Stols L, Kulkarni G, Harris B G, Donnelly M I
Environmental Research Division, Argonne National Laboratory, IL 60439, USA.
Appl Biochem Biotechnol. 1997 Spring;63-65:153-8. doi: 10.1007/BF02920421.
The malic enzyme gene of Ascaris suum, was cloned into the vector pTRC99a in two forms encoding alternative amino-termini. The resulting plasmids, pMEA1 and pMEA2, were introduced into Escherichia coli NZN111, a strain that is unable to grow fermentatively because of inactivation of the genes encoding pyruvate dissimilation. Induction of pMEA1, which encodes the native animoterminus, gave better overexpression of malic enzyme, approx 12-fold compared to uninduced cells. Under the appropriate culture conditions, expression of malic enzyme allowed the fermentative dissimilation of glucose by NZN111. The major fermentation product formed in induced cultures was succinic acid.
猪蛔虫的苹果酸酶基因以两种编码不同氨基末端的形式克隆到载体pTRC99a中。所得质粒pMEA1和pMEA2被导入大肠杆菌NZN111,该菌株由于编码丙酮酸异化的基因失活而无法进行发酵生长。编码天然氨基末端的pMEA1的诱导导致苹果酸酶的过表达效果更好,与未诱导的细胞相比约为12倍。在适当的培养条件下,苹果酸酶的表达使NZN111能够对葡萄糖进行发酵异化。诱导培养物中形成的主要发酵产物是琥珀酸。