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

作　　者：郑蝶 杨海泉[1,2] 夏媛媛 沈微[1,2] 陈献忠[1,2] ZHENG Die;YANG Haiquan;XIA Yuanyuan;SHEN Wei;CHEN Xianzhong(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与生物技术学报》2024年第2期38-46,共9页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(32001064)。

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OD_(600)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。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 OD_(600)30,the highest yield of N-acetylneuraminic acid reached 180 mmol/L,and the molar conversion rate was 65.45%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 was 42.5%.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.

关 键 词：N-乙酰神经氨酸 发酵液 全细胞催化 

分 类 号：Q819[生物学—生物工程]