Li Y, Liu B, Li X, Yang Y, Wang S
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing.
Chin J Biotechnol. 1992;8(2):99-106.
Our work has demonstrated the cloning of propionyl acylase gene and the expression of S. mycarofaciens mutant in S. lividans TK54. In this paper, we report the transformation of pIJM9 recombinant plasmid containing the propionyl acylase gene into spiramycin producer S. spiramyceticus. The results of colony hybridization and Southern hybridization showed that No. 61 transformant harbored the pIJM9 recombinant plasmid. TLC and bioautography showed that the Rf value of one component of the fermentation products of No. 61 transformant was similar to that of propionylspiramycin. The HPLC retention time of the components of the fermentation products of No. 61 transformant and that of propionylspiramycin were also similar. Mass spectrum analysis showed that there was propionylspiramycin II in the fermentation products of No. 61 transformant. According to these results, No. 61 transformant is shown to be a genetic engineered strain producing propionylspiramycin.
我们的工作已证明了丙酰化酶基因的克隆以及弗氏链霉菌突变体在变铅青链霉菌TK54中的表达。在本文中，我们报道了将含有丙酰化酶基因的pIJM9重组质粒转化到螺旋霉素产生菌生二素链霉菌中。菌落杂交和Southern杂交结果表明，61号转化体含有pIJM9重组质粒。薄层层析和生物自显影显示，61号转化体发酵产物的一个组分的比移值与丙酰螺旋霉素的比移值相似。61号转化体发酵产物组分的高效液相色谱保留时间与丙酰螺旋霉素的也相似。质谱分析表明，61号转化体的发酵产物中有丙酰螺旋霉素II。根据这些结果，61号转化体被证明是一株生产丙酰螺旋霉素的基因工程菌株。
