贝莱斯芽孢杆菌D103产纤维素酶优化及途径分析  

Optimization and pathway analysis of cellulase production by Bacillus velezensis D103

作　　者：黄悦姝 邓超 张馨月 高旺 曾楠 李炳学[1] HUANG Yue-shu;DENG Chao;ZHANG Xin-yue;GAO Wang;ZENG Nan;LI Bing-xue(School of Land and Environment,Shenyang Agricultural University,Shenyang 110866,China)

机构地区：[1]沈阳农业大学土地与环境学院,辽宁沈阳110866

出　　处：《生物技术》2023年第5期632-640,共9页Biotechnology

基　　金：国家重点研发计划项目(2017YFD0200807-3);辽宁省农村科技特派行动计划项目(2022-09);2020年度沈阳市科学技术计划项目(20-207-3-35)。

摘　　要：[目的]获得高纤维素酶活芽孢杆菌菌株,优化产酶条件并分析其产酶途径。[方法]应用刚果红染色法和羧甲基纤维素酶活法筛选纤维素降解菌株,利用PB试验及最陡爬坡试验优化降解菌的高纤维素酶活培养条件。对降解菌的全基因组数据进行生物信息学分析,探究其产纤维素酶途径。[结果]贝莱斯芽孢杆菌D103刚果红染色直径D/d值高达2.49,纤维素酶活性为41.73 U/mL。对D103培养条件优化后,BBD试验中最佳结果为酶活达到65.94 U/mL,提高53%。PB试验结果表明,培养时间对纤维素酶活影响最显著,响应面法分析,装液量与蛋白胨添加量交互影响最大。D103全基因组全长3857531 bp,GC含量46.7%,共编码基因3884个。D103全基因组中找到包括纤维素代谢及半纤维素代谢基因共16个。[结论]贝莱斯芽孢杆菌D103高产纤维素酶,其代谢通路中存在纤维素分析途径。[Objective]Obtaining a Bacillus strain with high cellulase activity,optimising enzyme production conditions and analysing the enzyme production pathway.[Method]The cellulose degrading strains were screened by Congo red staining and carboxymethyl cellulase activity method,and the high cellulase activity culture conditions of the degrading bacteria were optimised using PB test and steepest climb test.The whole genome data of the degrading bacteria were analyzed bioinformatically to investigate their cellulase production pathways.[Result]B.velezensis D103 had a Congo red stained diameter D/d value of 2.49 and a cellulase activity of 41.73 U/mL.After optimizing the culture conditions of D103,the best result in the BBD test was an enzyme activity of 65.94 U/mL,an increase of 53%.The results of the PB test showed that incubation time had the most significant effect on cellulase activity,and the response surface analysis showed that the amount of loading solution interacted most with the amount of peptone added.The whole genome of D103 is 3857531 bp in length,with a GC content of 46.7%,encoding a total of 3884 genes.Sixteen genes,including cellulose metabolism and hemicellulose metabolism,were found in the whole genome of D103.[Conclusion]B.velezensis D103 is high in cellulase production and has a cellulose analysis pathway in its metabolic pathway.

关 键 词：贝莱斯芽孢杆菌 纤维素酶活 羧甲基纤维素酶活法 响应面法 全基因组 纤维素降解基因 

分 类 号：S154.34[农业科学—土壤学]