4-丁内酰胺水解酶法制备γ-氨基丁酸

编辑 2019-05-28
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附加信息

  • DOI(*): 10.3969/j.issn.1006-8376.2011.01.010
  • 酶(*): 丁内酰胺水解酶
  • pH(*): 7.0
  • 温度(*): 40°C
  • 介质(*): 磷酸钠缓冲液
  • 催化剂类型(*): 发酵液
  • 酶物种来源或野生菌(*): 菌株HHSW -16
  • 产率(*): 99%
  • ee%(*):

目前,γ-氨基丁酸的生产主要采用化学合成法和发酵法,但均存在反应产物很多不利于分离,得率低的问题,从而不益用于食品和药物。故本文以2-吡咯烷酮为唯一碳源,筛选得到了一株具有较高4 -丁内酰胺水解酶的菌株,水解2-吡咯烷酮得到γ -氨基丁酸。然后分别对菌株的产酶培养条件和转化反应的条件进行研究优化,得到了最佳的培养和转化条件。结果表明: 编号为HHSW - 16 的菌株水解活性最高。优化后的培养基组成为: 葡萄糖11.50 g·L-1,牛肉膏6.35 g·L-1,酵母粉5.58 g·L-1,Na2HPO4·12H2O 5.0g·L-1,KH2PO4 1.5 g·L-1,MgSO4·7H2O 1.0 g·L -1,CaCl2·2H2O 10 mg·L -1,FeSO4·7H2O 8 mg·L-1,pH7.0。转化反应的最佳条件为: 温度40℃,pH 值7.0。此条件下,10 g·L -1底物加入1%湿菌体,酶促反应6 h 转化率达到99%以上。

1、不同菌株培养后的转化率和酶活力:

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2、碳源对转化率和酶活力的影响:

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3、氮源对转化率和酶活力的影响:

 9SP@{K`$J$7@WEQ`M7X8(4F.png

4、各因素质量浓度对酶活力的影响:

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5、初始pH 值对酶活力的影响:

 _V%WJO(0W@KYEM4)5E2@IN1.png

6、温度对转化反应的影响:

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7、pH 值对转化反应的影响:

 CH4B9$JOPI_X@KQ)S_ZCXN8.png

8、优化条件下的验证试验:

 O053PVY`6L2BE[H5PL}MB13.png

 

 

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