吡咯伯克霍尔德氏菌B1213产葡聚糖酶条件的优化

Optimization of Conditions of Glucanase Production by Burkholderia pyrrocinia B1213

以吡咯伯克霍尔德氏菌B1213为研究对象,优化其发酵产葡聚糖酶条件。首先,通过单因素实验对其培养条件(碳源种类、碳源粒度、碳源质量浓度、氮源种类、氮源质量浓度、转速、温度、装液量、接种量、初始pH值、表面活性剂种类、表面活性剂质量浓度和时间)进行初步优化,然后通过Plackett-Burman试验,筛选出4个显著影响B1213产葡聚糖酶的因素,再利用最陡爬坡试验确定其产葡聚糖酶的最大响应区域,最后通过Box-Behnken试验设计确定其最优产葡聚糖酶条件为:40~60目麦麸质量浓度50 g/L,酵母浸粉质量浓度8 g/L,初始pH 6.6,装液量30 mL/250 mL,接种量0.15%,曲拉通100质量浓度25 g/L,温度30℃,转速160 r/min和培养时间83 h。在此培养条件下B1213产葡聚糖酶活力达到1230 U/mL。研究结果为细菌B1213在白酒酿造中的应用提供理论基础。

In this study,the fermentation condition of glucanase production by Burkholderia pyrrocinia B1213 has been optimized.Firstly,cultural conditions(variety of carbon sources,particle size and mass concentration of carbon source,variety of nitrogen sources,mass concentration of nitrogen sources,shaking speed,temperature,liquid medium volume,inoculum volume,initial pH,variety of surfactants,mass concentration of surfactant and fermentation time)of B1213 were optimized by single-factor experiments.Then,four factors,signally affect production of glucanase by B1213 fermentation,were obtained by Plackett-Burman tests.The maximum response area of glucanase activity was determined by steepest ascent.Finally,Box-Behnken Design were used to ascertain the optimal condition of glucanase production.The optimal condition were:40-60 mesh of wheat bran 50 g/L,yeast extract 8 g/L,initial pH 6.6,liquid medium volume 30 mL/250 mL,inoculum volume 0.15%,Triton X-10025 g/L,incubating temperature 30℃,shaking speed 160 r/min and fermentation time 83 h.Under the optimal condition,the activity of glucanase produced by B1213 were achieved 1230 U/mL.This study provides academic value for application of bacteria B1213 in Baijiu brewing from the aspect of enzymes related to degradation of raw material for Baijiu brewing.