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重组大肠杆菌全细胞催化法生产N-乙酰神经氨酸

Production of N-Acetylneuraminic Acid from Fermentation Broth Using Recombinant Escherichia coli Whole Cells Biotransformation

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【作者】 郑蝶杨海泉夏媛媛沈微陈献忠

【Author】 ZHENG Die;YANG Haiquan;XIA Yuanyuan;SHEN Wei;CHEN Xianzhong;Key Laboratory of Industrial Biotechnology,Ministry of Education, Jiangnan University;School of Biotechnology,Jiangnan University;

【通讯作者】 陈献忠;

【机构】 江南大学工业生物技术教育部重点实验室江南大学生物工程学院

【摘要】 N-乙酰神经氨酸具有多种生物学功能,在食品添加剂和药物合成等方面有着重要的应用。为了降低成本,作者利用重组大肠杆菌,并以含有N-乙酰葡萄糖胺的发酵液为底物进行全细胞催化生产N-乙酰神经氨酸。结果表明,在温度为30℃、转化液初始pH为6.5、底物丙酮酸浓度为1.38 mol/L、Triton X-100添加质量分数为0.4%、菌体OD600为30的条件下在5 L发酵罐等条件下进行全细胞转化,可以得到N-乙酰神经氨酸180 mmol/L,摩尔转化率为65.45%。用阴离子树脂对产物进行初步分离纯化,经高效液相色谱分析证实纯化后产品为高纯度N-乙酰神经氨酸,其纯度为98.3%,结晶回收率为42.5%。作者建立并优化了一种以含有N-乙酰葡萄糖胺的发酵液为底物直接进行全细胞催化生产N-乙酰神经氨酸的工艺流程,既节约了成本,又具有可持续发展的优势。

【Abstract】 N-acetylneuraminic acid has various biological functions and has important applications in food additives and pharmaceutical synthesis, etc. In order to reduce its production cost, in this study, N-acetylneuraminic acid was produced by recombinant Escherichia coli whole cells using the fermentation broth containing N-acetylglucosamine as the substrate. The catalytic conditions of N-acetylneuraminic acid production were optimized. The optimum result was achieved at 30 ℃(the reaction temperature), initial pH 6.5, 1.38 mol/L pyruvate, 0.4% Triton X-100 added, and OD60030,the highest yield of N-acetylneuraminic acid reached 180 mmol/L, and the molar conversion rate was65.45% in 5 L fermentor. N-acetylneuraminic acid produced was separated and purified using anion resin. It was confirmed by high performance liquid chromatography that the sample was high purity N-acetylneuraminic acid. Its purity reached 98.3%, and the recovery rate of crystallization was42. 5 %. A method about N-acetylneuraminic acid produced from fermentation broth with N-acetylglucosamine using recombinant E. coli whole cells biotransformation was built and optimized, which not only saves costs but also has the advantage of sustainable development.

【基金】 国家自然科学基金项目(32001064)
  • 【文献出处】 食品与生物技术学报 ,Journal of Food Science and Biotechnology , 编辑部邮箱 ,2024年02期
  • 【分类号】TQ203.2;TQ922
  • 【下载频次】106
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