高产纤维素酶真菌的筛选鉴定与紫外诱变选育研究  

Screening and identification of high-yield cellulase fungi and study on UV mutagenesis breeding

作　　者：李婕 李敏 汪玲玲 周鹤 张欣莹 付文翔 邹伟 LI Jie;LI Min;WANG Ling-ling;ZHOU He;ZHANG Xin-ying;FU Wen-xiang;ZOU Wei

机构地区：[1]四川轻化工大学生物工程学院,四川宜宾644005 [2]四川轻化工大学浓香型白酒资源微生物与大数据实验室,四川宜宾644005

出　　处：《饲料研究》2023年第17期76-81,共6页Feed Research

基　　金：四川轻化工大学省级大学生创新创业训练计划项目(项目编号:S202010622085);四川轻化工大学研究生创新基金(项目编号:D10501594)。

摘　　要：试验旨在拟从腐朽木材和腐质土壤中获得1种高效的纤维素酶生产菌,利用紫外线诱变技术对其进行改造。采用刚果红染色法和酶活性试验,筛选出2株纤维素酶产率较高的菌(菌株HS5、菌株1)。经形态学和分子鉴定,菌株HS5为黄曲霉(Aspergillus flavus),菌株1为烟曲霉(Aspergillus fumigatus)。在固体产酶培养基中,以水稻秸秆为唯一的碳源,28℃培养4 d后,HS5滤纸酶(FPA)的酶活为306 U/g,羟甲基化纤维素(CMC)的酶活为592.2 U/g;菌株1的FPA酶活为214.2 U/g,CMC酶活为523.8 U/g,表现出了较好的降解纤维素的能力。紫外诱变处理得到产酶能力提高的突变菌株为UR-07和URM-13。与原始菌株相比,UR-07的FPA和CMC酶活分别比原始菌株HS5提高了15.86%、16.68%,URM-13的FPA和CMC酶活比菌株分别提高了26.94%、19.03%。研究表明,经紫外诱变后,产纤维素酶真菌的产酶能力有所提升且具有较好的遗传稳定性,两株菌株在纤维素酶的生产和纤维素类物质降解菌剂的研究中具有较高的潜力。The study was to obtain an efficient cellulase producing bacterium from decaying wood and decaying soil,and modify it using UV induction technology.Two cellulase producing bacteria(strain HS5,strain 1)with high cellulase yield were screened by Congo red method and enzyme activity test.After morphological and molecular identification,strain HS5 was Aspergillus flavus and strain 1 was Aspergillus fumigatus.After 4 d of cultivation at 28℃,the enzyme activity of HS5 filter paper enzyme(FPA)was 306 U/g and that of hydroxymethylated cellulose(CMC)was 592.2 U/g,the FPA activity of strain 1 was 214.2 U/g,and the CMC activity was 523.8 U/g,in solid enzyme producing medium with rice straw as the only carbon source,showing a good ability to degrade cellulose.After UV mutagenesis treatment,the mutant strains UR-07 and URM-13 with improved enzyme production capacity were finally obtained.Compared with the original strain,the FPA and CMC enzyme activities of UR-07 were increased by 15.86%and 16.68%,respectively,and the FPA and CMC enzyme activities of URM-13 were increased by 26.94%and 19.03%,respectively.The study indicates that the enzyme production ability of cellulase-producing fungi obtained after UV mutagenesis is improved and have good genetic stability,which provides a high potential strain for production of cellulase and the study of cellulose-degrading microbial agents.

关 键 词：烟曲霉 黄曲霉 纤维素酶 真菌选育 紫外诱变 

分 类 号：S816.7[农业科学—饲料科学]