Rendering one autolysis site in Bacillus subtilis neutral protease resistant to cleavage reveals a new fission

B. van den Burg, V.G.H. Eijsink, G. Vriend, O.R Veltman, G Venema

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7 Citations (Scopus)

Abstract

Autolytic degradation of the thermolysin-like proteinase of Bacillus subtilis (TLP-sub) is responsible for the irreversible inactivation of the enzyme at elevated temperatures. Previously we have reported five cleavage sites in Tip-sub [Van den Burg et al, (1990) Biochem. J. 272, 93-97]. In an attempt to render the enzyme less susceptible to autolytic breakdown, one of the fission sites, located between Leu-156 and Ile-157, was modified by replacing Ile-157, C-terminally located with respect to the fission site, by an Asp residue. Aspartic acid is less preferred at this position with respect to the substrate preference of TLP-sub, Modelling studies indicated that this mutation was unlikely to cause conformational changes in the enzyme. Although the 156-157 fission was not observed in the mutant enzyme, a new fission site, between Gly-148 and Val-149, was now observed.

Original languageEnglish
Pages (from-to)125-132
Number of pages8
JournalBiotechnology and applied biochemistry
Volume27
Issue number2
DOIs
Publication statusPublished - Apr-1998

Keywords

  • NUCLEOTIDE-SEQUENCE
  • AMINO-ACID
  • THERMOLYSIN
  • GENE
  • CLONING
  • EXPRESSION
  • STABILITY
  • STEAROTHERMOPHILUS
  • AMYLOLIQUEFACIENS
  • THERMOSTABILITY

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