• 한국식품영양학회지
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• 제6권3호
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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.

• 한국식품영양학회지
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• 제6권3호
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• pp.169-177
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• 1993

The conversion of glutamate by glutamine synthetase Is the endergonic reaction that demands ATP as its energy source. In order to supply efficiently ATP that is demanded in the conversion of glutamate to glutamine, the ATP- generating system by acetate kinase partially purified from Escherichia coli K-12 was coupled with glutamine synthetase partially purified 5. coli K-12 Pgln6. The optinum conditions of the coupled reaction were investigated. As the result, the highest conversion of glutamate to glutamine was shown In the reaction mixture containing 100mM glutamate, 100mM NHtCl, 50M acetyl phosphate, 5mM ADP, 40M MgCl2, 300mM potassium phosphate buffer (pH 7.5), 5mM MnCl2, Under this condition, the most effective concentrations of enzyme were 70unit/ml glutamine synthetase and 99unit/ml acetate kinase. Under the optinum conditions, 98% of 100mM glutamate was converted to glutamine within 6 hours.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 1991년도 춘계학술발표대회 논문집
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• pp.237-242
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• 1991

In order to increase the production of glutathione by maximizing the expression of recombinant gsh plasmids, two genes responsible for the biosynthesis of glutathione were cloned. A gshI gene was cloned onto pBR322 plasmid as 3.6Kb PstI DNA fragment from E. coli K-12 chromosomal DNA. Also gshII gene was cloned onto pUC13 plasmid as 2.2Kb PstI-BamHI DNA fragment. In order to improve the glutathione producing activity more efficiently, various recombinant plasmids containing tandem repeated gshI genes or both genes in various copy number onto the same vector were constructed. E. coli cells harboring pGH501 plasmid (pUC8-gshI$\cdot$I$\cdot$II) showed the highest glutathione synthesizing activity. The conditions for glutathione production with an ATP-generating system such as acetate kinase reaction of E. coli cells or glycolytic pathway of yeast cells were examined using the E. coli cells harboring the pGH501 plasmid. When the acetate kinase reaction of E. coli cells was used as an ATP generating system, 20mM of L-csteine was converted into glutathione with a yield of $100\%$.

• Toxicological Research
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• 제22권1호
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• pp.39-45
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• 2006

As the antioxidant and free radical scavenger, glutathione (GSH) participates in the preservation of cellular redox status and defense against reactive oxygen species and xenobiotics. Glutamate-cysteine ligase (GCL; also known as ${\gamma}$-glutamylcysteine synthetase, EC 6.3.2.2) is the rate limiting enzyme in GSH synthesis. In the present study, the accurate method for determination of GCL activity in crude liver extracts was developed by measuring both ${\gamma}$-glutamylcysteine and GSH from cysteine in the presence of glutamate, glycine and an ATP-generating system. We added glycine to promote the conversion of ${\gamma}$-glutamylcysteine to GSH, and to minimize the possibility of ${\gamma}$-glutamylcysteine metabolism to cysteine and oxoproline by ${\gamma}$-glutamylcyclotransferase. We established optimal conditions and substrate concentrations for the enzyme assay, and verified that inhibition of GCL by GSH did not interfere with this assay. Therefore, this assay of hepatic GCL under optimal conditions could provide a more accurate measurement of this enzyme activity in the crude liver extracts.

• 정보처리학회논문지B
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• 제11B권2호
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• pp.221-226
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• 2004

개미 집단 시스템(Ant Colony System ACS) 알고리즘은 조합 최적화 문제를 해결하기 위한 새로운 메타 휴리스틱 방법이다. 이것은 그리디 탐색뿐만 아니라 긍정적 피드백에 의한 탐색을 이용한 모집단에 근거한 접근법으로 조합 최적화 문제를 해결하기 위해 제안되었다. 최근까지 인접한 노드($v_i, v_j$)가 같은 색을 갖지 않도록 그래프 G의 노드 V에 색을 배정하는 문제인 그래프 착색 문제의 최적 해를 구하기 위하여 다양한 접근 방식들과 해법들이 제안되고 있다. 본 논문에서는 기존의 그래프 착색 문제의 해법으로 잘 알려진 그리디 알고리즘, 시뮬레이티드어넬링, 타부 탐색 등이 아닌 개미 집단 시스템 알고리즘으로 해법을 구하는 방법인 ANTCOL 알고리즘을 소개하고, ANTCOL을 해결하기 위해 제안된 기존의 생성 함수들(ANT_Random ANT_LF, ANT_SL, ANT_DSATUR, ANT_RLF)과, 본 논문에서 새롭게 제안된 방법으로 RLF에 무작위 기법을 적용한 XRLF를 생성 함수로 사용한 ANT_XRLF 방법과 ANT_XRLF에 재검색을 추가한 방법(ANT_XRLF_R)의 그래프 착색 결과 및 실행 시간을 비교, 분석하여 제안된 방법이 더 빠르게 수렴할 수 있음을 실험을 통해 알 수 있었다.