DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | Kim, Hak-Sung | - |
dc.contributor.advisor | 김학성 | - |
dc.contributor.author | Lee, Dong-Cheol | - |
dc.contributor.author | 이동철 | - |
dc.date.accessioned | 2011-12-12T07:52:19Z | - |
dc.date.available | 2011-12-12T07:52:19Z | - |
dc.date.issued | 1998 | - |
dc.identifier.uri | http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=144210&flag=dissertation | - |
dc.identifier.uri | http://hdl.handle.net/10203/27449 | - |
dc.description | 학위논문(박사) - 한국과학기술원 : 생물과학과, 1998.2, [ ix, 102 p. ] | - |
dc.description.abstract | D-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.language | eng | - |
dc.publisher | 한국과학기술원 | - |
dc.subject | Heterogeneous | - |
dc.subject | Glycerol | - |
dc.subject | Hydroxyphenylglycine | - |
dc.subject | Hydantoinase | - |
dc.subject | Modeling | - |
dc.subject | 모델링 | - |
dc.subject | 비균질 | - |
dc.subject | 글리세롤 | - |
dc.subject | 히드록시페닐글리신 | - |
dc.subject | 히단토인 | - |
dc.title | Process development for the production of optically pure D-p-hydroxyphenyl glycine using thermostable D-hydantoinase | - |
dc.title.alternative | 내열성 D-hydantoinase 를 이용하여 광학적으로 순수한 D-p-hydroxyphenylglycine을 제조하는 공정 개발 | - |
dc.type | Thesis(Ph.D) | - |
dc.identifier.CNRN | 144210/325007 | - |
dc.description.department | 한국과학기술원 : 생물과학과, | - |
dc.identifier.uid | 000945280 | - |
dc.contributor.localauthor | Kim, Hak-Sung | - |
dc.contributor.localauthor | 김학성 | - |
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