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营养物及补料方式对重组酵母产α-淀粉酶的影响

Effects of Nutrition and it’s Feeding Strategy on the Expression of α-amylase in Fedbatch Cultivation of Recombinant Yeast

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【作者】 张小里夏诏杰刘颖赵彬侠郑晓钢熊朝晖

【Author】 ZHANG Xiao-li, XIA Zhao-jie, LIU Ying, ZHAO Bin-xia, ZHENG Xiao-gang, XIONG Zhao-hui (Department of Chemical Engineering, Northwest University, Xi’an 710069, China)

【机构】 西北大学化工系! 陕西 西安 710069

【摘要】 研究了两段培养中基质浓度、配比及天然氮源添加方式对重组酵母发酵生产(-淀粉酶的影响。结果表明,重组酵母发酵适宜的初糖浓度为7g·L-1,葡萄糖和基础氮配比为7:4,生长期补料碳氮比为5:1,且产酶期供给天然氮源以少量多次脉冲补加酵母膏为宜。于2L发酵罐中培养,证明脉冲补加酵母膏、流动添加基本营养物的补料分批培养方式是有效的,指数流加基本营养物较之恒流量流加更能提高菌体浓度和(-淀粉酶表达水平。培养60小时细胞量达24.5g·L-1,酶活力达97.3U·mL-1。

【Abstract】 The effects of initial sugar concentration、ratio of glucose and yeast nitrogen base (without amino acid) and the different natural nitrogen feeding strategy in the expression stage on the production of (-amylase by recombinant yeast in shaking flask two-stage culture were investigated. The results showed that the proper initial sugar concentration was 7g·L-1, ratio of glucose and yeast nitrogen base was 7:4, ratio of carbon and nitrogen of the feeding nutrition was 5:1 respectively. The suitable feeding strategy of natural nitrogen source in the expression stage is " pulse feeding in a small amount of it". By the cultivation in 2L fermentor, it was confirmed that feeding basic nutrition and pulse feeding yeast extract were effective feeding strategy .The pulse feeding based on exponential feeding principle has more advantages than constant feeding in both enhancing cell mass and (-amylase productivity. The maximum cell density and (-amylase activity reached 24.5g·L-1 at 27h and 97.3U·mL-1 at 57h, respectively.

【基金】 国家自然科学基金资助项目!(编号CG983344)
  • 【文献出处】 高校化学工程学报 ,Journal of Chemical Engineering of Chinese Universities , 编辑部邮箱 ,2001年02期
  • 【分类号】TQ925
  • 【被引频次】10
  • 【下载频次】165
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