响应面法模拟优化蜡样芽孢杆菌W-3菌株产木聚糖酶发酵条件

doi:10.16431/j.cnki.1671-7236.2024.03.005

摘要/Abstract

摘要： 【目的】应用响应面法对产木聚糖酶的工艺进行优化,为进一步大规模生产提供参考依据。【方法】通过单因素试验分别考察碳源、氮源、pH、温度、接种量和转速6个因素对蜡样芽孢杆菌W-3产木聚糖酶的影响。通过Design Expert 10.0.3软件中的Box-Behnken进行响应面优化设计。通过对小麦麸皮、氯化铵、pH、温度、接种量和转速进行不同组合,系统地优化了发酵条件。利用二次响应面回归方法建立酶产量与各因素之间的数学模型,并通过回归分析确定各因素对酶产量的影响程度。【结果】小麦麸皮添加量、氯化铵添加量、pH和温度是影响发酵的主效应因素,单因子优化显示,蜡样芽孢杆菌W-3发酵产木聚糖酶条件为小麦麸皮35 g/L、氯化铵5 g/L、pH 8.0、温度70 ℃、接种量2%、转速210 r/min。在单因子优化的基础上进一步应用响应面优化,找到了更优的发酵参数,经过优化后的发酵条件为小麦麸皮33.0 g/L、氯化铵5.1 g/L、pH 8.3、温度73 ℃、接种量2%、转速210 r/min,该条件下蜡样芽孢杆菌W3发酵产生的木聚糖酶的酶活为523.24 U/mL。【结论】本试验通过响应面法优化了蜡样芽孢杆菌W-3发酵条件,在33.0 g/L小麦麸皮、5.1 g/L氯化铵、pH 8.3、73 ℃、2%接种量、210 r/min条件下,可获得较高的木聚糖酶的酶活(523.24 U/mL),比优化之前(225.53 U/mL)提高了2.32倍。

关键词: 蜡样芽孢杆菌W-3; 木聚糖酶; Box-Behnken设计; 响应面法

Abstract: 【Objective】 This experiment was aimed to optimize the production process of xylanase using response surface methodology,so as to provide a reference basis for further large-scale production.【Method】 Employing single-factor experiments,the influence of six factors,including carbon source,nitrogen source,pH,temperature,inoculum size and agitation speed,on xylanase production by Bacillus cereus (B. cereus) W-3 was examined meticulously.The Box-Behnken response surface optimization design in Design Expert 10.0.3 software was applied.The fermentation conditions were systematically optimized by varying combinations of wheat bran,ammonium chloride,pH,temperature,inoculum size and agitation speed.A mathematical model for enzyme production in relation to these factors was established using a quadratic response surface regression method.The impact levels of each factor on enzyme production were determined through regression analysis.【Result】 The addition of wheat bran,ammonium chloride,pH and temperature were the main effect factors affecting fermentation.Through single-factor optimization,it had been determined that B. cereus W-3 ferments to produce xylanase under the following conditions:Wheat bran concentration of 35 g/L,ammonium chloride concentration of 5 g/L,pH 8.0,a temperature of 70 ℃,an inoculum size of 2%,and an agitation speed of 210 r/min. Having built upon the single-factor optimization,further response surface optimization led to the discovery of superior fermentation parameters.The optimized fermentation conditions included 33.0 g/L wheat bran,5.1 g/L ammonium chloride,pH 8.3,a temperature of 73 ℃,an inoculum size of 2%,and an agitation speed of 210 r/min. Under the optimized fermentation conditions,xylanase enzymatic activity produced by B. cereus W-3 reached 523.24 U/mL.【Conclusion】 In this experiment,the B. cereus W-3 fermentation conditions was optimized by response surface methodology.Under the conditions of 33.0 g/L wheat bran,5.1 g/L ammonium chloride,pH 8.3,73 ℃,2% inoculum size and 210 r/min,a significantly higher xylanase activity of 523.24 U/mL could be obtained,representing a 2.32-fold increase compared to pre-optimization level (225.53 U/mL).

Key words: Bacillus cereus W-3; xylanase; Box-Behnken design; response surface methodology

中图分类号:

S852.61⁺6

肖遥, 李旭旭, 闫丽欢, 范新凤, 薛智权, 荆小院, 赵燕, 冯焱. 响应面法模拟优化蜡样芽孢杆菌W-3菌株产木聚糖酶发酵条件[J]. 中国畜牧兽医, 2024, 51(3): 936-944.

XIAO Yao, LI Xuxu, YAN Lihuan, FAN Xinfeng, XUE Zhiquan, JING Xiaoyuan, ZHAO Yan, FENG Yan. Simulation Optimization of Fermentation Conditions for Xylanase Production of Bacillus cereus W-3 by Response Surface Methodology[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(3): 936-944.

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