Purification and characterization of PBP4a, a new low-molecular-weight penicillin-binding protein from Bacillus subtilis.

Abstract:

:Penicillin-binding protein 4a (PBP4a) from Bacillus subtilis was overproduced and purified to homogeneity. It clearly exhibits DD-carboxypeptidase and thiolesterase activities in vitro. Although highly isologous to the Actinomadura sp. strain R39 DD-peptidase (B. Granier, C. Duez, S. Lepage, S. Englebert, J. Dusart, O. Dideberg, J. van Beeumen, J. M. Frère, and J. M. Ghuysen, Biochem. J. 282:781-788, 1992), which is rapidly inactivated by many beta-lactams, PBP4a is only moderately sensitive to these compounds. The second-order rate constant (k(2)/K) for the acylation of the essential serine by benzylpenicillin is 300,000 M(-1) s(-1) for the Actinomadura sp. strain R39 peptidase, 1,400 M(-1) s(-1) for B. subtilis PBP4a, and 7,000 M(-1) s(-1) for Escherichia coli PBP4, the third member of this class of PBPs. Cephaloridine, however, efficiently inactivates PBP4a (k(2)/K = 46,000 M(-1) s(-1)). PBP4a is also much more thermostable than the R39 enzyme.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Duez C,Vanhove M,Gallet X,Bouillenne F,Docquier J,Brans A,Frère J

doi

10.1128/JB.183.5.1595-1599.2001

keywords:

subject

Has Abstract

pub_date

2001-03-01 00:00:00

pages

1595-9

issue

5

eissn

0021-9193

issn

1098-5530

journal_volume

183

pub_type

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