Expression of the Klebsiella pneumoniae CG21 acetoin reductase gene in Clostridium acetobutylicum ATCC 824

Acetoin reductase catalyzes the production of 2,3-butanediol from acetoin. The gene encoding the acetoin reductase of Klebsiella pneumoniae CG21 was cloned and expressed in Escherichia coli and Clostridium acetobutylicum ATCC 824. The nucleotide sequence of the gene encoding the enzyme was determine...

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Published in:Journal of industrial microbiology & biotechnology 2001-10, Vol.27 (4), p.220-227
Main Authors: WARDWELL, S. A, YANG, Y. T, CHANG, H. Y, SAN, K. Y, RUDOLPH, F. B, BENNETT, G. N
Format: Article
Language:English
Subjects:
2,3-Butanediol
2,3-Butylene glycol
Acetoin - metabolism
acetoin reductase
acetone reductase
Alcohol Oxidoreductases - genetics
Alcohol Oxidoreductases - metabolism
Amino Acid Sequence
Base Sequence
Biochemistry
Biological and medical sciences
Biotechnology
Butylene Glycols - metabolism
Cloning
Cloning, Molecular
Clostridium - enzymology
Clostridium - genetics
Clostridium - growth & development
Clostridium acetobutylicum
Culture Media
DNA methylation
E coli
Enzymes
Escherichia coli - enzymology
Escherichia coli - genetics
Escherichia coli - growth & development
Fermentation
Fundamental and applied biological sciences. Psychology
Gas chromatography
Genes
Genetic engineering
Genetic technics
Industrial production
Klebsiella pneumoniae
Klebsiella pneumoniae - enzymology
Klebsiella pneumoniae - genetics
Metabolism
Methods. Procedures. Technologies
Microbiology
Microorganisms
Modification of gene expression level
Molecular Sequence Data
Plasmids
Sequence Analysis, DNA
Solvents
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container_issue 4
container_start_page 220
container_title Journal of industrial microbiology & biotechnology
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creator WARDWELL, S. A
YANG, Y. T
CHANG, H. Y
SAN, K. Y
RUDOLPH, F. B
BENNETT, G. N
description Acetoin reductase catalyzes the production of 2,3-butanediol from acetoin. The gene encoding the acetoin reductase of Klebsiella pneumoniae CG21 was cloned and expressed in Escherichia coli and Clostridium acetobutylicum ATCC 824. The nucleotide sequence of the gene encoding the enzyme was determined to be 768 bp long. Expression of the K. pneumoniae acetoin reductase gene in E. coli revealed that the enzyme has a molecular mass of about 31,000 Da based on sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The K. pneumoniae acetoin reductase gene was cloned into a clostridial/E. coli shuttle vector, and expression of the gene resulted in detectable levels of acetoin reductase activity in both E. coli and C. acetobutylicum. While acetoin, the natural substrate of acetoin reductase, is a typical product of fermentation by C. acetobutylicum, 2,3-butanediol is not. Analysis of culture supernatants by gas chromatography revealed that introduction of the K. pneumoniae acetoin reductase gene into C. acetobutylicum was not sufficient for 2,3-butanediol production even though the cultures were producing acetoin. 2,3-Butanediol was produced by cultures of C. acetobutylicum containing the gene only when commercial acetoin was added.
doi_str_mv 10.1038/sj.jim.7000179
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A ; YANG, Y. T ; CHANG, H. Y ; SAN, K. Y ; RUDOLPH, F. B ; BENNETT, G. N</creator><creatorcontrib>WARDWELL, S. A ; YANG, Y. T ; CHANG, H. Y ; SAN, K. Y ; RUDOLPH, F. B ; BENNETT, G. N</creatorcontrib><description>Acetoin reductase catalyzes the production of 2,3-butanediol from acetoin. The gene encoding the acetoin reductase of Klebsiella pneumoniae CG21 was cloned and expressed in Escherichia coli and Clostridium acetobutylicum ATCC 824. The nucleotide sequence of the gene encoding the enzyme was determined to be 768 bp long. Expression of the K. pneumoniae acetoin reductase gene in E. coli revealed that the enzyme has a molecular mass of about 31,000 Da based on sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The K. pneumoniae acetoin reductase gene was cloned into a clostridial/E. coli shuttle vector, and expression of the gene resulted in detectable levels of acetoin reductase activity in both E. coli and C. acetobutylicum. While acetoin, the natural substrate of acetoin reductase, is a typical product of fermentation by C. acetobutylicum, 2,3-butanediol is not. Analysis of culture supernatants by gas chromatography revealed that introduction of the K. pneumoniae acetoin reductase gene into C. acetobutylicum was not sufficient for 2,3-butanediol production even though the cultures were producing acetoin. 2,3-Butanediol was produced by cultures of C. acetobutylicum containing the gene only when commercial acetoin was added.</description><identifier>ISSN: 1367-5435</identifier><identifier>EISSN: 1476-5535</identifier><identifier>DOI: 10.1038/sj.jim.7000179</identifier><identifier>PMID: 11687934</identifier><language>eng</language><publisher>Heidelberg: Springer</publisher><subject>2,3-Butanediol ; 2,3-Butylene glycol ; Acetoin - metabolism ; acetoin reductase ; acetone reductase ; Alcohol Oxidoreductases - genetics ; Alcohol Oxidoreductases - metabolism ; Amino Acid Sequence ; Base Sequence ; Biochemistry ; Biological and medical sciences ; Biotechnology ; Butylene Glycols - metabolism ; Cloning ; Cloning, Molecular ; Clostridium - enzymology ; Clostridium - genetics ; Clostridium - growth &amp; development ; Clostridium acetobutylicum ; Culture Media ; DNA methylation ; E coli ; Enzymes ; Escherichia coli - enzymology ; Escherichia coli - genetics ; Escherichia coli - growth &amp; development ; Fermentation ; Fundamental and applied biological sciences. 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A</au><au>YANG, Y. T</au><au>CHANG, H. Y</au><au>SAN, K. Y</au><au>RUDOLPH, F. B</au><au>BENNETT, G. N</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of the Klebsiella pneumoniae CG21 acetoin reductase gene in Clostridium acetobutylicum ATCC 824</atitle><jtitle>Journal of industrial microbiology &amp; biotechnology</jtitle><addtitle>J Ind Microbiol Biotechnol</addtitle><date>2001-10-01</date><risdate>2001</risdate><volume>27</volume><issue>4</issue><spage>220</spage><epage>227</epage><pages>220-227</pages><issn>1367-5435</issn><eissn>1476-5535</eissn><abstract>Acetoin reductase catalyzes the production of 2,3-butanediol from acetoin. The gene encoding the acetoin reductase of Klebsiella pneumoniae CG21 was cloned and expressed in Escherichia coli and Clostridium acetobutylicum ATCC 824. The nucleotide sequence of the gene encoding the enzyme was determined to be 768 bp long. Expression of the K. pneumoniae acetoin reductase gene in E. coli revealed that the enzyme has a molecular mass of about 31,000 Da based on sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The K. pneumoniae acetoin reductase gene was cloned into a clostridial/E. coli shuttle vector, and expression of the gene resulted in detectable levels of acetoin reductase activity in both E. coli and C. acetobutylicum. While acetoin, the natural substrate of acetoin reductase, is a typical product of fermentation by C. acetobutylicum, 2,3-butanediol is not. Analysis of culture supernatants by gas chromatography revealed that introduction of the K. pneumoniae acetoin reductase gene into C. acetobutylicum was not sufficient for 2,3-butanediol production even though the cultures were producing acetoin. 2,3-Butanediol was produced by cultures of C. acetobutylicum containing the gene only when commercial acetoin was added.</abstract><cop>Heidelberg</cop><pub>Springer</pub><pmid>11687934</pmid><doi>10.1038/sj.jim.7000179</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record>
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subjects 2,3-Butanediol
2,3-Butylene glycol
Acetoin - metabolism
acetoin reductase
acetone reductase
Alcohol Oxidoreductases - genetics
Alcohol Oxidoreductases - metabolism
Amino Acid Sequence
Base Sequence
Biochemistry
Biological and medical sciences
Biotechnology
Butylene Glycols - metabolism
Cloning
Cloning, Molecular
Clostridium - enzymology
Clostridium - genetics
Clostridium - growth & development
Clostridium acetobutylicum
Culture Media
DNA methylation
E coli
Enzymes
Escherichia coli - enzymology
Escherichia coli - genetics
Escherichia coli - growth & development
Fermentation
Fundamental and applied biological sciences. Psychology
Gas chromatography
Genes
Genetic engineering
Genetic technics
Industrial production
Klebsiella pneumoniae
Klebsiella pneumoniae - enzymology
Klebsiella pneumoniae - genetics
Metabolism
Methods. Procedures. Technologies
Microbiology
Microorganisms
Modification of gene expression level
Molecular Sequence Data
Plasmids
Sequence Analysis, DNA
Solvents
title Expression of the Klebsiella pneumoniae CG21 acetoin reductase gene in Clostridium acetobutylicum ATCC 824
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