摘要
将甘草酸生物转化合成GAMG已成为国内外医药和食品行业研究的热点。该文在前期构建毕赤酵母(pichia pastoris)工程菌GS115-GUS的基础上,对重组毕赤酵母工程菌发酵条件进行了研究。通过单因素及正交试验建立工程菌最佳表达β-葡萄糖醛酸苷酶的诱导产酶方法:重组毕赤酵母GS115-GUS菌株在液体YPD培养基中活化24 h后,接种到BMGY中富集菌体,OD600达到6.0时,按照1∶1(V/V)的接种比率接入到pH为6.0的BMMY培养基中,加入1.5%诱导剂甲醇诱导产酶,且每隔24 h添加一次。该诱导产酶条件简单、易行、科学、合理,毕赤酵母高效表达GAMG的酶活高达1.97×103 U/mL,研究结果达到预期目标,为进一步研究重组酵母工程菌产β-葡萄糖醛酸苷酶的发酵水平、β-葡萄糖醛酸苷酶生物催化甘草酸制备GAMG以及工业化生产奠定基础。
Direct bio-transformation from GL to GAMG is becoming a hot research spot in pharmaceutical and food industry study at home and abroad. On the basis of the pre-built in recombinant pichia pastoris- GS115-GUS, research has been carried out about the fermentation conditions of recombinant pichia pastoris. By single factor test and orthogonal test on recombinant pichia pastoris, we obtained the best induction method of β-Glucuronidase： recombinant pichia pastoris was cultivated in YPD medium composition for 24 h, according to inovulation percentage 1：1 （V/V） to the BMMY which pH is 6.0, then adding 1.5% methanolt to induction, add it into the broth for each 24 h. The method is simple, easy, scientific and rational. The enzyme activity of high level expression of GAMG by Pichia pastoris is up to 1.97× 10^3 U/mL. The studying results achieved the anticipation target which lay the foundation for further study of fermentation levels of the recombinant pichia pastoris producing β-glucuronidas, direct biotransformation from GL to GAMG with β-glucuronidase and industrial production .
出处
《食品工业》
北大核心
2011年第2期67-70,共4页
The Food Industry
基金
2009年度国家大学生创新性实验计划项目"离子液体中酶法合成GAMG的研究"
教育部科学技术研究重点项目"固定化β-葡萄糖醛酸苷酶在离子液体中催化合成GAMG的研究"(209146)
石河子大学高层次人才科研启动资金专项"酶膜生物反应器制备GAMG的研究"(RCZX200805)资助
关键词
毕赤酵母
摇瓶发酵
发酵条件优化
甲醇诱导
Β-葡萄糖醛酸苷酶
Pichia Pastoris
flask-fermentation
optimization of fermentation conditions
methanol induction
β-Glucuronidase
