Martín L, Prieto M A, Cortés E, García J L
Department of Molecular Microbiology, CSIC, Madrid, Spain.
FEMS Microbiol Lett. 1995 Jan 15;125(2-3):287-92. doi: 10.1016/0378-1097(94)00510-x.
The pac gene encoding the penicillin G acylase (PGA) of Bacillus megaterium ATCC 14945 has been cloned in Escherichia coli HB101 (proA, leuB) using a selective minimal medium containing phenylacetyl-L-leucine instead of L-leucine. The nucleotide sequence of this gene has been determined and contains an open reading frame of 2406 nucleotides. The deduced amino acid sequence shows significant similarity with other beta-lactam acylases. Although the PGA of B. megaterium is extracellular, the enzyme produced in E. coli appears to have a cytoplasmic localization.
利用含有苯乙酰-L-亮氨酸而非L-亮氨酸的选择性基本培养基，已将编码巨大芽孢杆菌ATCC 14945青霉素G酰化酶（PGA）的pac基因克隆到大肠杆菌HB101（proA，leuB）中。已确定该基因的核苷酸序列，其包含一个2406个核苷酸的开放阅读框。推导的氨基酸序列与其他β-内酰胺酰化酶显示出显著的相似性。尽管巨大芽孢杆菌的PGA是胞外酶，但在大肠杆菌中产生的该酶似乎定位于细胞质中。
