食品科学 ›› 2018, Vol. 39 ›› Issue (4): 90-98.doi: 10.7506/spkx1002-6630-201804014

• 生物工程 • 上一篇    下一篇

双向单因素与田口法优化屎肠球菌产谷氨酸脱羧酶培养基

杨胜远1,李云2   

  1. (1.岭南师范学院化学化工学院,热带与南海资源协同创新中心,广东湛江 524048;2.韩山师范学院生命科学与食品科技学院,广东 潮州 521041)
  • 出版日期:2018-02-25 发布日期:2018-02-02
  • 基金资助:
    广东省自然科学基金项目(2014A030307039);岭南师范学院科研专项(ZL1602)

Optimization of Fermentation Medium for Glutamate Decarboxylase Production by Enterococcus faecium by Bidirectional One-Factor-at-a-Time and Taguchi Methods

YANG Shengyuan1, LI Yun2   

  1. (1. Tropical South China Sea’s Resources Collaborative Innovation Centre, College of Chemistry and Chemical Engineering, Lingnan Normal University, Zhanjiang 524048, China; 2. School of Life Sciences and Food Technology, Hanshan Normal University, Chaozhou 521041, China)
  • Online:2018-02-25 Published:2018-02-02

摘要: 采用双向单因素试验法及田口法,对屎肠球菌产谷氨酸脱羧酶(glutamate decarboxylase,GAD)的培养基进行了优化。结果表明,在筛选的4 种培养基中,蛋白胨-蔗糖-牛肉膏(peptone-sucrose-beef extract,PSB)培养基最利于屎肠球菌产GAD,但其配方中的CaCl2、K2HPO4和柠檬酸铵对GAD合成不利,蛋白胨、蔗糖、乙酸钠和谷氨酸一钠(monosodium glutamate,MSG)对GAD合成影响极显著(P<0.01),而牛肉膏对GAD合成影响不显著(P>0.10)。Minitab软件预测培养基的最优用量组合为蛋白胨15g/L、牛肉膏10g/L、蔗糖12.5g/L、乙酸钠6.0g/L、MSG10g/L、吐温801.0g/L,预测300mL发酵醪产GAD总活力为(399.30±12.51)U。经验证,GAD总活力为(384.26±10.32)U,与预测值没有显著性差异(P>0.05)。改良的PSB培养基的组分较优化前的PSB培养基显著减少,并更有利于GAD的合成,总酶活提高了45.71%。

关键词: 屎肠球菌, 谷氨酸脱羧酶, 培养基, 优化, 双向单因素法, 田口法

Abstract: The fermentation medium for glutamate decarboxylase (GAD) production by Enterococcus faecium was optimized by bidirectional one-factor-at-a-time and Taguchi methods. The results indicated that the peptone-sucrose-beef extract (PSB) medium was found to be the most suitable for GAD production among four tested media. However, CaCl2, K2HPO4 and ammonium citrate in the PSB medium were unfavourable for GAD production. Peptone, sucrose, sodium acetate and monosodium glutamate (MSG) had significant effects on GAD production (P < 0.01), which was not significantly affected by beef extract (P > 0.10). The optimal culture medium predicted by Minitab software consisted of 15 g/L peptone, 10 g/L beef extract, 12.5 g/L sucrose, 6.0 g/L sodium acetate, 10 g/L MSG, and 1.0 g/L Tween 80. The predicted total GAD activity obtained by using the optimized medium was (399.30 ± 12.51) U, which was not significantly different from the experimental value ((384.26 ± 10.32) U). The constituents of the modified PSB medium were much fewer compared with the initial one. However, the modified PSB medium was more suitable for GAD production, in which the total GAD activity produced by E. faecium was improved by 45.71% as compared to the initial one.

Key words: Enterococcus faecium, glutamate decarboxylase, medium, optimization, bidirectional one-factor-at-a-time method, Taguchi method

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