A comparative study on kinetics and substrate specificities of Phospholipase A1with Thermomyces lanuginosus lipase.
The mechanism of lipase binding to the lipid-water interface is crucial for substrate specificity and kinetic properties. In this study, the chain-length specificity, regiospecificity and substrate specificity of Phospholipase A1(PLA1) and its parent enzyme Thermomyces lanuginosus lipase (TLL) have been investigated using a classical emulsion system. The results show that both PLA1and TLL are 1,3-regioselective lipases. Additionally, the hydrolytic activity of PLA1is comparatively lower on short-chain triacylglyceride (TAG) and higher on phosphatidylcholine (PC) than the hydrolytic activity of TLL. Further, the results obtained with monolayer film techniques demonstrate that the C-terminal region regulates the binding of PLA1to PC. A hypothesis is presented according to which the α9 helix of C-terminal region in PLA1not only controls the opening of lid but also serves as a membrane anchor that assists in binding to PC. These findings bring new insight into rational design of novel lipases with intriguing functionalities.
College of Food Sciences and Engineering, South China University of Technology, Guangzhou 510640, PR China.
Ruipu
R
Xin
School of Chemistry and Chemical Engineering, Henan University of Technology, Zhengzhou 450001, PR China.
Faez Iqbal
FI
Khan
School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, PR China.
Zexin
Z
Zhao
College of Food Sciences and Engineering, South China University of Technology, Guangzhou 510640, PR China.
Zedong
Z
Zhang
School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, PR China.
Bo
B
Yang
College of Food Sciences and Engineering, South China University of Technology, Guangzhou 510640, PR China. Electronic address: yonghw@scut.edu.cn.
Yonghua
Y
Wang
D000595:Amino Acid Sequence; D004655:Emulsions; D057075:Enzyme Assays; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry* / Q000378:metabolism; D054466:Phospholipases A1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000072756:Protein Conformation, alpha-Helical; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004718:Saccharomycetales / Q000737:chemistry / Q000201:enzymology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013237:Stereoisomerism; D013379:Substrate Specificity; D014280:Triglycerides / Q000737:chemistry* / Q000378:metabolism
D004655:Emulsions; D005656:Fungal Proteins; D010713:Phosphatidylcholines; D011994:Recombinant Proteins; D014280:Triglycerides; D008049:Lipase; D054466:Phospholipases A1
Chain-length specificity; Interfacial binding; Lipase; Monolayer; Phospholipase A(1); Regiospecificity; Substrate specificity