Document Detail


Development and application of an arabinose-inducible expression system by facilitating inducer uptake in Corynebacterium glutamicum.
MedLine Citation:
PMID:  22685153     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Corynebacterium glutamicum is currently used for the industrial production of a variety of biological materials. Many available inducible expression systems in this species use lac-derived promoters from Escherichia coli that exhibit much lower levels of inducible expression and leaky basal expression. We developed an arabinose-inducible expression system that contains the L-arabinose regulator AraC, the P(BAD) promoter from the araBAD operon, and the L-arabinose transporter AraE, all of which are derived from E. coli. The level of inducible P(BAD)-based expression could be modulated over a wide concentration range from 0.001 to 0.4% L-arabinose. This system tightly controlled the expression of the uracil phosphoribosyltransferase without leaky expression. When the gene encoding green fluorescent protein (GFP) was under the control of P(BAD) promoter, flow cytometry analysis showed that GFP was expressed in a highly homogeneous profile throughout the cell population. In contrast to the case in E. coli, P(BAD) induction was not significantly affected in the presence of different carbon sources in C. glutamicum, which makes it useful in fermentation applications. We used this system to regulate the expression of the odhI gene from C. glutamicum, which encodes an inhibitor of α-oxoglutarate dehydrogenase, resulting in high levels of glutamate production (up to 13.7 mM) under biotin nonlimiting conditions. This system provides an efficient tool available for molecular biology and metabolic engineering of C. glutamicum.
Authors:
Yun Zhang; Xiuling Shang; Shujuan Lai; Guoqiang Zhang; Yong Liang; Tingyi Wen
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-06-08
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  78     ISSN:  1098-5336     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  2012 Aug 
Date Detail:
Created Date:  2012-07-30     Completed Date:  2012-11-28     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5831-8     Citation Subset:  IM    
Affiliation:
Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
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MeSH Terms
Descriptor/Qualifier:
Arabinose / metabolism*
Carbon / metabolism
Corynebacterium glutamicum / genetics*,  metabolism*
Escherichia coli / genetics
Escherichia coli Proteins / genetics
Flow Cytometry
Gene Expression Regulation, Bacterial / drug effects*
Genes, Reporter
Genetic Vectors
Genetics, Microbial / methods*
Green Fluorescent Proteins / genetics,  metabolism
Membrane Transport Proteins / genetics
Molecular Biology / methods
Promoter Regions, Genetic
Transcription Factors / genetics
Chemical
Reg. No./Substance:
0/Escherichia coli Proteins; 0/Membrane Transport Proteins; 0/Transcription Factors; 147-81-9/Arabinose; 147336-22-9/Green Fluorescent Proteins; 7440-44-0/Carbon
Comments/Corrections

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