A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics Study
Phospholipase B (EC 3.1.1.5) are a distinctive group of enzymes that catalyzes the hydrolysis of fatty acids esterified at the <i>sn-1</i> and <i>sn-2</i> positions forming free fatty acids and lysophospholipids. The structural information and catalytic mechanism of phospholi...
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MDPI AG
2020-02-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/2/231 |
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| author | Rabia Durrani Faez Iqbal Khan Shahid Ali Yonghua Wang Bo Yang |
| author_facet | Rabia Durrani Faez Iqbal Khan Shahid Ali Yonghua Wang Bo Yang |
| author_sort | Rabia Durrani |
| collection | DOAJ |
| description | Phospholipase B (EC 3.1.1.5) are a distinctive group of enzymes that catalyzes the hydrolysis of fatty acids esterified at the <i>sn-1</i> and <i>sn-2</i> 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 <i>Talaromyces marneffei</i> GD-0079 synthesized by genome mining library. The gene (TmPlb1) was expressed and the TmPLB1 was purified using <i>E. coli</i> 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 <sup>31</sup>P 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. |
| first_indexed | 2024-12-13T07:33:19Z |
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| last_indexed | 2024-12-13T07:33:19Z |
| publishDate | 2020-02-01 |
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| series | Biomolecules |
| spelling | doaj.art-f20f23584d234e0e83f76b07f90e99f62022-12-21T23:55:09ZengMDPI AGBiomolecules2218-273X2020-02-0110223110.3390/biom10020231biom10020231A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics StudyRabia Durrani0Faez Iqbal Khan1Shahid Ali2Yonghua Wang3Bo Yang4School of Biology and Biological Engineering South China University of Technology 382 East Outer Loop Rd, University Park, Guangzhou 510006, ChinaSchool of Electronic Science and Engineering, University of Electronic Science and Technology of China, Chengdu 610054, ChinaSchool 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, ChinaSchool 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, ChinaSchool of Biology and Biological Engineering South China University of Technology 382 East Outer Loop Rd, University Park, Guangzhou 510006, ChinaPhospholipase B (EC 3.1.1.5) are a distinctive group of enzymes that catalyzes the hydrolysis of fatty acids esterified at the <i>sn-1</i> and <i>sn-2</i> 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 <i>Talaromyces marneffei</i> GD-0079 synthesized by genome mining library. The gene (TmPlb1) was expressed and the TmPLB1 was purified using <i>E. coli</i> 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 <sup>31</sup>P 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.https://www.mdpi.com/2218-273X/10/2/231free fatty acids (ffas)nmr (nuclear magnetic resonance)phospholipase b<i>talaromyces marneffei</i>affinity chromatography |
| spellingShingle | Rabia Durrani Faez Iqbal Khan Shahid Ali Yonghua Wang Bo Yang A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics Study Biomolecules free fatty acids (ffas) nmr (nuclear magnetic resonance) phospholipase b <i>talaromyces marneffei</i> affinity chromatography |
| title | A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics Study |
| title_full | A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics Study |
| title_fullStr | A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics Study |
| title_full_unstemmed | A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics Study |
| title_short | A Thermolabile Phospholipase B from <i>Talaromyces marneffei</i> GD-0079: Biochemical Characterization and Structure Dynamics Study |
| title_sort | thermolabile phospholipase b from i talaromyces marneffei i gd 0079 biochemical characterization and structure dynamics study |
| topic | free fatty acids (ffas) nmr (nuclear magnetic resonance) phospholipase b <i>talaromyces marneffei</i> affinity chromatography |
| url | https://www.mdpi.com/2218-273X/10/2/231 |
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