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基因工程菌EGs2产β-葡聚糖酶条件优化及产酶特性
引用本文:李青青,陈启和,蒋孝燕,张秀艳,李婷,何国庆.基因工程菌EGs2产β-葡聚糖酶条件优化及产酶特性[J].农业生物技术学报,2007,15(4):708-712.
作者姓名:李青青  陈启和  蒋孝燕  张秀艳  李婷  何国庆
作者单位:1. 浙江大学食品科学与营养系,杭州,310029
2. 华中农业大学食品安全与微生物学系,武汉,430070
摘    要:摘要:用乳清粉作为主要培养基组成,采用正交试验对重组菌EGs2的发酵条件进行了优化,并对粗酶液的酶学性质进行了研究。重组菌EGs2的最佳发酵条件为:摇床转速170 r•min-1,接种量1 %,发酵培养基初始pH值6.0,种龄12 h。重组菌EGs2β-葡聚糖酶的表达与菌体生长呈正相关,发酵14h后,菌体生物量最高可达1.71 g•L -1,β-葡聚糖酶活力最高可达321.56 U•ml-1。所得粗酶液的最适反应温度和pH值分别为60 ℃和6.0,在70 ℃以下保温20 min后,残余酶活力均在80 %以上。在pH 5.0-9.0 放置48 h后仍能保持均90 %以上的残余酶活力。

关 键 词:β-葡聚糖酶  重组菌EGs2  发酵条件  酶学性质
文章编号:1006-1304(2007)04-0708-05
收稿时间:2006-12-25
修稿时间:2007-03-16

Optimization of Recombinant Escherichia coli EGs2 Fermentation Condition Affecting β-glucanase Production and Its Characteristics of Enzyme
LI Qing-qing,CHEN Qi-he,JIANG Xiao-yan,ZHANG Xiu-yan,LI Ting,HE Guo-qing.Optimization of Recombinant Escherichia coli EGs2 Fermentation Condition Affecting β-glucanase Production and Its Characteristics of Enzyme[J].Journal of Agricultural Biotechnology,2007,15(4):708-712.
Authors:LI Qing-qing  CHEN Qi-he  JIANG Xiao-yan  ZHANG Xiu-yan  LI Ting  HE Guo-qing
Institution:1.Food Science and Nutrition Departmen t, Zhefiang University, Hangzhou 310029,China; 2. Department of Food Science and Microbiology, Huazhong Agricultural University, Wuhan 430070, China
Abstract:Abstract: The fermentation conditions of mutant EGs2 (obtained from directed evolution for thermostable β-glucanase) for β-glucanase production were investigated with orthogonal experiment.Fermentation was conducted in 250 mL flask, each containing 30 mL of medium contained Whey 10 g•L-1,Yeast extract 5 g•L-1,NaCl 10 g•L-1,the temperature was 37 ℃.The optima culture conditions were as following: initial pH value 6.0, shaking speed 170 r•min-1,inoculation volume 1 %,inoculation time 12 h.β-glucanase production by mutant EGs2 was associated with cell growth and biomass,β-glucanase activity was increased significantly when cells entered growth phase.The bacterium began the stable phase after cultured for 14 h with the highest biomass 1.71 g•L -1 and β-glucanase activity 321.56 U•ml-1.When β-glucan was used as a substrate,the optimum temperature and pH were respectively 60 ℃ and 6.0.After 20 min incubation at 40 ℃,50 ℃,55 ℃,60 ℃,65 ℃,70 ℃, the residual activity remained at least 80%.After 48 h conservation at pH 5.0-9.0,the residual activity remained at least 90%.The enzyme was stable below 70 ℃ and at pH 5.0-9.0.
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