Probing the specificity of the S1 binding site of M222 mutants of subtilisin B. lentus with boronic acid inhibitors

Stabile, Michele R.; Lai, Wei-Shiang; DeSantis, Grace; Gold, Marvin; Jones, J. Bryan; Mitchinson, Colin; Bott, R.; Graycar, Thomas P.; Liu, Chung-Cheng

doi:10.1016/0960-894x(96)00466-0

Cited by 35 publications

(9 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…amylosacchariticus (327,328). The sequence was highly homologous to that of subtilisin E from B. subtilis 168 (269). The gene was expressed in B. subtilis ISW 1214 by using the vector pHY300PLK, with 20- fold-higher activity than that of the host and 4-fold-higher activity than that of B. subtilis subsp.…”

Section: Bacteriamentioning

confidence: 99%

“…The effect of substitution of Met222 with different amino acids revealed that small side chains yield the highest activity. The mutant enzymes Ser222, Ala222, and Leu222 were active and stable to peroxide for 1 h. Probing of the specificity of the S 1 binding site of Met222 Cys/Ser mutants of subtilisin from Bacillus lentus with boronic acid inhibitors revealed similar binding trends for the mutant and the parent (269). The disulfide bonds introduced into subtilisin away from its catalytic center were shown to possess increased autolytic stability (312).…”

Section: Bacteriamentioning

confidence: 99%

See 1 more Smart Citation

Molecular and Biotechnological Aspects of Microbial Proteases

Rao

Tanksale

Ghatge

et al. 1998

Microbiol Mol Biol Rev

1,743

721

View full text Add to dashboard Cite

SUMMARY Proteases represent the class of enzymes which occupy a pivotal position with respect to their physiological roles as well as their commercial applications. They perform both degradative and synthetic functions. Since they are physiologically necessary for living organisms, proteases occur ubiquitously in a wide diversity of sources such as plants, animals, and microorganisms. Microbes are an attractive source of proteases owing to the limited space required for their cultivation and their ready susceptibility to genetic manipulation. Proteases are divided into exo- and endopeptidases based on their action at or away from the termini, respectively. They are also classified as serine proteases, aspartic proteases, cysteine proteases, and metalloproteases depending on the nature of the functional group at the active site. Proteases play a critical role in many physiological and pathophysiological processes. Based on their classification, four different types of catalytic mechanisms are operative. Proteases find extensive applications in the food and dairy industries. Alkaline proteases hold a great potential for application in the detergent and leather industries due to the increasing trend to develop environmentally friendly technologies. There is a renaissance of interest in using proteolytic enzymes as targets for developing therapeutic agents. Protease genes from several bacteria, fungi, and viruses have been cloned and sequenced with the prime aims of (i) overproduction of the enzyme by gene amplification, (ii) delineation of the role of the enzyme in pathogenecity, and (iii) alteration in enzyme properties to suit its commercial application. Protein engineering techniques have been exploited to obtain proteases which show unique specificity and/or enhanced stability at high temperature or pH or in the presence of detergents and to understand the structure-function relationships of the enzyme. Protein sequences of acidic, alkaline, and neutral proteases from diverse origins have been analyzed with the aim of studying their evolutionary relationships. Despite the extensive research on several aspects of proteases, there is a paucity of knowledge about the roles that govern the diverse specificity of these enzymes. Deciphering these secrets would enable us to exploit proteases for their applications in biotechnology.

show abstract

Section: Bacteriamentioning

confidence: 99%

Section: Bacteriamentioning

confidence: 99%

Molecular and Biotechnological Aspects of Microbial Proteases

Rao

Tanksale

Ghatge

et al. 1998

Microbiol Mol Biol Rev

1,743

721

View full text Add to dashboard Cite

show abstract

“…O'Donovan et al [7] concluded a novel class of bacterial of boronic acids mutagen that may not act by direct covalent binding to DNA. Biomedical applications of boronic acid-containing polymers and biological of particular dichlorophenylboronic acids were studied [8][9][10]. The moiety of boronic acids was incorporated into nucleosides and amino acids as antiviral agents and anti-tumor [11].…”

Section: Introductionmentioning

confidence: 99%

Experimental (FT-IR, FT-Raman, UV–Vis, 1H and 13CNMR) and computational (density functional theory) studies on 3-bromophenylboronic acid

Karabacak

Kose

Ataç

et al. 2014

Journal of Molecular Structure

View full text Add to dashboard Cite

“…Significant effort was done to improve storage stability of detergent proteases by site-directed mutagenesis, where oxidation-labile residues were successfully replaced by more resistant amino acids. Although considerable improvement in Savinase ® storage stability was achieved (Von der Osten et al, 1993), these alterations in the enzyme structure resulted in substantial reduction of specific activity towards synthetic substrates compared to the wild type (Stabile et al, 1996). Another strategy to solve the problem is to physically "isolate" the enzymes from the detrimental causes.…”

Section: Introductionmentioning

confidence: 96%