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【目的】N-乙酰神经氨酸有多种生物学功能,其在治疗流感、神经性疾病、炎症和肿瘤等方面具有重要的医药价值。N-乙酰神经氨酸现有的生产方法产量低、成本高,难以满足医药工业大规模的需求,因而急需建立一种经济高效的生产方法。【方法】在前期研究中,构建了一株产N-乙酰神经氨酸的代谢工程菌(Escherichia coliΔnanTEK/pNA),本研究通过单因素试验对工程菌生物转化生产N-乙酰神经氨酸的过程进行优化,包括工程菌细胞培养条件(培养温度和时间)和全细胞生物转化的各种条件(转化温度、时间、表面活性剂等)。【结果】经过条件优化后,建立了全细胞催化法生产N-乙酰神经氨酸的工艺流程,使N-乙酰神经氨酸的产量提高到294.39 mmol/L。【结论】该方法具有操作简便、高效和经济的优势,为N-乙酰神经氨酸的规模化生产奠定了基础。
【Objective】 N-acetylneuraminic acid has a variety of biological functions and has important medical value in the treatment of influenza, neurological diseases, inflammation and cancer. The current production methods of N-acetylneuraminic acid are low in output and high in cost, which is difficult to meet the large-scale demand of the pharmaceutical industry. Therefore, an economical and efficient production method is urgently needed. 【Method】 In the previous study, a strain of Escherichia coliΔnanTEK / pNA producing N-acetylneuraminic acid was constructed. In this study, the process of bioconversion of engineering bacteria to N-acetylneuraminic acid Optimization was made to include various conditions (transformation temperature, time, surfactant, etc.) for engineering bacterial cell culture conditions (incubation temperature and time) and whole cell biotransformation. 【Result】 After optimization of the conditions, a whole-cell catalytic process for the production of N-acetylneuraminic acid was established, raising the yield of N-acetylneuraminic acid to 294.39 mmol / L. 【Conclusion】 The method is simple, efficient and economical. It laid the foundation for the large-scale production of N-acetylneuraminic acid.