Medium and copy number effects on the secretion of human proinsulin in Escherichia coli using the universal stress promoters uspA and uspB

被引:16
作者
Mergulhao, FJM
Monteiro, GA
Larsson, G
Sandén, AM
Farewell, A
Nystrom, T
Cabral, JMS
Taipa, MA
机构
[1] Inst Super Tecn, Ctr Biol & Chem Engn, P-1049001 Lisbon, Portugal
[2] Stockholm Ctr Phys Astron & Biotechnol, Swedish Ctr Bioprocess Technol, S-10691 Stockholm, Sweden
[3] Gothenburg Univ, Dept Gen & Marine Microbiol, S-41390 Gothenburg, Sweden
关键词
D O I
10.1007/s00253-003-1232-8
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The use of the uspA and uspB promoters (universal stress promoters) for heterologous protein production in Escherichia coli is described. Best results were obtained with a moderate copy number vector (1560 copies) bearing the uspA promoter, reaching 4.6 mg/g dry cell weight (DCW) of ZZ-proinsulin secreted to the periplasm and 1.9 mg/g DCW secreted to the culture medium. These values are about 1.7-fold higher than those previously reported with the same ZZ fusion tag and the SpA leader peptide showing that these stress promoters are potentially valuable for recombinant protein secretion in E. coli. It is further demonstrated that the use of M9 minimal medium is advantageous for protein secretion as compared to LB rich medium.
引用
收藏
页码:495 / 501
页数:7
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