四川农业大学学报

• 研究论文 • 上一篇    下一篇

响应面法优化巨大芽孢杆菌产纤维素酶发酵条件

  

  1. 四川农业大学生命科学与理学院,四川 雅安 625014
  • 收稿日期:2013-02-07 出版日期:2013-09-27 发布日期:2013-09-27

Optimization of Bacillus Megaterium cellulase production for submerged fermentation by response surface methodology

  • Received:2013-02-07 Online:2013-09-27 Published:2013-09-27

摘要:

【目的】为了解决纤维素酶发酵工业产量低、生产成本高等问题,加快纤维素酶工业化应用的步伐。【方法】采用5L发酵罐,利用响应面分析法(RSM)对基因工程菌WH320-pHIS1525-G7从接种量、木糖流速及流加时间进行产酶优化。【结果】确定其发酵工艺为:17.62%的接种量,温度37℃,pH7.0,罐压0.03~0.05Mpa,转速300r/min,0.41g/(L•h)的速度流加木糖11.33 h,发酵过程中控制溶氧浓度≥30%。【结论】在该发酵条件下测得纤维素酶活力为2.152U/mL,比优化前摇瓶发酵酶活力提高了2倍,可为工业化生产纤维素酶提供依据。

关键词: 巨大芽孢杆菌;中性纤维素酶;发酵条件;响应面分析法

Abstract:

【Objective】Cellulases have taken on satisfactory application performance and economic value in detergent industry because they show relative stability in neutral and basic conditions. However, the large-scale industrialized application of these enzymes was restricted by the low activity and high cost price. Improving their activity through fermentation engineering was considered an efficient approach to cut down the cost price.【Method】The 5L fermentation tank was used to optimize the fermentation conditions for producing neutral cellulase from a Bacillus Megaterium genetic engineering bacteria WH320-pHIS1525-G7 . Response surface method was used to optimize the fermentation conditions of Bacillus Megaterium genetic engineering bacteria WH320-pHIS1525-G7.【Result】The optimal fermentation conditions were 17.62% inoculation amount, fermentation temperature of 37℃, pH 7.0, tank pressure 0.03~0.05 Mpa, rotation speed of shaker 300 r/min, fed 11.33h xylose by 0.41g/(L•h) flow rate, dissolved oxygen≥30% during the fermentation process.【Conclusion】The cellulase activity was up to 2.152U/mL which was 2 times higher than the shaking culture. This research can provide the basis for the cellulase manufacture.

Key words: Bacillus Megaterium; neutral cellulase; fermentation conditions;response surface methodology