We employed random mutagenesis to determine the region of the initial unfolding of hyper-
alkaline-sensitive subtilisin, ALP I, that precedes the denaturation of the entire protein under …

An efficient random mutagenesis procedure coupled to a replica plate screen facilitated the
isolation of mutant subtilisins from Bacillus amyloliquefaciens that had altered autolytic …

It is scientifically and industrially important to clarify the stabilizing mechanism of proteases
in extraordinary environments. We used subtilisins ALP I and Sendai as models to study the …

The thermal stability of a cysteine‐free alkaline protease (Alp) secreted by the eukaryote
Aspergillus oryzae was improved both by the introduction of engineered twin disulfide …

This study was undertaken to characterize the effect of three point mutations made on aprA‐
subtilisin on the stability of the protein to both heat‐and detergent‐induced denaturation …

Extracellular proteases of the subtilisin-class depend upon calcium for stability. Calcium
binding stabilizes these proteins in natural extracellular environments, but is an Achilles' …

High-alkaline proteases are of great importance because of their proteolytic activity and
stability under high-alkaline condition. We have previously isolated a new protease (AprB) …

Crystallographic analysis of the highly alkaline M-protease from an alkaliphilic Bacillus
strain shows the occurrence of a unique salt bridge triad Arg19–Glu271–Arg275 (in …

Region‐specific random mutagenesis in the weak calcium binding site of subtilisin
Carlsberg and subsequent screening for variants with enhanced heat stability revealed two …

We have coevolved high activity and hyperstability in subtilisin by sequentially randomizing
12 amino acid positions in calcium-free subtilisin. The optimal amino acid for each …