酵母工程菌S.pombe yAS56的构建及魔芋红茶菌发酵工艺的研究  

Construction of engineered yeast S. pombe y AS56 and study of the fermentation techniques of konjac kombucha

作　　者：闯绍闯 龚文秀[1] 李清[1] 曹媛媛[1] 唐欣昀[1] 

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230036

出　　处：《食品工业科技》2015年第16期198-202,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金青年科学基金项目(41401269);安徽省教育厅自然基金项目(KJ2009A107)

摘　　要：为研究利用魔芋精粉制备魔芋红茶菌的可行性,从约氏梭菌(Clostridium josui)中克隆β-葡聚糖酶基因cel9C,构建粟酒裂殖酵母(Schizosaccharomyces pombe)重组表达质粒p ESP-2-2-cel9C,转化构建酵母工程菌,利用工程菌进行魔芋红茶菌发酵。采用析因实验设计和中心组合实验设计方法,以总酸为响应值,优化发酵条件,从7个因素中筛选出2个显著因素:装液量和时间。最佳条件为:绿茶0.4%、蔗糖7.5%、魔芋精粉0.25%、接种量10%、酿酒酵母/酵母工程菌接种细胞浓度比=0.5、装液量98.7 m L、时间124 h。在此条件下红茶菌总酸为(27.88±0.26)g/kg,与预测值相近,较优化前提高41.45%,表明利用魔芋精粉和酵母工程菌制备魔芋红茶菌具有可行性。In order to study the feasibility of using konjac powder for preparation of konjac kombucha,the gene cel9C coding 15-glucanase of Clostridium josui was cloned into the expression vector pESP-2-2 and the recombinant plasmid was transformed into Schizosaccharomyces pombe,to obtain the engineered yeast strain yAS56,and the engineered yeast was used in konjac kombucha fermentation. Using totat acid as the response value,the fractional factorial design and central composite design were applied to optimize the fermentation conditions. Two significant factors,loading volume and time,were selected among seven factors. The optimal conditions of fermentation were as following ： green tea 0.4% ,sucrose 7.5% ,konjac powder 0.25% ,inoculum size 10%, ratio of cells concentration of Saccharomyces cerevisiae/engineered yeast 0.5,loading volume 98.7 mL, time 124 h. The results of confirmation test with optimal parameters showed that the production of total acid was （27.88±0.26） gAg,close to the predictive value,being increased by 41.45% compared with the un-optimized conditions. This indicated that using konjac powder and engineered yeast strain for preparation of konjac kombucha was feasible.

关 键 词：cel9C基因 粟酒裂殖酵母 魔芋 红茶菌 优化 

分 类 号：TS275.4[轻工技术与工程—农产品加工及贮藏工程]