Biochemical and mutational analysis of a gingipain-like peptidase activity from Prevotella ruminicola B-1(4) and its role in ammonia production by ruminal bacteria

Madeira, HMF, Peng, L and Morrison, M (1997) Biochemical and mutational analysis of a gingipain-like peptidase activity from Prevotella ruminicola B-1(4) and its role in ammonia production by ruminal bacteria. Applied and Environmental Microbiology, 63 2: 670-675.

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Author Madeira, HMF
Peng, L
Morrison, M
Title Biochemical and mutational analysis of a gingipain-like peptidase activity from Prevotella ruminicola B-1(4) and its role in ammonia production by ruminal bacteria
Journal name Applied and Environmental Microbiology   Check publisher's open access policy
ISSN 0099-2240
Publication date 1997-02-01
Year available 1997
Sub-type Article (original research)
Open Access Status File (Publisher version)
Volume 63
Issue 2
Start page 670
End page 675
Total pages 6
Place of publication WASHINGTON
Publisher AMER SOC MICROBIOLOGY
Language eng
Abstract A chemical mutagenesis protocol was used with the ruminal bacterium Prevotella ruminicola strain B(1)J to generate mutant strains defective in peptidase activity, Compared with the wild-type parent strain, the isolated mutants possessed 1/10 of the enzyme activity responsible for cleavage of glycine-arginine-4-methoxy-beta-naphthylamide (Gly-Arg-MNA). A concomitant loss in activity against arginine-arginine-4-methoxy-beta-naphthylamide (Arg-Arg-MNA) was also observed. Both activities were similarly affected by various proteinase inhibitors, suggesting that the same enzyme is responsible for the Arg-Arg-MNA peptidase and Gly-Arg-MNA peptidase activities, Growth rates of wild-type and mutant strains grown in batch culture with various nitrogen sources did not differ, However, a role for the Gly-Arg-MNA peptidase activity was demonstrated in coculture experiments with gram-positive, ammonia-producing ruminal bacteria. The rate and extent of ammonia production were reduced by approximately 25% in cocultures containing the mutants when compared with that of wild-type-containing cultures. These reductions could not be accounted for simply by the decrease in ammonia production by the mutant strain alone. To our knowledge, this paper reports the first successful use of chemical mutagenesis with ruminal microorganisms.
Keyword Porphyromonas-Gingivalis
Cysteine Proteinase
Purification
Rumen
Argingipain
Culture
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: ResearcherID Downloads - Archived
 
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