• Journal of Microbiology and Biotechnology
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• 제21권9호
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• pp.972-978
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• 2011

In this study, we investigated the performance of an immobilized ${\beta}$-galactosidase inclusion bodies-containing Escherichia coli cell reactor, where the cells were immobilized in alginate beads, which were then used in repeated-batch operations for the hydrolysis of o-nitrophenyl-${\beta}$-D-galactoside or lactose over the long-term. In particular, in the Tris buffer system, disintegration of the alginate beads was not observed during the operation, which was observed for the phosphate buffer system. The o-nitrophenyl-${\beta}$-D-galactoside hydrolysis was operated successfully up to about 80 h, and the runs were successfully repeated at least eight times. In addition, hydrolysis of lactose was successfully carried out up to 240 h. Using Western blotting analyses, it was verified that the ${\beta}$-galactosidase inclusion bodies were sustained in the alginate beads during the repeated-batch operations. Consequently, we experimentally verified that ${\beta}$-galactosidase inclusion bodies-containing Escherichia coli cells could be used in a repeated-batch reactor as a biocatalyst for the hydrolysis of o-nitrophenyl-${\beta}$-D-galactoside or lactose. It is probable that this approach can be applied to enzymatic synthesis reactions for other biotechnology applications, particularly reactions that require long-term and stable operation.

• Journal of Microbiology and Biotechnology
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• 제19권6호
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• pp.582-587
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• 2009

Immobilized cells of Arthrohacter nitroguajacolicus ZJUTB06-99 capable of producing nitrilase were used for biotransformation of acrylonitrile to acrylic acid. Six different entrapment matrixes were chosen to search for a suitable support in terms of nitrilase activity. Ca-alginate proved to be more advantageous over other counterparts in improvement of the biocatalyst activity and bead mechanical strength. The effects of sodium alginate concentration, $CaCl_2$ concentration, bead diameter, and ratio by weight of cells to alginate, on biosynthesis of acrylic acid by immobilized cells were investigated. Maximum activity was obtained under the conditions of 1.5% sodium alginate concentration, 3.0% $CaCl_2$ concentration, and 2-mm bead size. The beads coated with 0.10% polyethylenimine (PEI) and 0.75% glutaraldehyde (GA) could tolerate more phosphate and decrease leakage amounts of cells from the gel. The beads treated with PEI/GA could be reused up to 20 batches without obvious decrease in activities, which increased about 100% compared with the untreated beads with a longevity of 11 batches.

• KSBB Journal
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• 제8권4호
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• pp.351-357
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• 1993

펄프 폐수처리를 위한 효과적인 생물학적 처리법 으로서 세 가지 고정화 방법과 자유균체를 이용한 방법을 사용하여 실험하였다. 균체로서는 Phaηe­r rochaete chrysosporium을 이용하였다. 고정화방법 중 Ca-alginate gel을 담체로 샤용한 방법이 가장 우수하였다. 이 방법은 안정성과 처리 효율변에서 좋은 결과를 보였으며, 약 400시간 동안 균체의 커다란 상태변화없이 반응시킬 수 있었다. 다른 고정화 방법들과 자유균체를 이용한 방법들 의 가장 큰 문제점은 생물 반응기 안에서의 공기 c channelling 현상에 의한 산소전달 저항과 균체끼리 의 clogging으로 인한 반응기 내부의 제어가 힘들게 된 것이었다.

• 대한화장품학회지
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• 제39권1호
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• pp.25-30
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• 2013

단백질을 연결시키는 효소인 transglutaminase는 모발에서 다양한 효과를 나타낼 수 있는 가능성이 있는데 이중에는 모발의 단단함을 증가시키는 작용도 포함된다. 여러 transglutaminase 효소 중 Streptomyces mobaraensis로부터 분리된 미생물 유래 효소를 손상된 모발에 사용한 후 인장강도를 평가한 결과 초기에 비해 15.64%까지 증가되는 것이 확인되었다. 이러한 효과는 transglutaminas가 샴푸를 사용하여 세정하는 과정에서의 모발 손상을 복구시킬 수 있다는 것을 의미한다. 또한 transglutaminase를 이용해 모발 표면의 특성을 개선 시킴으로써 모발의 윤기를 증가시키고 표면의 마찰력을 감소시키는데 유용하게 이용될 수 있었다.

• 한국수소및신에너지학회논문집
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• 제27권5호
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• pp.526-532
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• 2016

In this study, we synthesized a mediator immobilized biocatalyst([FCA/GOx]/PEI/CNT) by surface modification using ferrocene carboxylic acid(FCA), and evaluated its performance as anode catalyst for biofuel cell. Through the application of FCA on glucose oxidase (GOx), the free amine groups on the lysine residue of GOx surface reacted with carboxylic acid of FCA and make amide bond between GOx and FCA. As the result of that, the electron transfer of catalyst was increased up to 1.91 times($0.468mA{\cdot}cm^{-2}$) than the catalyst without surface modification (GOx/PEI/CNT), and high maxium power density of $1.79mA{\cdot}cm^{-2}$ was gained.

• KSBB Journal
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• 제27권1호
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• pp.61-66
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• 2012

In the screening process of organic solvent tolerant bacteria showing good growth in media containing several kinds of organic solvents, one strain was isolated and identified as Bacillus sp. BCNU 5006. The strain was able to tolerate many organic solvents including benzene, toluene, xylene, octane, dodecane, butanol and ethylbenzene. Likewise, it could also utilize these solvents as the sole source of carbon with significant enzyme production. The lipolytic enzyme stability of Bacillus sp. BCNU 5006 was studied in the presence of several kinds of solvents at a 25% (v/v) concentration. The highest enzyme stability was observed in the presence of octane (107%), followed by ethylbenzene (88%), decane (86%), and chloroform (85%). Especially, BCNU 5006 lipase was determined to be more stable than immobilized enzyme (Novozyme 435) in the presence of octane, chloroform and xylene. This organic solvent tolerant Bacillus sp. BCNU 5006 could be expected as a potential bioremediation agent and biocatalyst for biodegradation and provide on organic-solvent-based enzymatic synthetic method in industrial chemical processes.

• Journal of Microbiology and Biotechnology
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• 제18권5호
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• pp.946-951
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• 2008

The opd gene, encoding organophosphorus hydrolase (OPH) from Flavobacterium sp. capable of degrading a wide range of organophosphate pesticides, was surface- and intracellular-expressed in Synechococcus PCC7942, a prime example of photoautotrophic cyanobacteria. OPH was displayed on the cyanobacterial cell surface using the truncated ice nucleation protein as an anchoring motif. A minor fraction of OPH was displayed onto the outermost surface of cyanobacterial cells, as verified by immunostaining visualized under confocal laser scanning microscopy and OPH activity analysis; however, a substantial fraction of OPH was buried in the cell wall, as demonstrated by proteinase K and lysozyme treatments. The cyanobacterial outer membrane acts as a substrate (paraoxon) diffusion barrier affecting whole-cell biodegradation efficiency. After freeze-thaw treatment, permeabilized whole cells with intracellular-expressed OPH exhibited 14-fold higher bioconversion efficiency ($V_{max}/K_m$) than that of cells with surface-expressed OPH. As cyanobacteria have simple growth requirements and are inexpensive to maintain, expression of OPH in cyanobacteria may lead to the development of a low-cost and low-maintenance biocatalyst that is useful for detoxification of organophosphate pesticides.

• Journal of Microbiology
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• 제43권5호
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• pp.451-455
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• 2005

In this study, whole cells and a crude enzyme of Candida peltata were applied to an electrochemical bioreactor, in order to induce an increment of the reduction of xylose to xylitol. Neutral red was utilized as an electron mediator in the whole cell reactor, and a graphite-Mn(IV) electrode was used as a catalyst in the enzyme reactor in order to induce the electrochemical reduction of $NAD^+$ to NADH. The efficiency with which xylose was converted to xylitol in the electrochemical bioreactor was five times higher than that in the conventional bioreactor, when whole cells were employed as a biocatalyst. Meanwhile, the xylose to xylitol reduction efficiency in the enzyme reactor using the graphite-Mn (IV) electrode and $NAD^+$ was twice as high as that observed in the conventional bioreactor which utilized NADH as a reducing power. In order to use the graphite-Mn(IV) electrode as a catalyst for the reduction of $NAD^+$ to NADH, a bioelectrocatalyst was engineered, namely, oxidoreductase (e.g. xylose reductase). $NAD^+$ can function in this biotransformation procedure without any electron mediator or a second oxidoreductase for $NAD^+/NADH$ recycling

• Korean Journal of Chemical Engineering
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• 제35권12호
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• pp.2421-2429
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• 2018

Thiol-based self-assembled anchor linked to glucose oxidase (GOx) and gold nanoparticle (GNP) cluster is suggested to enhance the performance of glucose biosensor. By the adoption of thiol-based anchors, the activity of biocatalyst consisting of GOx, GNP, polyethyleneimine (PEI) and carbon nanotube (CNT) is improved because they play a crucial role in preventing the leaching out of GOx. They also promote electron collection and transfer, and this is due to a strong hydrophobic interaction between the active site of GOx and the aromatic ring of anchor, while the effect is optimized with the use of thiophenol anchor due to its simple configuration. Based on that, it is quantified that by the adoption of thiophenol as anchor, the current density of flavin adenine dinucleotide (FAD) redox reaction increases about 42%, electron transfer rate constant ($k_s$) is $9.1{\pm}0.1s^{-1}$ and the value is 26% higher than that of catalyst that does not use the anchor structure.

• Journal of the Korean Wood Science and Technology
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• 제47권6호
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• pp.751-760
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• 2019

Cellulase is an eco-friendly biocatalyst, and its demand is growing in many industrial applications such as food, textile, paper, and bioenergy. Strains with a high cellulase activities are the starting point for the economic production of cellulase. In a previous study, Acanthophysium sp. KMF001 with high cellulase production ability was selected among 54 wood-rotting fungi. In this study, we evaluated the cellulase productivity of Acanthophysium sp. KMF001 quantitatively and analyzed its taxonomic location using a genetic method. Acanthophysium sp. KMF001 showed high cellulase productivity similar to that of Acanthophysium bisporum and was much better than A. bisporum in specific enzyme activity. The 28S rRNA sequence of Acanthophysium sp. KMF001 was similar to that of Acanthophysium lividocaeruleum MB1825, with 98.40% homology. Phylogenetic analysis suggested that Acanthophysium sp. KMF001 is a new strain. In this study, we propose a new strain with high cellulase productivity.