产α-葡萄糖苷酶抑制剂罗汉果内生菌株的筛选鉴定及发酵条件优化  

Screening,identification and fermentation conditions optimization ofα-glucosidase inhibitor-producing endophytes from Siraitia grosvenorii

作　　者：周巧丽 付强 王琪 胡梦琪 金岳 赵丰丽[1,2] ZHOU Qiaoli;FU Qiang;WANG Qi;HU Mengqi;JIN Yue;ZHAO Fengli(College of Life Science,Guangxi Normal University,Guilin 541006,China;Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection,Ministry of Education,Guilin 541006,China)

机构地区：[1]广西师范大学生命科学学院,广西桂林541006 [2]珍稀濒危动植物生态与环境保护教育部重点实验室,广西桂林541006

出　　处：《中国酿造》2021年第4期105-108,共4页China Brewing

基　　金：桂林市科学研究与技术开发计划项目(20171015-2)。

摘　　要：采用对硝基苯基-α-D-吡喃葡萄糖苷(PNPG)法,从罗汉果内生菌中筛选高产α-葡萄糖苷酶抑制剂(α-GI)的菌株,对该菌株进行形态观察和分子生物学鉴定。以α-葡萄糖苷酶抑制率为评价指标,利用单因素试验及正交试验进行发酵条件优化,进一步用有机溶剂萃取发酵液后进行活性部位追踪。结果表明,从罗汉果内生菌中筛选出一株高产α-GI的菌株,编号为PD-14,被鉴定为贝莱斯芽孢杆菌(Bacillus velezensis),该菌株产α-GI的最佳发酵条件为初始pH值7,装液量100 mL/250 mL,发酵时间4 d,发酵温度30℃,接种量7%。此优化条件下,菌株PD-14发酵原液对α-葡萄糖苷酶的抑制率达到91.05%,是优化前的1.5倍。α-GI为胞外产物,萃取后筛选到活性部位为正丁醇相和水相,α-葡萄糖苷酶抑制率分别为77.61%和76.70%。The high-yieldα-glucosidase inhibitor(α-GI)strain was screened from Siraitia grosvenorii endophytes by p-nitrophenyl-α-D-glucopyranoside(PNPG)method,and the strain was identified by morphological observation and molecular biology.Taking the inhibition rate ofα-glucosidase as the evaluation index,the fermentation conditions were optimized by single factor test and orthogonal test.After further using organic solvents to extract the fermentation broth,the active site was tracked.The results showed that aα-GI high-producing strain was screened from Siraitia grosvenorii endophytes,numbered as PD-14,and identified as Bacillus velezensis.The optimal fermentation conditions forα-GI production by the strain were initial pH 7,loaded liquid 100 ml/250 ml,time 4 d,temperature 30℃,and inoculum 7%.Under the optimized conditions,the inhibition rate ofα-GI reached 91.05%,which was 1.5 times that of before optimization.Theα-GI was an extracellular product.After extraction,the active site n-butanol phase and water phase were screened,and theα-glucosidase inhibition rates were 77.61%and 76.70%,respectively.

关 键 词：罗汉果内生菌 Α-葡萄糖苷酶抑制剂 菌株筛选 发酵条件 优化 活性追踪 

分 类 号：Q939.9[生物学—微生物学]