荧光假单胞杆菌磷脂酶B界面催化分子机制初探

doi:10.11949/0438-1157.20200527

化工学报 ›› 2020, Vol. 71 ›› Issue (9): 4255-4259.DOI: 10.11949/0438-1157.20200527

荧光假单胞杆菌磷脂酶B界面催化分子机制初探

赵依威(),姜芳燕,李春,戴大章()

北京理工大学化学与化工学院，北京 100081

收稿日期:2020-05-08 修回日期:2020-07-18 出版日期:2020-09-05 发布日期:2020-09-05
通讯作者: 戴大章
作者简介:赵依威（1995—），男，硕士研究生，zhaoyiwei7139@163.com
基金资助:
国家自然科学基金项目(21276025)

Interfacial catalytic mechanism of Pseudomonas fluorescens phospholipase B

Yiwei ZHAO(),Fangyan JIANG,Chun LI,Dazhang DAI()

School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 100081, China

Received:2020-05-08 Revised:2020-07-18 Online:2020-09-05 Published:2020-09-05
Contact: Dazhang DAI

摘要/Abstract

摘要：

基于荧光假单胞杆菌磷脂酶B（Pf-PLB）一级序列，分析了Pf-PLB的相关理化性质及其二级结构，并采用同源建模的方法对Pf-PLB的三级结构进行了预测；基于其预测结构，提出“掀盖进入，侧向离去”假说模型，初步探讨了Pf-PLB界面催化的可能机制；采用定点突变的方法对Pf-PLB的结构进行改造，结合突变体Pf-PLB的催化特性对所提出的机制模型进行了验证。

关键词: 磷脂酶B, 结构预测, 催化机制

Abstract:

The physicochemical property and secondary structure of Pseudomonas fluorescens phospholipase B（Pf-PLB） were analyzed base on its primary structure, and the tertiary structure of Pf-PLB was predicated. A “lid-in, side-out” hypothesis model was put forward to illuminate the possible interfacial catalytic mechanism of Pf-PLB. The structure of Pf-PLB was modified by site-directed mutagenesis, and the proposed mechanism model was verified by combining with the catalytic properties of mutant Pf-PLB.

Key words: phospholipase B, structure prediction, catalytic mechanism

中图分类号:

TQ 028.8

赵依威, 姜芳燕, 李春, 戴大章. 荧光假单胞杆菌磷脂酶B界面催化分子机制初探[J]. 化工学报, 2020, 71(9): 4255-4259.

Yiwei ZHAO, Fangyan JIANG, Chun LI, Dazhang DAI. Interfacial catalytic mechanism of Pseudomonas fluorescens phospholipase B[J]. CIESC Journal, 2020, 71(9): 4255-4259.

图/表 7

图1 PL作用于磷脂分子位点示意图（X代表氢、胆碱、乙醇胺、丝氨酸、肌醇等）

Fig.1 The diagram of phospholipases acting on sites of the phosphatidylcholine

图2 ProtScale对Pf-PLB的疏水性的分析（正值为疏水，负值为亲水）

Fig.2 Analysis of the hydropathicity of Pf-PLB by ProtScale

图3 SOMPA对Pf-PLB进行二级结构预测

Fig.3 Prediction of secondary structure of Pf-PLB by SOMPA

图4 Pf-PLB与来源于E. coli和P. aeruginosa的FadL结构比较

Fig.4 Structure comparison between Pf-PLB and FadL from E. coli and P. aeruginosa

图5 Pf-PLB界面催化的可能机制L—低亲和力结合位点； H—高亲和力结合位点； N—N端结构域

Fig.5 Possible mechanism of Pf-PLB interfacial catalysis

图6 不同菌株所产Pf-PLB活性分析

Fig.6 Activity analysis of Pf-PLB from different strains

图7 Pf-PLB的Modeller模拟

Fig.7 Modeller simulation of Pf-PLB

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荧光假单胞杆菌磷脂酶B界面催化分子机制初探

Interfacial catalytic mechanism of Pseudomonas fluorescens phospholipase B

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