Characterization of a cloned subtilisin-like serine proteinase from a psychrotrophic Vibrio species

Jóhanna Arnórsdóttir, Rúna B. Smáradóttir, Ólafur Th Magnússon, Sigrídur H. Thorbjarnardóttir, Gudmundur Eggertsson, Magnús M. Kristjánsson*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

56 Citations (Scopus)


The gene encoding a subtilisin-like serine proteinase in the psychrotrophic Vibrio sp. PA44 has been successfully cloned, sequenced and expressed in Escherichia coli. The gene is 1593 basepairs and encodes a precursor protein of 530 amino acid residues with a calculated molecular mass of 55.7 kDa. The enzyme is isolated, however, as an active 40.6-kDa proteinase, without a 139 amino acid residue N-terminal prosequence. Under mild conditions the enzyme undergoes a further autocatalytic cleavage to give a 29.7-kDa proteinase that retains full enzymatic activity. The deduced amino acid sequence of the enzyme has high homology to proteinases of the proteinase K family of subtilisin-like proteinases. With respect to the enzyme characteristics compared in this study the properties of the wild-type and recombinant proteinases are the same. Sequence analysis revealed that especially with respect to the thermophilic homologues, aqualysin I from Thermus aquaticus and a proteinase from Thermus strain Rt41A, the cold-adapted Vibrio-proteinase has a higher content of polar/uncharged amino acids, as well as aspartate residues. The thermophilic enzymes had a higher content of arginines, and relatively higher number of hydrophobic amino acids and a higher aliphatic index. These factors may contribute to the adaptation of these proteinases to different temperature conditions.

Original languageEnglish
Pages (from-to)5536-5546
Number of pages11
JournalEuropean Journal of Biochemistry
Issue number22
Publication statusPublished - 2002

Other keywords

  • Cold adaptation
  • Proteinase K-like
  • Psychrotrophic
  • Subtilisin-like proteinase
  • Vibrio-proteinase


Dive into the research topics of 'Characterization of a cloned subtilisin-like serine proteinase from a psychrotrophic Vibrio species'. Together they form a unique fingerprint.

Cite this