摘要
应用紫外分光光度法、荧光分析法研究了羟丙基-β-环糊精(HP-β-CD)包合青霉素酶后酶活力的变化;荧光分析法检测不同温度下HP-β-CD对青霉素酶内在荧光的影响;将HP-β-CD与青霉素酶进行分子对接,分析青霉素酶与HP-β-CD相互作用的位点及作用方式;测定青霉素酶被HP-β-CD包合后对青霉素钠的分解速率.结果发现:HP-β-CD作用后的青霉素酶复合物的活力提高了2.12倍.荧光光谱研究发现:HP-β-CD对酶蛋白荧光有增敏作用,HP-β-CD包合青霉素酶的包合比为1∶1,包合反应自发进行.同步荧光结果显示:HP-β-CD与青霉素酶复合物同步荧光比青霉素酶强,酶蛋白荧光主要源于酪氨酸(Tyr)残基.分子对接实验结果显示:HP-β-CD与青霉素酶之间共形成了3对氢键,VAL39、ASN180和ASP183参与了氢键的形成.
Uv-vis and fluorescence analysis methods were applied to determine enzyme activity of penicillinase and HP-β-CD-penicillinase complex.Fluorescence analysis was used to study the influence of HP-β-CD on the inner fluorescent of penicillinase at different temperatures.The molecular docking was employed to study the interaction sites and modes between penicillinase and HP-β-CD.By comparing the influences on the hydrolysis rate of penicillin sodium brought by penicillinase and HP-β-CD-penicillinase complex,it could be found that the catalysis rate of HP-β-CD-penicillinase complex increased 2.12 times than penicillinase.The fluorescent spectrometry indicated that HP-β-CD could improve the sensitization of the fluorescence of penicillinase.When the inclusion rate of penicillinase and HP-β-CD was 1 ∶ 1,the inclusion reaction could act spontaneously.The synchronous fluorescence revealed that the complex of HP-β-CD and penicillinase was more intensity than that of penicillinase,and the fluorescence of penicillinase was mainly derived from tyrosine （Tyr） residue.The results of molecular docking showed that there were only three amino acid residues involved in the hydrogen bond formation,which were VAL39,ASN180 and ASP183.
出处
《华中师范大学学报（自然科学版）》
CAS
北大核心
2014年第4期532-537,共6页
Journal of Central China Normal University：Natural Sciences
基金
国家自然科学基金项目(31370379)
