青霉素酰化酶产生菌的分子育种及其产酶条件的研究

STUDY ON MOLECULAR BREEDING AND CULTURE CONDITIONS FOR HIGH PRODUCTION OF PEN1CILLIN-G-ACYLASE

用EcoR1分别酶切质粒pACYC184和Ec108染色体,经连接、转化、筛选,得到7株克隆了青霉素酰化酶基因的转化体-C600(pPAHD1-7)。重组质粒pPAHD1在不同受体细胞内的酶活表达水平相差20倍。据此构建的工程菌其青霉素酰化酶活比出发菌株提高4～5倍。本文还对工程菌的产酶条件进行了研究。

Plasmid pACYCl84 DNA and EC 108 chromosomal DNA were digested with restriction enzyme EcoRI respectivelv. Seven transformants containing penicillin-G-acylase gene were isolatated after ligation and transformation. The expression of the recombined plasmid discriminated in different recipient cells showing a 20 fold enzyme activity variation in the extreme.Under the appropriate circumstances the penicillin-G-acylase activity of the transformed Strain <Ecl08/pPAHD1) increased 4~ 5 fold in comparison with the original strain Ec108. With the specific characteristics of the fast growing rate, high level of penicillin-G-acylase productivity and low cost of the culture media, the transformed strain is considered to be potentially significant for pharmacutical industry.