红河传统发酵豆豉中高纤溶活性菌株的分离筛选及其纤溶酶基因分析副本.pdfVIP

现代食品科技 Modern Food Science and Technology 2014, Vol.30, No.8 红河传统发酵豆豉中高纤溶活性菌株的分离筛选 及其纤溶酶基因分析 1,2 1 1 1 1 1 龚福明 ，李晓然 ，宋园亮 ，罗义勇 ，张忠华 ，柳陈坚 （1.昆明理工大学生命科学与技术学院，云南昆明 650500）（2.德宏职业学院基础医学部，云南芒市 678400） 摘要：运用脱脂乳固体培养基及纤维蛋白固体培养基的两步分离筛选法，从云南红河传统发酵豆豉中分离筛选具有高产豆豉纤 溶酶活性的菌株，同时对它们的豆豉纤溶酶基因进行克隆分析，以期为新型功能性豆豉的研发提供备选菌株及理论依据。研究结果表 明，两步分离筛选法能有效地从云南红河传统发酵豆豉中筛选到高产豆豉纤溶酶的菌株Bacillus subtilis LC-2-1 ，对该高产菌株的豆豉 纤溶酶成熟肽基因分析及预测结果表明，菌株B subtilis LC-2-1 确实能分泌一种由 825 个碱基编码275 个氨基酸残基且分子量约为27.4 kDa 的豆豉纤溶酶，与纳豆激酶及其他豆豉纤溶酶相比差异显著，同源性仅为 85.1%。同时其豆豉纤溶酶活性分析结果则表明，菌株 B subtilis LC-2-1 所产纤溶酶活性较高，可达 79.84 U/mL。因此，本研究结果将为新型且具有溶血栓功能发酵豆豉的研发提供备选菌 株及理论依据。 关键词：传统发酵豆豉；豆豉纤溶酶；分离筛选；基因分析； 文章篇号：1673-9078(2014)8-76-81 Screening of High-yield Fibrinolytic Enzyme-producing Strains from Traditional Fermented Douchi (Honghe, Yunnan) and Analysis of the Gene Encoding the Fibrinolytic Enzyme 1, 2 1 1 1 1 1 GONG Fu-ming , LI Xiao-ran , SONG Yuan-liang , LUO Yi-yong , ZHANG Zhong-hua , LIU Chen-jian (1.Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, China) (2.Basic Medical Department, Dehong Vocational College, Mangshi, 678400, China) Abstract: High-yield fibrinolytic enzyme-producing bacterial strains were isolated and screened from traditionally fermented douchi from Honghe, by a two-step screening method, in which skim milk agar and fibrin agar were used. The bacterial gene encoding the fibrinolytic enzyme was cloned and analyzed to provide bacterial reference strains and a theoretical basis to develop a new version of functional douchi. Using these methods, a high-yield fibrinolytic enzyme-produci