4.2 Article

Heterologous protein production in Escherichia coli using the propionate-inducible pPro system by conventional and auto-induction methods

Journal

PROTEIN EXPRESSION AND PURIFICATION
Volume 61, Issue 2, Pages 197-203

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2008.06.008

Keywords

auto-induction; Escherichia coli; gene expression system; pPro system; prpBCDE promoter; T7 promoter

Funding

  1. National Institutes of Health [GM070763-1]

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We examined expression of two plant genes encoding coclaurine N-methyltransferase (CMT) and norcoclaurine synthase (NCS) in Escherichia coli from the Salmonella enterica prpBCDE promoter (Pp,B) and compared it to that from the strongest IPTG-inducible promoter, P-T7. In contrast to our previous study showing slightly higher production of green fluorescent protein (GFP) from the pPro system compared to that from the T7 system, production of two plant proteins CMT and NCS from P-prpB was 2- to 4-fold higher than that from P-T7. Unlike P-T7, expression from P-prB did not reduce cell growth even when highly induced, indicating that this propionate-inducible system is more efficient for overproduction of proteins that result in growth inhibition. In an auto-induction experiment, which does not require monitoring the culture or adding inducer during cell growth, the pPro system exhibited much higher protein production than the T7 system. These results strongly indicate that the pPro system is well-suited for overproduction of recombinant proteins. Published by Elsevier Inc.

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