ARTP-DES复合诱变选育高产纤维素酶突变菌株  

Breeding of mutant strains with high cellulase production by ARTP-DES compound mutagenesis

作　　者：王丰园 金海炎 张征田[1] 丁凌飞 鲁云风[1,3] WANG Feng-yuan;JIN Hai-yan;ZHANG Zheng-tian;DING Ling-fei;LU Yun-feng

机构地区：[1]南阳师范学院生命科学与农业工程学院,河南南阳473061 [2]车用生物燃料技术国家重点实验室,河南南阳473000 [3]河南省高校生物质资源化工程中心,河南南阳473061

出　　处：《饲料研究》2021年第21期92-97,共6页Feed Research

基　　金：河南省高等学校重点科研项目(项目编号:20A180022);车用生物燃料技术国家重点实验室开放课题(项目编号:KFKT2018002)。

摘　　要：为选育高产纤维素酶菌株,试验以拟康氏木霉菌株采用常温常压等离子体(ARTP)与硫酸二乙酯(DES)复合理化诱变,确定复合诱变条件为ARTP照射时间110 s,DES浓度为0.8%。采用刚果红染色法初筛突变菌株,得到19株突变株的透明圈与菌落直径的比值大于出发菌株。测定19株突变株纤维素酶活,获得一株纤维素酶活性较大的菌株AD-55,其滤纸酶活性、外切酶活性、内切酶活性分别为2.16、7.04、3.98 IU/mL。通过单因素与正交试验优化产酶培养基条件,得到最佳产酶培养条件为微晶纤维素1.25%、玉米浆粉1.20%、(NH_(4))_(2)SO_(4)0.20%。在此条件下,菌株AD-55的滤纸酶、外切酶、内切酶活性分别为2.89、8.82、5.10 IU/mL。菌株AD-55遗传较为稳定。In order to select a high cellulase producing strain, Trichoderma pseudokoningii was used as the starting strain, the compound physical and chemical mutagenesis of normal temperature and pressure plasma(ARTP) and diethyl sulfate(DES) was used. The compound mutagenesis conditions were determined as ARTP irradiation time of 110 s and DES concentration of 0.8%. Congo red staining was used to screen the mutant strains, and the ratio of transparent circle to colony diameter of 19 mutant strains was greater than that of the original strain. The cellulase activity of 19 mutant strains was determined, and a strain AD-55 with higher cellulase activity was obtained. The filter paper enzyme activity,exonuclease activity and endonuclease activity were 2.16, 7.04 and 3.98 IU/mL, respectively. Through single factor and orthogonal experiments, the optimum enzyme production medium conditions were microcrystalline cellulose 1.25%, corn pulp powder 1.20% and(NH_(4))_(2)SO_(4) 0.20%. Under the condition, the filter paper enzyme activity, exonuclease activity and endonuclease activity of strain AD-55 were 2.89, 8.82 and 5.10 IU/mL, respectively. The strain AD-55 was genetically stable.

关 键 词：常温常压等离子体 硫酸二乙酯 拟康氏木霉 纤维素酶 酶活性 

分 类 号：S811.6[农业科学—畜牧学]