金属离子对群体感应淬灭菌Serratia sp.Z4中AisZ酶的影响  

Effects of Metal Ions on Quorum-quenching Enzymes AisZ in Serratia sp.Z4

作　　者：赵一竹 刘栋 杨惠婷 李亚茹 葛世玫[1] ZHAO Yizhu;LIU Dong;YANG Huiting;LI Yaru;GE Shimei(College of Life and Environmental Science,Wenzhou University,Wenzhou,China 325035)

机构地区：[1]温州大学生命与环境科学学院,浙江温州325035

出　　处：《温州大学学报（自然科学版）》2023年第2期38-46,共9页Journal of Wenzhou University(Natural Science Edition)

基　　金：浙江省公益项目(LGF20E080022);温州市科研项目(H20210003)。

摘　　要：微生物通过信号分子的传递来进行群体感应交流从而调控多种生理行为活动.群体感应淬灭菌体内具有的群体感应淬灭酶可以降解信号分子,从而破坏群体感应系统,对微生物的多种活动产生影响.前期从群体感应淬灭菌Serratia sp.Z4(沙雷氏属)中克隆到一个群体感淬灭基因aisZ并在大肠杆菌Escherichia coli BL21(DE3)中进行了异源表达.现以aisZ基因为研究对象,以异源表达菌株E.coli BL21(DE3)/AisZ-pET-28a(+)为实验材料,通过在0.1 mmol/L的IPTG诱导菌株表达时,分别添加Zn^(2+)、Ca^(2+)和Co^(2+)等三种金属离子,从SDS-PAGA直接观察蛋白条带、生物传感器平板检测菌株对信号分子的降解、RT-qPCR检测aisZ的表达量三个方面来探究金属离子对淬灭酶AisZ的影响.结果表明,AisZ异源表达菌株在IPTG诱导时,加入0.2 mmol/L的Ca^(2+)或Co^(2+)都会抑制淬灭酶基因aisZ的表达,但Ca^(2+)和Co^(2+)在一定程度上可以提高淬灭酶的降解活性,这可能与淬灭酶的金属活性位点相关,还需要进一步研究.Microorganisms communicate by quorum sensing through signaling molecules to regulate a variety of physiological behavioral activities.The quorum-quenching bacteria harboring quorum quenching enzymes can degrade the signal molecules to disrupt the quorum sensing system and affect a variety of microbial activities.We have cloned a quorum quenching gene aisZ from the quorum quenching strain Serratia sp.Z4 and heterologously expressed it in Escherichia coli BL21(DE3).In this experiment,the aisZ gene was studied and its heterologous expression strain E.coli BL21(DE3)/AisZ-pET-28a(+)was used as the experimental material.The protein bands were directly observed from SDS-PAGA by adding three metal ions,such as Zn,Ca,and Co,respectively when the strain was induced to express AisZ by 0.1 mmol/L IPTG.The effect of metal ions on AisZ enzyme was investigated by direct observation of protein bands by SDS-PAGA,degradation of signal molecules by biosensor plates,and expression of aisZ by RT-qPCR.The results showed that the addition of 0.2 mmol/L Ca^(2+)or Co^(2+)to the AisZ heterologous expression strain during IPTG induction inhibited the expression of the quenchase gene but Ca^(2+)and Co^(2+)could enhance the degradation activity of quenchase to some extent,which might be related to the metal active site of quenchase and needs to be further investigated.

关 键 词：群体淬灭酶 蛋白诱导 C6-HSL 金属离子 RT-PCR 

分 类 号：Q935[生物学—微生物学]