葡萄糖氧化酶产生菌的诱变育种及发酵工艺优化  

Study on Fermentation Process Optimization and Mutation Breeding of Glucose Oxidase Producing Bacteria

作　　者：赵芬芬 黄月荣 赵辉 吴静 姜冬冬 ZHAO Fenfen;HUANG Yuerong;ZHAO Hui;WU Jing;JIANG Dongdong(Zhengzhou Yellow River Nursing Vocational College,Zhengzhou 450000,China;Zhengzhou Yuanchuang Gene Limited Company,Zhengzhou 450000,China)

机构地区：[1]郑州黄河护理职业学院,河南郑州450000 [2]郑州源创基因科技有限公司,河南郑州450000

出　　处：《工业微生物》2023年第5期137-140,共4页Industrial Microbiology

基　　金：河南省科学技术厅项目(232102310329);郑州黄河护理职业学院教育教学研究项目(HHJY2023011)。

摘　　要：文章为优化葡萄糖氧化酶产生菌的诱变育种和发酵工艺,将桔青霉CICC 40277作为生产菌株,通过对固、液培养基中菌种的生长和产酶活力进行比较,选取木薯叶为培养基质,经固体培养活化后,菌株的产酶活力最高达到66.48 U/g。然后采用2-脱氧-D-葡萄糖为菌株抑制剂,以甲基红为指示剂设计了一种高效、简便的筛选方法。通过紫外诱变育种,最终筛选得到一株高产菌株Uv-7,其产酶活力达到79.76 U/g,比原始菌株提高了20%。采用单因素试验对Uv-7菌株产葡萄糖氧化酶的最佳菌种活化条件进行优化,得到最适条件为:培养时间144 h、培养温度30℃、接种量20%(V/V);最适培养基组分为:以5.5 g木薯叶为固体基质,按照1∶1(W/V)的固液比添加盐溶液(g/L):MgSO_(4)·7H_(2)O 0.5、Na_(2)HPO_(4)2.0、KCl 0.3、CaCO_(3)3.0、葡萄糖30、酵母粉6,当pH为6.0时,菌株经活化后产葡萄糖氧化酶的活性提高到了101.52 U/g,与原始菌株相比提高了52.7%。In order to optimize the mutation breeding and fermentation process of glucose oxidase producing bacteria,Penicillium citri CICC 40277 was used as the production strain.By comparing the growth and enzyme production activity of the strains in solid and liquid culture media,cassava leaves were selected as the culture medium.After solid culture activation,the enzyme production activity of the strain reached the highest of 66.48 U/g,and an efficient and simple screening method was established using 2-deoxy-D-glucose as a strain inhibitor and the methul red as an indicator.Through UV mutation breeding,a high-yield strain Uv-7 was ultimately screened,with an enzyme production activity of 79.76 U/g,which was 20%higher than the original strain.The optimal activation conditions for glucose oxidase production by Uv-7 strain were optimized using a single factor experiment.The optimal conditions were obtained as follows:culture time 144 hours,culture temperature 30℃,and inoculation amount 20%(V/W);The optimal medium composition is using 5.5 g cassava leaves as a solid substrate,adding salt solution at a 1∶1(W/V)solid-liquid ratio:MgSO_(4)·7H_(2)O 0.5,Na_(2)HPO_(4)2.0,KCl 0.3,CaCO_(3)3.0,glucose 30,yeast powder 6.When the pH was 6.0,the activity of glucose oxidase produced by the activated strain increased to 101.52 U/g,which was 66.43%higher than the original strain.

关 键 词：葡萄糖氧化酶 桔青霉 木薯叶 固体发酵 紫外诱变 发酵条件优化 

分 类 号：TQ925[轻工技术与工程—发酵工程]