产耐高温果胶酶菌株筛选鉴定、产酶条件优化及酶学性质研究  

Screening and identification of high-temperature resistant pectinase-producing strains,optimization of enzyme production conditions and enzymatic properties

作　　者：陈杰 李豆南[1,2] 刘茂强 程度 田浩杰 邱树毅 唐维媛 王珂佳[2,4,5] CHEN Jie;LI Dounan;LIU Maoqiang;CHENG Du;TIAN Haojie;QIU Shuyi;TANG Weiyuan;WANG Kejia(College of Liquor and Food Engineering,Guizhou University,Guiyang 550025,China;Guizhou Province Key Laboratory of Fermentation Engineering and Biopharmacy,Guizhou University,Guiyang 550025,China;College of Food&Pharmaceutical Engineering,Guizhou Institute of Technology,Guiyang 550025,China;College of Life Sciences,Guizhou University,Guiyang 550025,China;Guizhou Light Industry Technical College,Guiyang 550025,China)

机构地区：[1]贵州大学酿酒与食品工程学院,贵州贵阳550025 [2]贵州大学贵州省发酵与生物制药重点实验室,贵州贵阳550025 [3]贵州理工学院食品药品制造工程学院,贵州贵阳5500254 [4]贵州大学生命科学学院,贵州贵阳550025 [5]贵州轻工职业技术学院,贵州贵阳550025

出　　处：《中国酿造》2023年第5期176-183,共8页China Brewing

基　　金：国家自然科学基金项目(32060518);酿酒生物技术及应用四川省重点实验室开放课题项目(NJ2022-06);贵州大学引进人才科研项目(贵大人基合字(2021)47号);科技平台及人才团队计划项目(黔科合平台人才[2018]5251);遵义市创新人才团队培养项目(遵义科人才[2020]9);贵州省发酵工程与白酒酿造人才基地(黔人领发[2018]3号);贵州省科技计划项目(黔科合支撑[2020]2Y054号)。

摘　　要：从酱香型白酒高温大曲中筛选产耐高温果胶酶菌株,结合菌落形态学观察、生理生化试验及分子生物学技术对其进行鉴定,并进行产酶条件优化及酶学性质研究。结果表明,筛选得到1株产耐高温果胶酶菌株FBKL4.013,经鉴定菌株FBKL4.013为高温放线糖莱斯菌(Laceyella sacchari)。其最佳产酶条件为培养温度50℃,培养时间54 h,装液量40 mL/250 mL,接种量10%,酵母粉、硫酸铵及可溶性淀粉添加量分别为1.0%、2.0%、8%。在此优化条件下,菌株FBKL4.013产果胶酶活力达到2565.43 U/mL,与初始相比提高了3.79倍。菌株FBKL4.013产果胶酶具备较好的耐高温特性,其酶动力学米氏常数(Km)和最大反应速度(Vm)值分别为8.40 mg/mL和0.53 mg/(mL·h)。A high-temperature resistant pectinase-producing strain was screened from high temperature Daqu,combined with colony morphological observation,physiological and biochemical tests and molecular biological technology,the enzyme production conditions were optimized and the enzyme properties were studied.The results showed that a high-temperature pectinase-producing strain FBKL4.013 was obtained,and the strain FBKL4.013 was identified as Laceyella sacchari.The optimal conditions for enzyme production were culture temperature 50℃,time 54 h,loading volume 40 ml/250 ml,inoculum 10%,addition of yeast powder,ammonium sulfate and soluble starch 1.0%,2.0%,8%,respectively.Under the optimized conditions,the pectinase activity of the strain FBKL4.013 was 2565.43 U/ml,which increased 3.79 times compared with the initial strain.The pectinase produced by the strain FBKL4.013 showed high temperature resistance,its enzyme kinetic Michaelis constants(Km)and maximum reaction velocity(Vm)values were 8.40 mg/ml and 0.53 mg/(ml·h),respectively.

关 键 词：果胶酶 高温大曲 筛选 鉴定 产酶条件优化 酶学性质 

分 类 号：TS201.2[轻工技术与工程—食品科学]