Cytidine triphosphate(CTP)is synthesized through a one-pot whole-cell catalysis pathway with cytidine as substrate.The co-expression and fusion expression systems of cytidine kinase(CDK)and acetate kinase(ACK),and the fusion expression system of cytidylate kinase(CMK)and ACK are respectively constructed in Escherichia coli BL21(DE3),in which the dual fusion catalytic system composed of SUMO-CDK-L2-ACK fusion strain and CMK-L2-ACK fusion strain is found to have better catalytic activity.A dual fusion strains whole-cell one-pot catalysis reaction system is established for the synthesis of CTP.The system is optimized in terms of temperature,pH,metal ions,acetyl phosphate dosage and bacterial powder dosage.It is indicated that in the optimized reaction system,the highest CTP yield reaches 108 mmol·L-1 in 75 min,and the total yield reaches 86.4％,which realizes the efficient one-pot synthesis of CTP.
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