title: Characterisitics and gene cloning of phospholipase D of the psychrophile, Shewanella sp.
abstract: Phospholipase D, with a molecular mass of 64 kDa, was purified from the psychrophile, Shewanella sp. The enzyme showed maximal activity at pH 7.8 and 40 degrees C in the presence of the Ca2+-ion, and its activity at 10 degrees C was 6.5% of maximum. The enzyme exhibited high activity to the non-micelle form of phosphatidylcholine in an aqueous solution containing water miscible alcohols such as methanol, ethanol, iso-propanol, and n-propanol. Nucleotide sequencing of the enzyme gene yielded a deduced amino acid sequence, which showed 36.2% identity to that of Streptomyces chromofuscus phospholipase D alone. The low sequence similarity to other phospholipase D enzymes suggests that the purified enzyme might be a novel phospholipase D.
authors.affiliation: Headquarters for Innovative Cooperation and Development, Kobe University, Japan. tsuruta@kobe-u.ac.jp
authors.forename: Hiroki
authors.initials: H
authors.lastname: Tsuruta
authors.forename: Ryuhei
authors.initials: R
authors.lastname: Hayashi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ohkawa
authors.forename: Nobuhide
authors.initials: N
authors.lastname: Ohkatsu
authors.forename: Masakazu
authors.initials: M
authors.lastname: Morimoto
authors.forename: Kaname
authors.initials: K
authors.lastname: Nishimoto
authors.forename: Sigeyuki
authors.initials: S
authors.lastname: Santou
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Aizono
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004795:Enzyme Stability; D015870:Gene Expression; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D002479:Inclusion Bodies / Q000737:chemistry; D019216:Metals, Heavy / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010739:Phospholipase D / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020592:Shewanella / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
chemical_list: D019216:Metals, Heavy; D011994:Recombinant Proteins; D010739:Phospholipase D