共表达纤维素酶菌株发酵条件研究
Study on fermentation conditions of co-expression cellulase strains
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摘要: 目的:为了解决在发酵过程中酶活较低、成本高等问题,加快纤维素酶工业化生产。方法:采用PlackettBurman进行实验设计,对影响基因工程菌p ET30b-BGL产酶因素:接种量(A)、培养温度(B)、装液量(C),诱导时间(D)、初始p H(E)、IPTG诱导浓度(F)进行筛选,进行方差分析,结果表明,培养温度、初始p H和装液量对产酶有重要影响。进一步用Box-Behnken的响应面方法,对这3个因素进行优化,结果:当温度为35℃,初始p H7.5,装液量为25m L(容积为100m L),p ET30b-BGL产酶的酶活最高达到2080.5U/m L,较优化前的1196.8U/m L提高了将近一倍。结论:本研究为进一步研究纤维素酶提供了依据。Abstract: Objective:To solve the low enzyme activity and high cost in the process of cellulase fermentation and ameliorate the industrial production. Methods :These factors Inoculums ' quality (A) , temperature (B) , liquid volume (C) , induce time (D) , initial p H (E) and IPTG concentrate (F) were used to screen and analyze by Plackett-Burman. From the results, Box-Behnken analyze was conducted, the temperature (B) , liquid volume (C) and initial p H (E) were used to optimized. Results:The enzyme activity of p ET30b-BGL had nearly 2-fold increase than before (1196.8U/m L to 2080.5U/m L) in the condition of 35℃, p H7.5 and 25 m L liquid volume (100m L in all) . Conclusion:This research could provide the basis for the further investigations of cellulase.