Recombinant Lipase from Gibberella zeae Exhibits Broad Substrate Specificity: A Comparative Study on Emulsified and Monomolecular Substrate
| Autor: | Bo Yang, Zexin Zhao, Ruixia Wei, Fanghua Wang, Yonghua Wang, Hui Zhang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine substrate specificity Phospholipase 01 natural sciences lcsh:Chemistry chemistry.chemical_compound stereospecificity Enzyme Stability lcsh:QH301-705.5 Spectroscopy Phospholipids Galactolipase activity biology Hydrolysis Temperature Stereoisomerism General Medicine Hydrogen-Ion Concentration Recombinant Proteins Computer Science Applications Gibberella zeae lipase monomolecular film technology molecular docking Molecular Docking Simulation Gibberella zeae Electrophoresis Polyacrylamide Gel Emulsions lipids (amino acids peptides and proteins) Stereochemistry Gibberella Lipolysis Catalysis Article Inorganic Chemistry 03 medical and health sciences Stereospecificity 010608 biotechnology Pressure Diglyceride Physical and Theoretical Chemistry Lipase Molecular Biology Organic Chemistry Substrate (chemistry) biology.organism_classification Kinetics 030104 developmental biology lcsh:Biology (General) lcsh:QD1-999 chemistry biology.protein Biocatalysis Glycolipids |
| Zdroj: | International Journal of Molecular Sciences; Volume 18; Issue 7; Pages: 1535 International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 18, Iss 7, p 1535 (2017) |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms18071535 |
| Popis: | Using the classical emulsified system and the monomolecular film technique, the substrate specificity of recombinant Gibberella zeae lipase (rGZEL) that originates from Gibberella zeae was characterized in detail. Under the emulsified reaction system, both phospholipase and glycolipid hydrolytic activities were observed, except for the predominant lipase activity. The optimum conditions for different activity exhibition were also determined. Compared with its lipase activity, a little higher ratio of glycolipid hydrolytic activity (0.06) than phospholipase activity (0.02) was found. rGZEL preferred medium chain-length triglycerides, while lower activity was found for the longer-chain triglyceride. Using the monomolecular film technique, we found that the preference order of rGZEL to different phospholipids was 1,2-diacyl-sn-glycero-3-phospho-l-serine (PS) > 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (PG) > 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) > l-α-phosphatidylinositol (PI) > cardiolipin (CL) > 3-sn-phosphatidic acid sodium salt (PA) > l-α-phosphatidylethanolamine (PE), while no hydrolytic activity was detected for sphingomyelin (SM). Moreover, rGZEL showed higher galactolipase activity on 1,2-distearoyimonoglactosylglyceride (MGDG). A kinetic study on the stereo- and regioselectivity of rGZEL was also performed by using three pairs of pseudodiglyceride enantiomers (DDGs). rGZEL presented higher preference for distal DDG enantiomers than adjacent ester groups, however, no hydrolytic activity to the sn-2 position of diglyceride analogs was found. Furthermore, rGZEL preferred the R configuration of DDG enantiomers. Molecular docking results were in concordance with in vitro tests. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|