ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2015
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2015 International Scholars Journals
Full Length Research Paper
Improved purification and some properties of a novel phospholipase C from Bacillus mycoides strain 970
Chang Gao Wang1, 2*, Ming Kai Chen3 and Tao Chen1
1Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
2Key Laboratory of Fermentation Engineering, Ministry of Education, Hubei University of Technology, Wuhan 430068, China.
3Renmin Hospital of Wuhan University, Wuhan 430060, China.
Accepted 15 December, 2014
Abstract
A novel microbial phospholipase C from Bacillus mycoides strain 970 was purified to apparent homogeneity by an improved purification process, in which DEAE-Cellulose adsorption was first used to remove bulk of dark brown impurities and then a combination of DEAE-Cellulose ion-exchange, Phenyl Sepharose hydrophobic interaction and Sephacryl S-100 gel filtration chromatography was used to separate the enzyme from remained contaminants. The enzyme appeared to be a single peptide of 75.1 kDa on SDS-PAGE. The optimum temperature and pH of the enzyme were 60°C and 7.0 - 7.5, respectively. The enzyme was stable at temperatures lower than 50°C and pH 5 - 9.5. This purified phospholipase C was characterized as a metallophospholipase C and Ca2+, Mg2+ , Zn2+, Ba2+ were essential for its PLC activity. The enzyme showed no lecithinase activity on egg yolk agar and also no hemolytic activity on blood agar.
Key words: Phospholipase C, purification, metalloenzyme, nonhemolytic activity, Bacillus mycoides.