大肠杆菌AJ 2595静息细胞由2',3'-二脱氧尿苷和相应嘌呤碱生产2',3'-二脱氧腺苷和2',3'-二脱氧肌苷。
Production of 2',3'-dideoxyadenosine and 2',3'-dideoxyinosine from 2',3'-dideoxyuridine and the corresponding purine bases by resting cells of Escherichia coli AJ 2595.
作者信息
Shirae H, Kobayashi K, Shiragami H, Irie Y, Yasuda N, Yokozeki K
机构信息
Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Japan.
出版信息
Appl Environ Microbiol. 1989 Feb;55(2):419-24. doi: 10.1128/aem.55.2.419-424.1989.
A novel microbial method for the production of 2',3'-dideoxynucleosides by transdideoxyribosylation has been developed. By screening microorganisms producing 2',3'-dideoxyadenosine (DDA) from 2',3'-dideoxyuridine (DDU) and adenine, Escherichia coli AJ 2595 was selected as the best producer. Optimal pH and temperature for the DDA-producing reaction were ca. 6.5 and 50 degrees C, respectively. Pi seemed to be an essential factor for the reaction, and its optimal concentration was ca. 25 mM. Moreover, polyethylene glycol had a notable effect on DDA production. Under the best conditions established, 52 mM DDA was obtained from 100 mM DDU and 100 mM adenine after 48 h of incubation from resting cells of E. coli AJ 2595. This strain could also produce 2',3'-dideoxynucleosides, such as 2',3'-dideoxyinosine (DDI), 2',3'-dideoxyguanosine, and 2',3'-dideoxythymidine, from DDU and the corresponding bases. In particular, this strain could produce DDI in high yield (ca. 32 mM from 100 mM DDU and 100 mM hypoxanthine) after 24 h of incubation. However, 2',3'-dideoxycytidine was not produced from DDU and cytosine by resting cells of E. coli AJ 2595.
已开发出一种通过转双脱氧核糖基化生产2′,3′-双脱氧核苷的新型微生物方法。通过筛选能从2′,3′-双脱氧尿苷（DDU）和腺嘌呤产生2′,3′-双脱氧腺苷（DDA）的微生物，大肠杆菌AJ 2595被选为最佳生产者。产生DDA反应的最佳pH和温度分别约为6.5和50℃。磷酸似乎是该反应的必需因素，其最佳浓度约为25 mM。此外，聚乙二醇对DDA的产生有显著影响。在确定的最佳条件下，大肠杆菌AJ 2595的静止细胞培养48小时后，从100 mM DDU和100 mM腺嘌呤中获得了52 mM DDA。该菌株还能从DDU和相应碱基产生2′,3′-双脱氧核苷，如2′,3′-双脱氧肌苷（DDI）、2′,3′-双脱氧鸟苷和2′,3′-双脱氧胸苷。特别是，该菌株在培养24小时后能高产率地产生DDI（从100 mM DDU和100 mM次黄嘌呤中约产生32 mM）。然而，大肠杆菌AJ 2595的静止细胞不能从DDU和胞嘧啶产生2′,3′-双脱氧胞苷。
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