青霉素酶基因异源表达及该酶分解牛奶中残留青霉素的研究  

Study on Heterologous Expression of Penicillinase Gene and the Penicillinase Degrading Residual Penicillin in Milk

作　　者：赵洪坤[1] 杜连祥[1] 李玉[1] 王晓娟[1] 路福平[1] 

机构地区：[1]天津科技大学生物工程学院天津市工业微生物重点实验室,天津300457

出　　处：《微生物学通报》2008年第6期893-897,共5页Microbiology China

摘　　要：以获得大量青霉素酶并将其用于分解牛奶中残留青霉素为目的,通过PCR方法从蜡样芽孢杆菌ATCC10987基因组中获得了青霉素酶基因,将该基因克隆至表达载体pET28a(+)中,并转化到E.coli BL21中;在IPTG诱导下对目的蛋白进行SDS-PAGE和酶活分析,结果显示最大酶活力可达到480.0 U/mL;利用Ni2+亲合层析柱纯化目的蛋白,纯化后的目的蛋白纯度超过90%;采用高碘酸钠氧化法制备固定化的青霉素酶,并利用该固定化酶将牛奶(含0.5U青霉素G/mL)中的青霉素分解到浓度小于4ppb程度。To obtain a number of penicillinases and degrade penicillin in milk by using the penicillinases, the gene encoding penicillinase was amplified by PCR from Bacillus cereus ATCC10987, cloned into pET28a（＋）, transformed into E, coli BL21; analysis of SDS-PAGE and penicillinase activity of the recombinant protein were done under induction of IPTG and the result showed that the maximum penicillinase activity reached 480 U/mL; the purity of penicillinase purified by Ni^2＋ Purification System was more than 90%; the immobilized penicillinases were obtained by sodium periodate method and the residual quantity of penicillin in milk（containing 0.5 U penicillin G/mL） was less than 4 ppb after degraded by the immobilized penicillinase.

关 键 词：青霉素酶 异源表达 纯化 固定化青霉素酶 

分 类 号：S859.84[农业科学—临床兽医学]