title: Large-scale production of phospholipase D from Streptomyces racemochromogenes and its application to soybean lecithin modification.
abstract: Phospholipase D (PLD) catalyzes transphosphatidylation, causing inter-conversion of the polar head group of phospholipids and phospholipid hydrolysis. Previously, we cloned PLD103, a PLD with high transphosphatidylation activity, from Streptomyces racemochromogenes strain 10-3. Here, we report the construction of an expression system for the PLD103 gene using Streptomyces lividans as the host bacterium to achieve large-scale production. The phosphatidylcholine (PC) hydrolysis activity of S. lividans transformed with the expression plasmid containing the PLD103 gene was approximately 90-fold higher than that of the original strain. The recombinant PLD103 (rPLD103) found in the supernatant of the transformant culture medium was close to homogeneous. The rPLD103 was indistinguishable from the native enzyme in molecular mass and enzymatic properties. Additionally, rPLD103 had high transphosphatidylation activity on PC as a substrate in a simple aqueous one-phase reaction system and was able to modify the phospholipid content of soybean lecithin. Consequently, the expression system produces a stable supply of PLD, which can then be used in the production of phosphatidyl derivatives from lecithin.
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: Shin-Ichiro
authors.initials: S
authors.lastname: Sakurai
authors.forename: Masataka
authors.initials: M
authors.lastname: Uchino
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: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D015870:Gene Expression; D007700:Kinetics; D054709:Lecithins / Q000737:chemistry*; D010739:Phospholipase D / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013025:Glycine max / Q000737:chemistry*; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D048170:Streptomyces lividans / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D054709:Lecithins; D010739:Phospholipase D