Process development for the production of optically pure D-p-hydroxyphenyl glycine using thermostable D-hydantoinase내열성 D-hydantoinase 를 이용하여 광학적으로 순수한 D-p-hydroxyphenylglycine을 제조하는 공정 개발

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dc.contributor.advisorKim, Hak-Sung-
dc.contributor.advisor김학성-
dc.contributor.authorLee, Dong-Cheol-
dc.contributor.author이동철-
dc.date.accessioned2011-12-12T07:52:19Z-
dc.date.available2011-12-12T07:52:19Z-
dc.date.issued1998-
dc.identifier.urihttp://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=144210&flag=dissertation-
dc.identifier.urihttp://hdl.handle.net/10203/27449-
dc.description학위논문(박사) - 한국과학기술원 : 생물과학과, 1998.2, [ ix, 102 p. ]-
dc.description.abstractD-Hydantoinase is an industrial enzyme widely used for the synthesis of optically active D-amino acids. In this study, we developed the practical processes for the production of N-carbamoyl-D-p-hydroxyphenylglycine, which is easily hydrolyzed to D-p-hydroxyphenylglycine under acidic conditions, from 5-(4-hydroxyphenyl)hydantoin using thermostable D-hydantoinase. A gene encoding thermostable D-hydantoinase of Bacillus stearothermophilus SD-1 has previously been cloned and constitutively expressed by its native promoter in Escherichia coli XL1-Blue (Lee et al. 1996. Ann. N. Y. Acad. Sci. 799: 401-405). At first, we attempted mass production of the D-hydantoinase by batch culture of the recombinant E. coli using glycerol as a carbon source. The plasmid content in cells increased in proportion to the culture temperature, which resulted in a two or three fold increase of the specific D-hydantoinase activity at 37 degree of Celsius compared with that at 30 degree of Celsius. The plasmid was stably maintained over 80 generations. When glycerol was initially added to a concentration of 100 g/L, the final biomass concentration reached about 50 g-dry cell weight/L in a 50 L-scale fermentation, resulting in the specific enzyme production of $3.8\times 10^{4}$ unit/g-dry cell weight in a soluble form. Glycerol-using batch cultivation of recombinant E. coli was found to be a cost-effective process for the mass production of industrially useful D-hydantoinase. Secondly, we optimized the process for the production of N-carbamoyl-D-p-hydroxyphenylglycine, which is readily hydrolyzed to D-p-hydroxyphenyl glycine under acidic conditions, from 5-(4-hydroxyphenyl)hydantoin using the mass-produced D-hydantoinase. The enzyme reaction was carried out in a heterogeneous system consisting of a high substrate concentration up to 300 g/L. In this reaction system, most of substrate is present in suspended particles. Optimal temperature and pH were determined to be 45 degree of Celsius and...eng
dc.languageeng-
dc.publisher한국과학기술원-
dc.subjectHeterogeneous-
dc.subjectGlycerol-
dc.subjectHydroxyphenylglycine-
dc.subjectHydantoinase-
dc.subjectModeling-
dc.subject모델링-
dc.subject비균질-
dc.subject글리세롤-
dc.subject히드록시페닐글리신-
dc.subject히단토인-
dc.titleProcess development for the production of optically pure D-p-hydroxyphenyl glycine using thermostable D-hydantoinase-
dc.title.alternative내열성 D-hydantoinase 를 이용하여 광학적으로 순수한 D-p-hydroxyphenylglycine을 제조하는 공정 개발-
dc.typeThesis(Ph.D)-
dc.identifier.CNRN144210/325007-
dc.description.department한국과학기술원 : 생물과학과, -
dc.identifier.uid000945280-
dc.contributor.localauthorKim, Hak-Sung-
dc.contributor.localauthor김학성-
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