高产纤溶酶菌株的诱变育种及产酶条件的响应面法优化  

Mutation of a strain with high production of fibrinolytic enzyme and process optimization of plasmin fermentation by response surface analysis

作　　者：刘林[1] 谢和[1] 

机构地区：[1]贵州大学生命科学学院,贵州贵阳550025

出　　处：《广东农业科学》2013年第7期103-107,共5页Guangdong Agricultural Sciences

摘　　要：以解淀粉芽孢杆菌GZJSI-12作为出发菌株,经紫外线和亚硝基胍复合诱变,筛选得到突变株GZJSI-12-7,其产纤溶酶活力为2 046.44 IU/mL,是出发菌株GZJSI-12的1.51倍。结合Plackett-Burman设计法和响应面分析法,对GZJSI-12-7合成纤溶酶的发酵条件进行优化。结果表明,初始MgSO4浓度、发酵温度和NaH2PO4浓度是影响纤溶酶发酵产量的主要因素,优化后的发酵条件为葡萄糖30 g/L、蛋白胨10 g/L、吐温-80 5 g/L、CaCl2 0.4 g/L、MgSO4 0.847 g/L、Na2HPO4 2 g/L、NaH2PO4 6.988 g/L、pH 9.0,接种量4%,种龄21 h,温度31.3℃,每250 mL装液量50 mL,转速150 r/min。在此条件下,纤溶酶产量为4 789.08 IU/mL。A mutant named GZJS1-12-7 was achieved from Bacillus amyloliquefaciens GZJSr12 with combined mutation by ordinal treatment of UV and nitrosoguanidine （NTG）. The plasmin activity of GZJSI-12-7 was 2 046.44 IU/mL, which was 1.51 times higher than that of GZJS1-12. Plackett-Burman and response surface analysis methods were applied to optimize conditions of fibrinolytic enzyme fermentation. The results indicated that fibrinolytic enzyme yield was mainly influenced by concentration of initial MgSO4, NaH2PO4 and temperature. The obtained optimal fermentation conditions were glucose 30 g/L, peptone 10 g/L, Tween-80 5 g/L, CaC12 0.4 g/L, MgSO4 0.847 g/L, Na2HPO4 2 g/L, NaH2PO4 6.988 g/L, pH 9.0, inoculation volume of 4%, age of inoeulum 21 h, temperature 31.3℃, liquid loading 50 mL per 250 mL and rotation speed of 150 r/min. Under these conditions, the fibrinolytic enzyme yield was 4 789.08 IU/mL.

关 键 词：纤溶酶 解淀粉芽孢杆菌 复合诱变 响应面分析法 

分 类 号：TQ464.8[化学工程—制药化工]