• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.329-332
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• 2001

It has been known that, in enzymatic synthesis of cephalexin, the conversion yield was reduced by high loading of ampicillin acylase. In order to elucidate this phenomena, pH-controlled synthesis of cephalexin was examined using a purified Acetobacter turbidans acylase. When the pH of the reaction mixture was maintained at $6.20{\pm}0.04$, the reduction of the maximal conversion rate was not observed even with high enzyme loading. The kinetic parameters also suggest that pH drop during the enzymatic synthesis of cephalexin was mainly attributed to the rapid hydrolysis of D-${\alpha}$-phenylglycine methyl ester to D-${\alpha}$-phenylglycine, rather than the disappearance of 7-amino-3-deacetoxycephalosporanic acid for cephalexin synthesis. At higher molar ratio of two substrates, [D-${\alpha}$-phenylglycine methyl ester]/[7-amino-3-deacetoxycephalosporanic acid], the conversion rate was also elevated under pH-controlled enzymatic synthesis, which implies that the main reason for the pH drop is due to the production of D-${\alpha}$-phenylglycine methyl easter, the effect of a water-methanol cosolvent system on the ester, the effect of a water-methanol cosolvent system on the conversion profile was also examined. Even the though the conversion rate was increased in 10% methanol solution, a higher than 16% methanol in the reaction mixture caused an inactivation of enzyme.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.297-301
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• 2004

The dTDP-D-glucose 4,6-dehydratase from Salmonella enterica was immobilized using covalent binding to cyanogen bromide activated sepharose. The immobilized enzyme was used to produce dTDP-4-keto-6-deoxy-D-glucose, a key sugar intermediate that can be used economically to produce diverse classes of unusual sugars appended in various antibiotics. The enzyme was immobilized on the sepharose after activation with cyanogen bromide. The maximum immobilization (80.03％) was achieved after 14 h of coupling. The covalently immobilized enzyme was stable, and an average of 78.4 ％ conversion was achieved until 120 h of immobilization when it was repeatedly used. Similar conversion was noticed for the first batch using the enzyme entrapped-hydrogel but activity was gradually decreased in the following batches. The production of dTDP-4-keto-6-deoxy-D-glucose by using an immobilized enzyme has high potential for commercial application.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1596-1602
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• 2009

A lipase-catalyzed esterification reaction of (S)-naproxen ethyl ester by CALB (Candida antarctica lipase B) enzyme was performed in supercritical carbon dioxide. Experiments were performed in a high-pressure cell for 10 h at a stirring rate of 150 rpm over a temperature range of 313.15 to 333.15 K and a pressure range of 50 to 175 bar. The productivity of (S)-naproxen ethyl ester was compared with the result in ambient condition. The total reaction time and conversion yields of the catalyzed reaction in supercritical carbon dioxide were compared with those at ambient temperature and pressure. The experimental results show that the conversion and reaction rate were significantly improved at critical condition. The maximum conversion yield was 9.9% (216 h) at ambient condition and 68.9% (3 h) in supercritical state. The effects of varying amounts of enzyme and water were also examined and the optimum condition was found (7 g of enzyme and 2% water content).

• KSBB Journal
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• 제6권3호
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• pp.309-319
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• 1991

효소를 이용하여 대구피 젤라틴 가수분해물을 연속적으로 생산하기 위해 한외여과막 반응기에서의 젤라틴 가수분해 최적 공정조건 및 한외여과막 반응기를 장시간 작동하였을 때의 효소활성 저하원인을 규명하였다. 효소농도가 0.4mg/ml까지 증가함에 따라 기절의 전환율은 증가하였으나 기질의 경우는 농도가 증가함에 따라 전환율이 감소하였다. 유량(Flux)이 클수록 전환율은 감소되었으며 잔류시간이 길수록 전환율을 증가하였다. 한외여과막 반응기 장치의 작동시 가질의 전환율은 반응시간 1시간 부근에서 최대 전환율을 나타내었으며 시간당(8시간 작동) 평균 전환율 감소는 2.1%였다. 한외여과막을 통한 효소 누출은 반응시간 10~60분 범위에서 최대였으며 6시간 이후에는 거의 누출되지 않았다. 전체 효소량 중 누출량은 $50^{\circ}C$에서 16%인데 비해 $35^{\circ}C$에서는 12%였다. 그러나 효소 누출과 기질 전환율 사이에 명백한 상호관계는 나타나지 않았다. 막에 의한 효소의 활성저하는 작동시간 120분까지 나타났으며, 이때 초기속도의 36%가 저하하였다. 연속식(CSTMR)에서 $K_m$ 값은 회분식의 그것보다 20배 큰 반면 $K_2$ 10.5배 낮았다. 최적 가수분해조건은(기질농도 10%, $50^{\circ}C$, pH 8.0, 유량 7.31ml/min, 잔류시간 82분, S / E = 50w / w) 하에서의 생산량은 효소 mg당 가수분해물이 285mg으로 회분식 44.8mg에 비해 6배 이상이었다.

• Archives of Pharmacal Research
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• 제21권1호
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• pp.1-5
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• 1998

Rebamipide, 2-(4-chlorobenzoylamino)-3-[2(1H)-quinolinone-4-yl]-propionic acid, a novel antipeptic ulcer agent, has been reported to prevent various acute experimental gastric mucosal lesions and to accelerate the healing of chronic ulcers. Therapeutic effect of rebamipide was investigated with regard to the inhibitory effect on xanthine oxidase activity and type conversion of the enzyme which play a profound role in oxygen radicals generation system. Intraperitoneal administration of rebamipide at 60 mg/kg body weight reduced the xanthine oxidase activity, lipid peroxide content in ammonia induced hemorrhagic lesion. These results suggest that the therapeutic effect of rebamipide on gastric mucosal lesion may be in part due to the inhibitory activity of xanthine oxidase and type conversion rate of the enzyme.

• 생명과학회지
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• 제11권3호
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• pp.230-234
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• 2001

The cyclodextrin glucanotransferase(CGTase) gene of Bacillus macerans was expressed in Saccharomyces cerevisiae and the recombinant CGTase was partially purified from the yeast culture supernatant. The optimal pH and temperature of the CGTase were found to be 6.0 and 5$0^{\circ}C$, respectively. The pH and temperature stabilities of the recombinant enzyme were significantly enhanced and the half life at 55$^{\circ}C$ was about 60 hr. When the recombinant CGTase was reacted with 5% soluble starch, the conversion yield of total cyclodextrin (CD) from starch was estimated to be 41% at 48 hr, whereas the wild type enzyme showed the yield of 12%. This improvement of conversion yield and thermal stability of CGTase may be useful for the development of low-cost CD production process.

• BMB Reports
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• 제37권6호
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• pp.642-647
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• 2004

The lysine residues of Bacillus licheniformis $\alpha$-amylase (BLA) were chemically modified using citraconic anhydride or succinic anhydride. Modification caused fundamental changes in the enzymes specificity, as indicated by a dramatic increase in maltosidase and a reduction in amylase activity. These changes in substrate specificity were found to coincide with a change in the cleavage pattern of the substrates and with a conversion of the native endo- form of the enzyme to a modified exo- form. Progressive increases in the productions of $\rho$-nitrophenol or glucose, when para nitrophenyl-maltoheptaoside or soluble starch, respectively, was used as substrate, were observed upon modification. The described changes were affected by the size of incorporated modified reagent: citraconic anhydride was more effective than succinic anhydride. Reasons for the observed changes are discussed and reasons for the effectivenesses of chemical modifications for tailoring enzyme specificities are suggested.

• Journal of Microbiology and Biotechnology
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• 제1권1호
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• pp.50-53
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• 1991

The reaction steps involved in the bioconversion of a chemically synthesized precursor, $D,L-2-amino-{\Delta}^2-thiazoline-4-carboxylic$ acid (D,L-ATC), to L-cysteine and the properties of the involved enzymes were investigated. It was found that the conversion consisted of two steps, i. e., D,L-ATC to S-carbamyl-L-cysteine (S-C-L-cysteine) and S-C-L-cysteine to L-cysteine, and the S-C-L-cysteine was an intermediate between them. While the enzymes involved in the reactions were induced by the addition of D,L-ATC as an inducer, S-C-L-cysteine induced only the enzyme involved in the latter step. The conversion of S-C-L-cysteine to L-cysteine could be also carried out in the presence of hydroxylamine and its rate was much faster than that by the corresponding enzyme. On the other hand, L-cysteine (or L-cystine) was decomposed to evolve $H_2S$ by the enzyme considered to be a kind of desulfhydrase. However, hydroxylamine was a perfect inhibitor for this enzyme.

• 멤브레인
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• 제8권3호
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• pp.170-176
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• 1998

가용성 전분으로부터 CGTase 효소에 의한 Cyclodextrin(CD) 동족체의 생산 실험을 dead-end형 막모듈을 설치한 막-효소 반응기에서 수행하였다. 회분 반응기에서의 CD 동족체 생산시 생산물 저해작용 및 생산된 CD의 다른 환원당으로의 분해에 의해 반응시간에 따른 전분의 CD 동족체로의 총 전환율이 최대 45%를 나타내었다가 급격히 감소하였다. 반면 분획분자량 10,000 달톤의 분리막이 설치된 막-효소 반응기에서의 CD 동족체 생산시 CD 동족체를 생산 즉시 반응기로부터 막을 통해 연속 분리할 수 있어 총 전환율이 35%로 일정하게 유지되었다. 10% 전분용액 및 2 atm 조작압력하에서 막-효소 반응기를 24시간 운전하였을 때 CD 동족체의 누적 생산량은 약 3.7 kg/m$^2$이었다.

• Journal of Microbiology and Biotechnology
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• 제24권10호
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• pp.1427-1437
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• 2014

Enzymatic hydrolysis of lignocellulosic biomass into fermentable sugars is a key step in the conversion of agricultural by-products to biofuels and value-added chemicals. Utilization of a robust microorganism for on-site production of biomass-degrading enzymes has gained increasing interest as an economical approach for supplying enzymes to biorefinery processes. In this study, production of multi-polysaccharide-degrading enzymes from Aspergillus aculeatus BCC199 by solid-state fermentation was improved through the statistical design approach. Among the operational parameters, yeast extract and soybean meal as well as the nonionic surfactant Tween 20 and initial pH were found as key parameters for maximizing production of cellulolytic and hemicellulolytic enzymes. Under the optimized condition, the production of FPase, endoglucanase, ${\beta}$-glucosidase, xylanase, and ${\beta}$-xylosidase was achieved at 23, 663, 88, 1,633, and 90 units/g of dry substrate, respectively. The multi-enzyme extract was highly efficient in the saccharification of alkaline-pretreated rice straw, corn cob, and corn stover. In comparison with commercial cellulase preparations, the BCC199 enzyme mixture was able to produce remarkable yields of glucose and xylose, as it contained higher relative activities of ${\beta}$-glucosidase and core hemicellulases (xylanase and ${\beta}$-xylosidase). These results suggested that the crude enzyme extract from A. aculeatus BCC199 possesses balanced cellulolytic and xylanolytic activities required for the efficient saccharification of lignocellulosic biomass feedstocks, and supplementation of external ${\beta}$-glucosidase or xylanase was dispensable. The work thus demonstrates the high potential of A. aculeatus BCC199 as a promising producer of lignocellulose-degrading enzymes for the biomass conversion industry.