碳氮源对枯草芽孢杆菌发酵产β-甘露聚糖酶的影响  

Effects of carbon and nitrogen sources on β-mannanase production from Bacillus subtilis

作　　者：龚劲松[1] 李恒[1] 刘恒霞[1] 陆震鸣[1] 蒋敏[1] 罗春勤 许正宏[1] 史劲松[1] 

机构地区：[1]江南大学药学院,江苏无锡214122 [2]永安缘和生物科技有限公司,四川成都611630

出　　处：《食品与发酵工业》2015年第10期34-39,共6页Food and Fermentation Industries

基　　金：江苏省产学研前瞻性联合研究项目(BY2012062);十二五国家科技支撑计划项目(2012BAD33B06)

摘　　要：为提高枯草芽孢杆菌产甘露聚糖酶的能力,对Bacillus subtilis YH12产β-甘露聚糖酶的培养基组分和培养条件进行了优化,重点考察了碳氮源对发酵产酶的影响。实验结果表明,以魔芋精粉和甘露寡糖作为碳源时能对菌株发酵产酶起到良好的诱导作用,魔芋甘露寡糖的诱导效果要优于魔芋精粉,从经济效益角度考虑选择魔芋精粉作为碳源和诱导底物;以胰蛋白胨作为发酵氮源时产酶效果最佳。研究同时发现,产酶的最佳碳氮比为2:1;Na^+、K^+和Mg^(2+)对酶活具有显著促进作用;对发酵条件的考察结果显示,选择初始pH和培养温度分别为7.5和30℃时产酶效果最好。产酶发酵过程及SDS-PAGE分析表明,发酵33 h时产酶量达到最高。在此条件下B.subtilis YH12发酵产酶高峰期β-甘露聚糖酶活力达到280 U/mL,相比初始水平提高约5倍。In order to improve the β-mannanase production from Bacillus subtilis YH12, the effect of medium components and culture conditions on fermentation was investigated, especially the carbon and nitrogen sources. Re- suits showed that moderate induction effect was observed with the konjacmannan oligosaccharide and konjac flour as the carbon source. Although konjacmannan oligosaccharide showed better improvement effect on enzyme formation than konjac flour, the latter was selected as the preferred carbon source and induction substrate due to its relatively high economic efficiency. Moreover, the tryptone was determined to be the optimum nitrogen source and meanwhile the optimal carbon-nitrogen ratio was 2： 1. Na ^＋ , K ^＋ , and Mg^2＋ significantly improved the β-mannanase production. The investigations on culture conditions showed an optimal culture pH and temperature of 7.5 and 30 ℃. The maxi- mum activity was achieved at 33 h through the fermentation curve and SDS-PAGE. Based on these data, the β-man- nanase production reached up to 280 U/mL, which was increased by 5-fold compared with the initial level.

关 键 词：枯草芽孢杆菌 Β-甘露聚糖酶 诱导 发酵条件 碳氮源 

分 类 号：TQ925[轻工技术与工程—发酵工程]