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Process characterization of a novel cross-regulation system for cloned protein production in Escherichia coli

Chen, Wilfred and Kallio, Pauli T. and Bailey, James E. (1995) Process characterization of a novel cross-regulation system for cloned protein production in Escherichia coli. Biotechnology Progress, 11 (4). pp. 397-402. ISSN 8756-7938. https://resolver.caltech.edu/CaltechAUTHORS:20180313-140924453

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Abstract

A novel cross-regulation expression system has been shown previously to be very effective for regulated recombinant protein production. Earlier studies established that this system offers better control of basal expression and higher maximal induced expression than more traditional vectors. Using production of cloned chloramphemcol acetyltransferase (CAT) as a model system, several factors determining performance of this system were examined. Specifically, the effects of varying induction times and inducer (IPTG) concentrations on cell growth and the rate of CAT product10n were examined. The CAT expression was maximally induced with at least 0.5 mM IPTG added at the midexponential growth phase. Specific CAT content (on a total protein basis) was correlated with the CAT mRNA level. CAT message levels were minimal preinduction and far above background postinduction, consistent with pr10r simulation results. Cessation of CAT accumulation as the culture entered the stationary phase coincided with a corresponding 10-fold decrease in the level of CAT mRNA which was likely caused by an increased mRNA degradation rate. Maintenance of significant CAT message levels with a concomitant 2-fold increase in CAT accumulation was achieved by extending cell growth in a fed-batch process.


Item Type:Article
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https://pubs.acs.org/doi/abs/10.1021/bp00034a006PublisherArticle
Additional Information:© 1995 American Chemical Society and American Institute of Chemical Engineers. This work was supported by the Advancement Industrial Concepts Division of the U.S. Department of Energy. W. Chen was supported in part by a Predoctoral Training Grant in Biotechnology from the National Institute of General Medical Sciences (National Research Service Award 5 T32 GM 08346-04, Pharmacology Science Program).
Funders:
Funding AgencyGrant Number
Department of Energy (DOE)UNSPECIFIED
NIH Predoctoral Fellowship5 T32 GM 08346-04
Issue or Number:4
Record Number:CaltechAUTHORS:20180313-140924453
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180313-140924453
Official Citation:Process characterization of a novel cross-regulation system for cloned protein production in Escherichia coli. Wilfred Chen, Pauli T. Kallio, and James E. Bailey. Biotechnology Progress 1995 11 (4), 397-402. DOI: 10.1021/bp00034a006
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:85282
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:13 Mar 2018 21:20
Last Modified:03 Oct 2019 19:29

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