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Constitutive production of human leptin by fed-batch culture of recombinant rpoS−Escherichia coli

Authors
 Ki Jun Jeong  ;  Jong Hyun Choi  ;  Moon-Hee Sung  ;  Sang Yup Lee  ;  Nae Choon Yoo  ;  Ki Chang Keum  ;  Won Min Yoo 
Citation
 PROTEIN EXPRESSION AND PURIFICATION, Vol.36(1) : 150-156, 2004 
Journal Title
 PROTEIN EXPRESSION AND PURIFICATION 
ISSN
 1046-5928 
Issue Date
2004
MeSH
Alanine Transaminase/genetics ; Bacterial Proteins/genetics* ; D-Alanine Transaminase ; Escherichia coli/genetics* ; Escherichia coli/metabolism ; Fermentation ; Gene Deletion ; Genetic Vectors/genetics ; Humans ; Leptin/biosynthesis* ; Leptin/genetics ; Promoter Regions, Genetic/genetics ; Sigma Factor/genetics*
Keywords
Constitutive promoter ; Protein expression ; Human leptin ; High cell density culture
Abstract
High-level production of human leptin by fed-batch culture of recombinant Escherichia coli using constitutive promoter system was investigated. For the constitutive expression of the obese gene encoding human leptin, the strong constitutive HCE promoter cloned from the d-amino acid aminotransferase gene of Geobacillus toebii was used. To develop an optimal host–vector system, several different recombinant E. coli strains were compared for leptin production. In flask cultures, E. coli FMJ123, which is a rpoS mutant strain, showed the highest level of leptin production (41% of total proteins). By comparing the expression levels of leptin in several different rpoS− and rpoS+ strains, it could be concluded that rpoS mutation positively affected constitutive production of leptin. For the large-scale production of human leptin, fed-batch cultures of recombinant E. coli FMJ123 were carried out using three different feeding solutions—chemically defined, yeast extract-containing, and casamino acid-containing feeding solutions. Among these, the use of casamino acid-containing feeding solution allowed production of leptin up to 2.1 g/L, which was 2.1- and 1.8-fold higher than that obtained with chemically defined and yeast extract-contained feeding solutions, respectively. These results suggest that the HCE promoter can be used for the efficient production of leptin, and most likely other recombinant proteins, in a constitutive manner.
Full Text
http://www.sciencedirect.com/science/article/pii/S1046592804001214
DOI
10.1016/j.pep.2004.04.007
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Radiation Oncology (방사선종양학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Plastic and Reconstructive Surgery (성형외과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
Keum, Ki Chang(금기창) ORCID logo https://orcid.org/0000-0003-4123-7998
Yoo, Nae Choon(유내춘)
Yoo, Won Min(유원민)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/111697
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