Ishida M, Sato K, Hashiguchi K, Ito H, Enei H, Nakamori S
Central Research Laboratories of Ajinomoto Co., Inc., Kanagawa, Japan.
Biosci Biotechnol Biochem. 1993 Oct;57(10):1755-6. doi: 10.1271/bbb.57.1755.
Decrease in L-threonine productivity caused not only by plasmid-free segregation but also by plasmid deletion was observed in a Brevibacterium flavum L-threonine producer when transformed with a recombinant plasmid carrying Escherichia coli thr operon. However, a recombinant strain, HT-16, was stabilized by the addition of trimethoprim (a selective marker on the vector) at concentration of 1000 micrograms/ml, which was rather higher than the minimum inhibitory one, into the stock culture medium. Strain HT-16 produced 64.4 g/liter of L-threonine, 30% higher than that of the host strain, after 92 h of cultivation in a small jar fermentor using acetic acid as a carbon source without trimethoprim.
在用携带大肠杆菌苏氨酸操纵子的重组质粒转化的黄色短杆菌L-苏氨酸生产菌中,观察到L-苏氨酸产量的下降不仅是由于无质粒分离,还由于质粒缺失。然而,通过在保藏培养基中添加浓度为1000微克/毫升的甲氧苄啶(载体上的一个选择标记,该浓度远高于最小抑菌浓度),重组菌株HT-16得以稳定。在使用醋酸作为碳源且无甲氧苄啶的小型罐式发酵罐中培养92小时后,菌株HT-16产生了64.4克/升的L-苏氨酸,比宿主菌株高30%。