高产纤维素酶菌株选育及其产酶工艺的研究
Breeding of High-Produing Cellulase Strain and Their Enzyme Production Process
王静蕾 1李嘉琪 1樊津池 1宋丽芬 1曲威1
作者信息
- 1. 中国农业大学烟台研究院,山东烟台 264670
- 折叠
摘要
为获取高产纤维素酶的菌株,本研究以潮湿纤维单胞菌(Cellulomonasuda)为研究对象,采用紫外诱变方式进行菌株突变,通过刚果红选择培养基筛选,以及测定纤维素酶活性后得到菌株W3,酶活高达37.6 U/mL,较初始菌株提升了49.4％.经传代十次后仍有较高酶活,结果说明该突变菌株遗传性状稳定且产纤维素酶能力强.对突变菌株进行单因素优化实验确定最优培养条件,后对关键因子通过正交实验确定最佳参数.最佳培养条件为:12.5 g/L蔗糖,2 g/L硝酸铵,2 g/L磷酸二氢钾,菌悬液接种量体积比3％,pH=6.0,发酵培养温度35℃.优化后酶活性达43.06 U/mL,极显著高于原始菌株(P=0.008 9＜0.01),较最初菌株酶活性提高了78.7％.研究结果可为高效降解纤维素提供一定理论依据和应用价值.
Abstract
In order to obtain the strain with high cellulase yield,this study used Cellulomonasuda as the re-search objec,the strain mutation by ultraviolet mutagenesis,the selected medium of Congo red ellulase activity to obtain strain W3,with enzyme activity up to 37.6 U/mL.Compared with the initial strain,the increase was 49.4％.The high enzyme activity after ten generations,indicating that the mutant strain had stable genetic charac-ter and strong cellulase production ability.The single factor optimization experiment was to the optimal culture con-ditions of the mutant strain,and then the best parameters were determined by orthogonal experiment for the key factors.The optimal culture conditions were as follows:12.5 g/L sucrose,2 g/L ammonium nitrate,2 g/L potassium dihydrogen phosphate,3％inoculation volume ratio of bacterial suspension,pH=6.0,fermentation culture tem-perature 35℃.The optimized enzyme activity reached 43.06 U/mL,which was 78.7％higher than that of the orig-inal strain.The results can provide the oretical basis and application value for the efficient degradation of cellulose.
关键词
纤维素酶/紫外诱变/发酵优化/正交实验Key words
Cellulase/Ultraviolet mutagenesis/Fermentation optimization/Orthogonal experiment引用本文复制引用
基金项目
烟台市校地融合发展项目(2021XDRHXMQT19)
中国农业大学烟台研究院"一流教研室"建设项目(202201Ys)
出版年
2024
NETL
NSTL
万方数据