Premium
Elevated Fis expression enhances recombinant protein production in Escherichia coli
Author(s) -
Richins Richard,
Htay Tin,
Kallio Pauli,
Chen Wilfred
Publication year - 1997
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19971020)56:2<138::aid-bit3>3.0.co;2-q
Subject(s) - chloramphenicol acetyltransferase , recombinant dna , biology , escherichia coli , mutant , protein biosynthesis , expression vector , fermentation , biochemistry , microbiology and biotechnology , chemistry , gene expression , gene , promoter
A genetic strategy to enhance recombinant protein production is discussed. A small DNA bending protein, Fis, which has been shown to activate rRNA synthesis upon a nutrient upshift, was overexpressed in E. coli strain W3110 carrying vector pUCR1. Overexpression of Fis during exponential growth was shown to activate rrn promoters to different extents. A 5‐fold improvement in chloramphenicol acetyltransferase (CAT) production in cultures with elevated Fis level was observed in shake‐flask cultivations. A similar improvement in the culture performance was also observed during fed‐batch fermentation; the specific CAT activity increased by more than 50% during the fed‐batch phase for cultures with elevated Fis expression. In contrast, no increase in specific CAT activity was detected for cultures carrying pUCR2, expressing a frame‐shift Fis mutant. Expression of Fis from a complementary vector, pKFIS, restored CAT production from W3110:pUCR2 to approximately the same level as cultures carrying pUCR1, indicating that the enhancement in CAT production was indeed Fis‐dependent. The framework presented here suggests that differential activation in recombinant protein production may be achieved with differential Fis overexpression. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 138–144, 1997.