• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.3
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• pp.179-182
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• 2001

Porcine liver esterase was immobilized in polyacrylamide gel for the enantioselective production of levofloxacin from ofloxacin butyl ester. The initial activity of immobilized esterase was found to be significantly affected by the polyacrylamide gel composition. The optimum concentrations of monomer and crosslinker were determined to be 20% and 8.3%, respectively. The activity of immobilized esterase was 55.4% compared to a free enzyme. Enantiomeric excess was maintained at 60%, almost the same level as that of free enzyme. In addition, the immobilized esterase could be used repeatedly up to 10 times without experiencing any severe loss of activity and enantioselectivity.

• Journal of Hydrogen and New Energy
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• v.33 no.5
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• pp.507-514
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• 2022

There is growing interest in sustainable energy sources that can reduce fossil fuel dependence and environmental pollution while meeting rapidly growing energy demands. Hydrogen have been investigated as one of the ideal alternative energies because it has relatively high efficiency without emitting pollutants. The light-sensitized enzymatic (LSE) system, which uses hydrogenase-enzymes, is one of the methods towards economically feasible system configurations that enhance the rate of hydrogen generation. Hydrogenase is an enzyme that catalyzes a reversible reaction that oxidizes molecular hydrogen or produces molecular hydrogen from protons and electrons. In this paper, utilization of [NiFe]-hydrogenase (from Pyrococcus furiosus) in photoelectrochemical hydrogen production system such as handling, immobilization, physicochemical and electrochemical analysis, process parameters, etc. was introduced.

• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.511-519
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• 2009

A chimeric gene encoding enhanced green fluorescent protein (EGFP) and a S-layer protein from Lactobacillus brevis KCTC3102, and/or two copies of the Fe-binding Z-domain, a synthetic analog of the B-domain of protein A, was constructed and expressed in Escherichia coli BL21(DE3). The S-layer fusion proteins produced in a 500-1 fermentor were likely to be stable in the range of pH 5 to 8 and $0^{\circ}C$ to $40^{\circ}C$. Their adhesive property enabled an easy and rapid immobilization of enzymes or antibodies on solid materials such as plastics, glass, sol-gel films, and intestinal epithelial cells. Owing to their affinity towards intestinal cells and immunoglobulin G, the S-layer fusion proteins enabled the adhesion of antibodies to human epithelial cells. In addition, feeding a mixture of the S-layer fusion proteins and antibodies against neonatal calf diarrhea (coronavirus, rotavirus, Escherichia coli, and Salmonella typhimurium) to Hanwoo calves resulted in 100% prevention of neonatal calf diarrhea syndrome (p<0.01), whereas feeding antibodies only resulted in 56% prevention.

• The Korean Journal of Food And Nutrition
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• v.6 no.3
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• pp.158-168
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• 1993

For the conversion of WAD to NADP, Immobilized Brevibacterium ammoniagenes cells with NAD kinase was coupled with ATP-generating system by acetate kinase. The membrane permeability of B. ammoniagenes was improved by toluene treatment of cells. The toluene treated B. ammoniagenes cells were immobilized for stable enzyme activity. Partially purified acetate kinase was used in the reaction system. The optimum conditions for the efficient conversion of UAD to WADP by energy-coupled system were investigated. B. ammoniagenes cells treated with toluene for the Improvement of membrane permeability showed 4.5 fold improved permeability in the conversion of NAD to NADP compared with Intact cells. 3% k-carrageenan as the immobilization matrix of B. ammoniagenes showed the best efficiency for the conversion of NAD to NADP The optimum conditions for the WAR to WARP conversion reaction coupled nth ATP-generating system were 10mM acetylphosphate, 5mM ADP 200mM inorganic phosphate, 10mM MgCl2, 250mg/ml Immobilized cells, 49.3mUnit/ml acetate kinase, pH 7.5 and 37$^{\circ}C$. Under the optimum conditions, 72% of 5mM(340mg/ml ) NAD was converted to UADP In 12 hours.

• The Korean Journal of Mycology
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• v.18 no.2
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• pp.84-88
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• 1990

Hydroxylation in the $11{\alpha}$-position of progesterone molecules was carried out using Rhizopus nigricans spores immobilized within various gel matrices, among which polyacrylamide and agar gel were proved to be the most effective. Reactions with the immobilized cells and in­tact free cells showed almost identical conversion rate of progesterone, optimal pH and reaction time for attaining maximal yield, from which were confirmed absence of any decay and modification of enzyme activities.

• KSBB Journal
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• v.8 no.4
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• pp.320-327
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• 1993

Latex microspheres of styrene/acryl copolymer with acrylamide functional group were used for the stable covalent immobilization of an enzyme applicable for enzymatic synthesis of glycoside. The latex microspheres were coated with polyethyleneimine to establish structural and functional properties relevant to the covalent Immobilization with a high retention of activity. Polythyleneimine-coated microspheres satisfactorily immobilized the invertase for methyl fructoside synthesis, and model reaction were formed into alginate-enclosed microspheres biocatalyst. Using the alginate-enclosed microspheres biocatalyst, the yield of model glycoside was obtained as high as 52.2% at concentration of aqueous 30%(v/v) methanol and 0.291mo1/1 sucrose solution with 2U/ml of activity. The present study showed that the latex microspheres were successfully applied to enzymatic synthesis of glycoside.

• Microbiology and Biotechnology Letters
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• v.9 no.3
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• pp.159-164
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• 1981

Reinforced Calcium-alginate gel entrappment method for enzyme immobilization is described with an example of penicillin amidase from Bacillus megaterium KFCC 10029, a partially constitutive mutant of B. megaterium ATCC 14945. Penicillin amidase recovered from the fermentation broth by adsorption on celite is mixed with alginate and gelatin solution, and cast into a pellet or noodle form by coagulation in calcium salt solution followed by crosslinking with glutaraldehyde. Optimum pH and temperature of the immobilized enzyme preparation were 8.0 and 6$0^{\circ}C$, respectively. Kinetic constants such as Km value and the inhibition constant of 6-APA and phenylacetic acid were 2.6 mM, 7.4 mM and 21.2 mM, respectively. The enzyme leakage from the adsorbent during operation was successfully prevented owing to the increase of physical strength of gel coat. The half lives in a column reactor were 6 and 30 days at the respective temperature of 4$0^{\circ}C$ and 3$0^{\circ}C$, which were the 6-8 fold increased values as compared with those of without entrappment. The results highly recommended the use of reinforced Calcium-alginate gel entrappment method for the enhancement of physical strength and the operational stability of alginate gel entrapped enzyme.

• International Journal of Oral Biology
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• v.37 no.4
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• pp.181-188
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• 2012

As the demand for large-scale analysis of gene expression using DNA arrays increases, the importance of the surface characterization of DNA arrays has emerged. We compared the efficiency of molecular biological applications on solid-phases with different surface polarities to identify the most optimal conditions. We employed thiol-gold reactions for DNA immobilization on solid surfaces. The surface polarity was controlled by creating a self-assembled monolayer (SAM) of mercaptohexanol or hepthanethiol, which create hydrophilic or hydrophobic surface properties, respectively. A hydrophilic environment was found to be much more favorable to solid-phase molecular biological manipulations. A SAM of mercaptoethanol had the highest affinity to DNA molecules in our experimetns and it showed greater efficiency in terms of DNA hybridization and polymerization. The optimal DNA concentration for immobilization was found to be 0.5 ${\mu}M$. The optimal reaction time for both thiolated DNA and matrix molecules was 10 min and for the polymerase reaction time was 150 min. Under these optimized conditions, molecular biology techniques including DNA hybridization, ligation, polymerization, PCR and multiplex PCR were shown to be feasible in solid-state conditions. We demonstrated from our present analysis the importance of surface polarity in solid-phase molecular biological applications. A hydrophilic SAM generated a far more favorable environment than hydrophobic SAM for solid-state molecular techniques. Our findings suggest that the conditions and methods identified here could be used for DNA-DNA hybridization applications such as DNA chips and for the further development of solid-phase genetic engineering applications that involve DNA-enzyme interactions.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.163-167
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• 2012

Since D-xylose is not fermentable in Saccharomyces cerevisiae, its conversion to D-xylulose is required for its application in biotechnological industries using S. cerevisiae. In order to convert D-xylose to D-xylulose by way of an enzyme immobilized system, D-xylose isomerase (XI) of Escherichia coli was fused with 10-arginine tag (R10) at its C-terminus for the simple purification and immobilization process using a cation exchanger. The fusion protein XIR10 was overexpressed in recombinant E. coli and purified to a high purity by a single step of cation exchange chromatography. The purified XIR10 was immobilized to a cation exchanger via the electrostatic interaction with the C-terminal 10-arginine tag. Both the free and immobilized XIR10 exhibited similar XI activities at various pH values and temperatures, indicating that the immobilization to the cation exchanger has a small effect on the enzymatic function of XIR10. Under optimized conditions for the immobilized XIR10, D-xylose was isomerized to D-xylulose with a conversion yield of 25%. Therefore, the results of this study clearly demonstrate that the electrostatic immobilization of XIR10 via the interaction between the 10-arginine tag and a cation exchanger is an applicable form of the conversion of D-xylose to D-xylulose.

• Applied Biological Chemistry
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• v.35 no.5
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• pp.410-414
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• 1992

Endoinulase from Streptomyces sp. S56 was immobilized by adsorption on DEAE-cellulose in 0.01 M citrate-sodium phosphate buffer, pH 6.0 and the properties of immobilized and free enzymes were investigated. The immobilized enzyme preparation, having 40 inulase activity units per dried matrix, revealed the maximal activity at $pH\;4.5{\sim}5.5$ and $55{\sim}60^{\circ}C$ and were most stable at pH 6 and 45^{\circ}C$. The immobilization caused a drop in optimum pH and affinity toward inulin, a slight increase in optimum temperature, an important increase in thermal stability and maximum reaction velocity. The immobilized endoinulase hydrolyzed the tuber extract of jerusalem artichoke and inulin, mainly into fructose and inulobise, degrading 63 and 78% of the total sugar respectively, within 48 hrs in batch reactor.