食品科学 ›› 2009, Vol. 30 ›› Issue (7): 181-185.doi: 10.7506/spkx1002-6630-200907041

• 生物工程 • 上一篇    下一篇

苯丙氨酸解氨酶重组大肠杆菌产酶条件研究

崔建东,张云峰,李 艳   

  1. 河北省发酵工程研究中心,河北科技大学生物科学与工程学院
  • 收稿日期:2008-06-09 修回日期:2008-10-30 出版日期:2009-04-01 发布日期:2010-12-29
  • 通讯作者: 崔建东 E-mail:cjd007cn@163.com
  • 基金资助:

    河北科技大学自然科学基金项目(XL200763)

Optimization of Production Conditions of Phenylalanine Ammonia-Lyase by Recombinant E.coli

CUI Jian-dong ZHANG Yun- feng LI Yan   

  1. (Hebei Fermentation Engineering Research Center, College of Bioscience and Bioengineering, Hebei University of Science and
    Technology, Shijiazhuang 050018, China)
  • Received:2008-06-09 Revised:2008-10-30 Online:2009-04-01 Published:2010-12-29
  • Contact: CUI Jian-dong E-mail:cjd007cn@163.com

摘要:

为了提高重组苯丙氨酸解氨酶(PAL)活力,研究培养基初始pH值、种龄、接种量、装液量和发酵时间对重组PAL活力的影响,并用响应面分析法优化了培养条件。单因素最适培养条件为:发酵培养基初始pH7.0、种龄10h、接种量10%、装液量50ml、发酵时间24h。响应面优化后的结果表明,种龄、接种量和发酵时间对重组PAL活力影响较大,但三者之间没有显著的交互作用。优化后的产酶条件是:种龄13.3h、接种量10.7%、发酵时间25.5h、在此条件下,PAL活力比不优化的培养条件的酶活增加了15%左右,重组PAL酶活达到14.95 U/ml。

关键词: 苯丙氨酸解氨酶, 重组大肠杆菌, 培养条件, 响应面分析

Abstract:

Effects of initial pH value of fermentation medium, inoculation time, inoculation volume, broth volume in 250-ml flask and fermentation time on the activity of phenylalanine ammonia-lyase (PAL) produced by recombinant E. coli JM109 were investigated through single-factor test, and then the main affecting factors confirmed in single-factor test were optimized through response surface analysis (RSA). The results showed that initial pH value of fermentation medium and broth volume in 250-ml flask are not main affecting factors, and they can be controlled at 6.5 to 7 and 50 ml, respectively; inoculation time, inoculation volume and fermentation time are main affecting factors, and their optimal levels are 13.3 h, 10.7 % and 25.5 h, respectively. By this optimization, the activity of PAL is increased by about 15%, reaching 14.95 U/ml.

Key words: phenylalanine ammonia-lyase, recombinant E. coli, culture conditions, response surface analysis

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