title: Cloning, expression, and identification of a novel extracellular cold-adapted alkaline protease gene of the marine bacterium strain YS-80-122.
abstract: As one of the most important groups of industrial enzymes, cold-adapted protease has been studied widely. An extracellular cold-adapted alkaline protease metalloproteinase (MP), produced by a marine bacterium strain YS-80-122, has been purified. The NH(2)-amino acid sequence of the purified alkaline protease MP was ANGTSSAFTQ, which was identical to that of the serralysin from Pseudomonas sp. "TAC II 18". The MP structural gene (lupA gene) was cloned by inverse PCR, and the open reading frame of 1,443 bp encoded a 463 amino acid protein (without signal peptide). Sequence alignment reveals that the alkaline protease MP belongs to the serralysin-type metalloproteases. The recombinant protein LupA was expressed in Escherichia coli, and Western blotting confirmed that the LupA was homologous to the cold-adapted alkaline protease MP.
authors.affiliation: Yellow Sea Fisheries Research Institute, Academy of Fishery Science of China, Qingdao 266071 Shandong Province, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.forename: Jianhua
authors.initials: J
authors.lastname: Hao
authors.forename: Chengye
authors.initials: C
authors.lastname: Yang
authors.forename: Mi
authors.initials: M
authors.lastname: Sun
mesh_terms: D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics*; D015153:Blotting, Western; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D005110:Extracellular Space / Q000201:enzymology*; D005798:Genes, Bacterial / Q000235:genetics*; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000378:metabolism; D012623:Seawater / Q000382:microbiology*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D011994:Recombinant Proteins; D008666:Metalloendopeptidases; C110245:cold alkaline protease