• 한국미생물·생명공학회지
• /
• 제51권4호
• /
• pp.333-352
• /
• 2023

Transaminase represents the most important biocatalysts used for the synthesis of chiral amines due to their stereoselectivity. They allow asymmetric synthesis with high yields and enantioselectivity from their corresponding ketones. Due to their environmentally friendly access for the preparation of chiral amines, they have attracted growing attention in recent times. Thus, the production of chiral compounds by transaminase catalysed reactions is considered as an important application in synthetic organic chemistry. Therefore, transaminase is considered to be an important enzyme in the pharmaceutical and chemical industries. ω-Transaminase holds great potential because of its wide substrate specificity thus making it a suitable enzyme to be used at an industrial scale. This review highlights the reaction mechanism, classification, substrate specificity, and biochemical properties. The review also showcases the application of ω-transaminase in organic chemistry with a focus on the production of active pharmaceutical ingredients (APIs).

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2003년도 생물공학의 동향(XIII)
• /
• pp.468-472
• /
• 2003

Recently, genetic engineering techniques have been used to display various heterologous peptides and proteins (enzyme, antibody, antigen, receptor and fluorescence protein, etc.) on the yeast cell surface. Living cells displaying various enzymes on their surface could be used repeatedly as 'whole cell biocatalysts' like immobilized enzymes. We constructed a yeast based whole cell biocatalyst displaying T. reesei cellobiohydrolase I (CBH I ) on the cell surface and endowed the yeast-cells with the ability to degrade cellulose. By using a cell surface engineering system based on ${\alpha}-agglutinin,$ CBH I was displayed on the cell surface as a fusion protein containing the N-terminal leader peptide encoding a Gly-Ser linker and the $Xpress^{TM}$ epitope. Localization of the fusion protein on the cell surface was confirmed by confocal microscopy. In this study, we report on the genetic immobilization of T. reesei CBH I on the S. cerevisiae and hydrolytic activity of cell surface displayed CBH I.

• Journal of Microbiology and Biotechnology
• /
• 제13권6호
• /
• pp.1001-1007
• /
• 2003

Metal ions contamination may inhibit microorganisms involved in the biodegradation of organic compounds and affect biodegradation rates. Therefore, it is likely that bioremediation of xenobiotics-contaminated soils and waste will require inoculation with efficient biodegrading microbial communities, with capabilities of being resistant to heavy metals as well. Two different transconjugants (Pseudomonas sp. KMl2TC and P. aeruginosa TC) were constructed by conjugation experiments. Results on MIC, induction and growth inhibition strongly indicated that arsenic-resistant plasmid, pKM20, could be mobilized, and the newly acquired phenotype of pKM20 was not only expressed but also well regulated, resulting in newly acquired resistances to $As^{5+},\;As^{3+},\;and\;Sb^{3+} in\;addition\;to\;Cd^{2+},\;Zn^{2+},\;and\;Hg^{2+}$. The phenol- degradation efficiencies of Pseudomonas sp. KMl2TC were maintained significantly even at high heavy metal concentrations at which these efficiencies of P. aeruginosa TC were completely impaired. The results in this study on the effects of heavy metals on phenol degradation, especially after conjugation, are the first ever reported. All the results described in this study encourage to establish a goal of making "designer biocatalysts" which could degrade certain xenobiotics in the area contaminated with multiple heavy metals.

• 공업화학
• /
• 제16권3호
• /
• pp.456-459
• /
• 2005

미생물 세포 유래의 에폭사이드 가수분해 효소 활성을 효율적으로 분석하기 위하여 UV 분광기 측정법을 최적화하였다. 세포 생촉매의 에폭사이드 가수분해 활성에 의해 p-nitrostyrene oxide (pNSO) 기질이 p-nitrostyrene diol로 전환된 양을 측정함으로써 에폭사이드 가수분해 활성을 평가하였다. Rhodosporidium toruloides를 생촉매로 사용하고 pNSO에 대한 입체선택적 가수분해 동력학을 UV 분광기 측정법으로 분석한 결과, $K_m$과 $V_m$ 값을 $2.457nmol/min{\cdot}mg$ 및 1.078 mM로 각각 결정할 수 있었다.

• Journal of Microbiology and Biotechnology
• /
• 제17권7호
• /
• pp.1083-1089
• /
• 2007

Because of their selectivity and catalytic efficiency, BVMOs are highly valuable biocatalysts for the chemoenzymatic synthesis of a broad range of useful compounds. In this study, we investigated the microbial Baeyer-Villiger oxidation and sulfoxidation of thioanisole and bicyclo[3.2.0]hept-2-en-6-one using whole Escherichia coli cells that recombined with each of the Baeyer-Villiger monooxygenases originated from Pseudomonas aeruginosa PAOl and two from Streptomyces coelicolor A3(2). The three BVMOs were identified in the microbial genome database by a recently described protein sequence motif; e.g., BVMO motif(FXGXXXHXXXW). The reaction products were identified as (R)-/(S)-sulfoxide and 2-oxabicyclo/3-oxabicyclo[3.3.0]oct-6-en-2-one by GC-MS analysis. Consequently, this study demonstrated that the three enzymes can indeed catalyze the Baeyer-Villiger reaction as a biocatalyst, and effective annotation tools can be efficiently exploited as a source of novel BVMOs.

• 한국수소및신에너지학회논문집
• /
• 제27권4호
• /
• pp.378-385
• /
• 2016

In this study, we synthesized a biocatalyst consisting of glucose oxidase (GOx), polyethyleneimine (PEI) and carbon nanotube (CNT) with addition of glutaraldehyde (GA)(GA/[GOx/PEI/CNT])for fabrication of glucose sensor. Main bonding of the GA/[GOx/PEI/CNT] catalyst was formed by crosslinking of functional end groups between GOx/PEI and GA. Catalytic activity of GA/[GOx/PEI/CNT] was quantified by UV-Vis and electrochemical measurements. As a result of that, high immobilization ratio of 199% than other catalyst (with only physical adsorption) and large sensitivity value of $13.4{\mu}A/cm^2/mM$ was gained. With estimation of the biosensor stability, it was found that the GA/[GOx/PEI/CNT] kept about 88% of its initial activity even after three weeks. It shows GA minimized the loss of GOx and improved sensing ability and stability compared with that using other biocatalysts.

• KSBB Journal
• /
• 제7권3호
• /
• pp.172-178
• /
• 1992

담체로 사용된 polyurethane foam은 Rizopus chinensis의 균사가 부착하여 안정하게 증식할 수 있게 하였다. 고정화를 위해 사용된 네 종류의 polyurethane foam중 GP-160이 고정하ㅗ 매체로 우수한 성질을 보였고, 입자의 크기는 7-8mm가 적당하였다. Rizopus chinensis의 현탁 배양과 polyurethane foam에서의 고정화 배양을 비교할 때, 전체 리파아제의 활성도는 큰 변화가 없었지만, 고정화 배양의 경우 extracellualar lipase의 생성을 억제하여 intracellular lipase의 활성도를 현탁 배양의 경우보다 약 2배가량 높일 수가 있었다. 고정화 세포의 열안정성을 조사하기 위하여 35~$50^{\circ}C$사이에서 열에의한 비활성화 에너지값을 구해본 결과, 그 값이 28.7kcal/mol로서 본 연구에서 제조된 고정화 세포의 생촉매가 배교적 좋은 열안정성을 갖고 있다.

• Journal of Microbiology and Biotechnology
• /
• 제16권2호
• /
• pp.295-298
• /
• 2006

Actinomycetes are ubiquitous Gram-positive soil bacteria and a group of the most important industrial microorganisms for the biosynthesis of many valuable secondary metabolites as well as the source of various bioconversion enzymes. Cytochrome P450 hydroxylase (CYP), a hemebinding protein, is known to be involved in the modification of various natural compounds, including polyketides, fatty acids, steroids, and some aromatic compounds. Previously, six different novel CYP genes were isolated from a rare actinomycetes called Sebekia benihana, and they were completely sequenced, revealing significant amino acid similarities to previously known CYP genes involved in Streptomyces secondary metabolism. In the present study, these six CYP genes were functionally expressed in Streptomyces lividans, using an $ermE^{*}$ promoter-containing Streptomyces expression vector. Among six CYP genes, two S. benihana CYP genes (CYP503 and CYP504) showed strong hydroxylation activities toward 7-ethoxycoumarin. Furthermore, the recombinant S. lividans containing both the S. benihana CYP506-ferredoxin genes as well as the S. coelicolor feredoxin reductase gene also demonstrated cyclosporin A hydroxylation activity, suggesting potential application of actinomycetes CYPs for the biocatalysts of natural product bioconversion.

• 공업화학
• /
• 제24권6호
• /
• pp.567-578
• /
• 2013

미생물연료전지는 혐기성 조건에서 미생물의 촉매 반응을 통해 유기물질의 화학에너지를 전기에너지로 변환하는 생물전기화학 장치이다. 미생물연료전지의 전력밀도 및 쿨롱효율은 산화전극 챔버 내 미생물의 종류, 시스템 구성요소 및 운전조건에 영향을 받는다. 미생물연료전지에서 달성할 수 있는 전력은 구성요소, 물리적 및 화학적 운전조건, 바이오 촉매 선택 등의 최적화로 디자인을 변형하여 현저하게 증가시킬 수 있다. 본 총설에서는 미생물연료전지의 구성, 운전 매개변수의 최적화 및 성능과 더불어 장래 응용에 대한 최근 연구를 중점적으로 고찰하고자 한다.

• Journal of Microbiology and Biotechnology
• /
• 제28권8호
• /
• pp.1346-1351
• /
• 2018

Oxidoreductases are effective biocatalysts, but their practical use is limited by the need for large quantities of NAD(P)H. In this study, a whole-cell biocatalyst for NAD(P)H cofactor regeneration was developed using the economical substrate glycerol. This cofactor regeneration system employs permeabilized Escherichia coli cells in which the glpD and gldA genes were deleted and the gpsA gene, which encodes $NAD(P)^+-dependent$ glycerol-3-phosphate dehydrogenase, was overexpressed. These manipulations were applied to block a side reaction (i.e., the conversion of glycerol to dihydroxyacetone) and to switch the glpD-encoding enzyme reaction to a gpsA-encoding enzyme reaction that generates both NADH and NADPH. We demonstrated the performance of the cofactor regeneration system using a lactate dehydrogenase reaction as a coupling reaction model. The developed biocatalyst involves an economical substrate, bifunctional regeneration of NAD(P)H, and simple reaction conditions as well as a stable environment for enzymes, and is thus applicable to a variety of oxidoreductase reactions requiring NAD(P)H regeneration.