[Objective] N-acetylneuraminic acid (Neu5Ac) has important medical value in the treatment of influenza,the neurological disease.The conventional methods of production have some shortcomings,such as high cost and low yield.So the available resource of Neu5Ac can not meet the large-scale demand of medical industry.[Methods] In earlier research,a metabolic engineering bacteria (Escherichia coli ΔnanTEK/pNA) has been constructed for the production of Neu5Ac,Whole-cell catalysis was used for the production of Neu5Ac with GlcNAc and pyruvate as the substrate.Culture condition (temperature and time) ofE.coli ΔnanTEK/pNA and subsequent biological transformation (temperature,time,Triton X-100 and so on) were studied by single factor experiment.[Results] After conditions optimization,the production of Neu5Ac was improved and 294.39 mmol/L of Neu5Ac was obtained.[Conclusion] The process can be used for industrial large-scale production of Neu5Ac in terms of convenience,efficiency and economy.%[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 coli ΔnanTEK/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.
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