Purification and characterization of cell wall hydrolase from alkalophilic Bacillus mutanolyticus YU5215

Seung Ho Ohk, Seung Woo Nam, Jin Man Kim, Yun Jung Yoo, Dong Hoon Bai

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4 Citations (Scopus)

Abstract

Streptococcus mutans has the capacity of inducing dental caries. Thus, to develop a novel way of preventing dental caries, a cell wall hydrolase-producing strain was isolated and its characteristics were investigated. Among 200 alkalophilic strains isolated from soil, 8 strains exhibited lytic activities against Streptococcus mutans. However, strain YU5215 with the highest cell wall hydrolase activity was selected for further study. Strain YU5215 was identified as a novel strain of Bacillus based on analyzing its 16S rDNA sequence and Bergey's Manual of Systematic Bacteriology, and thus designated as Bacillus mutanolyticus YU5215. The optimal conditions for the production of the cell wall hydrolase from Bacillus mutanolyticus YU5215 consisted of glucose (0.8%), yeast extract (1.2%), polypeptone (0.5%), K2HPO4 (0.1%), MgSO4·7H2O (0.02%), and Na2CO3 (1.0%) at pH 10.0. Bacillus mutanolyticus YU5215 was cultured at 30°C for 72 h to produce the cell wall hydrolase, which was then purified by acetone precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined as 22,700 Da by SDS-PAGE. When the cell wall peptidoglycan of Streptococcus mutans was digested with the lytic enzyme, no increase in the reducing sugars was observed, while the free amino acids increased, indicating that the lytic enzyme had an endopeptidase-like property. The amino terminus of the cell wall peptidoglycan digested by the lytic enzyme was determined as a glutamic acid, while the lytic site of the lytic enzyme in the Streptococcus mutans peptidoglycan was identified as the peptide linkage of L-Ala and D-Glu.

Original languageEnglish
Pages (from-to)1142-1149
Number of pages8
JournalJournal of microbiology and biotechnology
Volume14
Issue number6
Publication statusPublished - 2004 Dec

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

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