Cloning, expression, and identification of a novel extracellular cold-adapted alkaline protease gene of the marine bacterium strain YS-80-122

Appl Biochem Biotechnol. 2010 Nov;162(5):1497-505. doi: 10.1007/s12010-010-8927-y. Epub 2010 Mar 11.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptation, Physiological*
  • Amino Acid Sequence
  • Bacteria / enzymology*
  • Bacteria / genetics*
  • Blotting, Western
  • Chromatography, Gel
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / metabolism
  • Extracellular Space / enzymology*
  • Genes, Bacterial / genetics*
  • Metalloendopeptidases / chemistry
  • Metalloendopeptidases / genetics*
  • Metalloendopeptidases / isolation & purification
  • Metalloendopeptidases / metabolism
  • Molecular Sequence Data
  • Recombinant Proteins / metabolism
  • Seawater / microbiology*
  • Sequence Alignment
  • Sequence Analysis, DNA

Substances

  • Recombinant Proteins
  • Metalloendopeptidases
  • cold alkaline protease

Associated data

  • GENBANK/GU084389