食品与发酵工业 >

2021 , Vol. 47 >Issue 1: 43 - 49

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.024449

研究报告

毕赤酵母产伏马毒素B1羧酸酯酶发酵条件和培养基的优化

  • 赵一凡 ,
  • 常晓娇 ,
  • 杜稳 ,
  • 孙长坡 ,
  • 刘虎军
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  • (国家粮食和物资储备局科学研究院,北京,100037)
硕士,研究实习员(刘虎军助理研究员为通讯作者,E-mail:lhj@ags.ac.cn)

收稿日期: 2020-05-11

  修回日期: 2020-06-10

  网络出版日期: 2021-02-03

基金资助

国家自然科学基金项目(U1604234);中央级公益性科研院所基本科研业务费专项资金课题(ZX1903-4;JY2001)

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Optimization of fermentation condition and medium for fumonisin B1 carboxylesterase production in Pichia pastoris

  • ZHAO Yifan ,
  • CHANG Xiaojiao ,
  • DU Wen ,
  • SUN Changpo ,
  • LIU Hujun
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  • (Academy of National Food and Strategic Reserves Administration,Beijing 100037, China)

Received date: 2020-05-11

  Revised date: 2020-06-10

  Online published: 2021-02-03

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摘要

为提高毕赤酵母重组菌产伏马毒素B1(fumonisin B1,FB1)羧酸酯酶的摇瓶发酵水平,以酶活力为指标,对发酵条件和培养基进行优化。通过单因素试验对发酵条件进行优化。通过Plackett-Burman试验筛选出关键培养基成分,再进行正交试验建立试验数据样本,最后建立误差反向传播神经网络模型进行预测并寻找最优发酵培养基组成。经优化得到的发酵条件:诱导温度28 ℃,初始pH 6.0,每24 h补加体积分数为1%的甲醇,诱导96 h;优化后发酵培养基组成:蛋白胨 23 g/L、KH2PO4 44 g/L、PTM4 (Pichia trace minerals 4)微量元素溶液 1.5 mL/L、K2SO4 7.15 g/L、MgSO4·7H2O 5.85 g/L和酵母粉 5 g/L。FB1羧酸酯酶酶活力达到402 U/mL以上,较优化前提高了1.19倍。优化后显著提高了重组菌株的产酶能力,研究结果为FB1羧酸酯酶发酵罐优化提供了基础数据。

关键词： 伏马毒素; 羧酸酯酶; 发酵条件; 培养基优化; 人工神经网络

本文引用格式

赵一凡 , 常晓娇 , 杜稳 , 孙长坡 , 刘虎军 . 毕赤酵母产伏马毒素B1羧酸酯酶发酵条件和培养基的优化[J]. 食品与发酵工业, 2021 , 47(1) : 43 -49 . DOI: 10.13995/j.cnki.11-1802/ts.024449

Abstract

In order to improve the yield of fumonisin B1 (FB1) carboxylesterase produced by recombinant Pichia pastoris in shaking flask, the fermentation condition and medium were optimized with enzyme activity as index. The fermentation conditions were optimized through single factor experiment. The key components of fermentation medium were screened through Plackett-Burman experiment and then orthogonal experiment was carried out to establish the experimental data samples. Finally, the error Back Propagation (BP) neural network model was established to predict and search for the optimal composition of fermentation medium. After optimization, the optimal fermentation conditions were induction temperature 28 ℃, initial pH 6.0, 1% of methanol addition every 24 h, induction for 96 h. The optimal fermentation medium was peptone 23 g/L, KH2PO4 44 g/L, PTM4 1.5 mL/L, K2SO4 7.15 g/L, MgSO4·7H2O 5.85 g/L, yeast extract 5 g/L. FB1 carboxylesterase activity was over 402 U/mL, 1.19-fold higher than that of before optimization. Fermentation optimization in shaking flask can significantly improve the enzyme production capacity of recombinant strains. These results presented basic data for the optimization of FB1 carboxylesterase in fermenter.

Key words： fumonisins; carboxylesterase; fermentation conditions; medium optimization; artificial neural network

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