β-甘露聚糖酶产生菌DY-14的发酵优化及分离纯化

Optimization and purification of Beta-Mannanase by Bacillus subtilis DY-14

利用单因素实验和正交分析方法得到枯草芽孢杆菌DY-14发酵产酶的最佳培养基组分及条件:以魔芋粉为碳源(3%),蛋白胨为氮源(1.5%),初始pH 8.0,摇瓶装液量10 mL/100mL,接种量1%,37℃培养24h,最终产生的β-甘露聚糖酶活力可达3216 U/mL。DY-14发酵液经(NH4)2SO4盐析、DEAE-Sepharose阴离子交换层析、Sephadex G-75凝胶过滤,得到了电泳纯的β-甘露聚糖酶。

Single factor and orthogonal array design experiments were performed to obtain the optimum fermentation conditions for β-mannanase production,Bacillus subtilis DY-14 grew best when cultured at 37℃with the medium composed of 3% konjac powder as source,1.5% peptone as nitrogen source,and adjusting pH to 8.0.After 24 hours culturing,the enzyme could show the highest activity as 3216 U/mL.After salting out the DY-14 ferment liquid by（NH4）2SO4,use DEAE-Sepharose for anion exchange chromatography and Sephadex G-75 for gel filter,the electrophoresis pure of β-mannanase by Bacillus subtilis DY-14 was gained.