Application of Auto-induction System in the Synthesis of 2’-deoxycytidine

Abstract

Abstract: This experiment was carried out in order to synthesize 2’-deoxycytidine（dCyd）more efficiently. DCyd is a significant intermediate for many antiviral and anticancer drugs such as Gemcitabine（dFdC）, Dideoxycytidine（ddC）, etc. A convenient and efficient bioprocess was reported here to synthesize dCyd by E.coli BL21（DE3）containing gene of N-deoxyribosyltransferaseⅡ（NDT）over-expressed using ZYM-Fe-5052 auto-induction system. The results showed that auto-induction system could make the addition of inducer during cultivation unnecessary and obtain a higher cell density. The recombinant cells from auto-induction system were as efficient as that from LB system on the conversion yields of dCyd. When the ratio of 2’-deoxythymidine（dThd）and cytosine was 1∶3, the conversion yield could be 86.5%. The substrates ratio of 2’-deoxythymidine (dThd) and cytosine as 1∶1.5 and 1 mg/mL recombinant cells which were dissolved in phosphate buffer of pH6.5, then cultured at 30℃ for 1 h were selected as the suitable reaction conditions,and the conversion yield could be 71.9%. This method also has potential for the preparation of other nucleoside analogues.

Key words: Biocatalysis , N-deoxyribosyltransferase , 2’-deoxycytidine , Auto-induction system, Nucleoside analogues

Wang Xi,Duan Shenglin,Xiong Shuli,Zheng Guilan,Zhang Guiyou,Wang Hongzhong. Application of Auto-induction System in the Synthesis of 2’-deoxycytidine[J]. Biotechnology Bulletin, 2014, 0(11): 225-232.

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