摘要
为开发新型高产β-葡萄糖苷酶的微生物菌种资源,本实验从腐木中分离获得1株产β-葡萄糖苷酶的青霉菌株L1;经等离子-硫酸二乙酯复合诱变后利用七叶苷平板法初筛,摇瓶发酵复筛,最终获得1株可稳定遗传的突变菌株D-6,经单因素试验、Plackett-Burman试验、最陡爬坡试验和响应面试验确定了其发酵产酶最佳条件。结果表明,最佳产酶条件是:KH2PO46 g/L、MgSO4·7H2O1 g/L、CaCl20.5 g/L、FeS040.1 g/L,初始pH5.2,接种量5%(孢子浓度108个/mL),碳源添加量(X1)玉米秸秆45.74 g/L、氮源添加量(X2)(NH4)2SO47.23 g/L、装液量(X5)63 mL/250 mL发酵温度28℃摇床转速160 r/min,发酵时间132 h,D-6菌株的β-葡萄糖苷酶活力为142.92 U/mL,较出发菌株L1提高了274.4%。研究结果为产β-葡萄糖苷酶菌株发酵条件优化提供技术参考同时为该类菌株的开发和应用提供有效的菌种资源。
A Penicillium strain named L1 capable of producingβ-glucosidase was isolated from rotten wood with the aim to study and develop microbial origin(3-glucosidase resources.The strain was treated by atmospheric and room temperature plasma(ARTP)and diethyl sulfate(DES)mutation,followed by esculin agar plate screening and enzyme activity re-screening,aβ-glucosidase high-yield mutant D-6 with genetic stability was obtained.By one-factorat-a-time method,Plackett-Burman design,steepest ascent path and response surface methodology,the optimum conditions forβ-glucosidase production in fermentation were determined.The optimal fermentation conditions were KH2PO46 g/L,MgSO4·7H2O 1 g/L,CaCl20.5 g/L and FeSO40.1 g/L;pH value of 5.2,inoculum 5%(spore concentration 108/mL),corn straw(X1)45.74 g/L,(NH4)2SO47.23 g/L,packing volume(X5)63 mL/250 mL,temperature 28℃,shaking speed is 160 r/min,incubation 132 h.Under the above conditions,the(3-glucosidase activity of D-6 was 142.92 U/mL,with an increase of 274.4%compared with the original strain L1.The results obtained in the study were meaningful for technical reference and microbial resources ofβ-glucosidase producing strain.
作者
刘文静
程晗
陈崇艺
柴春月
田龙
周索
LIU Wenjing;CHENG Han;CHEN Chongyi;CHAI Chunyue;TIAN Long;ZHOU Suo(School of Life Science and Technology,Nanyang Normal University,Nanyang 473061,China;Henan Provincial Engineering and Technology Research Center for Mushroom Food,Nanyang 473061,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2019年第23期43-49,共7页
Food and Fermentation Industries
基金
南阳师范学院研究生创新基金项目(2018CX009/2019CX002)
关键词
Β-葡萄糖苷酶
青霉菌
七叶苷平板
复合诱变
响应面法
β-glycosidase
Penicillium
esculin agar plates
compound mutation
response surface methodology
