Tian J, Song H, Wu B, Wang Z, Li J
Department of Microbiology, China Agricultural University, Beijing 100094.
Wei Sheng Wu Xue Bao. 2000 Feb;40(1):26-31.
Plasmid pJMC2 was constructed by cloning the parDE fragment of RK2 into pTZ18U-PHB which harbored phaCAB from Alcaligenes eutrophus and was transferred into E. coli HMS174 and E. coli JM107 separately. It is very stable in its hosts cultured in medium without ampicillin. E. coli HMS 174(pTZ18U-PHB) and E. coli JM107(pTZ18U-PHB) produced P(3HB-co-3HB) in a low phosphate concentration medium(18 mmol/L). The proportion of 3-hydroxyvalerate(3HV) in the polymer was 5%-8%. A fed-batch culture of E. coli HMS174(pJMC2) was conduct in a 5 L automatically controlled fermentor, the final dry cell weight, P(3HB-co-3HV) content, and th 3HV proportion were 42.5 g/L, 70% and 4.9% respectively.
通过将RK2的parDE片段克隆到携带嗜碱假单胞菌phaCAB的pTZ18U - PHB中构建了质粒pJMC2,并分别将其转入大肠杆菌HMS174和大肠杆菌JM107。在不含氨苄青霉素的培养基中培养时,它在宿主中非常稳定。大肠杆菌HMS 174(pTZ18U - PHB)和大肠杆菌JM107(pTZ18U - PHB)在低磷酸盐浓度培养基(18 mmol/L)中产生聚(3 - 羟基丁酸酯 - 共 - 3 - 羟基戊酸酯)[P(3HB - co - 3HB)]。聚合物中3 - 羟基戊酸酯(3HV)的比例为5% - 8%。在5 L自动控制发酵罐中对大肠杆菌HMS174(pJMC2)进行分批补料培养,最终干细胞重量、P(3HB - co - 3HV)含量和3HV比例分别为42.5 g/L、70%和4.9%。