of Open Access Journals (Sweden)

    Enas N. Danial

    2015-04-01

    Full Text Available This study evaluated the biological importance of immobilized urease enzyme over the free urease. The support material used for urease immobilization was alginate. Generally, the immobilization of urease in alginate gel showed a marked increase in Km and Vmax. However, the immobilized urease showed higher thermal stability than that of free enzyme. The rate of thermal inactivation of the immobilized enzyme decreased due to entrapment in gel matrix. Also, the activity of the immobilized urease was more stable in retention than that of the free enzyme during the storage in solution, although the activity of the immobilized enzyme was lower in comparison with the free enzyme. A stable immobilized system and long storage life are convenient for applications that would not be feasible with a soluble enzyme system. These results highlighted the technical and biochemical benefits of immobilized urease over the free enzyme.

  1. Immobilized Lactobacillus acidophilus produced from whey and alginate

    Directory of Open Access Journals (Sweden)

    P. R. Rosa

    2013-06-01

    Full Text Available An analysis was made of the use of whey fermentation by Lactobacillus acidophilus LA-5 for encapsulated probiotic bacteria cell production. Fermentation was done in a 2-liter Biostat B Fermentor at 28±1 ºC without air supply and agitation maintained at 200 rpm. Different processing conditions were studied using Center Composite Design applied to Surface Response Methodology. Maximum cell yield (2.7 x10(10 NMP/mL for 36 hours was achieved with 30.85 g/L of lactose, a pH value of 6.45 and 1.04 g/L of inoculum. Cell growth was evaluated using reconstituted and fresh whey after 144 hours of fermentation in pre-optimized conditions. Cell concentration after fermentation was 10(10 MPN/mL in all the assays. The Verhulst model proved to be satisfactory to fit the experimental results, providing a stationary cell concentration of 6.0 g/L and a specific growth rate of 0.09 h-1. Cells were collected by centrifugation at 15000g for 5 minutes at 4 ºC, immobilized in 2% alginate, and dried to a constant weight at 50 ºC. Immobilized probiotic cells presented 10(11 MPN/g, a time required to kill 90% of the organisms (D value of 18 h (70 ºC, an activation energy of 76.04 kJ/mol for thermal inactivation, and an in vitro resistance to low pH (D value of 62.5 min at 37 ºC, pH 2.5.

  2. Excess Weapons Plutonium Immobilization in Russia

    Energy Technology Data Exchange (ETDEWEB)

    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&D 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

  3. Accumulation of uranium by immobilized persimmon tannin

    International Nuclear Information System (INIS)

    We have discovered that the extracted juice of unripe astringent persimmon fruit, designated as kakishibu or shibuol, has an extremely high affinity for uranium. To develop efficient adsorbents for uranium, we tried to immobilize kakishibu (persimmon tannin) with various aldehydes and mineral acids. Persimmon tannin immobilized with glutaraldehyde can accumulate 1.71 g (14 mEq U) of uranium per gram of the adsorbent. The uranium accumulating capacity of this adsorbent is several times greater than that of commercially available chelating resins (2-3 mEq/g). Immobilized persimmon tannin has the most favorable features for uranium recovery; high selective adsorption ability, rapid adsorption rate, and applicability in both column and batch systems. The uranium retained on immobilized persimmon tannin can be quantitatively and easily eluted with a very dilute acid, and the adsorbent can thus be easily recycled in the adsorption-desorption process. Immobilized persimmon tannin also has a high affinity for thorium. 23 refs., 13 figs., 7 tabs

  4. Immobilization of Methyltrioxorhenium on Mesoporous Aluminosilicate Materials

    Directory of Open Access Journals (Sweden)

    Martina Stekrova

    2014-03-01

    Full Text Available The presented report focuses on an in-depth detailed characterization of immobilized methyltrioxorhenium (MTO, giving catalysts with a wide spectra of utilization. The range of mesoporous materials with different SiO2/Al2O3 ratios, namely mesoporous alumina (MA, aluminosilicates type Siral (with Al content 60%–90% and MCM-41, were used as supports for immobilization of MTO. The tested support materials (aluminous/siliceous exhibited high surface area, well-defined regular structure and narrow pore size distribution of mesopores, and therefore represent excellent supports for the active components. Some of the supports were modified by zinc chloride in order to obtain catalysts with higher activities for instance in metathesis reactions. The immobilization of MTO was optimized using these supports and it was successful using all supports. The success of the immobilization of MTO and the properties of the prepared heterogeneous catalysts were characterized using X-ray Fluorescence (XRF, atomic absorption spectroscopy (AAS, X-ray powder diffraction (XRD, scanning electron microscopy (SEM, physical adsorption of N2, ultraviolet-visible spectroscopy (UV-Vis, infrared spectroscopy (FTIR, Fourier Transform Infrared Spectroscopy (FTIR using pyridine as a probe molecule and X-ray photoelectron spectroscopy (XPS. Furthermore, the catalytic activity of the immobilized MTO on the tested supports was demonstrated on metathesis reactions of various substrates.

  5. Plasmid stability in immobilized and free recombinant Escherichia coli JM105(pKK223-200): importance of oxygen diffusion, growth rate, and plasmid copy number.

    OpenAIRE

    de Taxis du Poët, P; Arcand, Y; Bernier, R.; Barbotin, J N; Thomas, D.

    1987-01-01

    Stability of the plasmid pKK223-200 in Escherichia coli JM105 was studied for both free and immobilized cells during continuous culture. The relationship between plasmid copy number, xylanase activity, which was coded for by the plasmid, and growth rate and culture conditions involved complex interactions which determined the plasmid stability. Generally, the plasmid stability was enhanced in cultured immobilized cells compared with free-cell cultures. This stability was associated with modif...

  6. An Efficient, Recyclable, and Stable Immobilized Biocatalyst Based on Bioinspired Microcapsules-in-Hydrogel Scaffolds.

    Science.gov (United States)

    Zhang, Shaohua; Jiang, Zhongyi; Shi, Jiafu; Wang, Xueyan; Han, Pingping; Qian, Weilun

    2016-09-28

    Design and preparation of high-performance immobilized biocatalysts with exquisite structures and elucidation of their profound structure-performance relationship are highly desired for green and sustainable biotransformation processes. Learning from nature has been recognized as a shortcut to achieve such an impressive goal. Loose connective tissue, which is composed of hierarchically organized cells by extracellular matrix (ECM) and is recognized as an efficient catalytic system to ensure the ordered proceeding of metabolism, may offer an ideal prototype for preparing immobilized biocatalysts with high catalytic activity, recyclability, and stability. Inspired by the hierarchical structure of loose connective tissue, we prepared an immobilized biocatalyst enabled by microcapsules-in-hydrogel (MCH) scaffolds via biomimetic mineralization in agarose hydrogel. In brief, the in situ synthesized hybrid microcapsules encapsulated with glucose oxidase (GOD) are hierarchically organized by the fibrous framework of agarose hydrogel, where the fibers are intercalated into the capsule wall. The as-prepared immobilized biocatalyst shows structure-dependent catalytic performance. The porous hydrogel permits free diffusion of glucose molecules (diffusion coefficient: ∼6 × 10(-6) cm(2) s(-1), close to that in water) and retains the enzyme activity as much as possible after immobilization (initial reaction rate: 1.5 × 10(-2) mM min(-1)). The monolithic macroscale of agarose hydrogel facilitates the easy recycling of the immobilized biocatalyst (only by using tweezers), which contributes to the nonactivity decline during the recycling test. The fiber-intercalating structure elevates the mechanical stability of the in situ synthesized hybrid microcapsules, which inhibits the leaching and enhances the stability of the encapsulated GOD, achieving immobilization efficiency of ∼95%. This study will, therefore, provide a generic method for the hierarchical organization of (bio

  7. Fiber optic biosensor of immobilized firefly luciferase

    Institute of Scientific and Technical Information of China (English)

    蔡谨; 孟文芳; 吉鑫松

    2002-01-01

    Luciferase from firefly lantern extract was immobilized on CNBr-activated Sepharose 4B. The kinetic properties of immobilized luciferase were extensively studied. The Km′ for D-luciferin is 11.9 μmol/L, the optimum pH and temperature for Sepharose-bound enzyme were 7.8 and 25℃ respectively. A luminescence fiber optic biosensor, making use of immobilized crude luciferase, was developed for assay of ATP. The peak light intensity was linear with respect to ATP concentration in range of 10-9-10-5 mol/L. A biological application was also demonstrated with the determination of serum ATP from rats bred in low versus normal oxygen environments.

  8. Fiber optic biosensor of immobilized firefly luciferase

    Institute of Scientific and Technical Information of China (English)

    蔡谨; 吉鑫松; 等

    2002-01-01

    Luciferase from firefly lantern extract was immobilized on CNBr-activated Sepharose 4B,The kinetic properties of immobilized luciferase were extensively studied.The Km' for D-luciferin is 11.9umol/L,the optimum pH and temperature for Sepharose-bound enzyme were 7.8 and 25℃ respectively.A luminescence fiber optic biosensor,making use of immobilized crude luciferase was developed for assay of ATP.The peak light intensity was linear with respect to ATP concentration in range of 10-9-10-5mol/L.A biological application was also demonstrated with the determination of serum ATP from rats bred in low versus normal oxygen environments.

  9. [Water binding of adsorptive immobilized lipases].

    Science.gov (United States)

    Loose, S; Meusel, D; Muschter, A; Ruthe, B

    1990-01-01

    It is supposed that not only the total water content of lipase preparations but more their state of water binding is of technological importance in enzymatic interesterification reactions in systems nearly free from water. The isotherms at 65 degrees C of two microbial lipases immobilized on various adsorbents as well as different adsorbents themselves are shown. The water binding capacity in the range of water content of technological interest decreases from the anion exchange resin Amberlyst A 21 via nonpolar adsorbent Amberlite XAD-2 to kieselguhr Celite 545. It is demonstrated that water binding by lipases is depending on temperature but is also affected by adsorptive immobilization. Adsorptive immobilized lipases show hysteresis, which is very important for preparing a definite water content of the enzyme preparations. PMID:2325750

  10. [Use of immobilization in the study of glyceraldehyde 3-phosphate dehydrogenase. Immobilized monomers].

    Science.gov (United States)

    Muronets, V I; Ashmarina, L I; Asriiants, R A; Nagradova, N K

    1982-06-01

    Active immobilized monomers of glyceraldehyde 3-phosphate dehydrogenase were prepared by means of dissociation of the tetrameric enzyme molecule covalently bound to Sepharose via a single subunit. The conditions were elaborated to achieve the inactivation and solubilization of the non-covalently bound subunits leaving the monomer coupled to the matrix intact. This procedure differs from the previously developed method of matrix-bound oligomeric enzymes dissociation in a detail which was found to be essentially important. The widely used method includes complete denaturation of all subunits during treatment with urea followed by reactivation of the immobilized one, whereas only the non-covalently bound subunits suffer denaturation under the conditions developed in the present work. The immobilized monomers of glyceraldehyde 3-phosphate dehydrogenase exhibit Vmax and Km (for NAD and substrate) values similar to those found for the immobilized tetramer. Reassociation of the immobilized monomers with soluble enzyme subunits obtained in the presence of urea produces matrix-bound tetrameric species. Immobilized trimers ae formed upon incubation of matrix-bound monomers in a diluted apoenzyme solution. The immobilized monomeric, trimeric and tetrameric enzyme species were used to study the role of subunit interactions in cooperative phenomena exhibited by the dehydrogenase. PMID:7115810

  11. Selection of support materials for immobilization of Burkholderia cepacia PCL3 in treatment of carbofuran-contaminated water.

    Science.gov (United States)

    Laocharoen, S; Plangklang, P; Reungsang, A

    2013-01-01

    This study investigated the utilization of agricultural matrices as the support materials for cell immobilization to improve the technique of bioremediation. Coir, bulrush, banana stem and water hyacinth stem in both delignified and undelignified forms were used to immobilize Burkholderia cepacia PCL3 in bioremediation of carbofuran at 5 mg l(-1) in synthetic wastewater. Undelignified coir was found to be the most suitable support material for cell immobilization, giving the short half-life of carbofuran of 3.40 d (2.8 times shorter than the treatments with free cells). In addition, it could be reused three times without a loss in ability to degrade carbofuran. The growth and degradation ability of free cells were completely inhibited at the initial carbofuran concentrations of 250 mg l(-1), while there was no inhibitory effect of carbofuran on the immobilized cells. The results indicated a great potential for using the agricultural matrices as support material for cell immobilization to improve the overall efficiency of carbofuran bioremediation in contaminated water by B. cepacia PCL3.

  12. Selection of support materials for immobilization of Burkholderia cepacia PCL3 in treatment of carbofuran-contaminated water.

    Science.gov (United States)

    Laocharoen, S; Plangklang, P; Reungsang, A

    2013-01-01

    This study investigated the utilization of agricultural matrices as the support materials for cell immobilization to improve the technique of bioremediation. Coir, bulrush, banana stem and water hyacinth stem in both delignified and undelignified forms were used to immobilize Burkholderia cepacia PCL3 in bioremediation of carbofuran at 5 mg l(-1) in synthetic wastewater. Undelignified coir was found to be the most suitable support material for cell immobilization, giving the short half-life of carbofuran of 3.40 d (2.8 times shorter than the treatments with free cells). In addition, it could be reused three times without a loss in ability to degrade carbofuran. The growth and degradation ability of free cells were completely inhibited at the initial carbofuran concentrations of 250 mg l(-1), while there was no inhibitory effect of carbofuran on the immobilized cells. The results indicated a great potential for using the agricultural matrices as support material for cell immobilization to improve the overall efficiency of carbofuran bioremediation in contaminated water by B. cepacia PCL3. PMID:24527620

  13. Decolorization and partial mineralization of a polyazo dye by Bacillus firmus immobilized within tubular polymeric gel

    OpenAIRE

    Ogugbue, Chimezie Jason; Morad, Norhashimah; Sawidis, Thomas; Oranusi, Nathaniel A.

    2011-01-01

    The degradation of C.I. Direct red 80, a polyazo dye, was investigated using Bacillus firmus immobilized by entrapment in tubular polymeric gel. This bacterial strain was able to completely decolorize 50 mg/L of C.I. Direct red 80 under anoxic conditions within 12 h and also degrade the reaction intermediates (aromatic amines) during the subsequent 12 h under aerobic conditions. The tubular gel harboring the immobilized cells consisted of anoxic and aerobic regions integrated in a single unit...

  14. Immobilized reactor for rapid destruction of recalcitrant organics and inorganics in tannery wastewater

    Institute of Scientific and Technical Information of China (English)

    A. Ganesh Kumar; G. Sekaran; S. Swarnalatha; B. Prasad Rao

    2005-01-01

    The wastewater discharged from tanneries lack biodegradability due to the presence of recalcitrant compounds at significant concentration. The focal theme of the present investigation was to use chemo-autotrophic activated carbon oxidation(CAACO) reactor, an immobilized cell reactor using chemoautotrophs for the treatment of tannery wastewater. The treatment scheme comprised of anaerobic treatment, sand filtration, and CAACO reactor, which remove COD, BOD, TOC, VFA and sulphides respectively by 86%, 95%, 81%,71% and 100%. Rice bran mesoporous activated carbon prepared indigenously and was used for immobilization of chemoautotrophs. The degradation of xenobiotic compounds by CAACO was confirmed through HPLC and FT-IR techniques.

  15. Immobilization of spent resin with epoxy resin

    International Nuclear Information System (INIS)

    immobilization of spent resin using epoxy resin has been conducted. The spent resin was mixtured with epoxy resin in variation of concentration, i.e., 30, 40, 50, 60, 70 weight percent of spent resin. The mixture were pour into the plastic tube, with a diameter of 40 mm and height of 40 mm. The density, compressive strength and leaching rate were respectively measured by quanta chrome, paul weber apparatus and gamma spectrometer. The results showed that the increasing of waste concentration would be decreased the compressive strength, and increased density by immobilized waste. The leaching rate of 137Cs from waste product was not detected in experiment (author)

  16. Radiation Synthesis of Nanogel for Bioactives Immobilization

    International Nuclear Information System (INIS)

    Both hydrophilic and hydrophobic core nanogel are currently being developed for immobilization and delivery purposes in Malaysian Nuclear Agency. Hydrophilic nanogel is produced by using inverse micelles irradiation of polyethelyne glycol diacrylate (PEGDA). The hydrophobic nanogel is produced via irradiation of acrylated form of palm oil. These nanogels will be used to immobilize bio actives such as curcumin, tyhmoquinone, oryzanol and chitosan. Preliminary investigation of the nanogel size using dynamic light scattering (DLS) shows that nanogel with sizes below 100nm can be obtained. (author)

  17. Use of PVA-gel immobilized cells: a new strategy for biotechnological production of Xylitol from sugarcane bagasse hidrolysate/ Uso de células imobilizadas em gel de PVA: uma nova estratégia para produção biotecnológica de Xilitol a partir de bagaço de cana-de-açúcar

    Directory of Open Access Journals (Sweden)

    Júlio César dos Santos

    2005-06-01

    Full Text Available Sugarcane bagasse is one of the most abundant residues in Brazil due to the large number of sugaralcohol industries. This biomass contains a high concentration of carbohydrates, which can be converted into products of high economic value, such as xylitol. Xylitol, a polyol with anticariogenic properties, is similar in sweetening power to sucrose, and has high potential for use in the food and pharmaceutical industries. Several studies have been carried out to produce xylitol by biotechnological processes. However, there is little information on the use of immobilized cells in these bioprocesses. The objective of this review was to present a new possibility to produce xylitol by biotechnological processes, using sugarcane bagasse hydrolysate and immobilized cells in PVA-gel.O bagaço de cana-de-açúcar é um dos resíduos mais abundantes no Brasil devido ao grande número de indústrias sucroalcooleiras. Esta biomassa contém elevado teor de carboidratos, podendo ser utilizada na produção de compostos de interesse econômico como o xilitol. O xilitol é um poliol de cinco carbonos que apresenta poder adoçante semelhante ao da sacarose e propriedades anti-cariogênicas, tendo elevado potencial de uso nas indústrias alimentícias e farmacêuticas. Diversos estudos buscando o desenvolvimento de processos de produção de xilitol por via biotecnológica têm sido realizados, entretanto pouco tem sido escrito sobre a utilização de células imobilizadas no bioprocesso. A presente revisão tem como objetivo apresentar uma possibilidade de produção de xilitol a partir de hidrolisado hemicelulósico de bagaço de canade-açúcar, em sistema com células imobilizadas em gel de álcool polivinílico.

  18. Intensified nitrogen removal in immobilized nitrifier enhanced constructed wetlands with external carbon addition.

    Science.gov (United States)

    Wang, Wei; Ding, Yi; Wang, Yuhui; Song, Xinshan; Ambrose, Richard F; Ullman, Jeffrey L

    2016-10-01

    Nitrogen removal performance response of twelve constructed wetlands (CWs) to immobilized nitrifier pellets and different influent COD/N ratios (chemical oxygen demand: total nitrogen in influent) were investigated via 7-month experiments. Nitrifier was immobilized on a carrier pellet containing 10% polyvinyl alcohol (PVA), 2.0% sodium alginate (SA) and 2.0% calcium chloride (CaCl2). A batch experiment demonstrated that 73% COD and 85% ammonia nitrogen (NH4-N) were degraded using the pellets with immobilized nitrifier cells. In addition, different carbon source supplement strategies were applied to remove the nitrate (NO3-N) transformed from NH4-N. An increase in COD/N ratio led to increasing reduction in NO3-N. Efficient nitrification and denitrification promoted total nitrogen (TN) removal in immobilized nitrifier biofortified constructed wetlands (INB-CWs). The results suggested that immobilized nitrifier pellets combined with high influent COD/N ratios could effectively improve the nitrogen removal performance in CWs. PMID:27396293

  19. Nitrification performance of nitrifying bacteria immobilized in waterborne polyurethane at low ammonia nitrogen concentrations

    Institute of Scientific and Technical Information of China (English)

    Yamei Dong; Zhenjia Zhang; Yongwei Jin; Zhirong Li; Jian Lu

    2011-01-01

    Suspended and waterborne polyurethane immobilized nitrifying bacteria have been adopted for evaluating the effects of environmental changes, such as temperature, dissolved oxygen (DO) concentration and pH, on nitrification characteristics under conditions of low ammonia concentrations.The results showed that nitrification was prone to complete with increasing pH, DO and temperature.Sensitivity analysis demonstrated the effects of temperature and pH on nitrification feature of suspended bacteria were slightly greater than those of immobilized nitrifying bacteria.Immobilized cells could achieve complete nitrification at low ammonia concentrations when DO was sufficient.Continuous experiments were carried out to discuss the removal of ammonia nitrogen from synthetic micropollute source water with the ammonia concentration of about 1 mg/L using immobilized nitrifying bacteria pellets in an up-flow inner circulation reactor under different hydraulic retention times (HRT).The continuous removal rate remains above 80% even under HRT 30 min.The results verified that the waterborne polyurethane immobilized nitrifying bacteria pellets had great potential applications for micro-pollution source water treatment.

  20. Improved immobilization of laccase on a glassy carbon electrode by oriented covalent attachment

    Directory of Open Access Journals (Sweden)

    Liu Xin

    2014-01-01

    Full Text Available A laccase from Thermus thermophilus HB27 was reported to be potentially useful in the design of a temperature controlled biofuel cell. For enhancing its application in different thermal conditions, we engineered a laccase-oriented immobilized electrode. A site-directed mutant N323C of the laccase was constructed. A photometric assay was employed in order to compare the catalytic properties of wild-type laccase and mutant. The mutant was attached to a glass carbon electrode by covalent cross-linking. The electrochemical properties of the immobilized laccase were investigated by cyclic voltammetry. This immobilization allowed the active electrode to function at temperatures up to 95°C. The thermal and pH dependence profiles were similar to those of the soluble enzyme investigated by spectrophotometry.

  1. Kinetic effects on signal normalization in oligonucleotide microchips with labeled immobilized probes.

    Science.gov (United States)

    Pan'kov, S V; Chechetkin, V R; Somova, O G; Antonova, O V; Moiseeva, O V; Prokopenko, D V; Yurasov, R A; Gryadunov, D A; Chudinov, A V

    2009-10-01

    Among various factors affecting operation of oligonucleotide microchips, the variations in concentration and in homogeneous distribution of immobilized probes over the cells are one of the most important. The labeling of immobilized probes ensures the complete current monitoring on the probe distribution and is reliable and convenient. Using hydrogel-based oligonucleotide microchips, the applicability of Cy3-labeled immobilized probes for quality control and signal normalization after hybridization with Cy5-labeled target DNA was investigated. This study showed that proper signal normalization should be different in thermodynamic conditions and in transient regime with hybridization far from saturation. This kinetic effect holds for both hydrogel-based and surface oligonucleotide microchips. Besides proving basic features, the technique was assessed on a sampling batch of 50 microchips developed for identifying mutations responsible for rifampicin and isoniazid resistance of Mycobacterium tuberculosis.

  2. Immobilization and direct electrochemistry of copper-containing enzymes on active carbon

    Institute of Scientific and Technical Information of China (English)

    SUN Dongmei; CAI Chenxin; XING Wei; LU Tianhong

    2004-01-01

    Two typical and important copper-containing enzymes, laccase (Lac) and tyrosinase (Tyr), have been immobilized on the surface of active carbon with simple adsorption method. The cyclic voltammetric results indicated that the active carbon could promote the direct electron transfer of both Lac and Tyr and a pair of well-defined and nearly symmetric redox peaks appeared on the cyclic voltammograms of Lac or Tyr with the formal potential, E0′, independent on the scan rate. The further experimental results showed that the immobilized copper-containing oxidase displayed an excellent electrocatalytic activity to the electrochemical reduction of O2. The immobilization method presented here has several advantages, such as simplicity, easy to operation and keeping good activity of enzyme etc., and could be further used to study the direct electrochemistry of other redox proteins and enzymes and fabricate the catalysts for biofuel cell.

  3. Element Partitioning in Glass-Ceramic Designed for Actinides Immobilization

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    <正>Glass-ceramics were designed for immobilization of actinides. In order to immobilizing more wastes in the matrix and to develop the optimum formulation for the glass-ceramic, it is necessary to study the

  4. Preparation of Laccase Immobilized Cryogels and Usage for Decolorization

    Directory of Open Access Journals (Sweden)

    Murat Uygun

    2013-01-01

    Full Text Available Poly(methyl methacrylate-co-glycidyl methacrylate (poly(MMA-co-GMA cryogels were synthesized by radical cryopolymerization technique. Then, laccase enzyme was covalently attached to the cryogel and characterized by using swelling studies and SEM and EDX analyses. Kinetic properties and optimum conditions of the immobilized and free laccase were studied and it was found that of the immobilized laccase was lower than that of free laccase. of the immobilized laccase was increased upon immobilization. Optimum pH was found to be 4.0 for each type of laccase, while optimum temperature was shifted to the warmer region after the immobilization. It was also found that thermal stability of the immobilized laccase was higher than that of free laccase. Immobilized laccase could be used for 10 times successive reuse with no significant decrease in its activity. Also, these laccase immobilized cryogels were successfully used for the decolorization of seven different dyes.

  5. Limb immobilization induces a coordinate down-regulation of mitochondrial and other metabolic pathways in men and women.

    Directory of Open Access Journals (Sweden)

    Arkan Abadi

    Full Text Available Advancements in animal models and cell culture techniques have been invaluable in the elucidation of the molecular mechanisms that regulate muscle atrophy. However, few studies have examined muscle atrophy in humans using modern experimental techniques. The purpose of this study was to examine changes in global gene transcription during immobilization-induced muscle atrophy in humans and then explore the effects of the most prominent transcriptional alterations on protein expression and function. Healthy men and women (N = 24 were subjected to two weeks of unilateral limb immobilization, with muscle biopsies obtained before, after 48 hours (48 H and 14 days (14 D of immobilization. Muscle cross sectional area (approximately 5% and strength (10-20% were significantly reduced in men and women (approximately 5% and 10-20%, respectively after 14 D of immobilization. Micro-array analyses of total RNA extracted from biopsy samples at 48 H and 14 D uncovered 575 and 3,128 probes, respectively, which were significantly altered during immobilization. As a group, genes involved in mitochondrial bioenergetics and carbohydrate metabolism were predominant features at both 48 H and 14 D, with genes involved in protein synthesis and degradation significantly down-regulated and up-regulated, respectively, at 14 D of muscle atrophy. There was also a significant decrease in the protein content of mitochondrial cytochrome c oxidase, and the enzyme activity of cytochrome c oxidase and citrate synthase after 14 D of immobilization. Furthermore, protein ubiquitination was significantly increased at 48 H but not 14 D of immobilization. These results suggest that transcriptional and post-transcriptional suppression of mitochondrial processes is sustained throughout 14 D of immobilization, while protein ubiquitination plays an early but transient role in muscle atrophy following short-term immobilization in humans.

  6. Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae)

    OpenAIRE

    Seniwati

    2012-01-01

    Enzyme immobilization is a recovery technique that has been studied in several years, using support as a media to help enzyme dissolutions to the reaction substrate. Immobilization method used in this study was adsorption method, using specific lipase from Aspergillus oryzae. Lipase was partially purified from the culture supernatant of Aspergillus oryzae. Enzyme was immobilized by adsorbed on silica gel. Studies on free and immobilized lipase systems for determination of op...

  7. Immobilization of Isolated Lipase From Moldy Copra (Aspergillus Oryzae)

    OpenAIRE

    Seniwati Dali; A. B. D. Rauf Patong; M.Noor Jalaluddin; Pirman; Baharuddin Hamzah

    2011-01-01

    Enzyme immobilization is a recovery technique that has been studied in several years, using support as a media to help enzyme dissolutions to the reaction substrate. Immobilization method used in this study was adsorption method, using specific lipase from Aspergillus oryzae. Lipase was partially purified from the culture supernatant of Aspergillus oryzae. Enzyme was immobilized by adsorbed on silica gel. Studies on free and immobilized lipase systems for determination of optimum pH, optimum ...

  8. Kinetic studies on degradation of Reactive Red 120 dye in immobilized packed bed reactor by Bacillus cohnii RAPT1.

    Science.gov (United States)

    Padmanaban, V C; Geed, Sachin RameshRao; Achary, Anant; Singh, R S

    2016-08-01

    The degradation of Reactive Red 120 using Bacillus cohnii RAPT1 immobilized on polyurethane was studied. Initial experiments indicated that the percentage removal of dye in immobilized batch was significantly higher than batch (without immobilization). The optimum process parameters such as effect of dye concentration, time of immobilization on Poly Urethane Foam, initial inoculum size, pH and temperature for removal of dye were investigated and was found as 200ppm, 36h, 300*10(6) colony forming units/ml, 8.0 and 35°C respectively. Under optimum conditions, 100% removal of dye was obtained within 4h. The kinetics of biodegradation for the batch with free cells and immobilised packed batch was found to be IInd order with kinetic constant and initial rate of reaction as 0.0408, 0.084L/(mgday) and 1632, 3360 (mg/Lday) respectively. PMID:26968121

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

    Science.gov (United States)

    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. PMID:27211619

  10. Surface engineering of Ti-O films by photochemical immobilization of gelatin

    International Nuclear Information System (INIS)

    Crystalline Ti-O films were prepared by unbalanced magnetron sputtering and the structure was confirmed by XRD. An organic layer of 3-aminopropylphosphonic acid (APP) was first introduced on the Ti-O films by self-assembling. The stability of the APP on Ti-O films was confirmed by XPS and FTIR analysis. Simultaneously, azido group was introduced in gelatin molecule to act as photoreactive point. The derivated gelatin was spin-coated onto the self-assembled layer and immobilized by UV irradiating. Chemical patterned surface was obtained by using a photomask when irradiating and confirmed by sirius red staining and surface profile analysis. Measured by surface profilometer, the thickness of the immobilized gelatin was about 5-20 nm. The adhering of human endothelial EVC304 cells on APP modified surface was enhanced in the cell culture test. Moreover, the adherence and growth of cells were prior on gelatin-immobilized region visually seen on the patterned surface. This result indicated gelatin-immobilized Ti-O surface can serve as a biocompatible biomaterial for endothelialization

  11. Surface engineering of Ti-O films by photochemical immobilization of gelatin

    Energy Technology Data Exchange (ETDEWEB)

    Weng, Y.J.; Ren, J.R. [Department of Materials Engineering, Southwest Jiaotong University, Chengdu 610031 (China); Key Lab for Advanced Technologies of Materials, Ministry of Education, Southwest Jiaotong University, Chengdu, 610031 (China); Huang, N. [Department of Materials Engineering, Southwest Jiaotong University, Chengdu 610031 (China) and Key Lab for Advanced Technologies of Materials, Ministry of Education, Southwest Jiaotong University, Chengdu, 610031 (China)], E-mail: nhuang@263.net; Wang, J.; Chen, J.Y.; Leng, Y.X.; Liu, H.Q. [Department of Materials Engineering, Southwest Jiaotong University, Chengdu 610031 (China); Key Lab for Advanced Technologies of Materials, Ministry of Education, Southwest Jiaotong University, Chengdu, 610031 (China)

    2008-12-01

    Crystalline Ti-O films were prepared by unbalanced magnetron sputtering and the structure was confirmed by XRD. An organic layer of 3-aminopropylphosphonic acid (APP) was first introduced on the Ti-O films by self-assembling. The stability of the APP on Ti-O films was confirmed by XPS and FTIR analysis. Simultaneously, azido group was introduced in gelatin molecule to act as photoreactive point. The derivated gelatin was spin-coated onto the self-assembled layer and immobilized by UV irradiating. Chemical patterned surface was obtained by using a photomask when irradiating and confirmed by sirius red staining and surface profile analysis. Measured by surface profilometer, the thickness of the immobilized gelatin was about 5-20 nm. The adhering of human endothelial EVC304 cells on APP modified surface was enhanced in the cell culture test. Moreover, the adherence and growth of cells were prior on gelatin-immobilized region visually seen on the patterned surface. This result indicated gelatin-immobilized Ti-O surface can serve as a biocompatible biomaterial for endothelialization.

  12. Immobilized Ruthenium Catalyst for Carbon Dioxide Hydrogenation

    Institute of Scientific and Technical Information of China (English)

    Ying Min YU; Jin Hua FEI; Yi Ping ZHANG; Xiao Ming ZHENG

    2006-01-01

    Three kinds of cross linked polystyrene resin (PS) supported ruthenium complexes were developed as catalysts for the synthesis of formic acid from carbon dioxide hydrogenation. Many factors, such as the functionalized supports, solvents and ligands, could influence their activities and reuse performances greatly. These immobilized catalysts also offer the industrial advantages such as easy separation.

  13. Plutonium Immobilization Can Loading Preliminary Specifications

    Energy Technology Data Exchange (ETDEWEB)

    Kriikku, E.

    1998-11-25

    This report discusses the Plutonium Immobilization can loading preliminary equipment specifications and includes a process block diagram, process description, equipment list, preliminary equipment specifications, plan and elevation sketches, and some commercial catalogs. This report identifies loading pucks into cans and backfilling cans with helium as the top priority can loading development areas.

  14. Immobilization of horseradish peroxidase onto kaolin.

    Science.gov (United States)

    Šekuljica, Nataša Ž; Prlainović, Nevena Ž; Jovanović, Jelena R; Stefanović, Andrea B; Djokić, Veljko R; Mijin, Dušan Ž; Knežević-Jugović, Zorica D

    2016-03-01

    Kaolin showed as a very perspective carrier for the enzyme immobilization and it was used for the adsorption of horseradish peroxidase (HRP). The effects of the enzyme concentration and pH on the immobilization efficiency were studied in the reaction with pyrogallol and anthraquinone dye C.I. Acid Violet 109 (AV 109). In addition, Fourier transform infrared spectroscopy, scanning electron microscopy and analysis by Brunauer-Emmett-Teller were performed for kaolin, thermally activated kaolin and the immobilized enzyme. It has been shown that 0.1 IU of HRP-kaolin decolorized 87 % of dye solution, under the optimal conditions (pH 5.0, temperature 24 °C, dye concentration 40 mg/L and 0.2 mM of H2O2) within 40 min. The immobilized HRP decolorization follows the Ping Pong Bi-Bi mechanism with dead-end inhibition by the dye. The biocatalyst retained 35 ± 0.9 % of the initial activity after seven cycles of reuse in the decolorization reaction of AV 109 under optimal conditions in a batch reactor. The obtained kinetic parameters and reusability study confirmed improvement in performances of k-HRP compared to free, indicating that k-HRP has a great potential for environmental purposes. PMID:26747440

  15. Immobilizing Biomolecules Near the Diffraction Limit

    DEFF Research Database (Denmark)

    Skovsen, Esben; Petersen, Maria Teresa Neves; Gennaro, Ane Kold Di;

    2009-01-01

    Our group has previously shown that biomolecules containing disulfide bridges in close proximity to aromatic residues can be immobilized, through covalent bonds, onto thiol derivatized surfaces upon UV excitation of the aromatic residue(s). We have also previously shown that our new technology ca...

  16. Application of radiopolymerization for immobilization of enzymes

    International Nuclear Information System (INIS)

    Hydrophilic glass-forming monomers were used in an application of irradiation technology for the immobilization of cellulase and cellobiase. Experiments to observe the effect of additives such as silicates and polyethylene glycol in the enzyme entrapment are reported on. In all cases, enzymatic activity was maintained for more than fifteen batch enzyme reactions. (Author)

  17. Silanization and antibody immobilization on SU-8

    Science.gov (United States)

    Joshi, Manoj; Pinto, Richard; Rao, V. Ramgopal; Mukherji, Soumyo

    2007-01-01

    SU-8, an epoxy based negative photoresist, has emerged as a structural material for microfabricated sensors due to its attractive mechanical properties like low Young's modulus and chemical properties like inertness to various chemicals used in microfabrication. It can be used to fabricate MEMS structures of high aspect ratio. However, the use of SU-8 in BioMEMS application has been limited by the fact that immobilization of biomolecules on SU-8 surfaces has not been reported. In this study, the epoxy groups on the SU-8 surface were hydrolyzed in the presence of sulphochromic solution. Following this, the surface was treated with [3-(2-aminoethyl) aminopropyl]-trimethoxysilane (AEAPS). The silanized SU-8 surface was used to incubate human immunoglobulin (HIgG). The immobilization of HIgG was proved by allowing FITC tagged goat anti-human IgG to react with HIgG. This process of antibody immobilization was used to immobilize HIgG on microfabricated SU-8 cantilevers.

  18. Phosphopeptide enrichment by immobilized metal affinity chromatography

    DEFF Research Database (Denmark)

    Thingholm, Tine E.; Larsen, Martin R.

    2016-01-01

    Immobilized metal affinity chromatography (IMAC) has been the method of choice for phosphopeptide enrichment prior to mass spectrometric analysis for many years and it is still used extensively in many laboratories. Using the affinity of negatively charged phosphate groups towards positively...

  19. Covalently immobilized gelatin gradients within three-dimensional porous scaffolds

    Institute of Scientific and Technical Information of China (English)

    WU JinDan; TAN HuaPing; LI LinHui; GAO ChangYou

    2009-01-01

    A stable gelatin gradient providing continuous increment of signaling for cell adhesion and proliferation was fabricated within 3D poly(L-lactic acid) (PLLA) scaffolds. The porous PLLA scaffold fabricated by NaCI particle leaching was vertically fixed on a glass vial. 1,6-Hexanediamine/propanol solution was continuously injected into the vial by a micropump to aminolyze the PLLA scaffold. As a result of reaction time difference,the introduced-NH2 groups increased continuously along with the longitude of the PLLA scaffold in the z-direction. After covalent immobilization of gelatin by glutaraldehyde coupling,the gelatin gradient scaffold was thus obtained. In vitro chondrocyte culture showed that the cells had higher viability and more extending morphology in the gelatin gradient scaffold than that in the uniform gelatin control.

  20. Immobilization of Active Bacteriophages on Polyhydroxyalkanoate Surfaces.

    Science.gov (United States)

    Wang, Chanchan; Sauvageau, Dominic; Elias, Anastasia

    2016-01-20

    A rapid, efficient technique for the attachment of bacteriophages (phages) onto polyhydroxyalkanoate (PHA) surfaces has been developed and compared to three reported methods for phage immobilization. Polymer surfaces were modified to facilitate phage attachment using (1) plasma treatment alone, (2) plasma treatment followed by activation by 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS), (3) plasma-initiated acrylic acid grafting, or (4) plasma-initiated acrylic acid grafting with activation by EDC and sulfo-NHS. The impact of each method on the surface chemistry of PHA was investigated using contact angle analysis and X-ray photoelectron spectroscopy. Each of the four treatments was shown to result in both increased hydrophilicity and in the modification of the surface functional groups. Modified surfaces were immersed in suspensions of phage T4 for immobilization. The highest level of phage binding was observed for the surfaces modified by plasma treatment alone. The change in chemical bond states observed for surfaces that underwent plasma treatment is suspected to be the cause of the increased binding of active phages. Plasma-treated surfaces were further analyzed through phage-staining and fluorescence microscopy to assess the surface density of immobilized phages and their capacity to capture hosts. The infective capability of attached phages was confirmed by exposing the phage-immobilized surfaces to the host bacteria Escherichia coli in both plaque and infection dynamic assays. Plasma-treated surfaces with immobilized phages displayed higher infectivity than surfaces treated with other methods; in fact, the equivalent initial multiplicity of infection was 2 orders of magnitude greater than with other methods. Control samples - prepared by immersing polymer surfaces in phage suspensions (without prior plasma treatment) - did not show any bacterial growth inhibition, suggesting they did not bind

  1. Ethanol fermentation of mahula (Madhuca latifolia L.) flowers using free and immobilized yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Swain, M R; Kar, S; Sahoo, A K; Ray, R C

    2007-01-01

    There is a growing interest to find alternate bioresources for production of ethanol, apart from cane/sugar beet molasses and starchy crops like sweet sorghum, cassava and sweet potato. Mahula (Madhuca latifolia L.) is a forest tree abundantly available in the Indian subcontinent and its flowers are very rich in fermentable sugars (28.1-36.3 g 100 g(-1)). Batch fermentation of fresh and 12-month-stored flowers with free (whole cells) and immobilized cells of Saccharomyces cerevisiae (strain CTCRI) was carried out in 2-l Erlenmeyer flasks. The ethanol yields were 193 and 148 g kg(-1) (using free cells) and 205 and 152 g kg(-1) (using immobilized cells) from fresh and 12-month-stored mahula flowers, respectively. PMID:16580830

  2. Design features of the laboratory-scale radiochemical immobilization system

    International Nuclear Information System (INIS)

    Under the High-Level Waste Immobilization Program, the Pacific Northwest Laboratory (PNL) is studying various ways to solidify high-level nuclear wastes. A variety of waste forms and processes are being investigated, with the most highly developed process being spray calcination coupled with in-can melting. This report describes a remote laboratory-scale system that was designed for the purpose of investigating the effects of different operating conditions and waste compositions on the product and on the effluents generated. It is termed laboratory-scale because of its nominal 1 L/h feed rate as compared to well over 300 L/h for full-scale equipment at PNL. The equipment currently consists of a feed system, a spray calciner, an in-can melter, and an effluent control system. It is operated in a shielded radiochemical hot cell using radioactive high-level liquid waste (HLLW) to answer questions on the deposition of radiochemicals during actual waste processing. The effluent control system can be modified in order to test different effluent systems, one of which has been proposed by the Savannah River Laboratories (SRL) for use in the Savannah River Plant vitrification system. The laboratory-scale system can also be used to test alternative immobilization processes, since spray calcination is a common processing step in many alternative waste form flowsheets. Thus, only the addition of a specific forming step such as pelletizing or sintering is necessary

  3. Tolerance and immobilization of cobalt by some bacteria from ferromanganese crusts of the afanasiy Nikitin Seamounts

    Digital Repository Service at National Institute of Oceanography (India)

    Krishnan, K.P.; Fernandes, C.E.G.; Fernandes, S.O.; LokaBharathi, P.A.

    hours at 23(? 1)oC. The cells were then scraped and suspended in sterile physiological saline (100ml). The suspension was centrifuged at 5000 r.p.m for 15 minutes. Cell pellets were washed twice with sterile saline and resuspened. Direct cell counts... at 5000 r.p.m for 15 minutes and a representative portion of the cell pellet was examined microscopically under a bright field Tolerance and immobilization of cobalt 5 (Olympus BHF ?342) microscope. Another portion of the cell pellet was dried overnight...

  4. Immobilization of Escherichia coli containing ω‐transaminase activity in LentiKats®

    DEFF Research Database (Denmark)

    Cárdenas‐Fernández, Max; Lima Afonso Neto, Watson; López, Carmen;

    2012-01-01

    Whole Escherichia coli cells overexpressing ω‐transaminase (ω‐TA) and immobilized cells entrapped in LentiKats® were used as biocatalysts in the asymmetric synthesis of the aromatic chiral amines 1‐phenylethylamine (PEA) and 3‐amino‐1‐phenylbutane (APB). Whole cells were permeabilized...... whole cell biocatalysis, the reaction with IPA was one order of magnitude faster than with Ala. No reaction was detected when permeabilized E. coli cells containing ω‐TA were employed using Ala as the amino donor. Additionally, the synthesis of APB from 4‐phenyl‐2‐butanone and IPA was studied. Whole...

  5. Immobilization of amyloglucosidase using two forms of polyurethane polymer.

    Science.gov (United States)

    Storey, K B; Duncan, J A; Chakrabarti, A C

    1990-03-01

    Amyloglucosidase was covalently immobilized using two hydrophilic prepolymers: Hypol FHP 2002 (creates foams) and Hypol FHP 8190H (creates gels). The foamable prepolymer was superior as a support for enzyme immobilization. The percent activity immobilized in the polyurethane foams was 25 +/- 1.5%. Large substrates (greater than 200,000 daltons in mol wt) were hydrolyzed as effectively as smaller ones by the immobilized enzyme. The Km value of the foam-immobilized enzyme increased from 0.76 mg/mL (free) to 0.86 mg/mL (immobilized), whereas the Vmax dropped from 90.9 (free) to 12.4 nmol glucose/min/mL (immobilized). The long-term (2 mo) storage stability of amyloglucosidase was enhanced by immobilization in foams (70% activity retained; free enzyme only retained 50%). Immobilization also improved the enzyme stability to various denaturing agents (sodium chloride, urea, and ethanol). The immobilized enzyme exhibited increased stability compared to the free enzyme at high temperatures (95 degrees C). Both glycogen and starch could be utilized by the immobilized enzyme, indicating that this technique could prove useful for starch hydrolysis. PMID:2112366

  6. Immobilization of horseradish peroxidase on modified chitosan beads.

    Science.gov (United States)

    Monier, M; Ayad, D M; Wei, Y; Sarhan, A A

    2010-04-01

    A method has been developed to immobilize horseradish peroxidase (HRP) on modified chitosan beads by means of graft copolymerization of polyethylacrylate in presence of potassium persulphate and Mohr's salt redox initiator. The activity of free and immobilized HRP was studied. FTIR spectroscopy and scanning electron microscopy were used to characterize HRP immobilization. The efficiency of the immobilization was investigated by examining the relative enzymatic activity of free enzyme before and after the HRP immobilization. The obtained values were found to reach 98.4%. The results show that the optimum temperature of immobilized HRP was 45 degrees C, which was identical to that of free enzyme, and the immobilized HRP exhibited a higher relative activity than that of free HRP over 45 degrees C. The optimal pH for immobilized HRP was 10, which was higher than that of the free HRP (pH 9.0), and the immobilization resulted in stabilization of enzyme over a broader pH range. The apparent kinetic constant value (K(m)) of immobilized HRP was 3.784 mmol ml(-1), which was higher than that of free HRP. On the other hand, the activity of immobilized HRP decreased slowly against time when compared to that of the free HRP, and could retain 65.8% residual activity after 6 consecutive cycles. PMID:20060854

  7. Principles, techniques, and applications of biocatalyst immobilization for industrial application.

    Science.gov (United States)

    Eş, Ismail; Vieira, José Daniel Gonçalves; Amaral, André Corrêa

    2015-03-01

    Immobilization is one of the most effective and powerful tools used in industry, which has been studied and improved since the last century. Various immobilization techniques and support materials have been used on both laboratory and industrial scale. Each immobilization technique is applicable for a specific production mostly depending on the cost and sensibility of process. Compared to free biocatalyst systems, immobilization techniques often offer better stability, increased activity and selectivity, higher resistance, improved separation and purification, reuse of enzymes, and consequently more efficient process. Recently, many reviews have been published about immobilization systems; however, most of them have focused on a specific application or not emphasized in details. This review focuses on most commonly used techniques in industry with many recent applications including using bioreactor systems for industrial production. It is also aimed to emphasize the advantages and disadvantages of the immobilization techniques and how these systems improve process productivity compared to non-immobilized systems.

  8. Screening of supports for immobilization of commercial porcine pancreatic lipase

    Directory of Open Access Journals (Sweden)

    Robison Scherer

    2011-12-01

    Full Text Available The aim of this work is to report the performance of different supports for the immobilization of commercial porcine pancreatic lipase. The immobilization tests were carried out in several types of Accurel, activated alumina, kaolin, montmorillonite, ion exchange resins and zeolites. The characterization of the supports showed differences in terms of specific area and morphology. The characteristics of the supports influenced the amount of enzyme adsorbed, yield of immobilization and esterification activity of the resulting immobilized catalyst. The clays KSF and natural and pillared montmorillonites presented potential for use as support for lipase immobilization in terms of yield and esterification activity. Yields of immobilization of 76.32 and 52.01% were achieved for clays KSF and natural montmorillonite, respectively. Esterification activities of 754.03, 595.51, 591.88 and 515.71 U.g-1 were obtained for lipases immobilized in Accurel MP-100, Amberlite XAD-2, mordenite and pillared montmorillonite, respectively.

  9. Mapping and identification of HeLa cell proteins separated by immobilized pH-gradient two-dimensional gel electrophoresis and construction of a two-dimensional polyacrylamide gel electrophoresis database

    DEFF Research Database (Denmark)

    Shaw, AC; Rossel Larsen, M; Roepstorff, P;

    1999-01-01

    The HeLa cell line, a human adenocarcinoma, is used in many research fields, since it can be infected with a wide range of viruses and intracellular bacteria. Therefore, the mapping of HeLa cell proteins is useful for the investigation of parasite host cell interactions. Because of the recent...

  10. ROLE OF GLUTAMATE DEHYDROGENASE AND GLUTAMINE SYNTHETASE IN CHLORELLA VULGARIS DURING ASSIMILATION OF AMMONIUM WHEN JOINTLY IMMOBILIZED WITH THE MICROALGAE-GROWTH-PROMOTING BACTERIUM AZOSPIRILLUM BRASILENSE(1).

    Science.gov (United States)

    De-Bashan, Luz E; Magallon, Paola; Antoun, Hani; Bashan, Yoav

    2008-10-01

    Enzymatic activities of glutamate dehydrogenase (GDH) and glutamine synthetase (GS) participating in the nitrogen metabolism and related ammonium absorption were assayed after the microalga Chlorella vulgaris Beij. was jointly immobilized with the microalgae-growth-promoting bacterium Azospirillum brasilense. At initial concentrations of 3, 6, and 10 mg · L(-1)  NH4 (+) , joint immobilization enhances growth of C. vulgaris but does not affect ammonium absorption capacity of the microalga. However, at 8 mg · L(-1)  NH4 (+) , joint immobilization enhanced ammonium absorption by the microalga without affecting the growth of the microalgal population. Correlations between absorption of ammonium per cell and per culture showed direct (negative and positive) linear correlations between these parameters and microalga populations at 3, 6, and 10 mg · L(-1)  NH4 (+) , but not at 8 mg · L(-1)  NH4 (+) , where the highest absorption of ammonium occurred. In all cultures, immobilized and jointly immobilized, having the four initial ammonium concentrations, enzymatic activities of Chlorella are affected by A. brasilense. Regardless of the initial concentration of ammonium, GS activity in C. vulgaris was always higher when jointly immobilized and determined on a per-cell basis. When jointly immobilized, only at an initial concentration of 8 mg · L(-1)  NH4 (+) was GDH activity per cell higher.

  11. Sequential Fermentation with Selected Immobilized Non-Saccharomyces Yeast for Reduction of Ethanol Content in Wine.

    Science.gov (United States)

    Canonico, Laura; Comitini, Francesca; Oro, Lucia; Ciani, Maurizio

    2016-01-01

    The average ethanol content of wine has increased over the last two decades. This increase was due to consumer preference, and also to climate change that resulted in increased grape maturity at harvest. In the present study, to reduce ethanol content in wine, a microbiological approach was investigated, using immobilized selected strains of non-Saccharomyces yeasts namely Starmerella bombicola, Metschnikowia pulcherrima, Hanseniaspora osmophila, and Hanseniaspora uvarum to start fermentation, followed by inoculation of free Saccharomyces cerevisiae cells. The immobilization procedures, determining high reaction rates, led a feasible sequential inoculation management avoiding possible contamination under actual winemaking. Under these conditions, the immobilized cells metabolized almost 50% of the sugar in 3 days, while S. cerevisiae inoculation completed all of fermentation. The S. bombicola and M. pulcherrima initial fermentations showed the best reductions in the final ethanol content (1.6 and 1.4% v/v, respectively). Resulting wines did not have any negative fermentation products with the exception of H. uvarum sequential fermentation that showed significant amount of ethyl acetate. On the other hand, there were increases in desirable compounds such as glycerol and succinic acid for S. bombicola, geraniol for M. pulcherrima and isoamyl acetate and isoamyl alcohol for H. osmophila sequential fermentations. The overall results indicated that a promising ethanol reduction could be obtained using sequential fermentation of immobilized selected non-Saccharomyces strains. In this way, a suitable timing of second inoculation and an enhancement of analytical profile of wine were obtained. PMID:27014203

  12. Preparation and characterization of latex films photo-immobilized with IFN-α.

    Science.gov (United States)

    Wu, Lifang; Hu, Kaikai; Zhang, Li; Chen, Wuya; Chen, Xiaohui; You, Rong; Yin, Liang; Guan, Yan-Qing

    2016-09-01

    We developed a biomaterial by photo-immobilizing interferon-α (IFN-α) on the surface of latex condom films for the prevention and treatment of cervicitis, cervical cancers and diseases caused by cervical virus. The IFN-α modification by photoactive N-(4-azidobenzoyloxy) succinimide was characterized on a nano-scale by spectroscopy analysis and micro morphology. The anti-bacterial, anti-cancer, and anti-viral effects of the modified bioactive latex films were evaluated by antibacterial susceptibility testing, Gram staining, flow cytometry, immunofluorescence, and Western blotting. Our results showed that the photo-immobilized IFN-α latex films effectively inhibited the growth of both Neisseria gonorrhoeae and human cervical cancer HeLa cells. Moreover, the expression of anti-viral proteins, including P56, MxA, and 2', 5'-OAS, in the human cervical epithelial cell line NC104 was significantly increased by photo-immobilized IFN-α latex films. Taken together, these results suggest that photo-immobilized IFN-α latex films may have therapeutic effects against cervicitis, cervical cancers, and cervical virus. PMID:27137809

  13. Sequential Fermentation with Selected Immobilized Non-Saccharomyces Yeast for Reduction of Ethanol Content in Wine

    Science.gov (United States)

    Canonico, Laura; Comitini, Francesca; Oro, Lucia; Ciani, Maurizio

    2016-01-01

    The average ethanol content of wine has increased over the last two decades. This increase was due to consumer preference, and also to climate change that resulted in increased grape maturity at harvest. In the present study, to reduce ethanol content in wine, a microbiological approach was investigated, using immobilized selected strains of non-Saccharomyces yeasts namely Starmerella bombicola, Metschnikowia pulcherrima, Hanseniaspora osmophila, and Hanseniaspora uvarum to start fermentation, followed by inoculation of free Saccharomyces cerevisiae cells. The immobilization procedures, determining high reaction rates, led a feasible sequential inoculation management avoiding possible contamination under actual winemaking. Under these conditions, the immobilized cells metabolized almost 50% of the sugar in 3 days, while S. cerevisiae inoculation completed all of fermentation. The S. bombicola and M. pulcherrima initial fermentations showed the best reductions in the final ethanol content (1.6 and 1.4% v/v, respectively). Resulting wines did not have any negative fermentation products with the exception of H. uvarum sequential fermentation that showed significant amount of ethyl acetate. On the other hand, there were increases in desirable compounds such as glycerol and succinic acid for S. bombicola, geraniol for M. pulcherrima and isoamyl acetate and isoamyl alcohol for H. osmophila sequential fermentations. The overall results indicated that a promising ethanol reduction could be obtained using sequential fermentation of immobilized selected non-Saccharomyces strains. In this way, a suitable timing of second inoculation and an enhancement of analytical profile of wine were obtained. PMID:27014203

  14. Immobilization of calcium and phosphate ions improves the osteoconductivity of titanium implants.

    Science.gov (United States)

    Sunarso; Toita, Riki; Tsuru, Kanji; Ishikawa, Kunio

    2016-11-01

    In this work, to elevate weak osteoconductivity of titanium (Ti) implant, we prepared a Ti implant having both calcium and phosphate ions on its surface. To modify calcium and phosphate ions onto Ti, phosphate ions were first immobilized by treating the Ti with a NaH2PO4 solution, followed by CaCl2 treatment to immobilize calcium ions, which created the calcium and phosphate ions-modified Ti (Ca-P-Ti). X-ray photoelectron spectroscopy and thin-layer X-ray diffraction measurement confirmed that both phosphate and calcium ions were co-immobilized onto the Ti surface on the molecular level. Three-hour after seeding MC3T3-E1 murine pre-osteoblast cells on substrates, cell number on Ca-P-Ti was much larger than that of Ti and phosphate-modified Ti (P-Ti), but was similar to that of calcium-modified Ti (Ca-Ti). Also, MC3T3-E1 cells on Ca-P-Ti expressed larger amount of vinculin, a focal adhesion protein, than those on other substrates, probably resulting in larger cell size as well as greater cell proliferation on Ca-P-Ti than those on other substrates. Alkaline phosphatase activity of cells on Ca-P-Ti was greater than those on Ti and P-Ti, but was almost comparable to that of Ca-Ti. Moreover, the largest amount of bone-like nodule formation was observed on Ca-P-Ti. These results provide evidence that calcium and phosphate ions-co-immobilization onto Ti increased the osteoconductivity of Ti by stimulating the responses of pre-osteoblast cells. This simple modification would be promising technique for bone tissue implant including dental and orthopedic implants. PMID:27524023

  15. Immobilization of Dystrophin and Laminin α2-Chain Deficient Zebrafish Larvae In Vivo Prevents the Development of Muscular Dystrophy.

    Directory of Open Access Journals (Sweden)

    Mei Li

    Full Text Available Muscular dystrophies are often caused by genetic alterations in the dystrophin-dystroglycan complex or its extracellular ligands. These structures are associated with the cell membrane and provide mechanical links between the cytoskeleton and the matrix. Mechanical stress is considered a pathological mechanism and muscle immobilization has been shown to be beneficial in some mouse models of muscular dystrophy. The zebrafish enables novel and less complex models to examine the effects of extended immobilization or muscle relaxation in vivo in different dystrophy models. We have examined effects of immobilization in larvae from two zebrafish strains with muscular dystrophy, the Sapje dystrophin-deficient and the Candyfloss laminin α2-chain-deficient strains. Larvae (4 days post fertilization, dpf of both mutants have significantly lower active force in vitro, alterations in the muscle structure with gaps between muscle fibers and altered birefringence patterns compared to their normal siblings. Complete immobilization (18 hrs to 4 dpf was achieved using a small molecular inhibitor of actin-myosin interaction (BTS, 50 μM. This treatment resulted in a significantly weaker active contraction at 4 dpf in both mutated larvae and normal siblings, most likely reflecting a general effect of immobilization on myofibrillogenesis. The immobilization also significantly reduced the structural damage in the mutated strains, showing that muscle activity is an important pathological mechanism. Following one-day washout of BTS, muscle tension partly recovered in the Candyfloss siblings and caused structural damage in these mutants, indicating activity-induced muscle recovery and damage, respectively.

  16. Plasma Treated High-Density Polyethylene (HDPE Medpor Implant Immobilized with rhBMP-2 for Improving the Bone Regeneration

    Directory of Open Access Journals (Sweden)

    Jin-Su Lim

    2014-01-01

    Full Text Available We investigate the bone generation capacity of recombinant human bone morphogenetic protein-2 (rhBMP-2 immobilized Medpor surface through acrylic acid plasma-polymerization. Plasma-polymerization was carried out at a 20 W at an acrylic acid flow rate of 7 sccm for 5 min. The plasma-polymerized Medpor surface showed hydrophilic properties and possessed a high density of carboxyl groups. The rhBMP-2 was immobilized with covalently attached carboxyl groups using 1-ethyl-3-(3-dimethylaminopropyl carbodiimide and N-hydroxysuccinimide. Carboxyl groups and rhBMP-2 immobilization on the Medpor surface were identified by Fourier transform infrared spectroscopy. The activity of Medpor with rhBMP-2 immobilized was examined using an alkaline phosphatase assay on MC3T3-E1 cultured Medpor. These results showed that the rhBMP-2 immobilized Medpor increased the level of MC3T3-E1 cell differentiation. These results demonstrated that plasma surface modification has the potential to immobilize rhBMP-2 on polymer implant such as Medpor and can be used for the binding of bioactive nanomolecules in bone tissue engineering.

  17. Enzyme-Immobilized Microfluidic Process Reactors

    Directory of Open Access Journals (Sweden)

    Hideaki Maeda

    2011-07-01

    Full Text Available Microreaction technology, which is an interdisciplinary science and engineering area, has been the focus of different fields of research in the past few years. Several microreactors have been developed. Enzymes are a type of catalyst, which are useful in the production of substance in an environmentally friendly way, and they also have high potential for analytical applications. However, not many enzymatic processes have been commercialized, because of problems in stability of the enzymes, cost, and efficiency of the reactions. Thus, there have been demands for innovation in process engineering, particularly for enzymatic reactions, and microreaction devices represent important tools for the development of enzyme processes. In this review, we summarize the recent advances of microchannel reaction technologies especially for enzyme immobilized microreactors. We discuss the manufacturing process of microreaction devices and the advantages of microreactors compared to conventional reaction devices. Fundamental techniques for enzyme immobilized microreactors and important applications of this multidisciplinary technology are also included in our topics.

  18. Process Technology for Immobilized Lipasecatalyzed Reactions

    DEFF Research Database (Denmark)

    Xu, Yuan

    -down experimental work is described in this thesis. The methodology uses economic targets to test options characterized via a set of tools. In order to validate the methodology, two processes based on immobilized lipase-catalysis have been studied: transesterification and esterification of vegetable oils...... for the production of biodiesel. The two processes are focused on the conversion of the two main components of vegetable oil materials, glyceride esters and free fatty acids respectively, into fatty acid alkyl esters. Although biodiesel is conventionally prepared via chemical-catalyzed transesterification...... of vegetable oils with methanol to produce fatty acid methyl esters (FAME), this work has been focused on the production of fatty acid ethyl esters (FAEE) with bioethanol due to the expected improved sustainability of this type of biodiesel. A key reaction characteristic of the immobilized lipase...

  19. Glucose oxidase immobilization onto carbon nanotube networking

    CERN Document Server

    Karachevtsev, V A; Zarudnev, E S; Karachevtsev, M V; Leontiev, V S; Linnik, A S; Lytvyn, O S; Plokhotnichenko, A M; Stepanian, S G

    2012-01-01

    When elaborating the biosensor based on single-walled carbon nanotubes (SWNTs), it is necessary to solve such an important problem as the immobilization of a target biomolecule on the nanotube surface. In this work, the enzyme (glucose oxidase (GOX)) was immobilized on the surface of a nanotube network, which was created by the deposition of nanotubes from their solution in 1,2-dichlorobenzene by the spray method. 1-Pyrenebutanoic acid succinimide ester (PSE) was used to form the molecular interface, the bifunctional molecule of which provides the covalent binding with the enzyme shell, and its other part (pyrene) is adsorbed onto the nanotube surface. First, the usage of such a molecular interface leaves out the direct adsorption of the enzyme (in this case, its activity decreases) onto the nanotube surface, and, second, it ensures the enzyme localization near the nanotube. The comparison of the resonance Raman (RR) spectrum of pristine nanotubes with their spectrum in the PSE environment evidences the creat...

  20. Immobilization of Polymeric Luminophor on Nanoparticles Surface

    Science.gov (United States)

    Bolbukh, Yuliia; Podkoscielna, Beata; Lipke, Agnieszka; Bartnicki, Andrzej; Gawdzik, Barbara; Tertykh, Valentin

    2016-04-01

    Polymeric luminophors with reduced toxicity are of the priorities in the production of lighting devices, sensors, detectors, bioassays or diagnostic systems. The aim of this study was to develop a method of immobilization of the new luminophor on a surface of nanoparticles and investigation of the structure of the grafted layer. Monomer 2,7-(2-hydroxy-3-methacryloyloxypropoxy)naphthalene (2,7-NAF.DM) with luminophoric properties was immobilized on silica and carbon nanotubes in two ways: mechanical mixing with previously obtained polymer and by in situ oligomerization with chemisorption after carrier's modification with vinyl groups. The attached polymeric (or oligomeric) surface layer was studied using thermal and spectral techniques. Obtained results confirm the chemisorption of luminophor on the nanotubes and silica nanoparticles at the elaborated synthesis techniques. The microstructure of 2,7-NAF.DM molecules after chemisorption was found to be not changed. The elaborated modification approach allows one to obtain nanoparticles uniformly covered with polymeric luminophor.

  1. Technologies for immobilization and disposal of tritium

    International Nuclear Information System (INIS)

    This study was done within a program one of whose objectives was to know the state of the technology development for tritium separation in the moderator circuit at HWR and to define the possible technologies to be applied to the Argentine nuclear power plants. Within this framework the strategies adopted by each country and the available technologies for a safe disposal of tritium, not only in its gaseous state tritium but also as tritiated water were analyzed. It is considered that if the selected separation method is such that the tritium is in its gaseous state, the hydride formation for long periods of immobilization should be studied. whereas if it were triated water immobilization should be studied to choose the technology between cementation and drying agents, in both cases the final disposal site will have to be selected. (author). 8 refs

  2. Thermoresponsive and bioactive poly(vinyl ether)-based hydrogels synthesized by radiation copolymerization and photochemical immobilization

    International Nuclear Information System (INIS)

    A thermoresponsive hydrogel was synthesized by radiation copolymerization of ethylene glycol vinyl ether (EGVE) and butyl vinyl ether (BVE) in the presence of cross-linking agent diethylene glycol divinyl ether. The gel was modified by a cell adhesion factor RGD by photochemical immobilization technique. While the unmodified hydrogel shows fully hydrated form at low temperatures (+4 oC) and it extensively dehydrates at 37 oC, the biomodified hydrogel still kept its thermoresponsive character after immobilization. The effectiveness of immobilization was checked with FTIR-ATR and XPS. The use of bioactive thermoresponsive hydrogels in cell culture applications was investigated. For this purpose, cell culture experiments were realized by L929 mouse fibroblasts. Cell attachment experiments revealed the effect of immobilized RGD with higher values of cell attachment (∼85%), which were obtained especially in the absence of serum. The thermoresponsive character of the hydrogel was useful for the application of low-temperature treatment in order to recover the attached viable cells from the surface of the hydrogel without using trypsin. When the culture temperature was decreased from 37 to 10 deg. C for 30 min ∼80% of the cells were detached from the hydrogel surface

  3. Immobilization of Rocky Flats graphite fines residues

    International Nuclear Information System (INIS)

    The Savannah River Technology Center (SRTC) is developing an immobilization process for graphite fines residues generated during nuclear materials production activities at the Rocky Flats Environmental Technology Site (Rocky Flats). The continued storage of this material has been identified as an item of concern. The residue was generated during the cleaning of graphite casting molds and potentially contains reactive plutonium metal. The average residue composition is 73 wt% graphite, 15 wt% calcium fluoride (CaF2), and 12 wt% plutonium oxide (PuO2). Approximately 950 kg of this material are currently stored at Rocky Flats. The strategy of the immobilization process is to microencapsulate the residue by mixing with a sodium borosilicate (NBS) glass frit and heating at nominally 700 C. The resulting waste form would be sent to the Waste Isolation Pilot Plant (WIPP) for disposal. Since the PuO2 concentration in the residue averages 12 wt%, the immobilization process was required to meet the intent of safeguards termination criteria by limiting plutonium recoverability based on a test developed by Rocky Flats. The test required a plutonium recovery of less than 4 g/kg of waste form when a sample was leached using a nitric acid/CaF2 dissolution flowsheet. Immobilization experiments were performed using simulated graphite fines with cerium oxide (CeO2) as a surrogate for PuO2 and with actual graphite fines residues. Small-scale surrogate experiments demonstrated that a 4:1 frit to residue ratio was adequate to prevent recovery of greater than 4 g/kg of cerium from simulated waste forms. Additional experiments investigated the impact of varying concentrations of CaF2 and the temperature/heating time cycle on the cerium recovery. Optimal processing conditions developed during these experiments were subsequently demonstrated at full-scale with surrogate materials and on a smaller scale using actual graphite fines

  4. Immobilizing Biomolecules Near the Diffraction Limit

    DEFF Research Database (Denmark)

    Skovsen, Esben; Neves-Petersen, Maria Teresa; Kold, Ane;

    2009-01-01

    Our group has previously shown that biomolecules containing disulfide bridges in close proximity to aromatic residues can be immobilized, through covalent bonds, onto thiol derivatized surfaces upon UV excitation of the aromatic residue(s). We have also previously shown that our new technology ca...... a substrate, which can be generated by a UV diffraction pattern. Such patterns can have sub-micron feature sizes and could therefore be of great relevance for present and future nanotechnological applications....

  5. Uranium Immobilization by Sulfate-reducing Biofilms

    International Nuclear Information System (INIS)

    Hexavalent uranium [U(VI)] was immobilized using biofilms of the sulfate-reducing bacterium (SRB) Desulfovibrio desulfuricans G20. The biofilms were grown in flat-plate continuous-flow reactors using lactate as the electron donor and sulfate as the electron acceptor. U(VI) was continuously fed into the reactor for 32 weeks at a concentration of 126 ?M. During this time, the soluble U(VI) was removed (between 88 and 96% of feed) from solution and immobilized in the biofilms. The dynamics of U immobilization in the sulfate-reducing biofilms were quantified by estimating: (1) microbial activity in the SRB biofilm, defined as the hydrogen sulfide (H2S) production rate and estimated from the H2S concentration profiles measured using microelectrodes across the biofilms; (2) concentration of dissolved U in the solution; and (3) the mass of U precipitated in the biofilm. Results suggest that U was immobilized in the biofilms as a result of two processes: (1) enzymatically and (2) chemically, by reacting with microbially generated H2S. Visual inspection showed that the dissolved sulfide species reacted with U(VI) to produce a black precipitate. Synchrotron-based U L3-edge X-ray absorption near edge structure (XANES) spectroscopy analysis of U precipitated abiotically by sodium sulfide indicated that U(VI) had been reduced to U(IV). Selected-area electron diffraction pattern and crystallographic analysis of transmission electron microscope lattice-fringe images confirmed the structure of precipitated U as being that of uraninite

  6. Combustion synthesis of radioactive waste immobilization

    Institute of Scientific and Technical Information of China (English)

    ZHANG Ruizhu; GUO Zhimeng; LU Xin; JIA Chengchang; LIN Tao

    2005-01-01

    Using chromium oxide (CrO3) as an oxidant, the immobilization of simulating radioactive waste in perovskite (CaTiO3) structure by a combustion synthesis (CS) method was tested. The products were characterized by Archimedes liquid displacement technique, microhardness technique, X-ray diffraction, and scanning electron microscopy. The leaching rate was measured by the method of MCC-1 or MCC-2.The primary results show that the CS method can be used to solidify the immobilizate waste effectively.

  7. Chromium (VI) biosorption by immobilized Aspergillus niger in continuous flow system with special reference to FTIR analysis.

    Science.gov (United States)

    Chhikara, S; Hooda, A; Rana, L; Dhankhar, R

    2010-09-01

    Aspergillus niger was treated with acid and immobilized in calcium alginate matrix. The dynamic removal of Cr (VI) ion was studied using continuously fed column packed with immobilized biosorbent beads. Column experiments were carried out to study the effect of various bed heights (20, 30, 40 cm) under different flow rates (5, 7.5, 10 ml min(-1)) on efficiency of biosorption. The maximum time (1020 minutes; 17 hr) before breakthrough point was observed in case of 40 cm bed height with flow rate of 5ml min(-1). FTIR analysis of acid treated immobilized A. niger was used fora qualitative and preliminary analysis of chemical functional groups present on its cell wall which provided the information on nature of cell wall and Cr (VI) interaction during the process of biosorption. The IR spectra of biosorbent recorded before and after chromium biosorption had shown some changes in the band patterns, which were finally analyzed and was found that chemical interaction such as ion-exchange between carboxyl (-COOH), hydroxyl (-OH) and amine (-NH2) group of biosorbent and Chromium ion were mainlyinvolved in biosorption of Cr (VI) onto A. niger cell wall surface. The biosorbed metal was eluted from biosorbent by using 0.1 M H2SO4 as eluant. Immobilized biosorbent could be reused for five consecutive biosorption and desorption cycles without apparent loss of efficiency after its reconditioning. Considering all above factors together this paper discusses the efficient chromium biosorption process carried out by immobilized A. niger biosorbent. PMID:21387903

  8. Graft linker immobilization for spatial control of protein immobilization inside fused microchips.

    Science.gov (United States)

    Shirai, Kentaro; Renberg, Björn; Sato, Kae; Mawatari, Kazuma; Konno, Tomohiro; Ishihara, Kazuhiko; Kitamori, Takehiko

    2009-12-01

    Fused silica glass microchips have several attractive features for lab-on-a-chip applications; they can be machined with excellent precision down to nanospace; are stable; transparent and can be modified with a range of silanization agents to change channel surface properties. For immobilization, however, ligands must be added after bonding, since the harsh bonding conditions using heat or hydrofluoric acid would remove all prior immobilized ligands. For spatial control over immobilization, UV-mediated immobilization offers several advantages; spots can be created in parallel, the feature size can be made small, and spatial control over patterns and positions is excellent. However, UV sensitive groups are often based on hydrophobic chemical moieties, which unfortunately result in greater non-specific binding of biomolecules, especially proteins. Here, we present techniques in which any -CH(x) (x=1,2,3) containing surface coating can be used as foundation for grafting a hydrophilic linker with a chemical anchor, a carboxyl group, to which proteins and amine containing molecules can be covalently coupled. Hence, the attractive features of many well-known protein and biomolecule repelling polymer coatings can be utilized while achieving site-specific immobilization only to pre-determined areas within the bonded microchips.

  9. Immobilization of enzyme on a polymer surface

    Science.gov (United States)

    Shen, Lei; Cheng, Kenneth Chun Kuen; Schroeder, McKenna; Yang, Pei; Marsh, E. Neil G.; Lahann, Joerg; Chen, Zhan

    2016-06-01

    We successfully immobilized enzymes onto polymer surfaces via covalent bonds between cysteine groups of the enzyme and dibromomaleimide functionalities present at the polymer surface. In this work, we used nitroreductase (NfsB) as a model enzyme molecule. The polymers were prepared by chemical vapor deposition (CVD) polymerization, resulting in surfaces with dibromomaleimide groups. NfsB variants were engineered so that each NfsB molecule only has one cysteine group on the enzyme surface. Two different NfsB constructs were studied, with cysteines at the positions of H360 and V424, respectively. A combination of sum frequency generation (SFG) vibrational and attenuated total reflectance Fourier transformed infrared (ATR-FTIR) spectroscopies were used to deduce the orientation of the immobilized enzymes on the surface. It was found that the orientation of the immobilized enzymes is controlled by the position of the cysteine residue in the protein. The NfsB H360C construct exhibited a similar orientational behavior on the polymer surface as compared to that on the self-assembled monolayer surface, but the NsfB V424C construct showed markedly different orientations on the two surfaces.

  10. Immobilization of mycoplana sp. MVMB2 isolated from petroleum contaminated soil onto papaya stem (carica papaya l.) and its application on degradation of phenanthrene

    Energy Technology Data Exchange (ETDEWEB)

    Brinda Lakshmi, Mahalingam; Muthukumar, Karuppan; Velan, Manickam [Environmental Biotechnology Laboratory, Department of Chemical Engineering, A.C. College of Technology, Anna University, Chennai (India)

    2012-08-15

    This study presents the degradation of phenanthrene using immobilized Mycoplana sp. MVMB2 isolated from contaminated soil. Papaya stem pretreated by two stage processes, treating with acid or alkali and drying, was used for the immobilization of Mycoplana sp. Alkali pretreated papaya stem was found to be most effective in cell uptake compared to acid treated one. The maximum immobilization capacity at various physiochemical conditions for the alkali pretreated papaya stem was found to be at 320 min time, pH 6.5, 30 C temperature, and 18.6 x 10{sup 6} cells/mL initial concentrations. The adsorption mechanism of Mycoplana sp. MVMB2 on pretreated papaya stem was assessed using various kinetic and isotherm models. The immobilization of Mycoplana sp. MVMB2 on to pretreated papaya stem was corroborated by scanning electron microscopy and Fourier transformed IR spectroscopy analysis. The performance of immobilized cells in batch reactor showed more than 95% phenanthrene degradation within 72 h, whereas, free cells were found to require 120 h. The immobilized cells also showed better degradation performance in the packed column study. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  11. Co-immobilization of glucose oxidase and xylose dehydrogenase displayed whole cell on multiwalled carbon nanotube nanocomposite films modified electrode for simultaneous voltammetric detection of D-glucose and D-xylose.

    Science.gov (United States)

    Li, Liang; Liang, Bo; Li, Feng; Shi, Jianguo; Mascini, Marco; Lang, Qiaolin; Liu, Aihua

    2013-04-15

    In this paper, we first report the construction of Nafion/glucose oxidase (GOD)/xylose dehydrogenase displayed bacteria (XDH-bacteria)/multiwalled carbon nanotubes (MWNTs) modified electrode for simultaneous voltammetric determination of D-glucose and D-xylose. The optimal conditions for the immobilized enzymes were established. Both enzymes retained their good stability and activities. In the mixture solution of D-glucose and D-xylose containing coenzyme NAD⁺ (the oxidized form of nicotinamide adenine dinucleotide), the Nafion/GOD/XDH-bacteria/MWNTs modified electrode exhibited quasi-reversible oxidation-reduction peak at -0.5 V (vs. saturated calomel electrode, SCE) originating from the catalytic oxidation of D-glucose, and oxidation peak at +0.55 V(vs. SCE) responding to the oxidation of NADH (the reduced form of nicotinamide adenine dinucleotide) by the carbon nanotubes, where NADH is the resultant product of coenzyme NAD⁺ involved in the catalysis of D-xylose by XDH-displayed bacteria. For the proposed biosensor, cathodic peak current at -0.5 V was linear with the concentration of D-glucose within the range of 0.25-6 mM with a low detection limit of 0.1 mM D-glucose (S/N=3), and the anodic peak current at +0.55 V was linear with the concentration of d-xylose in the range of 0.25∼4 mM with a low detection limit of 0.1 mM D-xylose (S/N=3). Further, D-xylose and D-glucose did not interfere with each other. 300-fold excess saccharides including D-maltose, D-galactose, D-mannose, D-sucrose, D-fructose, D-cellobiose, and 60-fold excess L-arabinose, and common interfering substances (100-fold excess ascorbic acid, dopamine, uric acid) as well as 300-fold excess D-xylitol did not affect the detection of D-glucose and D-xylose (both 1 mM). Therefore, the proposed biosensor is stable, specific, reproducible, simple, rapid and cost-effective, which holds great potential in real applications. PMID:23202346

  12. Enzyme immobilization and biocatalysis of polysiloxanes

    Science.gov (United States)

    Poojari, Yadagiri

    Lipases have been proven to be versatile and efficient biocatalysts which can be used in a broad variety of esterification, transesterification, and ester hydrolysis reactions. Due to the high chemo-, regio-, and stereo-selectivity and the mild conditions of lipase-catalyzed reactions, the vast potential of these biocatalysts for use in industrial applications has been increasingly recognized. Polysiloxanes (silicones) are well known for their unique physico-chemical properties and can be prepared in the form of fluids, elastomers, gels and resins for a wide variety of applications. However, the enzymatic synthesis of silicone polyesters and copolymers is largely unexplored. In the present investigations, an immobilized Candida antarctica lipase B (CALB) on macroporous acrylic resin beads (Novozym-435 RTM) has been successfully employed as a catalyst to synthesize silicone polyesters and copolymers under mild reaction conditions. The silicone aliphatic polyesters and the poly(dimethylsiloxane)--poly(ethylene glycol) (PDMS-PEG) copolymers were synthesized in the bulk (without using a solvent), while the silicone aromatic polyesters, the silicone aromatic polyamides and the poly(epsilon-caprolactone)--poly(dimethylsiloxane)--poly(epsilon-caprolactone) (PCL-PDMS-PCL) triblock copolymers were synthesized in toluene. The synthesized silicone polyesters and copolymers were characterized by Gel Permeation Chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetric Analysis (TGA), Differential Scanning Calorimetry (DSC) and Wide Angle X-ray Diffraction (WAXD). This dissertation also describes a methodology for physical immobilization of the enzyme pepsin from Porcine stomach mucosa in silicone elastomers utilizing condensation-cure room temperature vulcanization (RTV) of silanol-terminated poly(dimethylsiloxane) (PDMS). The activity and the stability of free pepsin and pepsin immobilized in silicone elastomers were studied with respect to p

  13. Magnetic Sephadex as a carrier for enzyme immobilization and drug targeting

    Energy Technology Data Exchange (ETDEWEB)

    Torchilin, V.P.; Papisov, M.I.; Smirnov, V.N.

    1985-04-01

    Magnetic materials were suggested as carriers for protein immobilization about 10 years ago. The main advantage of these carriers is their ability to be concentrated near magnetic terminals upon the application of the external magnetic field. This property is used in technological processes for selective catalyst removal from the reaction mixture (3), in immunological studies for the separation of cells to which magnetic particles are specifically bound modified with antibodies against cell surface components, in experiments for the drug targeting in vivo into appropriate tissues under the action of external magnetic field. There exist a number of methods to obtain porous magnetic carriers, containing immobilized matter not only on the surface, but also in the volume of a particle. Normally, these preparations are obtained by the granule formation from the suspension of ferromagnetic particles in the solution or melt of appropriate high-molecular-weight compound. The drawback of the above-mentioned methods is the pronounced aggregation of ferromagnetic particles. The aggregation does not permit to use concentrated enough suspensions of magnetic particles and causes the formation of the product with a variety of sizes and magnetic properties. We made an attempt to synthesize the magnetic carrier for protein immobilization on the basis of commercial Sephadex porous spheres. Sephadex granules were made magnetic by adsorptional fixation of ferromagnetic particles in its pores. The properties of the ''native'' and ''magnetic'' Sephadexes as carriers for protein immobilization were compared by parallel immobilization on both carriers of alpha-chymotrypsin and T I-albumin. In in vivo experiments we studied the ability of magnetic Sephadex to be concentrated in a desired region of the circulation under the action of external magnetic field.

  14. Lipase immobilization and production of fatty acid methyl esters from canola oil using immobilized lipase

    International Nuclear Information System (INIS)

    Lipase enzyme from Aspergillus oryzae (EC 3.1.1.3) was immobilized onto a micro porous polymeric matrix which contains aldehyde functional groups and methyl esters of long chain fatty acids (biodiesel) were synthesized by transesterification of crude canola oil using immobilized lipase. Micro porous polymeric matrix was synthesized from styrene-divinylbenzene (STY-DVB) copolymers by using high internal phase emulsion technique and two different lipases, Lipozyme TL-100L® and Novozym 388®, were used for immobilization by both physical adsorption and covalent attachment. Biodiesel production was carried out with semi-continuous operation. Methanol was added into the reactor by three successive additions of 1:4 M equivalent of methanol to avoid enzyme inhibition. The transesterification reaction conditions were as follows: oil/alcohol molar ratio 1:4; temperature 40 oC and total reaction time 6 h. Lipozyme TL-100L® lipase provided the highest yield of fatty acid methyl esters as 92%. Operational stability was determined with immobilized lipase and it indicated that a small enzyme deactivation occurred after used repeatedly for 10 consecutive batches with each of 24 h. Since the process is yet effective and enzyme does not leak out from the polymer, the method can be proposed for industrial applications. -- Research highlights: → Lipozyme TL-100L and Novozym 388 were immobilized onto micro porous polymeric matrix by both physical adsorption and covalent linking. → Immobilized enzymes were used for synthesis of fatty acid methyl esters by transesterification of canola oil and methanol using semi-continuous operation system. → According to chromatographic analysis, Lipase Lipozyme TL-100L resulted in the highest yield of methyl ester as 92%.

  15. Activation of accumulated nitrite reduction by immobilized Pseudomonas stutzeri T13 during aerobic denitrification.

    Science.gov (United States)

    Ma, Fang; Sun, Yilu; Li, Ang; Zhang, Xuening; Yang, Jixian

    2015-01-01

    The excellent removal efficiency of nitrate by the aerobic denitrifier, Pseudomonas stutzeri T13, was achieved in free cells system. However, poor nitrite reduction prevents efficient aerobic denitrification because of the nitrite accumulation. This problem could be conquered by immobilizing the cells on supports. In this study, strain T13 was immobilized by mycelial pellets (MPs), polyurethane foam cubes (PFCs) and sodium alginate beads (SABs). Higher removal percentages of TN in MP (43.78%), PFC (42.31%) and SAB (57.25%) systems were achieved compared with the free cell system (29.7%). Furthermore, the optimal condition for immobilized cell systems was as follows: 30°C, 100rpm shaking speed and pH 7. The shock-resistance of SAB system was relatively poor, which could collapse under either alkaline (pH=9) or high rotating (200rpm) conditions. The recycling experiments demonstrated that the high steady TN removal rate could be maintained for seven cycles in both MP and PFC systems. PMID:25827250

  16. Design of a papain immobilized antimicrobial food package with curcumin as a crosslinker.

    Directory of Open Access Journals (Sweden)

    Cynthya Maria Manohar

    Full Text Available 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.

  17. A whisker-like carbon composite for the immobilization of laccase and its bioelectrochemistry

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A novel mesoporous carbon/whisker-like carbon (MCWC) composite was used for the immobilization of laccase (Lac) and its bioelectrochemical behaviors were studied.It was confirmed by XPS that Lac was strongly adsorbed on the surface of the MCWC composite.The cyclic voltammetric results showed that the immobilized Lac underwent a direct quasi-reversible electrochemical reaction.The value of the electron transfer rate constant ks was estimated to be 0.770 s-1,indicating a reasonably fast electron transfer between the immobilized Lac and the underlying electrode.The surface concentration (I-) of Lac was estimated to be 2.730 × 10-12 mol/cm2.Further experimental results showed that the immobilized Lac displayed an appreciable electrocatalytic activity to the electrochemical reduction of O2.These properties could be attributed to the particular structure of loosely packed nanometer-scale carbon whiskers and the existence of a large amount of oxygen-containing groups.The immo- bilization method and the novel carrier (MCWC) may find new applications in fabricating the biocatalysts for biofuel cells.

  18. Decolorization of the metal textile dye Lanaset Grey G by immobilized white-rot fungi.

    Science.gov (United States)

    Daâssi, Dalel; Mechichi, Tahar; Nasri, Moncef; Rodriguez-Couto, Susana

    2013-11-15

    In this paper we studied the ability of four Tunisian-isolated fungi (i.e. Coriolopsis gallica, Bjerkandera adusta, Trametes versicolor and Trametes trogii) immobilized into Ca-alginate beads to decolorize the metal textile dye Lanaset Grey G (LG). The effect of different operational conditions, such as initial dye concentration, temperature, pH, beads/medium ratio and agitation, on dye decolorization by the immobilized fungi was investigated. Maximal decolorization percentages of 88.7%, 89.3%, 82.1% and 81.3% for C. gallica, B. adusta, T. versicolor and T. trogii were attained, respectively, when operating at an initial LG concentration of 150 mg/L, pH values of 5.0-6.0, temperatures of 40-45 °C and a beads/medium ratio of 20% (w/v) in static conditions after 72 h of incubation. Subsequently, the re-usability of the immobilized fungi was evaluated. After three decolorization cycles, the decolorization percentage of free cell cultures dropped to values below 36%, while decolorization percentages of about 75%, 70%, 60% and 68% were obtained by the immobilized cultures of C. gallica, B. adusta, T. versicolor and T. trogii, respectively.

  19. Immobilization study of biosorption of heavy metal ions onto activated sludge

    Institute of Scientific and Technical Information of China (English)

    WU Hai-suo; ZHANG Ai-qiang; WANG Lian-sheng

    2004-01-01

    Activated sludge was immobilized into Ca-alginate beads via entrapment, and biosorption of three heavy metal ions, copper(Ⅱ), zinc(Ⅱ), and chromimum(Ⅱ), from aqueous solution in the concentration range of 10-100 mg/L was studied by using both entrapped activated sludge and inactivated free biomass at pH≤ 5. A biphasic metal adsorption pattern was observed in all immobilized biomass experiments. The biosorption of metal ions by the biosorbents increased with the initial concentration increased in the medium. The adsorption rate of immobilized pretreated activated sludge(PAS) was much lower than that of free PAS due to the increase in mass transfer resistance resulting from the polymeric matrix. Biosorption equilibrium of beads was established in about 20 h and the adsorbed heavy metal ions did not change further with time. No significant effect of temperature was observed in the test for free biomass while immobilized PAS appeared to be strong temperature dependent in the test range of 10 and 40℃.Besides, the content of activated sludge in the calcium alginate bead has an influence on the uptake of heavy metals. The sorption equilibrium was well modeled by Langmuir isotherm, implying monomolecular adsorption mechanism. Carboxyl group in cell wall played an important role in surface adsorption of heavy metal ions on PAS.

  20. Chromium immobilization by extra- and intraradical fungal structures of arbuscular mycorrhizal symbioses.

    Science.gov (United States)

    Wu, Songlin; Zhang, Xin; Sun, Yuqing; Wu, Zhaoxiang; Li, Tao; Hu, Yajun; Lv, Jitao; Li, Gang; Zhang, Zhensong; Zhang, Jing; Zheng, Lirong; Zhen, Xiangjun; Chen, Baodong

    2016-10-01

    Arbuscular mycorrhizal (AM) fungi can enhance plant Cr tolerance through immobilizing Cr in mycorrhizal roots. However, the detailed processes and mechanisms are unclear. The present study focused on cellular distribution and speciation of Cr in both extraradical mycelium (ERM) and mycorrhizal roots exposed to Cr(VI) by using field emission scanning electron microscopy equipped with energy dispersive X-ray spectrometer (FE-SEM-EDS), scanning transmission soft X-ray microscopy (STXM) and X-ray absorption fine structure (XAFS) spectroscopy techniques. We found that amounts of particles (possibly extracellular polymeric substances, EPS) were produced on the AM fungal surface upon Cr(VI) stress, which contributed greatly to Cr(VI) reduction and immobilization. With EDS of the surface of AM fungi exposed to various Cr(VI) levels, a positive correlation between Cr and P was revealed, suggesting that phosphate groups might act as counter ions of Cr(III), which was also confirmed by the XAFS analysis. Besides, STXM and XAFS analyses showed that Cr(VI) was reduced to Cr(III) in AM fungal structures (arbuscules, intraradical mycelium, etc.) and cell walls in mycorrhizal roots, and complexed possibly with carboxyl groups or histidine analogues. The present work provided evidence of Cr immobilization on fungal surface and in fungal structures in mycorrhizal roots at a cellular level, and thus unraveled the underlying mechanisms by which AM symbiosis immobilize Cr. PMID:27209517

  1. Comparative study between yeasts immobilized on alumina beads and on membranes prepared by two routes

    Directory of Open Access Journals (Sweden)

    Kiyohara Pedro K.

    2003-01-01

    Full Text Available Alumina channeled beads and rough surface membranes prepared from aqueous sols of fibrillar pseudoboehmite are able to immobilize yeasts for ethanol fermentation of sugar solutions. This paper describes comparative results of assays carried out with yeasts immobilized onto alpha-alumina beads and membranes prepared under two different conditions of processing and firing. The fermentation tests evaluated by the decrease of fermentable sugars, referred as Brix degrees per hour, indicated that the yeasts immobilized on beads had similar performance, probably because their surfaces, even being morphologically different, presented the same value of open porosity. One type of membrane (asymmetrical; precursor: pseudoboehmite; firing temperature 1,150ºC; crystal structure; alpha-alumina had better performance than the other type (asymmetrical; precursor: fibrillar pseudoboehmite plus aluminum hydroxiacetate mixture; 1,150ºC; alpha-alumina because the yeast cells entered into their porous interior through the surface slits, were immobilized and their growth was easier than on the external surface.

  2. Design of a papain immobilized antimicrobial food package with curcumin as a crosslinker.

    Science.gov (United States)

    Manohar, Cynthya Maria; Prabhawathi, Veluchamy; 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

  3. Immobilized synthetic pathway for biodegradation of toxic recalcitrant pollutant 1,2,3-trichloropropane.

    Science.gov (United States)

    Dvorak, Pavel; Bidmanova, Sarka; Damborsky, Jiri; Prokop, Zbynek

    2014-06-17

    The anthropogenic compound 1,2,3-trichloropropane (TCP) has recently drawn attention as an emerging groundwater contaminant. No living organism, natural or engineered, is capable of the efficient aerobic utilization of this toxic industrial waste product. We describe a novel biotechnology for transforming TCP based on an immobilized synthetic pathway. The pathway is composed of three enzymes from two different microorganisms: engineered haloalkane dehalogenase from Rhodococcus rhodochrous NCIMB 13064, and haloalcohol dehalogenase and epoxide hydrolase from Agrobacterium radiobacter AD1. Together, they catalyze consecutive reactions converting toxic TCP to harmless glycerol. The pathway was immobilized in the form of purified enzymes or cell-free extracts, and its performance was tested in batch and continuous systems. Using a packed bed reactor filled with the immobilized biocatalysts, 52.6 mmol of TCP was continuously converted into glycerol within 2.5 months of operation. The efficiency of the TCP conversion to the intermediates was 97%, and the efficiency of conversion to the final product glycerol was 78% during the operational period. Immobilized biocatalysts are suitable for removing TCP from contaminated water up to a 10 mM solubility limit, which is an order of magnitude higher than the concentration tolerated by living microorganisms. PMID:24787668

  4. Chromium immobilization by extra- and intraradical fungal structures of arbuscular mycorrhizal symbioses.

    Science.gov (United States)

    Wu, Songlin; Zhang, Xin; Sun, Yuqing; Wu, Zhaoxiang; Li, Tao; Hu, Yajun; Lv, Jitao; Li, Gang; Zhang, Zhensong; Zhang, Jing; Zheng, Lirong; Zhen, Xiangjun; Chen, Baodong

    2016-10-01

    Arbuscular mycorrhizal (AM) fungi can enhance plant Cr tolerance through immobilizing Cr in mycorrhizal roots. However, the detailed processes and mechanisms are unclear. The present study focused on cellular distribution and speciation of Cr in both extraradical mycelium (ERM) and mycorrhizal roots exposed to Cr(VI) by using field emission scanning electron microscopy equipped with energy dispersive X-ray spectrometer (FE-SEM-EDS), scanning transmission soft X-ray microscopy (STXM) and X-ray absorption fine structure (XAFS) spectroscopy techniques. We found that amounts of particles (possibly extracellular polymeric substances, EPS) were produced on the AM fungal surface upon Cr(VI) stress, which contributed greatly to Cr(VI) reduction and immobilization. With EDS of the surface of AM fungi exposed to various Cr(VI) levels, a positive correlation between Cr and P was revealed, suggesting that phosphate groups might act as counter ions of Cr(III), which was also confirmed by the XAFS analysis. Besides, STXM and XAFS analyses showed that Cr(VI) was reduced to Cr(III) in AM fungal structures (arbuscules, intraradical mycelium, etc.) and cell walls in mycorrhizal roots, and complexed possibly with carboxyl groups or histidine analogues. The present work provided evidence of Cr immobilization on fungal surface and in fungal structures in mycorrhizal roots at a cellular level, and thus unraveled the underlying mechanisms by which AM symbiosis immobilize Cr.

  5. Endothelialization of polyurethanes: Surface silanization and immobilization of REDV peptide.

    Science.gov (United States)

    Butruk-Raszeja, Beata A; Dresler, Magdalena S; Kuźmińska, Aleksandra; Ciach, Tomasz

    2016-08-01

    The paper presents method for chemical immobilization of arginine-glutamic acid-aspartic acid-valine (REDV) peptide on polyurethane surface. The peptide has been covalently bonded using silanes as a spacer molecules. The aim of this work was to investigate the proposed modification process and assess its biological effectiveness, especially in contact with blood and endothelial cells. Physicochemical properties were examined in terms of wettability, atomic composition and density of introduced functional groups and peptide molecules. Experiments with blood showed that material coating reduced number of surface-adhered platelets and fibrinogen molecules. In contrast to polyurethane (PU), there were no blood components deposited on REDV-modified materials (PU-REDV); fibrinogen adsorption on PU-REDV surface has been strongly reduced compared to PU. Analysis of cell adhesion after 1, 2, 3, 4, and 5 days of culture showed a significant increase of the cell-coated area on PU-REDV compared to PU. However, an intense cell growth appeared also on the control surface modified without the addition of REDV. Thus, the positive effect of REDV peptide on the adhesion of HUVEC could not be unequivocally confirmed. PMID:27110909

  6. Covalent immobilization of Pseudomonas cepacia lipase on semiconducting materials

    Science.gov (United States)

    Fernandez, Renny Edwin; Bhattacharya, Enakshi; Chadha, Anju

    2008-05-01

    Lipase from Pseudomonas cepacia was covalently immobilized on crystalline silicon, porous silicon and silicon nitride surfaces. The various stages of immobilization were characterized using FTIR (Fourier transform infrared) spectroscopy. The surface topography of the enzyme immobilized surfaces was investigated using scanning electron microscopy (SEM). The quantity of the immobilized active enzyme was estimated by the para-nitrophenyl palmitate (pNPP) assay. The immobilized lipase was used for triglyceride hydrolysis and the acid produced was detected by a pH sensitive silicon nitride surface as a shift in the C- V (capacitance-voltage) characteristics of an electrolyte-insulator-semiconductor capacitor (EISCAP) thus validating the immobilization method for use as a biosensor.

  7. Covalent immobilization of Pseudomonas cepacia lipase on semiconducting materials

    International Nuclear Information System (INIS)

    Lipase from Pseudomonas cepacia was covalently immobilized on crystalline silicon, porous silicon and silicon nitride surfaces. The various stages of immobilization were characterized using FTIR (Fourier transform infrared) spectroscopy. The surface topography of the enzyme immobilized surfaces was investigated using scanning electron microscopy (SEM). The quantity of the immobilized active enzyme was estimated by the para-nitrophenyl palmitate (pNPP) assay. The immobilized lipase was used for triglyceride hydrolysis and the acid produced was detected by a pH sensitive silicon nitride surface as a shift in the C-V (capacitance-voltage) characteristics of an electrolyte-insulator-semiconductor capacitor (EISCAP) thus validating the immobilization method for use as a biosensor

  8. Insulin action in human thighs after one-legged immobilization

    DEFF Research Database (Denmark)

    Richter, Erik; Kiens, Bente; Mizuno, M.;

    1989-01-01

    Insulin action was assessed in thighs of five healthy young males who had one knee immobilized for 7 days by a splint. The splint was not worn in bed. Subjects also used crutches to prevent weight bearing of the immobilized leg. Immobilization decreased the activity of citrate synthase and 3-OH......-acyl-CoA-dehydrogenase in the vastus lateralis muscle by 9 and 14%, respectively, and thigh volume by 5%. After 7 days of immobilization, a two-step euglycemic hyperinsulinemic clamp procedure combined with arterial and bilateral femoral venous catheterization was performed. Insulin action on glucose uptake and tyrosine...... release of the thighs at mean plasma insulin concentrations of 67 (clamp step I) and 447 microU/ml (clamp step II) was decreased by immobilization, whereas immobilization did not affect insulin action on thigh exchange of free fatty acids, glycerol, O2, or potassium. Before and during the clamp step I...

  9. Immobilization of activated sludge using improved polyvinyl alcohol (PVA) gel

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The microbial immobilization method using polyvinyl alcohol (PVA) gel as an immobilizing material was improved and used for entrapment of activated sludge. The OUR (oxygen uptake rate) was used to characterize the biological activity of immobilized activated sludge. Three kinds of PVA-immobilized particles of activated sludge, that is, PVA-boric acid beads, PVA-sodium nitrate beads and PVA-orthophosphate beads was prepared, and their biological activity was compared by measuring the OUR value. The bioactivity of both autotrophic and heterotrophic microorganisms of activated sludge was determined using different synthetic wastewater media (containing 250 mg/L COD and 25 mg/L NH4+-N). The experimental results showed that the bioactivity and stability of the three kinds of immobilized activated sludge was greatly improved after activation. With respect of the bioactivity and the mechanical stability, the PVA-orthophosphate method may be a promising and economical technique for microbial immobilization.

  10. Covalent immobilization of Pseudomonas cepacia lipase on semiconducting materials

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez, Renny Edwin [Microelectronics and MEMS Laboratory, Department of Electrical Engineering, Indian Institute of Technology Madras, Chennai (India)], E-mail: rennyedwin@gmail.com; Bhattacharya, Enakshi [Microelectronics and MEMS Laboratory, Department of Electrical Engineering, Indian Institute of Technology Madras, Chennai (India)], E-mail: enakshi@ee.iitm.ac.in; Chadha, Anju [Department of Biotechnology, National Centre for Catalysis Research, Indian Institute of Technology Madras, Chennai (India)], E-mail: anjuc@iitm.ac.in

    2008-05-30

    Lipase from Pseudomonas cepacia was covalently immobilized on crystalline silicon, porous silicon and silicon nitride surfaces. The various stages of immobilization were characterized using FTIR (Fourier transform infrared) spectroscopy. The surface topography of the enzyme immobilized surfaces was investigated using scanning electron microscopy (SEM). The quantity of the immobilized active enzyme was estimated by the para-nitrophenyl palmitate (pNPP) assay. The immobilized lipase was used for triglyceride hydrolysis and the acid produced was detected by a pH sensitive silicon nitride surface as a shift in the C-V (capacitance-voltage) characteristics of an electrolyte-insulator-semiconductor capacitor (EISCAP) thus validating the immobilization method for use as a biosensor.

  11. The immobilization of lipase on PVDF-co-HFP membrane

    Science.gov (United States)

    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.

  12. Limb immobilization alters functional electrophysiological parameters of sciatic nerve

    OpenAIRE

    J.S.M. Alves; J.H. Leal-Cardoso; F.F.U. Santos-Junior; Carlos, P.S.; Silva, R. C.; Lucci, C.M.; S. N. BAO; Ceccatto, V.M.; R. Barbosa

    2013-01-01

    Immobilization, used in clinical practice to treat traumatologic problems, causes changes in muscle, but it is not known whether changes also occur in nerves. We investigated the effects of immobilization on excitability and compound action potential (CAP) and the ultrastructure of the rat sciatic nerve. Fourteen days after immobilization of the right leg of adult male Wistar rats (n=34), animals were killed and the right sciatic nerve was dissected and mounted in a moist chamber. Nerves were...

  13. Ex vivo model of an immobilized-enzyme reactor.

    OpenAIRE

    Bernstein, H; Langer, R

    1988-01-01

    Immobilized-enzyme reactors are beginning to be studied for a variety of therapeutic applications. To facilitate the design of these devices for different clinical situations and a diverse patient population, mathematical models may be valuable. An immobilized-heparinase (EC 4.2.2.7) reactor was selected as a model system. The device removes heparin from blood that has been anticoagulated to prevent thrombus formation. Heparinase was immobilized to cross-linked agarose particles. A mathematic...

  14. Preparation of Magnetic Chitosan Nanoparticles and Immobilization of Laccase

    Institute of Scientific and Technical Information of China (English)

    FANG Hua; HUANG Jun; DING Liyun; LI Mingtian; CHEN Zhao

    2009-01-01

    The magnetic chitosan nanoparticles were prepared by reversed-phase suspension method using Span-80 as an emulsifier, glutaraldehyde as cross-linking reagent. And the nanoparticles were characterized by TEM, FT-IR and hysteresis loop. The results show that the nanoparticles are spherical and almost superparamagnetic. The laccase was immobilized on nanoparticles by adsorption and subsequently by cross-linking with glutaraldehyde. The immobilization conditions and charac-terizations of the immobilized laccase were investigated. The optimal immobilization conditions were as follows: 10 mL of phosphate buffer (0.1 M, pH 7.0) containing 50 mg of magnetic chitosan nanoparticles, 1.0 mg·mL-1 of laccase and 1% (v/v) glutaraldehyde, immobilization temperature of 4 ℃ and immobilization time of 4 h. The immobilized laccase exhibited an appreciable catalytic capability (480 units·g-1 support) and had good storage stability and operation stability. The Km of immobilized and free laccase for ABTS were 140.6 and 31.1 μM in phosphate buffer (0.1 M, pH 3.0) at 37 ℃, respectively. The immobilized laccase is a good candidate for the research and development of biosensors based on laccase catalysis.

  15. Laccase immobilized on magnetic carriers for biotechnology applications

    Energy Technology Data Exchange (ETDEWEB)

    Rotkova, Jana [Department of Biological and Biochemical Sciences, University of Pardubice, Strossova 239, 530 03 Pardubice (Czech Republic); Sulakova, Romana [Department of Technology of Organic Compounds, Doubravice 41, 533 53 Pardubice (Czech Republic); Korecka, Lucie; Zdrazilova, Pavla; Jandova, Miroslava [Department of Biological and Biochemical Sciences, University of Pardubice, Strossova 239, 530 03 Pardubice (Czech Republic); Lenfeld, Jiri; Horak, Daniel [Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovskeho nam. 2, 162 06 Praha (Czech Republic); Bilkova, Zuzana [Department of Biological and Biochemical Sciences, University of Pardubice, Strossova 239, 530 03 Pardubice (Czech Republic)], E-mail: Zuzana.Bilkova@upce.cz

    2009-05-15

    Laccase catalyzing the oxidation of p-diphenols has been applied in many industrial and biotechnology areas. Immobilized form of laccase has overcome the problem with contamination of the final product. Nevertheless sensitive enzymes immobilized to the matrix can be inactivated by the environmental conditions. The aim of this research was to prepare carrier with improved activity and responsible stability even under extreme reaction conditions. Laccase immobilized through carbohydrate moieties on magnetic hydrazide bead cellulose with a final activity of 0.63 I.U./1 ml of settled carrier confirmed that carriers with oriented immobilized enzyme might be useful in routine biocatalytic applications.

  16. Biochemical studies on immobilized fungal β-glucosidase

    Directory of Open Access Journals (Sweden)

    S. A. Ahmed

    2013-12-01

    Full Text Available β-Glucosidase from Aspergillus niger was immobilized on sponge by covalent binding through a spacer group (glutaraldehyde. Sponge-immobilized enzyme had the highest immobilization yield (95.67% and retained 63.66% of the original activity exhibited by the free enzyme. The optimum pH of the immobilized enzyme remains almost the same as for the free enzyme (pH 4.0. The optimum temperature for β-glucosidase activity was increased by 10 ºC after immobilization. The activation energy (Ea of the immobilized β-glucosidase was lower than the free enzyme (3.34 and 4.55 kcal/mol, respectively. Immobilized β-glucosidase exhibited great thermal stability and retained all the initial activity after incubation at 55 ºC for 2 h; however, the free enzyme retained 89.25% under the same condition. The calculated half-life (t½ value of heat inactivation of immobilized enzyme at 60, 65 and 70 ºC was 213.62, 72.95 and 56.80 min, respectively, whereas at these temperatures the free enzyme was less stable (half-life of 200.0, 55.31 and 49.5 min, respectively. The deactivation rate constant at 65 ºC for the immobilized β-glucosidase is 9.5x10-3/ min, which was lower than that of the free form (12.53x10-3/ min. The immobilization process improved the pH stability of the enzyme (immobilized and free enzyme retained 69.35 and 39.86%, respectively, of their initial activity after 45 min at pH 7.5. The effect of some chemical substances on the activity of the immobilized and free β-glucosidase has been investigated. In the presence of sodium dodecyl sulfate (SDS and p-chloromercuri benzoate (p-CMB the immobilized enzyme retained 36.13 and 45.34%, respectively, of the initial activity, which is higher than that of free enzyme (13.71 and 1.61%, respectively. The Michaelis constant (Km value of the free enzyme was 40.0 mM, while the apparent Km value for the immobilized enzyme was 46.51 mM. The maximum reaction rate (v max of immobilized β-glucosidase was smaller

  17. Production and Immobilization of Partially Purified Lipase From Penicillium chrysogenum

    OpenAIRE

    Shafei, M. S.; Allam, R F

    2010-01-01

    An extracellular lipase from Penicillium chrysogenum produced maximal activity 225 U/mL after four days at pH 6.5. It was partially purified 4.1 fold by ammonium sulphate precipitation (70%). The enzyme was immobilized on various carriers viz. alginate, k-carrageenan and polyacrylamide gel. The immobilization yield of enzyme immobilized in kcarrageenan and polyacrylamide gel (63.41% and 48.93% respectively) was low in comparison to that immobilized with alginate (81.57%). Different concentra...

  18. Immobilization of thorium over fibroin by polyacrylonitrile (PAN)

    Energy Technology Data Exchange (ETDEWEB)

    Aslani, M.A.A.; Akyil, S.; Eral, M. [Ege Univ. Inst. of Nuclear Sciences, Bornova-Izmir (Turkey)

    1997-12-31

    This report describes a process for immobilization of thorium over fibroin, which was used as a bio-adsorbant, by polyacrylonitrile. The amounts of thorium in aqueous solutions which may be leached in various aqueous ambients were detected by a spectrophotometer. The results show that polyacrylonitrile processes are feasible to immobilize spent fibroins. The leachability of the materials immobilized with polyacrylonitrile can meet the requirements of storage and final disposal. The leachability of thorium ions from immobilized spent fibroin was rather low for 8 months.

  19. Immobilization of Lipase by Covalent Binding on Crosslinked Ally Dextran

    Institute of Scientific and Technical Information of China (English)

    WangChen; SongGuoqiang; 等

    1998-01-01

    Lipase was immobilized by covalent binding on crosslinked allyl dextran using SESA as coupling agent.It is shown that this immobilization approach is an efficient one for lipase.The activity of the immobilized lipase can reach to 300-450U/g(dry weight).It exhibits good temperature stability,can retain 88% activity after being incubated at 70℃ for 2h.Special effects will be expected from our immobilized lipase in its applications in organic media due to the nature of the support.

  20. Arsenic mobilization and immobilization in paddy soils

    Science.gov (United States)

    Kappler, A.; Hohmann, C.; Zhu, Y. G.; Morin, G.

    2010-05-01

    Arsenic is oftentimes of geogenic origin and in many cases bound to iron(III) minerals. Iron(III)-reducing bacteria can harvest energy by coupling the oxidation of organic or inorganic electron donors to the reduction of Fe(III). This process leads either to dissolution of Fe(III)-containing minerals and thus to a release of the arsenic into the environment or to secondary Fe-mineral formation and immobilisation of arsenic. Additionally, aerobic and anaerobic iron(II)-oxidizing bacteria have the potential to co-precipitate or sorb arsenic during iron(II) oxidation at neutral pH that is usually followed by iron(III) mineral precipitation. We are currently investigating arsenic immobilization by Fe(III)-reducing bacteria and arsenic co-precipitation and immobilization by anaerobic iron(II)-oxidizing bacteria in batch, microcosm and rice pot experiments. Co-precipitation batch experiments with pure cultures of nitrate-dependent Fe(II)-oxidizing bacteria are used to quantify the amount of arsenic that can be immobilized during microbial iron mineral precipitation, to identify the minerals formed and to analyze the arsenic binding environment in the precipitates. Microcosm and rice pot experiments are set-up with arsenic-contaminated rice paddy soil. The microorganisms (either the native microbial population or the soil amended with the nitrate-dependent iron(II)-oxidizing Acidovorax sp. strain BoFeN1) are stimulated either with iron(II), nitrate, or oxygen. Dissolved and solid-phase arsenic and iron are quantified. Iron and arsenic speciation and redox state in batch and microcosm experiments are determined by LC-ICP-MS and synchrotron-based methods (EXAFS, XANES).

  1. An Amperometric Biosensor for Glucose Determination Prepared from Glucose Oxidase Immobilized in Polyaniline-Polyvinylsulfonate Film

    OpenAIRE

    Halit Arslan; Selvin Ustabaş; Fatma Arslan

    2011-01-01

    In this study, a novel amperometric glucose biosensor with immobilization of glucose oxidase on electrochemically polymerized polyaniline-polyvinylsulphonate (Pani-Pvs) films has been accomplished via the entrapment technique. Electropolymerization of aniline on the Pt surface of the Pt electrode was carried out at constant potential (0.75 V, vs. Ag/AgCl) using an electrochemical cell containing aniline and polyvinylsulphonate. Firstly, the optimum working conditions for preparing polyaniline...

  2. Efficient biodegradation of chlorophenols in aqueous phase by magnetically immobilized aniline-degrading Rhodococcus rhodochrous strain

    OpenAIRE

    Hou, Jianfeng; Liu, Feixia; Wu, Nan; Ju, Jiansong; Yu, Bo

    2016-01-01

    Background Chlorophenols are environmental contaminants, which are highly toxic to living beings due to their carcinogenic, mutagenic and cytotoxic properties. Bacterial degradation has been considered a cost-effective and eco-friendly method of removing chlorophenols, compared to the traditional physical–chemical processes. Results In this study, we first developed an efficient process for the biodegradation of chlorophenols by magnetically immobilized Rhodococcus rhodochrous cells. R. rhodo...

  3. Detoxification of Corncob Acid Hydrolysate with SAA Pretreatment and Xylitol Production by Immobilized Candida tropicalis

    OpenAIRE

    Li-Hong Deng; Yong Tang; Yun Liu

    2014-01-01

    Xylitol fermentation production from corncob acid hydrolysate has become an attractive and promising process. However, corncob acid hydrolysate cannot be directly used as fermentation substrate owing to various inhibitors. In this work, soaking in aqueous ammonia (SAA) pretreatment was employed to reduce the inhibitors in acid hydrolysate. After detoxification, the corncob acid hydrolysate was fermented by immobilized Candida tropicalis cell to produce xylitol. Results revealed that SAA pretr...

  4. Immobilization of degradative bacteria in polyurethane-based foams: embedding efficiency and effect on bacterial activity

    Energy Technology Data Exchange (ETDEWEB)

    Wilde, E.W. [Westinghouse Savannah River Company, Aiken, SC (United States); Radway, J.C.; Hazen, T.C.; Hermann, P. [Matrix R and D Corp., Dover, NH (United States)

    1996-09-03

    The immobilization of TCE-degrading bacterium Burkholderia cepacia was evaluated using hydrophilic polyurethane foam. The influence of several foam formulation parameters upon cell retention was examined. Surfactant type was a major determinant of retention, with a lecithin- based compound retaining more cells than pluronic or silicone based surfactants. Excessive amounts of surfactant led to increased washout of bacteria. Increasing the biomass concentration from 4.8% to 10.5% caused fewer cells to be washed out. Embedding at reduced temperature did not significantly affect retention, while the use of a silane binding agent gave inconsistent results. The optimal formulation retained all but 0.2% of total embedded cells during passage of 2 liters of water through columns containing 2 g of foam. All foam formulations tested reduced the culturability of embedded cells by several orders of magnitude. However, O{sub 2} and CO{sub 2} evolution rates of embedded cells were never less than 50% of unembedded cells. Nutrient amendments stimulated an increase in cell volume and ribosomal activity as indicated by hybridization studies using fluorescently labeled ribosomal probes. these results indicated that, although immobilized cells were nonculturable, they were metabolically active and thus could be used for biodegradation of toxic compounds.

  5. Asymmetric Synthesis of (-)-1-Trimethylsilyl-ethanol with Immobilized Saccharomyces Cerevisiae Cells in Water/Organic Solvent Biphasic System%水/有机溶剂双相中固定化啤酒酵母细胞催化三甲基硅乙酮不对称还原

    Institute of Scientific and Technical Information of China (English)

    娄文勇; 宗敏华; 范晓丹

    2003-01-01

    Asymmetric synthesis of (-)-1-trimethylsilyl-ethanol with immobilized Saccharomyces cerevisiae cells in water/organic solvent biphasic system was studied. The effects of shake speed, hydrophobicity of organic solvent,volume ratio of water phase to organic phase, pH value of aqueous phase and reaction temperature on the initial reaction rate, maximum yield and enantiomeric excess (ee) of the product were systematically explored. All the above-mentioned factors had significant influence on the reaction. n-Hexane was found to be the best organic solvent for the reaction. The optimum shake speed, volume ratio of water phase to organic phase, pH value and reaction temperature were 150 r.min- 1, 1/2, 8 and 30℃ respectively, under which the maximum yield and enantiomeric excess of the product were as high as 96.8% and 95.7%, which are 15% and 16% higher than those of the corresponding reaction performed in aqueous phase. To our best knowledge, this is the most satisfactory result obtained.

  6. Tritium immobilization and packaging using metal hydrides

    International Nuclear Information System (INIS)

    Tritium recovered from CANDU heavy water reactors will have to be packaged and stored in a safe manner. Tritium will be recovered in the elemental form, T2. Metal tritides are effective compounds in which to immobilize the tritium as a stable non-reactive solid with a high tritium capacity. The technology necessary to prepare hydrides of suitable metals, such as titanium and zirconium, have been developed and the properties of the prepared materials evaluated. Conceptual designs of packages for containing metal tritides suitable for transportation and long-term storage have been made and initial testing started. (author)

  7. Detoxification of Corncob Acid Hydrolysate with SAA Pretreatment and Xylitol Production by Immobilized Candida tropicalis

    Directory of Open Access Journals (Sweden)

    Li-Hong Deng

    2014-01-01

    Full Text Available Xylitol fermentation production from corncob acid hydrolysate has become an attractive and promising process. However, corncob acid hydrolysate cannot be directly used as fermentation substrate owing to various inhibitors. In this work, soaking in aqueous ammonia (SAA pretreatment was employed to reduce the inhibitors in acid hydrolysate. After detoxification, the corncob acid hydrolysate was fermented by immobilized Candida tropicalis cell to produce xylitol. Results revealed that SAA pretreatment showed high delignification and efficient removal of acetyl group compounds without effect on cellulose and xylan content. Acetic acid was completely removed, and the content of phenolic compounds was reduced by 80%. Furthermore, kinetic behaviors of xylitol production by immobilized C. tropicalis cell were elucidated from corncob acid hydrolysate detoxified with SAA pretreatment and two-step adsorption method, respectively. The immobilized C. tropicalis cell showed higher productivity efficiency using the corncob acid hydrolysate as fermentation substrate after detoxification with SAA pretreatment than by two-step adsorption method in the five successive batch fermentation rounds. After the fifth round fermentation, about 60 g xylitol/L fermentation substrate was obtained for SAA pretreatment detoxification, while about 30 g xylitol/L fermentation substrate was obtained for two-step adsorption detoxification.

  8. A comparison of redox polymer and enzyme co-immobilization on carbon electrodes to provide membrane-less glucose/O2 enzymatic fuel cells with improved power output and stability.

    Science.gov (United States)

    Rengaraj, Saravanan; Kavanagh, Paul; Leech, Dónal

    2011-12-15

    Glassy carbon and graphite electrodes modified with films of enzyme and osmium redox polymer, cross linked with poly (ethylene glycol) diglycidyl ether, were used for elaboration of a glucose/O(2) enzymatic fuel cell. The redox polymers [Os(4,4'-dimethoxy-2,2'-bipyridine)(2)(polyvinylimidazole)(10)Cl](+) and [Os(4,4'-dichloro-2,2'-bipyridine)(2)(polyvinylimidazole)(10)Cl](+) were selected to facilitate transfer of electrons from the glucose oxidase (GOx) active site to the T1 Cu site of multicopper oxygenases of Trametes hirsuta laccase (ThLacc) and Myrothecium verrucaria bilirubin oxidase (MvBOD). Maximum power density at pH 5.5 of 3.5 μW cm(-2) at a cell voltage of 0.35 V was obtained for an assembled membrane-less fuel cell based on ThLacc on glassy carbon as cathode, in the presence of 0.1 M glucose, 37 °C, with lower power observed at pH 7.4 and 4.5. Replacement of the ThLacc cathode with that of MvBOD produced 10 μW cm(-2) at 0.25 V under pseudo-physiological conditions. Replacement of glassy carbon with graphite as base electrode material resulted in increased redox polymer loading, leading to an increase in power output to 43 μW cm(-2) at 0.25 V under similar conditions. Improved stabilization of biofilms was achieved through covalent anchoring of enzyme and redox polymer on graphite electrodes, derivatized via electrochemical reduction of the diazonium cation generated in situ from p-phenylenediamine. Enzymatic fuel cells using this approach retained 70% power at 24 h, whereas fuel cells prepared without chemical anchoring to graphite retained only 10% of power over the same interval. PMID:22005596

  9. Animal Bone Char Solubilization with Itaconic Acid Produced by Free and Immobilized Aspergillus terreus Grown on Glycerol-Based Medium

    NARCIS (Netherlands)

    Vassilev, N.; Medina, A.; Eichler-Lobermann, B.; Flor-Peregrin, E.; Vassileva, M.

    2012-01-01

    Cells of Aspergillus terreus, free and immobilized in polyurethane foam, were employed in itaconic acid fermentation processes on glycerol-based media. The purpose was to assess their suitability for animal bone char solubilization and the development of a biotechnological alternative to P fertilize

  10. Plutonium immobilization in glass and ceramics

    Energy Technology Data Exchange (ETDEWEB)

    Knecht, D.A. [Lockheed Martin Idaho Technologies, Idaho Falls (United States); Murphy, W.M. [Southwest Research Institute, San Antonio, TX (United States)

    1996-05-01

    The Materials Research Society Nineteenth Annual Symposium on the Scientific Basis for Nuclear Waste Management was held in Boston on November 27 to December 1, 1995. Over 150 papers were presented at the Symposium dealing with all aspects of nuclear waste management and disposal. Fourteen oral sessions and on poster session included a Plenary session on surplus plutonium dispositioning and waste forms. The proceedings, to be published in April, 1996, will provide a highly respected, referred compilation of the state of scientific development in the field of nuclear waste management. This paper provides a brief overview of the selected Symposium papers that are applicable to plutonium immobilization and plutonium waste form performance. Waste forms that were described at the Symposium cover most of the candidate Pu immobilization options under consideration, including borosilicate glass with a melting temperature of 1150 {degrees}C, a higher temperature (1450 {degrees}C) lanthanide glass, single phase ceramics, multi-phase ceramics, and multi-phase crystal-glass composites (glass-ceramics or slags). These Symposium papers selected for this overview provide the current status of the technology in these areas and give references to the relevant literature.

  11. Immobilization of Polymeric Luminophor on Nanoparticles Surface.

    Science.gov (United States)

    Bolbukh, Yuliia; Podkoscielna, Beata; Lipke, Agnieszka; Bartnicki, Andrzej; Gawdzik, Barbara; Tertykh, Valentin

    2016-12-01

    Polymeric luminophors with reduced toxicity are of the priorities in the production of lighting devices, sensors, detectors, bioassays or diagnostic systems. The aim of this study was to develop a method of immobilization of the new luminophor on a surface of nanoparticles and investigation of the structure of the grafted layer. Monomer 2,7-(2-hydroxy-3-methacryloyloxypropoxy)naphthalene (2,7-NAF.DM) with luminophoric properties was immobilized on silica and carbon nanotubes in two ways: mechanical mixing with previously obtained polymer and by in situ oligomerization with chemisorption after carrier's modification with vinyl groups. The attached polymeric (or oligomeric) surface layer was studied using thermal and spectral techniques. Obtained results confirm the chemisorption of luminophor on the nanotubes and silica nanoparticles at the elaborated synthesis techniques. The microstructure of 2,7-NAF.DM molecules after chemisorption was found to be not changed. The elaborated modification approach allows one to obtain nanoparticles uniformly covered with polymeric luminophor. PMID:27090657

  12. Limb immobilization alters functional electrophysiological parameters of sciatic nerve.

    Science.gov (United States)

    Alves, J S M; Leal-Cardoso, J H; Santos-Júnior, F F U; Carlos, P S; Silva, R C; Lucci, C M; Báo, S N; Ceccatto, V M; Barbosa, R

    2013-08-01

    Immobilization, used in clinical practice to treat traumatologic problems, causes changes in muscle, but it is not known whether changes also occur in nerves. We investigated the effects of immobilization on excitability and compound action potential (CAP) and the ultrastructure of the rat sciatic nerve. Fourteen days after immobilization of the right leg of adult male Wistar rats (n=34), animals were killed and the right sciatic nerve was dissected and mounted in a moist chamber. Nerves were stimulated at a baseline frequency of 0.2 Hz and tested for 2 min at 20, 50, and 100 Hz. Immobilization altered nerve excitability. Rheobase and chronaxy changed from 3.13 ± 0.05 V and 52.31 ± 1.95 µs (control group, n=13) to 2.84 ± 0.06 V and 59.71 ± 2.79 µs (immobilized group, n=15), respectively. Immobilization altered the amplitude of CAP waves and decreased the conduction velocity of the first CAP wave (from 93.63 ± 7.49 to 79.14 ± 5.59 m/s) but not of the second wave. Transmission electron microscopy showed fragmentation of the myelin sheath of the sciatic nerve of immobilized limbs and degeneration of the axon. In conclusion, we demonstrated that long-lasting leg immobilization can induce alterations in nerve function. PMID:23969978

  13. Limb immobilization alters functional electrophysiological parameters of sciatic nerve

    Directory of Open Access Journals (Sweden)

    J.S.M. Alves

    2013-08-01

    Full Text Available Immobilization, used in clinical practice to treat traumatologic problems, causes changes in muscle, but it is not known whether changes also occur in nerves. We investigated the effects of immobilization on excitability and compound action potential (CAP and the ultrastructure of the rat sciatic nerve. Fourteen days after immobilization of the right leg of adult male Wistar rats (n=34, animals were killed and the right sciatic nerve was dissected and mounted in a moist chamber. Nerves were stimulated at a baseline frequency of 0.2 Hz and tested for 2 min at 20, 50, and 100 Hz. Immobilization altered nerve excitability. Rheobase and chronaxy changed from 3.13±0.05 V and 52.31±1.95 µs (control group, n=13 to 2.84±0.06 V and 59.71±2.79 µs (immobilized group, n=15, respectively. Immobilization altered the amplitude of CAP waves and decreased the conduction velocity of the first CAP wave (from 93.63±7.49 to 79.14±5.59 m/s but not of the second wave. Transmission electron microscopy showed fragmentation of the myelin sheath of the sciatic nerve of immobilized limbs and degeneration of the axon. In conclusion, we demonstrated that long-lasting leg immobilization can induce alterations in nerve function.

  14. Evaluation of Fungal Laccase Immobilized on Natural Nanostructured Bacterial Cellulose.

    Science.gov (United States)

    Chen, Lin; Zou, Min; Hong, Feng F

    2015-01-01

    The aim of this work was to assess the possibility of using native bacterial nanocellulose (BC) as a carrier for laccase immobilization. BC was synthesized by Gluconacetobacter xylinus, which was statically cultivated in a mannitol-based medium and was freeze-dried to form BC sponge after purification. For the first time, fungal laccase from Trametes versicolor was immobilized on the native nanofibril network-structured BC sponge through physical adsorption and cross-linking with glutaraldehyde. The properties including morphologic and structural features of the BC as well as the immobilized enzyme were thoroughly investigated. It was found that enzyme immobilized by cross-linking exhibited broader pH operation range of high catalytic activity as well as higher running stability compared to free and adsorbed enzyme. Using ABTS as substrate, the optimum pH value was 3.5 for the adsorption-immobilized laccase and 4.0 for the crosslinking-immobilized laccase. The immobilized enzyme retained 69% of the original activity after being recycled seven times. Novel applications of the BC-immobilized enzyme tentatively include active packaging, construction of biosensors, and establishment of bioreactors.

  15. Immobilization of fission products in phosphate ceramic waste forms

    Energy Technology Data Exchange (ETDEWEB)

    Singh, D. [Argonne National Lab., IL (United States)

    1996-10-01

    The goal of this project is to develop and demonstrate the feasibility of a novel low-temperature solidification/stabilization (S/S) technology for immobilizing waste streams containing fission products such as cesium, strontium, and technetium in a chemically bonded phosphate ceramic. This technology can immobilize partitioned tank wastes and decontaminate waste streams containing volatile fission products.

  16. Evaluation of fungal laccase immobilized on natural nanostructured bacterial cellulose

    Directory of Open Access Journals (Sweden)

    Lin eChen

    2015-11-01

    Full Text Available The aim of this work was to assess the possibility of using native bacterial nanocellulose (BC as a carrier for laccase immobilization. BC was synthesized by Gluconacetobacter xylinus, which was statically cultivated in a mannitol-based medium and was freeze-dried to form BC sponge after purification. For the first time, fungal laccase from Trametes versicolor was immobilized on the native nanofibril network-structured BC sponge through physical adsorption and cross-linking with glutaraldehyde. The properties including morphologic and structural features of the BC as well as the immobilized enzyme were thoroughly investigated. It was found that enzyme immobilized by cross-linking exhibited broader pH operation range of high catalytic activity as well as higher running stability compared to free and adsorbed enzyme. Using ABTS as substrate, the optimum pH value was 3.5 for the adsorption-immobilized laccase and 4.0 for the crosslinking-immobilized laccase. The immobilized enzyme retained 69% of the original activity after being recycled 7 times. Novel applications of the BC-immobilized enzyme tentatively include active packaging, construction of biosensors, and establishment of bioreactors.

  17. Evaluation of Fungal Laccase Immobilized on Natural Nanostructured Bacterial Cellulose.

    Science.gov (United States)

    Chen, Lin; Zou, Min; Hong, Feng F

    2015-01-01

    The aim of this work was to assess the possibility of using native bacterial nanocellulose (BC) as a carrier for laccase immobilization. BC was synthesized by Gluconacetobacter xylinus, which was statically cultivated in a mannitol-based medium and was freeze-dried to form BC sponge after purification. For the first time, fungal laccase from Trametes versicolor was immobilized on the native nanofibril network-structured BC sponge through physical adsorption and cross-linking with glutaraldehyde. The properties including morphologic and structural features of the BC as well as the immobilized enzyme were thoroughly investigated. It was found that enzyme immobilized by cross-linking exhibited broader pH operation range of high catalytic activity as well as higher running stability compared to free and adsorbed enzyme. Using ABTS as substrate, the optimum pH value was 3.5 for the adsorption-immobilized laccase and 4.0 for the crosslinking-immobilized laccase. The immobilized enzyme retained 69% of the original activity after being recycled seven times. Novel applications of the BC-immobilized enzyme tentatively include active packaging, construction of biosensors, and establishment of bioreactors. PMID:26617585

  18. Stability improvement of immobilized lactoperoxidase using polyaniline polymer.

    Science.gov (United States)

    Jafary, Fariba; Kashanian, Soheila; Sharieat, Ziadin Samsam; Jafary, Farzaneh; Omidfar, Kobra; Paknejad, Maliheh

    2012-12-01

    Enzyme engineering via immobilization techniques is perfectly compatible against the other chemical or biological approximate to improve enzyme functions and stability. In this study lactoperoxidase was immobilized onto polyaniline polymer activated with glutaraldehyde as a bifunctional agent, to improve enzyme properties. Polyaniline polymer was used due its unique physical and chemical properties to immobilize lactoperoxidase (LPO). The optimum activity of immobilized LPO was observed at pH 6 and 55 °C, which has been increased about 10 °C for the immobilized enzyme. The immobilized enzyme maintained absolutely active for 60 days whereas the native enzyme lost 80 % of its initial activity within this period of time. Moreover, the immobilized enzyme can be reused for several times without loss of activity. The kinetic parameter studies showed slight differences between free and immobilized enzymes. The K(m) and K(m.app) were calculated to be 0.6 and 0.4; also V(max) and V(max.app) were 1.3 and 0.9 respectively.

  19. DOE materials program supporting immobilization of radioactive wastes

    International Nuclear Information System (INIS)

    A summary is presented of the DOE program for developing waste-form criteria, immobilization processes, and generation and evaluation of performance characterization data. Interrelationships are discussed among repository design, materials requirements, immobilization process definition, quality assurance, and risk analysis as part of the National Environmental Policy Act and regulatory processes

  20. Evaluation of Fungal Laccase Immobilized on Natural Nanostructured Bacterial Cellulose

    Science.gov (United States)

    Chen, Lin; Zou, Min; Hong, Feng F.

    2015-01-01

    The aim of this work was to assess the possibility of using native bacterial nanocellulose (BC) as a carrier for laccase immobilization. BC was synthesized by Gluconacetobacter xylinus, which was statically cultivated in a mannitol-based medium and was freeze-dried to form BC sponge after purification. For the first time, fungal laccase from Trametes versicolor was immobilized on the native nanofibril network-structured BC sponge through physical adsorption and cross-linking with glutaraldehyde. The properties including morphologic and structural features of the BC as well as the immobilized enzyme were thoroughly investigated. It was found that enzyme immobilized by cross-linking exhibited broader pH operation range of high catalytic activity as well as higher running stability compared to free and adsorbed enzyme. Using ABTS as substrate, the optimum pH value was 3.5 for the adsorption-immobilized laccase and 4.0 for the crosslinking-immobilized laccase. The immobilized enzyme retained 69% of the original activity after being recycled seven times. Novel applications of the BC-immobilized enzyme tentatively include active packaging, construction of biosensors, and establishment of bioreactors. PMID:26617585

  1. Preparation and characterization of immobilized lipase on magnetic hydrophobic microspheres

    DEFF Research Database (Denmark)

    Guo, Zheng; Bai, Shu; Sun, Yan

    2003-01-01

    than the gravitational sedimentation. Candida cylindracea lipase (CCL) was immobilized to the porous carrier at up to 6750 IU/g carrier, remarkably higher than the previous studies. The pH and temperature dependencies of the immobilized CCL were investigated and the optimum temperature and p...

  2. Affinity-based in situ product removal coupled with co-immobilization of oily substrate and filamentous fungus.

    Science.gov (United States)

    Dukler, A; Freeman, A

    1998-01-01

    In situ product removal (ISPR) involves actions taken for the fast removal of a product from the producing cell. ISPR is implemented to improve yield and productivity via minimization of product inhibition, minimization of product losses due to degradation or evaporation, and reduction of the number of subsequent downstream processing steps. Here we describe the implementation of affinity-based, specific ISPR as a crucial component of an integrative approach to problems associated with the biocatalytic production of a product exhibiting poor water solubility from an oily, water-insoluble precursor. Our integrative ISPR-based approach consists of co-immobilization of the oily substrate emulsion and the biocatalyst within bilayered alginate beads. A particulate-specific adsorbent, exhibiting high binding capacity of the product, is suspended in the reaction medium with periodical replacements. According to this approach, ISPR implementation is expected to shift the equilibration of product distribution between the co-immobilized oily substrate and the outer medium via specific product immobilization onto the added adsorbent. The product may subsequently be readily recovered via single-step final purification. This integrative approach was successfully demonstrated by the affinity-based ISPR of gamma-decalactone (4-decanolide). gamma-Decalactone was produced from castor oil via its beta-oxidation by the filamentous fungus Tyromyces sambuceus, co-immobilized with emulsified substrate within bilayered alginate beads. Product immobilization onto medium-suspended epichlorohydrin-crosslinked beta-cyclodextrin resulted in higher yield and easy pure product recovery. PMID:10076845

  3. Non-constrictive bead immobilization leading to decreased and uniform shear stress in microfluidic bead-based ELISA

    CERN Document Server

    Mitra, Kinshuk; Chidambaram, Preethi; Maharry, Aaron P; Xu, Ronald X; Tweedle, Michael F

    2014-01-01

    Microfluidic biosensors have been utilized for sensing a wide range of antigens using numerous configurations. Bead based microfluidic sensors have been a popular modality due to the plug and play nature of analyte choice and the favorable geometry of spherical sensor scaffolds. While constriction of beads against fluid flow remains a popular method to immobilize the sensor, it results in poor fluidic regimes and shear conditions around sensor beads that can affect sensor performance. We present an alternative means of sensor bead immobilization using poly-carbonate membrane. This system results in several orders of magnitude lower variance of flow radially around the sensor bead. Shear stress experienced by our non-constrictive immobilized bead was three orders of magnitude lower. We demonstrate ability to quantitatively sense EpCAM protein, a marker for cancer stem cells and operation under both far-red and green wavelengths with no auto-fluorescence.

  4. Design-Only Conceptual Design Report: Plutonium Immobilization Plant

    Energy Technology Data Exchange (ETDEWEB)

    DiSabatino, A.; Loftus, D.

    1999-01-01

    This design-only conceptual design report was prepared to support a funding request by the Department of Energy Office of Fissile Materials Disposition for engineering and design of the Plutonium Immobilization Plant, which will be used to immobilize up to 50 tonnes of surplus plutonium. The siting for the Plutonium Immobilization Plant will be determined pursuant to the site-specific Surplus Plutonium Disposition Environmental Impact Statement in a Plutonium Deposition Record of Decision in early 1999. This document reflects a new facility using the preferred technology (ceramic immobilization using the can-in-canister approach) and the preferred site (at Savannah River). The Plutonium Immobilization Plant accepts plutonium from pit conversion and from non-pit sources and, through a ceramic immobilization process, converts the plutonium into mineral-like forms that are subsequently encapsulated within a large canister of high-level waste glass. The final immobilized product must make the plutonium as inherently unattractive and inaccessible for use in nuclear weapons as the plutonium in spent fuel from commercial reactors and must be suitable for geologic disposal. Plutonium immobilization at the Savannah River Site uses: (1) A new building, the Plutonium Immobilization Plant, which will convert non-pit surplus plutonium to an oxide form suitable for the immobilization process, immobilize plutonium in a titanate-based ceramic form, place cans of the plutonium-ceramic forms into magazines, and load the magazines into a canister; (2) The existing Defense Waste Processing Facility for the pouring of high-level waste glass into the canisters; and (3) The Actinide Packaging and Storage Facility to receive and store feed materials. The Plutonium Immobilization Plant uses existing Savannah River Site infra-structure for analytical laboratory services, waste handling, fire protection, training, and other support utilities and services. The Plutonium Immobilization Plant

  5. Electron beam technology for production of preparations of immobilized enzymes

    International Nuclear Information System (INIS)

    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 is done due to free-radical linking of enzyme and carrier with formation of mycelium-like structures. Immobilization improves heat resistance of enzyme up to 60 centigrade without substrate and up to 80 centigrade in presence of substrate, widens range pH activity in comparison with non-immobilized forms. Immobilized proteases does not contain peroxides and 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

  6. STUDIES ON IMMOBILIZED GLUCOSE OXIDASE BY DIETHYLAMINOETHYL CELLULOSE COMPLEXES

    Institute of Scientific and Technical Information of China (English)

    WANG Lingzhi; YUAN Hong; FANG Shibi; JIANG Yingyan

    1993-01-01

    The properties of immobilized glucose oxidase (GOD) by the complexes of diethylaminoethyl cellu -lose(DEAEC) with different polymers, such as polymethylacrylic acid (PMAA), polyacrylic acid (PAA), polystyrene sulfonic acid (PSSA), polyvinylalcohol (PVA), polyethylene oxide (PEO)and styrene-maleic acid copolymer (PSMA) were investigated. The activity of immobilized GOD was obviously influenced by the component of the DEAEC complexes. The relative activity of the immobilized GOD reached to maximum and over 90% of the native GOD. when the DEAEC-PMAA DEAEC-PAA complexes were used as a carrier with the molar ratio of DEAEC and polyacid of about one. Michaelis constants (Km) of the immobilized enzymes of DEAEC-GOD-PMAA and DEAEC-GOD-PAA were determined to be 1.25 and 1.00, respectively. Moreover, the immobilized GOD has a good storage stability and cyclic life.

  7. Surface engineering of stainless steel materials by covalent collagen immobilization to improve implant biocompatibility.

    Science.gov (United States)

    Müller, Rainer; Abke, Jochen; Schnell, Edith; Macionczyk, Frank; Gbureck, Uwe; Mehrl, Robert; Ruszczak, Zbigniev; Kujat, Richard; Englert, Carsten; Nerlich, Michael; Angele, Peter

    2005-12-01

    It was shown recently that the deposition of thin films of tantalum and tantalum oxide enhanced the long-term biocompatibility of stainless steel biomaterials due to an increase in their corrosion resistance. In this study, we used this tantalum oxide coating as a basis for covalent immobilization of a collagen layer, which should result in a further improvement of implant tissue integration. Because of the high degradation rate of natural collagen in vivo, covalent immobilization as well as carbodiimide induced cross-linking of the protein was performed. It was found that the combination of the silane-coupling agent aminopropyl triethoxysilane and the linker molecule N,N'-disulphosuccinimidyl suberate was a very effective system for collagen immobilizing. Mechanical and enzymatic stability testing revealed a higher stability of covalent bound collagen layers compared to physically adsorbed collagen layers. The biological response induced by the surface modifications was evaluated by in vitro cell culture with human mesenchymal stem cells as well as by in vivo subcutaneous implantation into nude mice. The presence of collagen clearly improved the cytocompatibility of the stainless steel implants which, nevertheless, significantly depended on the cross-linking degree of the collagen layer. PMID:15967497

  8. Photosynthesis, growth and hydrocarbon production of Botryococcus braunii immobilized by entrapment and adsorption in polyurethane foams

    Energy Technology Data Exchange (ETDEWEB)

    Bailliez, C.; Largeau, C.; Casadevall, E.; Yang Lianwan; Berkaloff, C.

    1988-09-01

    Direct entrapment of the hydrocarbon-rich alga Botryococcus braunii was examined using eleven polyurethane prepolymers. A high toxicity was observed in several foams. With five of the tested prepolymers, nevertheless, a large part of the algal population can survive entrapment and substantial photosynthetic capacity, ca. 40-60% relative to free controls, was retained one day after immobilization. However, prolonged batches under standard conditions revealed a long-term toxicity; as a result the photosynthetic capacity and hydrocarbon production of the entrapped cultures were strongly reduced relative to free controls. Immobilization of B. braunii was also achieved, with a loading yield of ca. 70%, via adsorption of FHP 4000 and FHP 5000 foams. Subsequent batch cultures under shaken and airlift conditions revealed a substantial release, ca. 30% of free cells, at the end of the cultures. However, the release from these adsorbed cultures was no higher than from directly entrapped B. braunii. Furthermore, no toxic effects were noted in the adsorbed cultures; they showed active growth, hihg photosynthetic capacity and produced quite large amounts of hydrocarbons, the chemical structure and the relative abundance of which were not altered by immobilization. Taking into account cell leakage, it appears that adsorbed cultures exhibit a similar, and sometimes even higher metabolic activity than free controls; thus, under air-lift conditions, high biomass and hydrocarbon productivities can be achieved.

  9. Biodegradation of toluene using Candida tropicalis immobilized on polymer matrices in fluidized bed bioreactors.

    Science.gov (United States)

    Song, JiHyeon; Namgung, HyeongKyu; Ahmed, Zubair

    2012-11-30

    A yeast strain, Candida tropicalis, was whole-cell-immobilized on polymer matrices of polyethylene glycol (PEG) and polyethylene glycol/activated carbon/alginate (PACA). The polymer matrices were used as fluidized materials in bubble-column bioreactors for the biodegradation of toluene. Simultaneously, another bubble-column bioreactor using granular activated carbon (GAC) and a conventional compost biofilter were operated for comparison. In the compost biofilter, the toluene removal efficiency gradually deteriorated due to the limitation of microbial activity. The toluene removal in the GAC bioreactor was relatively high because of an increase of toluene mass transfer. However, low toluene removal efficiencies were observed in the PEG bioreactor, presumably because the synthetic polymer alone was not suitable for yeast cell immobilization. In the PACA bioreactor, toluene removal was found to be greater than 95% overall. The CO(2) yield coefficient calculated at the highest toluene loading condition for the PACA bioreactor was found to be higher than those observed in the other bioreactors. Furthermore, almost complete elimination capacities were observed in the PACA bioreactor at short-term toluene loading up to 180 g/m(3)/h. In conclusion, the immobilization of C. tropicalis in the PACA matrix resulted in enhanced toluene biodegradation because of the increases of both mass transfer and microbial activity.

  10. Continuous ethanol production using yeast immobilized on sugar-cane stalks

    Energy Technology Data Exchange (ETDEWEB)

    Vasconcelos, J.N. de [Alagoas Univ., Maceio, AL (Brazil). Dept. de Engenharia Quimica]. E-mail: jnunes@ctec.ufal.br; Lopes, C.E. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Antibioticos; Franca, F.P. de [Universidade Federal, Rio de Janeiro, RJ (Brazil). Escola de Quimica. Dept. de Engenharia Bioquimica

    2004-09-01

    Sugar-cane stalks, 2.0 cm long, were used as a support for yeast immobilization envisaging ethanol production. The assays were conducted in 38.5 L fermenters containing a bed of stalks with 50% porosity. The operational stability of the immobilized yeast, the efficiency and stability of the process, as well as the best dilution rate were evaluated. Molasses from demerara sugar production was used in the medium formulation. It was diluted to obtain 111.75 {+-} 1.51 g/L without any further treatment. Sulfuric acid was used to adjust the pH value to around 4.2. Every two days Kamoran HJ (10 ppm) or with a mixture containing penicillin (10 ppm) and tetracycline (10 ppm), was added to the medium. Ethanol yield and efficiency were 29.64 g/L.h and 86.40%, respectively, and the total reducing sugars conversion was 74.61% at a dilution rate of 0.83 h{sup -1}. The yeast-stalk system was shown to be stable for over a 60 day period at extremely variable dilution rates ranging from 0.05 h{sup -1} to 3.00 h{sup -1}. The concentration of immobilized cell reached around 109 cells/gram of dry sugar-cane stalk when the fermenter was operating at the highest dilution rate (3.00 h{sup -1}). (author)

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

    Science.gov (United States)

    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.

  12. Continuous ethanol production using yeast immobilized on sugar-cane stalks

    Directory of Open Access Journals (Sweden)

    J. N. de Vasconcelos

    2004-09-01

    Full Text Available Sugar-cane stalks, 2.0 cm long, were used as a support for yeast immobilization envisaging ethanol production. The assays were conducted in 38.5 L fermenters containing a bed of stalks with 50% porosity. The operational stability of the immobilized yeast, the efficiency and stability of the process, as well as the best dilution rate were evaluated. Molasses from demerara sugar production was used in the medium formulation. It was diluted to obtain 111.75 ± 1.51 g/L without any further treatment. Sulfuric acid was used to adjust the pH value to around 4.2. Every two days Kamoran HJ (10 ppm or with a mixture containing penicillin (10 ppm and tetracycline (10 ppm, was added to the medium. Ethanol yield and efficiency were 29.64 g/L.h and 86.40%, respectively, and the total reducing sugars (TRS conversion was 74.61% at a dilution rate of 0.83 h-1. The yeast-stalk system was shown to be stable for over a 60 day period at extremely variable dilution rates ranging from 0.05 h-1 to 3.00 h-1. The concentration of immobilized cell reached around 109 cells/gram of dry sugar-cane stalk when the fermenter was operating at the highest dilution rate (3.00 h-1.

  13. Ethanol fermentation in a magnetically fluidized bed reactor with immobilized Saccharomyces cerevisiae in magnetic particles.

    Science.gov (United States)

    Liu, Chun-Zhao; Wang, Feng; Ou-Yang, Fan

    2009-01-01

    Ethanol fermentation by immobilized Saccharomyces cerevisiae cells in magnetic particles was successfully carried out in a magnetically stabilized fluidized bed reactor (MSFBR). These immobilized magnetic particles solidified in a 2 % CaCl(2) solution were stable and had high ethanol fermentation activity. The performance of ethanol fermentation of glucose in the MSFBR was affected by initial particle loading rate, feed sugar concentration and dilution rate. The ethanol theoretical yield, productivity and concentration reached 95.3%, 26.7 g/L h and 66 g/L, respectively, at a particle loading rate of 41% and a feed dilution rate of 0.4 h(-1) with a glucose concentration of 150 g/L when the magnetic field intensity was kept in the range of 85-120 Oe. In order to use this developed MSFBR system for ethanol production from cheap raw materials, cane molasses was used as the main fermentation substrate for continuous ethanol fermentation with the immobilized S. cerevisiae cells in the reactor system. Molasses gave comparative ethanol productivity in comparison with glucose in the MSFBR, and the higher ethanol production was observed in the MSFBR than in a fluidized bed reactor (FBR) without a magnetic field. PMID:18760598

  14. Nitric Acid-Treated Carbon Fibers with Enhanced Hydrophilicity for Candida tropicalis Immobilization in Xylitol Fermentation

    Directory of Open Access Journals (Sweden)

    Le Wang

    2016-03-01

    Full Text Available Nitric acid (HNO3-treated carbon fiber (CF rich in hydrophilic groups was applied as a cell-immobilized carrier for xylitol fermentation. Using scanning electron microscopy, we characterized the morphology of the HNO3-treated CF. Additionally, we evaluated the immobilized efficiency (IE of Candida tropicalis and xylitol fermentation yield by investigating the surface properties of nitric acid treated CF, specifically, the acidic group content, zero charge point, degree of moisture and contact angle. We found that adhesion is the major mechanism for cell immobilization and that it is greatly affected by the hydrophilic–hydrophilic surface properties. In our experiments, we found 3 hto be the optimal time for treating CF with nitric acid, resulting in an improved IE of Candida tropicalis of 0.98 g∙g−1 and the highest xylitol yield and volumetric productivity (70.13% and 1.22 g∙L−1∙h−1, respectively. The HNO3-treated CF represents a promising method for preparing biocompatible biocarriers for multi-batch fermentation.

  15. Targeted Delivery of Hyaluronan-Immobilized Magnetic Ceramic Nanocrystals.

    Science.gov (United States)

    Wu, Hsi-Chin; Wang, Tzu-Wei; Hsieh, Shun-Yu; Sun, Jui-Sheng; Kang, Pei-Leun

    2016-01-01

    Effective cancer therapy relies on delivering the therapeutic agent precisely to the target site to improve the treatment outcome and to minimize side effects. Although surgery, chemotherapy, and radiotherapy are the standard methods commonly used in clinics, hyperthermia has been developed as a new and promising strategy for cancer therapy. In this study, magnetic bioceramic hydroxyapatite (mHAP) nanocrystals have been developed as heat mediator for intracellular hyperthermia. Hyaluronic acid (HA) modified mHAP nanocrystals are synthesized by a wet chemical precipitation process to achieve active targeting. The results demonstrate that the HA targeting moiety conjugated by a poly(ethylene glycol) (PEG) spacer arm is successfully immobilized on the surface of mHAP. The HA-modified mHAP possesses relatively good biocompatibility, an adequate biodegradation rate and superparamagnetic properties. The HA-modified mHAP could be localized and internalized into HA receptor-overexpressed malignant cells (e.g., MDA-MB-231 cell) and used as the heat generating agent for intracellular hyperthermia. The results from this study indicate that biocompatible HA-modified mHAP shows promise as a novel heat mediator and a specific targeting nanoagent for intracellular hyperthermia cancer therapy. PMID:27301176

  16. Immobilization of proteins onto microbeads using a DNA binding tag for enzymatic assays.

    Science.gov (United States)

    Kojima, Takaaki; Mizoguchi, Takuro; Ota, Eri; Hata, Jumpei; Homma, Keisuke; Zhu, Bo; Hitomi, Kiyotaka; Nakano, Hideo

    2016-02-01

    A novel DNA-binding protein tag, scCro-tag, which is a single-chain derivative of the bacteriophage lambda Cro repressor, has been developed to immobilize proteins of interest (POI) on a solid support through binding OR consensus DNA (ORC) that is tightly bound by the scCro protein. The scCro-tag successfully bound a transglutaminase 2 (TGase 2) substrate and manganese peroxidase (MnP) to microbeads via scaffolding DNA. The resulting protein-coated microbeads can be utilized for functional analysis of the enzymatic activity using flow cytometry. The quantity of bead-bound proteins can be enhanced by increasing the number of ORCs. In addition, proteins with the scCro-tag that were synthesized using a cell-free protein synthesis system were also immobilized onto the beads, thus indicating that this bead-based system would be applicable to high-throughput analysis of various enzymatic activities.

  17. Process Technology for Immobilized LipaseProcess Technology for Immobilized Lipase-catalyzed

    DEFF Research Database (Denmark)

    Xu, Yuan

    -down experimental work is described in this thesis. The methodology uses economic targets to test options characterized via a set of tools. In order to validate the methodology, two processes based on immobilized lipase-catalysis have been studied: transesterification and esterification of vegetable oils...... for the production of biodiesel. The two processes are focused on the conversion of the two main components of vegetable oil materials, glyceride esters and free fatty acids respectively, into fatty acid alkyl esters. Although biodiesel is conventionally prepared via chemical-catalyzed transesterification...... of vegetable oils with methanol to produce fatty acid methyl esters (FAME), this work has been focused on the production of fatty acid ethyl esters (FAEE) with bioethanol due to the expected improved sustainability of this type of biodiesel. A key reaction characteristic of the immobilized lipase...

  18. Immobilization of -Amylase onto Luffa operculata Fibers

    Directory of Open Access Journals (Sweden)

    Ricardo R. Morais

    2013-01-01

    Full Text Available A commercial amylase (amy was immobilized by adsorption onto Luffa operculata fibers (LOFs. The derivative LOF-amy presented capacity to hydrolyze starch continuously and repeatedly for over three weeks, preserving more than 80% of the initial activity. This system hydrolyzed more than 97% of starch during 5 min, at room temperature. LOF-amy was capable to hydrolyze starch from different sources, such as maize (93.96%, wheat (85.24%, and cassava (79.03%. A semi-industrial scale reactor containing LOF-amy was prepared and showed the same yield of the laboratory-scale system. After five cycles of reuse, the LOF-amy reactor preserved over 80% of the initial amylase activity. Additionally, the LOF-amy was capable to operate as a kitchen grease trap component in a real situation during 30 days, preserving 30% of their initial amylase activity.

  19. Stability of immobilized yeast alcohol dehydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    Ooshima, H.; Genko, Y.; Harano, Y.

    1981-12-01

    The effects of substrate on stabilities of native (NA) and three kinds of immobilized yeast alcohol dehydrogenase (IMA), namely PGA (the carrier; porous glass), SEA (agarose gel) prepared covalently, and AMA (anion-exchange resin) prepared ionically, were studied. The following results were obtained. 1) The deactivations of NA and IMA free from the substrate or in the presence of ethanol obey the first-order kinetics, whereas, in the presence of butyraldehyde, their deactivation behaviors are explained on the basis of coexistence of two components of YADHs, namely the labile E1 and the comparatively stable E2, with different first-order deactivation constants. (2) A few attempts for stabilization of IMA were carried out from the viewpoint of the effects of crosslinkages among the subunits of YADH for PGA and the multibonding between the carrier and enzyme for SEA. The former is effective for the stabilization, whereas the latter is not. (Refs. 19).

  20. Ceramic Hosts for Fission Products Immobilization

    International Nuclear Information System (INIS)

    Natural spinel, perovskite and zirconolite rank among the most leach resistant of mineral forms. They also have a strong affinity for a large number of other elements and including actinides. Specimens of natural perovskite and zirconolite were radioisotope dated and found to have survived at least 2 billion years of natural process while still remain their loading of uranium and thorium . Developers of the Synroc waste form recognized and exploited the capability of these minerals to securely immobilize TRU elements in high-level waste . However, the Synroc process requires a relatively uniform input and hot pressing equipment to produce the waste form. It is desirable to develop alternative approaches to fabricate these durable waste forms to immobilize the radioactive elements. One approach is using a high temperature process to synthesize these mineral host phases to incorporate the fission products in their crystalline structures. These mineral assemblages with immobilized fission products are then isolated in a durable high temperature glass for periods measured on a geologic time scale. This is a long term research concept and will begin with the laboratory synthesis of the pure spinel (MgAl2O4), perovskite (CaTiO3) and zirconolite (CaZrTi2O7) from their constituent oxides. High temperature furnace and/or thermal plasma will be used for the synthesis of these ceramic host phases. Nonradioactive strontium oxide will be doped into these ceramic phases to investigate the development of substitutional phases such as Mg1-xSrxAl2O4, Ca1-xSrxTiO3 and Ca1-xSrxZrTi2O7. X-ray diffraction will be used to establish the crystalline structures of the pure ceramic hosts and the substitution phases. Scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDX) will be performed for product morphology and fission product surrogates distribution in the crystalline hosts. The range of strontium doping is planned to reach the full substitution of the divalent

  1. Low Temperature Waste Immobilization Testing Vol. I

    Energy Technology Data Exchange (ETDEWEB)

    Russell, Renee L.; Schweiger, Michael J.; Westsik, Joseph H.; Hrma, Pavel R.; Smith, D. E.; Gallegos, Autumn B.; Telander, Monty R.; Pitman, Stan G.

    2006-09-14

    The Pacific Northwest National Laboratory (PNNL) is evaluating low-temperature technologies to immobilize mixed radioactive and hazardous waste. Three waste forms—alkali-aluminosilicate hydroceramic cement, “Ceramicrete” phosphate-bonded ceramic, and “DuraLith” alkali-aluminosilicate geopolymer—were selected through a competitive solicitation for fabrication and characterization of waste-form properties. The three contractors prepared their respective waste forms using simulants of a Hanford secondary waste and Idaho sodium bearing waste provided by PNNL and characterized their waste forms with respect to the Toxicity Characteristic Leaching Procedure (TCLP) and compressive strength. The contractors sent specimens to PNNL, and PNNL then conducted durability (American National Standards Institute/American Nuclear Society [ANSI/ANS] 16.1 Leachability Index [LI] and modified Product Consistency Test [PCT]) and compressive strength testing (both irradiated and as-received samples). This report presents the results of these characterization tests.

  2. Study of CRP immobilization on nanostructured silicon

    Energy Technology Data Exchange (ETDEWEB)

    Simion, Monica, E-mail: moni304ro@yahoo.com [National Institute for Research and Development in Microtechnologies (IMT - Bucharest), 32B Erou Iancu Nicolae Street, 72996 Bucharest (Romania); Ruta, Lavinia L.; Matache, Mihaela [University of Bucharest, Department of Chemistry, Division of Organic Chemistry, 90-92 Panduri Street, 050663 Bucharest (Romania); Kleps, Irina; Miu, Mihaela [National Institute for Research and Development in Microtechnologies (IMT - Bucharest), 32B Erou Iancu Nicolae Street, 72996 Bucharest (Romania); Paraschivescu, Codruta C. [University of Bucharest, Department of Chemistry, Division of Organic Chemistry, 90-92 Panduri Street, 050663 Bucharest (Romania); Bragaru, Adina; Ignat, Teodora [National Institute for Research and Development in Microtechnologies (IMT - Bucharest), 32B Erou Iancu Nicolae Street, 72996 Bucharest (Romania)

    2010-05-25

    C-reactive protein (CRP) is a phylogenetically highly conserved plasma protein, which is widely used as an indicator of inflammatory states due to rapid increase of its plasma concentration up to 1000 times compared to normal values. Detection of CRP levels in a rapid, simultaneous and multiplex format is therefore of great interest for diagnostics. Microarray technology could provide such a multiplex format of CRP levels detection. Different nanostructured porous silicon (PS) surfaces were obtained and used for the immobilization of CRP and anti-human CRP antibodies in order to achieve an optimum microarray assay. Comparative analysis of the attachment degree and preservation of the biomolecules activity on the silicon surfaces and functionalized glass slides is also described.

  3. Ceramic Hosts for Fission Products Immobilization

    Energy Technology Data Exchange (ETDEWEB)

    Peter C Kong

    2010-07-01

    Natural spinel, perovskite and zirconolite rank among the most leach resistant of mineral forms. They also have a strong affinity for a large number of other elements and including actinides. Specimens of natural perovskite and zirconolite were radioisotope dated and found to have survived at least 2 billion years of natural process while still remain their loading of uranium and thorium . Developers of the Synroc waste form recognized and exploited the capability of these minerals to securely immobilize TRU elements in high-level waste . However, the Synroc process requires a relatively uniform input and hot pressing equipment to produce the waste form. It is desirable to develop alternative approaches to fabricate these durable waste forms to immobilize the radioactive elements. One approach is using a high temperature process to synthesize these mineral host phases to incorporate the fission products in their crystalline structures. These mineral assemblages with immobilized fission products are then isolated in a durable high temperature glass for periods measured on a geologic time scale. This is a long term research concept and will begin with the laboratory synthesis of the pure spinel (MgAl2O4), perovskite (CaTiO3) and zirconolite (CaZrTi2O7) from their constituent oxides. High temperature furnace and/or thermal plasma will be used for the synthesis of these ceramic host phases. Nonradioactive strontium oxide will be doped into these ceramic phases to investigate the development of substitutional phases such as Mg1-xSrxAl2O4, Ca1-xSrxTiO3 and Ca1-xSrxZrTi2O7. X-ray diffraction will be used to establish the crystalline structures of the pure ceramic hosts and the substitution phases. Scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDX) will be performed for product morphology and fission product surrogates distribution in the crystalline hosts. The range of strontium doping is planned to reach the full substitution of the divalent

  4. Triple touch sperm immobilization vs. single touch sperm immobilization in ICSI - a randomised trial

    Directory of Open Access Journals (Sweden)

    Velaers An

    2012-08-01

    Full Text Available Abstract Background Although different techniques for sperm immobilization have been described, their value has not been assessed in an adequately powered randomized study. The aim of this study was to compare two types of sperm immobilization methods prior to ICSI and to test the hypothesis that triple touch immobilization (TTIm would lead to a higher (5% -65% up to 70% fertilization rate (FR than single touch immobilization (STIm. Methods A total of 3056 metaphase II (MII oocytes, from 290 patients, were randomly assigned to the STIm group (n = 1528 oocytes; 145 cycles or to the TTIm group (n = 1528 oocytes; 138 cycles. A total of 1478 oocytes (STIm group and 1476 oocytes (TTIm group were used in the statistical analysis. The primary outcome variable was FR. Secondary outcome variables included: number of good quality embryos (GQE on day 2 and day 3, implantation rate (IR and implantation with foetal heart beat rate (FHB. Statistical analysis was done using the Fisher Exact test with a significance level of 0.05. Results The results showed no differences in FR between both groups. The proportion of good quality embryos on day 3, was significantly higher in the STIm group (37.5% compared to the TTIm group (31.8%; p = 0.02. Conclusions In this RCT, the hypothesis that the post-ICSI FR would be higher after TTIm than after STIm was not confirmed and the number of good quality embryos on day 3 was significantly lower in the TTIm group than in the STIm group. These data suggest that more ‘aggressive’ TTIm technique has no advantages compared to the STIm technique.

  5. Resveratrol immobilization and release in polymeric hydrogels

    International Nuclear Information System (INIS)

    Resveratrol (3, 4', 5-trihydroxystilbene) is a polyphenolic produced by a wide variety of plants in response to injury and found predominantly in grape skins. This active ingredient has been shown to possess benefits for the health, such as the antioxidant capacity which is related to the prevention of several types of cancer and skin aging. However, the oral bioavailability of resveratrol is poor and makes its topical application interesting. The purpose of this study was to immobilize resveratrol in polymeric hydrogels to obtain a release device for topical use. The polymeric matrices composed of poli(N-vinyl-2-pyrrolidone) (PVP), poly(ethyleneglycol) (PEG) and agar or PVP and glycerol irradiated at 20 kGy dose were physical-chemically characterized by gel fraction and swelling tests and its preliminary biocompatibility by in vitro test of cytotoxicity using the technique of neutral red uptake. Due to low solubility of resveratrol in water, the addition of 2% ethanol to the matrices was verified. All matrices showed a high crosslinking degree, capacity of swelling and the preliminary cytotoxicity test showed nontoxicity effect. The devices were obtained by resveratrol immobilization in polymeric matrices, carried out in a one-or-two-steps process, that is, before or after irradiation, respectively. The one step resveratrol devices were characterized by gel fraction, swelling tests and preliminary biocompatibility, and their properties were maintained even after the resveratrol incorporation. The devices containing 0,05% of resveratrol obtained by one-step process and 0,1% of resveratrol obtained by two-steps process were submitted to the release test during 24 h. Resveratrol quantification was done by high performance liquid chromatography (HPLC). The results obtained in the kinetics of release showed that only the devices obtained by two-step process release the resveratrol, which demonstrate antioxidant capacity after the release. (author)

  6. Preparation of guava wine using immobilized yeast

    Directory of Open Access Journals (Sweden)

    Surajbhan Sevda

    2014-12-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Saccharomyces cerevisiae (NCIM 3287 was immobilized in sodium alginate beads to produce a biocatalyst for use in guava wine making. The immobilized biocatalyst was found to be suitable for guava juice fermentation at ambient temperatures. The study included effects of alginate concentrations, initial bead loadings and bead diameters on fermentation. The optimized parameters were  2% (w/v alginate concentration, 60g/l initial bead loading and           3 mm bead diameter for effective fermentation of guava must. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;}

  7. Production of laccase by Coriolus versicolor and its application in decolorization of dyestuffs: (Ⅱ) Decolorization of dyes by laccase containing fermentation broth with or without self-immobilized mycelia

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    The capability of decolorization for commercial dyes byCoriolus versicolor fermentation broth containing laccase with orwithout immobilized mycelium was evaluated. With cell-freefermentation broth containing laccase, high decolorization ratiowas achieved for acid orange 7, but not for the other dyesconcerned. The immobilized mycelium was proved to be more efficientthan the cell-free system. All the four dyestuffs studied werefound being decolourized with certain extent by immobilizedmycelium. The repeated-batch decolorization was carried out withsatisfactory results. The experimental data showed that thecontinuous decolorization of wastewater from a printing and dyeingindustry was possible by using the self-immobilized C. Versicolor.

  8. Release of Paramecium immobilization antigen to the non-nutrient medium.

    Science.gov (United States)

    Wyroba, E

    1980-01-01

    The instability of Paramecium aurelia surface components has been shown: after 60 min incubation of dense cell suspension ((1-2 . 10(6) cells per ml) in Tris-Ca buffer at 4 degrees C or 23 degrees C the surface coat was partially stripped off and the proteins were released to the medium. The electrophoretic analysis of the released proteins shows one major band of mol. wt. 280,000--300,000 and some minor bands. The major released protein is the immobilization antigen as proved using immunodiffusion test and antibody precipitation technique.

  9. Enhanced stability of catalase covalently immobilized on functionalized titania submicrospheres.

    Science.gov (United States)

    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). PMID:23827593

  10. Skeletal muscle HSP expression in response to immobilization and remobilization.

    Science.gov (United States)

    Venojärvi, M; Kvist, M; Jozsa, L; Kalimo, H; Hänninen, O; Atalay, M

    2007-04-01

    Heat shock proteins play an important regulatory role in the cellular defence. Oxidative stress is one of the factors inducing heat shock protein expression. This study tested the effects of 4 weeks of immobilization and subsequent remobilization on heat shock protein expression and oxidative stress in the lateral gastrocnemius and plantaris muscles of the rat. Active mobilization or free mobilization protocols were used for remobilization. In active mobilization, strenuous uphill treadmill running, twice a day, was started immediately after the immobilization and lasted for six days. Rats in the free mobilization group moved freely in their cages immediately after the immobilization. Expression of heat shock proteins was upregulated during the recovery from immobilization, especially in the lateral gastrocnemius muscle in the active mobilization group. However, markers of oxidative stress, such as protein carbonyls and 4-hydroxynonenal protein adducts, or activities of the antioxidant enzymes glutathione peroxidase and glutathione reductase, did not change after the immobilization and subsequent recovery. In summary, following immobilization, both intensive and spontaneous exercise upregulated the heat shock protein expressions in the lateral gastrocnemius muscle and partly in the plantaris muscle, which may contribute to the recovery from immobilization atrophy. PMID:17024631

  11. Production and Immobilization of Partially Purified Lipase From Penicillium chrysogenum

    Directory of Open Access Journals (Sweden)

    Shafei, M. S.

    2010-01-01

    Full Text Available An extracellular lipase from Penicillium chrysogenum produced maximal activity 225 U/mL after four days at pH 6.5. It was partially purified 4.1 fold by ammonium sulphate precipitation (70%. The enzyme was immobilized on various carriers viz. alginate, k-carrageenan and polyacrylamide gel. The immobilization yield of enzyme immobilized in kcarrageenan and polyacrylamide gel (63.41% and 48.93% respectively was low in comparison to that immobilized with alginate (81.57%. Different concentrations of alginate were tried to study their effect on lipase production. Maximum immobilization yield was observed with 3% alginate. The optimal pH of the partially purified lipase was 7.5 and the optimum temperature was 35 °C. At 60 °C the immobilized enzyme retained 62.79% of its activity. Broader pH tolerance and higher heat stability could be achieved by this method. Immobilized lipase retained 72.09% relative activity after six hydrolysis cycles.

  12. Acetylcholinesterase immobilized onto PEI-coated silica nanoparticles.

    Science.gov (United States)

    Tumturk, Hayrettin; Yüksekdag, Hazer

    2016-01-01

    Polyethyleneimine (PEI) coated-silica nanoparticles were prepared by the Stöber method. The formation and the structure of the nanoparticles were characterized by ATR-FT-IR spectroscopy and transmission electron microscopy (TEM). TEM images of the silica and PEI-coated nanoparticles revealed that they were well dispersed and that there was no agglomeration. The acetylcholineesterase enzyme was immobilized onto these nanoparticles. The effects of pH and temperature on the storage stability of the free and immobilized enzyme were investigated. The optimum pHs for free and immobilized enzymes were determined as 7.0 and 8.0, respectively. The optimum temperatures for free and immobilized enzymes were found to be 30.0 and 35.0°C, respectively. The maximum reaction rate (Vmax) and the Michaelis-Menten constant (Km) were investigated for the free and immobilized enzyme. The storage stability of acetylcholinesterase was increased when immobilized onto the novel PEI-coated silica nanoparticles. The reuse numbers of immobilized enzyme were also studied. These hybrid nanoparticles are desirable as carriers for biomedical applications.

  13. Thermal stability of the immobilized fructosyltransferase from Rhodotorula sp

    Directory of Open Access Journals (Sweden)

    E Aguiar-Oliveira

    2011-09-01

    Full Text Available The thermal stability of the extracellular fructosyltransferase (FTase from Rhodotorula sp., recovered from cultivation medium by ethanol precipitation and immobilized onto niobium ore, was studied by Arrhenius plot, half - life profile, half - inactivation temperature (T50 and thermodynamic parameters. The Arrhenius plot showed two different behaviors with different deactivation energies (Ead only after immobilization, the transition occurring in the temperature interval between 51 and 52ºC. T50 for the free enzyme was estimated to be around 62ºC and, after immobilization, 66ºC. After 15 minutes at 52ºC, it was also possible to observe enzymatic activation for both the free and immobilized forms, but greater activation was achieved at pH 4.5 with the immobilized enzyme. Between 47 - 51ºC the immobilized enzyme was more stable than the free enzyme, with pH 6.0 being the more stable condition for the immobilized enzyme. However, above 52ºC the free form was more stable.

  14. Integrated development and testing plan for the plutonium immobilization project

    International Nuclear Information System (INIS)

    This integrated plan for the DOE Office of Fissile Materials Disposition (MD) describes the technology development and major project activities necessary to support the deployment of the immobilization approach for disposition of surplus weapons-usable plutonium. The plan describes details of the development and testing (D and T) tasks needed to provide technical data for design and operation of a plutonium immobilization plant based on the ceramic can-in-canister technology (''Immobilization Fissile Material Disposition Program Final Immobilization Form Assessment and Recommendation'', UCRL-ID-128705, October 3, 1997). The plan also presents tasks for characterization and performance testing of the immobilization form to support a repository licensing application and to develop the basis for repository acceptance of the plutonium form. Essential elements of the plant project (design, construction, facility activation, etc.) are described, but not developed in detail, to indicate how the D and T results tie into the overall plant project. Given the importance of repository acceptance, specific activities to be conducted by the Office of Civilian Radioactive Waste Management (RW) to incorporate the plutonium form in the repository licensing application are provided in this document, together with a summary of how immobilization D and T activities provide input to the license activity. The ultimate goal of the Immobilization Project is to develop, construct, and operate facilities that will immobilize from about 18 to 50 tonnes (MT) of U.S. surplus weapons usable plutonium materials in a manner that meets the ''spent fuel'' standard (Fissile Materials Storage and Disposition Programmatic Environmental Impact Statement Record of Decision, ''Storage and Disposition Final PEIS'', issued January 14, 1997, 62 Federal Register 3014) and is acceptable for disposal in a geologic repository. In the can-in-canister technology, this is accomplished by encapsulating the

  15. Enhanced biodegradation of pyrene and indeno(1,2,3-cd)pyrene using bacteria immobilized in cinder beads in estuarine wetlands.

    Science.gov (United States)

    Huang, Ru-Ying; Tian, Wei-Jun; Liu, Qing; Yu, Hui-Bo; Jin, Xin; Zhao, Yang-Guo; Zhou, Yu-Hang; Feng, Gong

    2016-01-15

    Two strains (Pseudomonas taiwanensis PYR1 and Acinetobacter baumannii INP1) were isolated from PAH-contaminated Liaohe estuarine wetland using enrichment. The cells of PYR1 and INP1 were immobilized in cinder beads for pyrene and indeno(1,2,3-cd)pyrene biodegradation in wetland. Biodegradation of pyrene and indeno(1,2,3-cd)pyrene in soils from wetland was carried out in pots using free cells as well as those immobilized in cinder beads to ascertain the role of bioaugmentation. Supported by the cinder beads, the immobilized cells degraded 70.7% and 80.9% of pyrene and indeno(1,2,3-cd)pyrene respectively after 30 days. While the free cells degraded only 58.2% and 55.3%. Additionally, microbial analysis with high-throughput sequencing revealed the changes of microbial communities in soil without and with cinder beads immobilized with strains. The result indicated that Gammaproteobacteria were dominant PAH-degrading groups during bioaugmentation. This effective approach can be used to treat other PAH-contaminated wetlands by immobilizing different species of bacteria in cinder beads.

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

    Science.gov (United States)

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

    2015-02-01

    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.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. Electronic supplementary information (ESI) available: TEM image of CDs, BET XRD

  17. Immobilization and functional reconstitution of antibody Fab fragment by solid-phase refolding.

    Science.gov (United States)

    Kumada, Yoichi; Hamasaki, Kyoto; Nakagawa, Aya; Sasaki, Eiju; Shirai, Tatsunori; Okumura, Masahiro; Inoue, Manami; Kishimoto, Michimasa

    2013-12-31

    In this study, we demonstrated the successful preparation of a Fab antibody-immobilized hydrophilic polystyrene (phi-PS) plate via one- and two-step solid-phase refolding methods. Both polystyrene-binding peptide (PS-tag)-fused Fd fragment of heavy chain (Fab H-PS) and full-length of light-chain (Fab L-PS) were individually produced in insoluble fractions of Escherichia coli cells, and they were highly purified in the presence of 8M of urea. Antigen-binding activities of Fab antibody immobilized were correctly recovered by the one-step solid-phase refolding method that a mixture of Fab H-PS and Fab L-PS was immobilized in the presence of 0.5-2M urea, followed by surface washing of the phi-PS plate with PBST. These results indicate that by genetic fusion of a PS-tag, a complex between Fab H and Fab L was efficiently immobilized on the surface of a phi-PS plate even in the presence of a low concentration of urea, and was then correctly refolded to retain its high antigen-binding activity via removal of the urea. A two-step solid-phase refolding method whereby Fab H-PS and Fab L-PS were successively refolded on the surface of a phi-PS plate also resulted in Fab antibody formation on the plate. Furthermore, both the binding affinity and the specificity of the Fab antibody produced by the two-step method were highly maintained, according to the results of sandwich ELISA and competitive ELISA using Fab antibody-immobilized plate via two-step solid-phase refolding. Thus, the solid-phase refolding method demonstrated in this study should be quite useful for the preparation of a Fab antibody-immobilized PS surface with high efficiency from individually produced Fab H-PS and Fab L-PS. This method will be applicable to the preparation of a large Fab antibody library on the surface of a PS plate for use in antibody screening.

  18. Immobilization of free-ranging moose (Alces alces with medetomidine-ketamine and remobilization with atipamezole

    Directory of Open Access Journals (Sweden)

    Jon M. Arnemo

    1995-12-01

    Full Text Available Seventeen free-ranging moose {Alces alces (2 adult males, 13 adult females and 2 female calves were immobilized with a combination of medetomidine hydochloride (MED and ketamine hydrochloride (KET in early autumn (August-September. Drugs were administrated with plastic projectile syringes fired from a dart gun, either from a car or by approaching the animals on foot. MED at 30 mg/adult and 15 mg/calf in combination with KET at 400 mg/adult and 200 mg/calf induced complete immobilization with sternal or lateral recumbency and loss of the corneal reflex in all individuals. The mean ± SD time from darting to when the animals were found was 18.3 ± 8.7 min for adults and the mean distance covered by these animals between darting and recumbency was 320 + 200 m. No side effects of clinical significance were detected and registration of the rectal temperature (38.8 ± 0.5°C, heart rate (44 ± 7 beats/min, respiratory rate (31 ±20 breaths/min and relative arterial oxygen saturation (89 ± 3 %, n=8 during immobilization in adults showed that these physiological parameters were within the safe ranges established for moose. Blood samples from adults were analyzed for 17 haematological and 33 serum biochemical constituents and the results were compared to corresponding values found in moose immobilized with etorphine (ETO. Although the lower levels (p<0.05 found for haematocrit, red blood cells, haemoglobin and Cortisol in the MED-KET group may indicate a difference in the stress response, the low muscle enzyme levels in both groups show that these immobilizing drugs and capture methods induce very little physical stress in moose. A hyperglycaemic response was found in MED-KET treated animals. Atipamezole hydrochloride (ATI rapidly remobilized all animals and the time elapsing from ATI administration to standing was 3-9 ± 1.8 min after i.v./s.c. treatment (n=7 and 6.9 ± 3.4 min after i.m./s.c. injection (n=8. No side effects were detected after

  19. 固定化连续发酵研究及应用进展%Advance in the Research on and the Application of the Continuous Fermentation with Immobilized Technology

    Institute of Scientific and Technical Information of China (English)

    贾淑丽; 侯红萍

    2012-01-01

    Immobilized continuous fermentation refers to the placement of immobilized cells or immobilized enzyme in certain bioreactor for continuous fermentation.In this paper,the research progress in bioreactor,fermenting technology,and dynamics for immobilized continuous fermentation were elaborated,and the application of immobilized continuous fermentation in different fields was also introduced.%固定化连续发酵是将固定化细胞或固定化酶置于某一生物反应器内进行连续发酵的技术。阐述了固定化连续发酵生物反应器、发酵工艺及动力学的研究,并介绍了该技术在各个领域的应用进展。

  20. Cadmium and Mercury Uptake by Immobilized Pleurotus sapidus

    OpenAIRE

    YALÇINKAYA, Yağmur

    2002-01-01

    Pleurotus sapidus} basidiospores immobilized onto Ca-alginate beads were used for the removal of cadmium and mercury ions from aqueous solutions. The biosorption of Cd(II) and Hg(II) ions on the alginate beads and both immobilized live and heat inactivated fungal mycelia of Pleurotus sapidus} was studied from aqueous solutions in the concentration range of 30-500 mg L - 1. The biosorption of Cd(II) and Hg(II) ions by the alginate and both live and heat inactivated immobilized prepara...