重组大肠杆菌全细胞催化法生产N-乙酰神经氨酸

doi:10.12441/spyswjs.20220217002

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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
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                        10.12441/spyswjs.20220217002
                    
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中图分类号:Q 819
基金项目:国家自然科学基金项目（32001064）

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

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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 OD₆₀₀ 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.

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郑蝶,杨海泉,夏媛媛,沈微,陈献忠.重组大肠杆菌全细胞催化法生产N-乙酰神经氨酸[J].食品与生物技术学报,2024,43(2):38-46.

ZHENG Die, YANG Hai-quan, XIA Yuan-yuan, SHEN Wei, CHEN Xian-zhong. Production of N-Acetylneuraminic Acid from Fermentation Broth Using Recombinant Escherichia coli Whole Cells Biotransformation[J]. Journal of Food Science and Biotechnology,2024,43(2):38-46.

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在线发布日期: 2024-03-15
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