Enhanced thermal stability of an alkaline protease, AprP, isolated from a Pseudomonas sp by mutation at an autoproteolysis site, Ser-331

Citation
Jw. Jang et al., Enhanced thermal stability of an alkaline protease, AprP, isolated from a Pseudomonas sp by mutation at an autoproteolysis site, Ser-331, BIOT APP B, 34, 2001, pp. 81-84
Citations number
11
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biotecnology & Applied Microbiology","Biochemistry & Biophysics
Journal title
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
ISSN journal
0885-4513 → ACNP
Volume
34
Year of publication
2001
Part
2
Pages
81 - 84
Database
ISI
SICI code
0885-4513(200110)34:<81:ETSOAA>2.0.ZU;2-4
Abstract
The thermal stability of the alkaline protease extracellular subtilisin-typ e serine protease (AprP) from Pseudomonas sp. KFCC 10818 was improved by al tering an amino acid residue at an autoproteolytic cleavage site. N-termina l sequence analysis of the autoproteolytic products of the protein revealed the presence of two cleavage sites, Ser-307 and Ser-331. To increase the t hermal stability of the enzyme, serine residues of these sites were replace d with aspartate. The S331D mutant enzyme was successfully purified and cha racterized whereas the S307D mutant was not. The half-lives of the S331D mu tant at 55 degreesC and 60 degreesC were 1.5 and 2.4 times longer than that of the wild-type enzyme, respectively. In addition, the catalytic efficien cy was also enhanced.