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

• DOI: 10.1128/jb.178.23.6778-6781.1996
• Publication Date: 2016-11-10T13:42:19Z
• Authors: O Kwon; D K Bhattacharyya; R Meganathan
• Keywords: Homotetramer; Molecular mass; Coenzyme A

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