Improvement of thermal stability of subtilisin J by changing the primary autolysis site.

Abstract

The thermostability of subtilisin J, an extracellular serine protease secreted from Bacillus stearothermophilus, has been improved by changing the primary autolysis site of the Asp-49 mutant protein. Previously we have shown that the Asp-49 mutant protein has proteolytic activity, but so unstable that it was primarily autolyzed in Tyr-58-Gln-59 peptide bond during cultivation (Jang et al. Biochim. Biophys. Acta. 1162, 233-235 1993). In the present study, to mitigate the autolytic degradation and increase the thermostability, we deleted the Tyr-58 residue using the Asp-49 mutant as a template. This mutant (Asp-49/delta Tyr-58 mutant) protein showed an improved resistance to heat treatment without changing the catalytic efficiency of the enzyme. These results show that change of primary autolysis site can stabilize the subtilisin.

• Authors: K H Bae, J S Jang, K S Park, S H Lee, S M Byun
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 207 Issue 1 Pg. 20-4 (Feb 06 1995) ISSN: 0006-291X [Print] United States
• PMID: 7857265 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Recombinant Proteins
• subtilisin J
• Serine
• Serine Endopeptidases
• Subtilisins

Topics

• Amino Acid Sequence
• Autolysis
• Enzyme Stability
• Geobacillus stearothermophilus (enzymology)
• Hot Temperature
• Kinetics
• Molecular Sequence Data
• Mutagenesis, Site-Directed
• Recombinant Proteins (biosynthesis, chemistry, metabolism)
• Sequence Homology, Amino Acid
• Serine
• Serine Endopeptidases (chemistry)
• Subtilisins (biosynthesis, chemistry, metabolism)
• Thermodynamics