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Title of Journal: Appl Microbiol Biotechnol

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Abbravation: Applied Microbiology and Biotechnology

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Springer-Verlag

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DOI

10.1016/0300-9629(79)90015-X

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1432-0614

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Characterization of the T7 promoter system for exp

Authors: Yali Xu Stefan Rosenkranz ChiaoLing Weng Jeno M Scharer Murray MooYoung C Perry Chou
Publish Date: 2006/01/13
Volume: 72, Issue: 3, Pages: 529-536
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Abstract

The pac gene encoding penicillin acylase PAC was overexpressed under the regulation of the T7 promoter in Escherichia coli PAC with its complex formation mechanism serves as a unique target protein for demonstration of several key strategies for enhancing recombinant protein production The current T7 system for pac overexpression was fraught with various technical hurdles Upon the induction with a conventional inducer of isopropylβdthiogalactopyranoside IPTG the production of PAC was limited by the accumulation of PAC precursors proPAC as inclusion bodies and various negative cellular responses such as growth inhibition and cell lysis The expression performance could be improved by the coexpression of degP encoding a periplasmic protein with protease and chaperone activities In addition to IPTG arabinose was shown to be another effective inducer Interestingly arabinose not only induced the current T7 promoter system for pac expression but also facilitated the posttranslational processing of proPAC for maturation resulting in significant enhancement for the production of PAC Glycerol appeared to have an effect similar to but not as significant as arabinose for enhancing the production of PAC The study highlights the importance of developing suitable genetically engineered strains with culture conditions for enhancing recombinant protein production in E coli

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