title: Molecular and catalytic properties of monoacetylphloroglucinol acetyltransferase from Pseudomonas sp. YGJ3.
abstract: Monoacetylphloroglucinol (MAPG) acetyltransferase, catalyzing the conversion of MAPG to 2,4-diacetylphloroglucinol (DAPG), was purified from Pseudomonas sp. YGJ3 grown without Cl(-). Cl(-) and pyoluteorin repressed expression of the enzyme. SDS-polyacrylamide gel electrophoresis showed that the purified enzyme (M(r)=330 kDa) was composed of three subunits of 17, 38, and 43 kDa, and protein sequencing identified these as PhlB, PhlA, and PhlC respectively. The enzyme catalyzed the reversible disproportionation of 2 moles of MAPG to phloroglucinol (PG) and DAPG. The equilibrium constant K (=[DAPG][PG]/[MAPG](2)) was estimated to be about 1.0 at 25 °C. A KpnI 20-kb DNA fragment was cloned from the genomic DNA of strain YGJ3, and a 12,598-bp long DNA region containing the phl gene cluster phlACBDEFGHI was sequenced. PCR cloning and expression of the phl genes in Escherichia coli confirmed that expression of phlACB genes produced MAPG ATase.
authors.affiliation: Department of Biomolecular Science, Faculty of Engineering, Gifu University, Gifu, Japan.
authors.forename: Asuka
authors.initials: A
authors.lastname: Hayashi
authors.forename: Hiroki
authors.initials: H
authors.lastname: Saitou
authors.forename: Tomomi
authors.initials: T
authors.lastname: Mori
authors.forename: Ikue
authors.initials: I
authors.lastname: Matano
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sugisaki
authors.forename: Kiyofumi
authors.initials: K
authors.lastname: Maruyama
mesh_terms: D000123:Acetyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D055162:Biocatalysis; D003001:Cloning, Molecular; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D010696:Phloroglucinol / Q000031:analogs & derivatives* / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; C494385:monoacetylphloroglucinol; C059817:2,4-diacetylphloroglucinol; D010696:Phloroglucinol; D000123:Acetyltransferases