《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (4): 1217-1226.doi: 10.16431/j.cnki.1671-7236.2019.04.031

• 基础兽医 • 上一篇    下一篇

一株枯草芽孢杆菌产抗菌肽培养基筛选及发酵工艺优化的研究

张志焱1, 赵倩1, 于佳民1, 刘长庆2, 徐海燕1, 谷巍1   

  1. 1. 山东省动物微生态制剂省级重点实验室, 山东宝来利来生物工程股份有限公司, 泰安 271000;
    2. 泰安市农业局, 泰安 271000
  • 收稿日期:2018-08-20 出版日期:2019-04-20 发布日期:2019-04-22
  • 作者简介:张志焱(1969-),女,研究员,研究方向:动物微生态制剂的研究与开发,E-mail:905817462@qq.com
  • 基金资助:

    "十三五"国家重点研发计划项目"动物疫病生物防治性制剂研制与产业化"(2017YFD0501000)

Selection of Culture Medium and Optimization of Fermention Conditions for Bacillus subtilis Producing Antimicrobial Peptides

ZHANG Zhiyan1, ZHAO Qian1, YU Jiamin1, LIU Changqing2, XU Haiyan1, GU Wei1   

  1. 1. Shandong Key Laboratory of Animal Micro-Ecological Agent, Shandong Baolai-Leelai Bioengineering Co., Ltd., Tai'an 271000, China;
    2. Tai'an Agricultural Bureau, Tai'an 271000, China
  • Received:2018-08-20 Online:2019-04-20 Published:2019-04-22

摘要:

试验旨在对一株产抗菌肽枯草芽孢杆菌BL0006的发酵培养基和发酵条件进行优化,以提高发酵液中抗菌肽的效价。首先通过单因子试验筛选出培养基的最适碳源、氮源与无机盐分别为葡萄糖、蛋白胨和K2HPO4;在此基础上采用最陡爬坡路径逼近最大响应区域,再利用Box-Behnken试验设计及响应面分析法得到培养基最佳配方:通过摇瓶单因素试验对发酵条件、温度、接种量、转速进行了优化,优化后的发酵培养基为葡萄糖47.7 g/L、蛋白胨29.0 g/L、K2HPO4 3.3 g/L,摇瓶发酵工艺为接种量30 mL/L、初始pH 7.5、摇床转速220 r/min、培养温度37℃。优化后抗菌肽的效价提高到了4.25×104 U/mL,是优化前的3.51倍。综合试验结果,本研究筛选的培养基和优化后的发酵工艺可达到高产、降低成本、缩短发酵时间的效果。

关键词: 枯草芽孢杆菌; 抗菌肽; 培养基; 发酵工艺; 优化

Abstract:

The aim of this study was to optimize the fermentation medium and fermentation conditions of Bacillus subtilis BL0006 and improve the titer of antimicrobial peptide.Through the single factor experiment,the optimum carbon source,nitrogen source and inorganic salt were selected as glucose,peptone and K2HPO4.The path of steepest ascent was undertaken to approach the optimal region of the three significant factors.Box-Behnken design and response surface analysis were adopted to get the best formula of culture medium that could improve the titer of antimicrobial peptide.The fermentation conditions,temperature,inoculation quantity and rotational speed were optimized by shaking flask single factor test.The optimized fermentation medium was glucose 47.7 g/L,peptone 29.0 g/L,K2HPO4 3.3 g/L.The optimized fermentation process was inoculum size 30 mL/L,pH 7.5,220 r/min and 37℃.Under the optimized conditions,the titer of antimicrobial peptide was 4.25×104 U/mL and was 3.51 times as high as before optimization.In conclusion,the selected medium and the optimized fermentation process could achieve high yield,reduce cost and shorten fermentation time.

Key words: Bacillus subtilis; antibacterial peptide; medium; fermentation process; optimization

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