产β-D-葡萄糖苷酶乳酸菌的筛选、鉴定及系统发育分析

摘要

In search for a glycosidase that can enhance flavor in fruit wine.Four strains of β-D-glucosidase were screened from 63 strains of Lactobacillus with Esculin color-developing plate method.The p-nitro-phenyl-β-D-glucopyranoside (PNPG) were used as a substrate and method of differential centrifugation graded precipitation was utilized to the initial location of β-D-glucosidase.In addition,traditional identification methods,such as morphology,physiological and biochemical tests,and sequence analysis of 16S rDNA were used to identify the four strains.The result indicated that the β-D-glucosidases generated by four strains were all endoenzyme;FL12 and Hsb were Lactobacillus plantarum,and CM3 was Lactobacillus pentosus and T61 was Lactobacillus brevis.%为寻找一种适用于果酒增香的糖苷酶.利用七叶苷显色平板法,从63株乳酸菌中筛选得到4株产β-D-葡萄糖苷酶的菌株.再以对硝基苯基β-D-葡萄糖苷(PN PG)为底物,利用差速离心分级沉淀对β-D-葡萄糖苷酶进行初步定位.此外通过形态特征、生理生化实验等传统鉴定方法,结合16S rDNA序列分析对其进行鉴定.结果表明:所研究的4株菌株所产β-D-葡萄糖苷酶均为胞内酶;FL12和Hsb鉴定为植物乳杆菌(Lactobacillus plantarum),CM3鉴定为戊糖乳杆菌(Lactobacillus pentosus),T61鉴定为短乳杆菌(Lactobacillus brevis).