title: Physiological and biochemical characteristics of poly gamma-glutamate synthetase complex of Bacillus subtilis.
abstract: An enzymatic system for poly gamma-glutamate (PGA) synthesis in Bacillus subtilis, the PgsBCA system, was investigated. The gene-disruption experiment showed that the enzymatic system was the sole machinery of PGA synthesis in B. subtilis. We succeeded in achieving the enzymatic synthesis of elongated PGAs with the cell membrane of the Escherichia coli clone producing PgsBCA in the presence of ATP and D-glutamate. The enzyme preparation solubilized from the membrane with 8 mM Chaps catalyzed ADP-forming ATP hydrolysis only in the presence of glutamate; the D-enantiomer was the best cosubstrate, followed by the L-enantiomer. Each component of the system, PgsB, PgsC, and PgsA, was translated in vitro and the glutamate-dependent ATPase reaction was kinetically analyzed. The PGA synthetase complex, PgsBCA, was suggested to be an atypical amide ligase.
authors.affiliation: Department of Bioresources Science, Faculty of Agriculture, Kochi University, Nankoku, Japan. ashiuchi@cc.kochi-u.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Ashiuchi
authors.forename: C
authors.initials: C
authors.lastname: Nawa
authors.forename: T
authors.initials: T
authors.lastname: Kamei
authors.forename: J J
authors.initials: JJ
authors.lastname: Song
authors.forename: S P
authors.initials: SP
authors.lastname: Hong
authors.forename: M H
authors.initials: MH
authors.lastname: Sung
authors.forename: K
authors.initials: K
authors.lastname: Soda
authors.forename: H
authors.initials: H
authors.lastname: Misono
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D003902:Detergents / Q000378:metabolism; D017353:Gene Deletion; D015870:Gene Expression; D005970:Glutamate Synthase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011099:Polyglutamic Acid / Q000096:biosynthesis* / Q000378:metabolism
chemical_list: D003902:Detergents; D009097:Multienzyme Complexes; D011099:Polyglutamic Acid; D000255:Adenosine Triphosphate; D005970:Glutamate Synthase; D000251:Adenosine Triphosphatases