枯草芽胞杆菌甘露聚糖酶发酵条件优化  

Optimization of Fermentation Conditions of Mannanase by Bacillus subtilis

作　　者：李盛前 侯颖[1,3] 范鑫诺 陈阳阳 宫强[1,2] 李阳 徐建强[4] LI Sheng-qian;HOU Ying;FAN Xin-nuo;CHEN Yang-yang;GONG Qiang;LI Yang;XU Jian-qiang(Coll.of Food&Bio-Engin.,Henan Uni.of Sci.&Tech.,Luoyang 471023;Henan Prov.Res.Centre of Food Microbial Engin.&Tech.,Luoyang 471023;Key Lab.of Microbial Res.Devel.&Appl.at Uni.Level,Luoyang 471023;Coll.of Horticul.&Plant Protect.,Henan Uni.of Sci.&Tech.,Luoyang 471023)

机构地区：[1]河南科技大学食品与生物工程学院,河南洛阳471023 [2]河南省食品微生物工程技术研究中心,河南洛阳471023 [3]微生物资源开发与利用河南科技大学重点实验室,河南洛阳471023 [4]河南科技大学园艺与植物保护学院,河南洛阳471023

出　　处：《微生物学杂志》2022年第6期39-45,共7页Journal of Microbiology

基　　金：国家自然科学基金项目(31400098);河南省科技攻关计划项目(202102110071,172102310435)。

摘　　要：旨在以枯草芽胞杆菌Bacillus subtilis J为生产菌株,发酵生产β-甘露聚糖酶,通过优化产酶条件,以达到提高β-甘露聚糖酶产量的目的。利用DNS比色法检测β-甘露聚糖酶活力,采用单因素试验,研究碳氮源种类及碳氮源浓度、温度、pH、接种量和装液量对菌株Bacillus subtilis J发酵产β-甘露聚糖酶的影响,结合响应面试验设计确定菌株Bacillus subtilis J发酵产甘露聚糖酶的最优发酵培养条件。单因素试验和响应面试验得到最优的发酵条件为魔芋粉28 g/L,胰蛋白胨21 g/L,K_(2)HPO_(4)6 g/L,MgSO_(4)·7H_(2)O 1 g/L,温度31℃,pH值8.5,接种量1%(体积分数),装液量50 mL/250 mL,发酵周期24 h。利用优化后的培养基生产β-甘露聚糖酶,其酶活力达到84.38 U/mL,是初始发酵培养基产酶活力的3.36倍。通过对发酵条件的优化,大幅度提高了β-甘露聚糖酶的产量,为其工业生产提供参考。The purpose of this study is to improve the yield of β-mannanase through optimization of fermentation conditions of Bacillus subtilis J as production strain. The activity of β-mannanase was detected by DNS colorimetry, and the effects of type of carbon source and concentration of carbon and nitrogen sources, temperature, pH, inoculum volume and liquid filling volume on β-mannanase production by B. subtilis J were studied by single factor experiment, combined with the optimal fermentation conditions of mannanase by the strain were determined by response surface methodology. The optimal fermentation conditions by single factor and response surface experiments obtained as follows: konjak(Amophophallus rivieri) powder 28 g/L, tryptone 21 g/L, K_(2)HPO_(4)6 g/L, MgSO_(4)·7H_(2)O 1 g/L, 31 ℃, pH 8.5, inoculum volume 1%, liquid filling volume 50 mL/250 mL, and ferment for 24 h. The yield of β-mannanase produced by B. subtilis J in optimized conditions reached to 84.38 U/mL, 3.36 times of that in the initial fermentation medium. By optimizing the fermentation conditions, the yield of β-mannanase was greatly increased, and provided data reference for the industrial production.

关 键 词：Β-甘露聚糖酶 甘露聚糖 枯草芽胞杆菌 发酵条件优化 响应面试验 

分 类 号：Q939.97[生物学—微生物学]