PMID: 8955296Dec 1, 1996Paper

Menaquinone (vitamin K2) biosynthesis: overexpression, purification, and properties of o-succinylbenzoyl-coenzyme A synthetase from Escherichia coli

Journal of Bacteriology
O KwonR Meganathan

Abstract

The coenzyme A (CoA)- and ATP-dependent conversion of o-succinylbenzoic acid [OSB; 4-(2'-carboxyphenyl)-4-oxobutyric acid], to o-succinylbenzoyl-CoA is carried out by the enzyme o-succinylbenzoyl-CoA synthetase. o-Succinylbenzoyl-CoA is a key intermediate in the biosynthesis of menaquinone (vitamin K2) in both gram-negative and gram-positive bacteria. The enzyme has been overexpressed and purified to homogeneity. The purified enzyme was found to have a native molecular mass of 185 kDa as determined by gel filtration column chromatography on Sephacryl S-200. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis established a subunit molecular mass of 49 kDa. Thus, the enzyme is a homotetramer. The enzyme showed a pH optimum of 7.5 to 8.0 and a temperature optimum of 30 to 40 degrees C. The Km values for OSB, ATP, and CoA were 16, 73.5, and 360 microM, respectively. Of the various metal ions tested, Mg2+ was found to be the most effective in stimulating the enzyme activity. Studies with substrate analogs showed that neither benzoic acid nor benzoylpropionic acid (succinylbenzene) is a substrate for the enzyme. Thus, it appears that both the benzoyl carboxyl group and the succinyl side chain are required for activation of the ...Continue Reading

References

Jul 1, 1991·Zeitschrift Für Naturforschung. C, a Journal of Biosciences·H J Sieweke, E Leistner
Jan 1, 1990·Methods in Enzymology·F W StudierJ W Dubendorff
Feb 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·S Tabor, C C Richardson
Sep 1, 1982·Microbiological Reviews·R Bentley, R Meganathan

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Citations

Mar 11, 1998·Natural Product Reports·P M Dewick
Oct 11, 2017·Bioinformatics and Biology Insights·Nicolas CarelsPatricia Azambuja
Jul 16, 2008·Biochemistry·Rui WuDebra Dunaway-Mariano
Aug 1, 2009·EcoSal Plus·R Meganathan, Ohsuk Kwon
Dec 18, 2007·Journal of Bacteriology·James P ColemanEverett C Pesci
Sep 8, 1998·Microbiology and Molecular Biology Reviews : MMBR·M K Berlyn

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