title: Purification, biochemical characterization, and cloning of phospholipase D from Streptomyces racemochromogenes strain 10-3.
abstract: We previously isolated Streptomyces racemochromogenes strain 10-3, which produces a phospholipase D (PLD) with high transphosphatidylation activity. Here, we purified and cloned the PLD (PLD103) from the strain. PLD103 exerted the highest hydrolytic activity at a slightly alkaline pH, which is in contrast to the majority of known Streptomyces PLDs that have a slightly acidic optimum pH. PLD103 shares only 71-76% amino acid sequence identity with other Streptomyces PLDs that have a slightly acidic optimum pH; thus, the diversity in the primary structure might explain the discrepancy observed in the optimum pH. The purified PLD displayed high transphosphatidylation activity in the presence of glycerol, L: -serine, and 2-aminoethanol hydrochloride with a conversion rate of 82-97% in a simple one-phase system, which was comparable to the rate of other Streptomyces PLDs in a complicated biphasic system.
authors.affiliation: Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri, Hokkaido 099-2493, Japan. y3nakaza@bioindustry.nodai.ac.jp
authors.forename: Yozo
authors.initials: Y
authors.lastname: Nakazawa
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Sagane
authors.forename: Teppei
authors.initials: T
authors.lastname: Kikuchi
authors.forename: Masataka
authors.initials: M
authors.lastname: Uchino
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nagai
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Sato
authors.forename: Kazuki
authors.initials: K
authors.lastname: Toeda
authors.forename: Katsumi
authors.initials: K
authors.lastname: Takano
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D010739:Phospholipase D / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D017385:Sequence Homology; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D010739:Phospholipase D