Overproduction and purification of pseudomonas fluorescens esterase Ⅲ from escherichia coli = 대장균에서 발현된 pseudomonas fluorescens esterase Ⅲ의 대량생산 및 순수정제

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The esterase III encoded by estC gene of Pseudomonas fluorescens was overexpressed in Escherichia coli BL21(DE3) by using the T7 expression system. The open reading frame of estC gene amplified by polymerase chain reaction was subcloned into T7 promoter-containing plasmid, pRSET. The resultant plasmid, pREIII was used for transformation of E. coli cells. The expression level of esteraseIII in E. coli harboring pREIII was 20 times higher than that harboring pUE892 which contains estC under lac promoter. The protein was successfully expressed without forming any inclusion body in side the E. coli cells. The amount of the enzyme was estimated to be 7-14\% of total cellular protein as judged by densitometer after SDS-gel electrophoresis. Following sequential DEAE-Sepharose ion exchange chromatography and Superose 12 gel filtration chromatography, the protein was purified to homogeneity. After purification, activity yield was 34\% and thr specific activity was 107000.
Advisors
Yoo, Ook-Joonresearcher유욱준researcher
Description
한국과학기술원 : 생명과학과,
Publisher
한국과학기술원
Issue Date
1994
Identifier
69124/325007 / 000923179
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생명과학과, 1994.2, [ vi, 49 p. ]

Keywords

단백질 분리.

URI
http://hdl.handle.net/10203/28417
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=69124&flag=dissertation
Appears in Collection
BS-Theses_Master(석사논문)
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