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 language | English |
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Pages (from-to) | 125-132 |
Number of pages | 8 |
Journal | Biotechnology and applied biochemistry |
Volume | 27 |
Issue number | 2 |
DOIs | |
Publication status | Published - Apr-1998 |
Keywords
- NUCLEOTIDE-SEQUENCE
- AMINO-ACID
- THERMOLYSIN
- GENE
- CLONING
- EXPRESSION
- STABILITY
- STEAROTHERMOPHILUS
- AMYLOLIQUEFACIENS
- THERMOSTABILITY