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Open AccessArticle

A Thermolabile Phospholipase B from Talaromyces marneffei GD-0079: Biochemical Characterization and Structure Dynamics Study

1
School of Biology and Biological Engineering South China University of Technology 382 East Outer Loop Rd, University Park, Guangzhou 510006, China
2
School of Electronic Science and Engineering, University of Electronic Science and Technology of China, Chengdu 610054, China
3
School of Food Science and Engineering, Guangdong Research Center of Lipid Science and Applied Engineering Technology, State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou 510641, China
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Biomolecules 2020, 10(2), 231; https://doi.org/10.3390/biom10020231
Received: 2 December 2019 / Revised: 20 January 2020 / Accepted: 28 January 2020 / Published: 4 February 2020
(This article belongs to the Section Enzymology)
Phospholipase B (EC 3.1.1.5) are a distinctive group of enzymes that catalyzes the hydrolysis of fatty acids esterified at the sn-1 and sn-2 positions forming free fatty acids and lysophospholipids. The structural information and catalytic mechanism of phospholipase B are still not clear. Herein, we reported a putative phospholipase B (TmPLB1) from Talaromyces marneffei GD-0079 synthesized by genome mining library. The gene (TmPlb1) was expressed and the TmPLB1 was purified using E. coli shuffle T7 expression system. The putative TmPLB1 was purified by affinity chromatography with a yield of 13.5%. The TmPLB1 showed optimum activity at 35 °C and pH 7.0. The TmPLB1 showed enzymatic activity using Lecithin (soybean > 98% pure), and the hydrolysis of TmPLB1 by 31P NMR showed phosphatidylcholine (PC) as a major phospholipid along with lyso-phospholipids (1-LPC and 2-LPC) and some minor phospholipids. The molecular modeling studies indicate that its active site pocket contains Ser125, Asp183 and His215 as the catalytic triad. The structure dynamics and simulations results explained the conformational changes associated with different environmental conditions. This is the first report on biochemical characterization and structure dynamics of TmPLB1 enzyme. The present study could be helpful to utilize TmPLB1 in food industry for the determination of food components containing phosphorus. Additionally, such enzyme could also be useful in Industry for the modifications of phospholipids. View Full-Text
Keywords: free fatty acids (FFAs); NMR (nuclear magnetic resonance); phospholipase B; Talaromyces marneffei; affinity chromatography free fatty acids (FFAs); NMR (nuclear magnetic resonance); phospholipase B; Talaromyces marneffei; affinity chromatography
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MDPI and ACS Style

Durrani, R.; Khan, F.I.; Ali, S.; Wang, Y.; Yang, B. A Thermolabile Phospholipase B from Talaromyces marneffei GD-0079: Biochemical Characterization and Structure Dynamics Study. Biomolecules 2020, 10, 231.

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