产果胶酶枯草芽孢杆菌的鉴定、发酵条件优化及产物酶学性质的研究  

Identification,Optimization of Fermentation Conditions of Bacillus subtilis Which Produces Pectinase and the Enzymatic Properties

作　　者：白兰芳[1] 高慧[2] 刘明启[1] 戴贤君[1] 关荣发[1] 

机构地区：[1]中国计量学院生命科学学院,浙江杭州310018 [2]浙江省畜牧技术服务中心,浙江杭州310021

出　　处：《中国畜牧杂志》2011年第19期63-68,共6页Chinese Journal of Animal Science

基　　金：浙江省自然科学基金资助项目(Y3090503);杭州市科技局资助项目(20110232B81)

摘　　要：以果胶为唯一碳源筛选到1株产果胶酶的菌株JLSP-13,通过菌落形态观察、生理生化指标试验和16SrDNA保守序列分析,鉴定JLSP-13为枯草芽孢杆菌,命名为Bacillus subtilis JLSP-13。采用响应面法(RSM)优化其产酶条件,并系统研究产物酶学性质。结果表明:B.subtilis JLSP-13的最佳发酵时间为25.02 h,添加可溶性淀粉和果胶的最佳浓度分为0.61%和0.042%,B.subtilis JLSP-13果胶酶(BpgA)活性最大预测值达38.8 U/mL,验证值为37.6 U/mL,是基础培养基的2.6倍。BpgA的最适温度为60℃,Tm为82.9℃,其热稳定性较好;最适pH为6.0,在pH 6.0～8.0范围内较稳定;Km和Vmax分别为5.128 mg/mL、12.92 mg(/mL.min),BpgA能快速降低果胶底物的黏度。The strain JLSP-13 was isolated from soil samples using pectin as sole carbon source. The population shape and individual form was observed and identified from the aspects of physiological and biochemical index and 16S rDNA sequence analysis. The fermentation condition for pectinase production was optimized by response surface methodology （RSM）. The isolated strain was identified as Bacillus subtilis JLSP-13. The maximum of Bacillus subtilis pectinase （BpgA） yield （37.6 U/mL） obtained at 0.042% pectin, 0.61% soluble starch and 25.02 h fermentation, which was close to the predicted one （38.8 U/mL）, and was 2.6 times as high as that of basic medium. The optimum temperature and pH of BpgA were 60℃ and pH 6.0, respectively. BpgA exhibited Km and Vmax values of 5.128 mg/mL and 12.92 mg/（mL＇min）, respectively. The BpgA caused a rapid decrease in viscosity of pectin.

关 键 词：果胶酶 枯草芽孢杆菌 鉴定 响应面法 酶学性质 

分 类 号：S816.703[农业科学—饲料科学]