Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol
Abstract The recombinant Yleh from a tropical marine yeast Yarrowia lipolytica NCIM 3589 exhibited a high epoxide hydrolase activity of 9.34 ± 1.80 µmol min-1 mg-1 protein towards 1,2-epoxyoctane (EO), at pH 8.0 and 30 °C. The reaction product was identified as 1,2-Octanediol (OD) by GC-MS using EO...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2023-07-01
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| Series: | AMB Express |
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| Online Access: | https://doi.org/10.1186/s13568-023-01584-1 |
| _version_ | 1797769333564243968 |
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| author | Vijaya P. Godase V. Ravi Kumar Ameeta Ravi Kumar |
| author_facet | Vijaya P. Godase V. Ravi Kumar Ameeta Ravi Kumar |
| author_sort | Vijaya P. Godase |
| collection | DOAJ |
| description | Abstract The recombinant Yleh from a tropical marine yeast Yarrowia lipolytica NCIM 3589 exhibited a high epoxide hydrolase activity of 9.34 ± 1.80 µmol min-1 mg-1 protein towards 1,2-epoxyoctane (EO), at pH 8.0 and 30 °C. The reaction product was identified as 1,2-Octanediol (OD) by GC-MS using EO and H2O18 as substrate, affirming the functionality of Yleh as an epoxide hydrolase. For EO, the K m, V max, and k cat/K m values were 0.43 ± 0.017 mM, 0.042 ± 0.003 mM min-1, and 467.17 ± 39.43 mM-1 min-1, respectively. To optimize the reaction conditions for conversion of racemic EO by Yleh catalyst to enantiopure (R)-1,2-octanediol, initially, Response Surface Methodology was employed. Under optimized reaction conditions of 15 mM EO, 150 µg purified Yleh at 30 °C a maximal diol production of 7.11 mM was attained in a short span of 65 min with a yield of 47.4%. Green technology using deep eutectic solvents for the hydrophobic substrate (EO) were tested as co-solvents in Yleh catalyzed EO hydrolysis. Choline chloride-Glycerol, produced 9.08 mM OD with an increased OD yield of 60.5%. Thus, results showed that deep eutectic solvents could be a promising solvent for Yleh-catalyzed reactions making Yleh a potential biocatalyst for the biosynthesis of enantiopure synthons. |
| first_indexed | 2024-03-12T21:07:28Z |
| format | Article |
| id | doaj.art-de3c1f8d45ec49bfa33e95c43f5acaa0 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-03-12T21:07:28Z |
| publishDate | 2023-07-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-de3c1f8d45ec49bfa33e95c43f5acaa02023-07-30T11:25:23ZengSpringerOpenAMB Express2191-08552023-07-0113111510.1186/s13568-023-01584-1Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediolVijaya P. Godase0V. Ravi Kumar1Ameeta Ravi Kumar2Biochemistry Research Laboratory, Department of Biotechnology (Formerly Institute of Bioinformatics and Biotechnology), Savitribai Phule Pune UniversityChemical Engineering and Process Development Division, National Chemical LaboratoryBiochemistry Research Laboratory, Department of Biotechnology (Formerly Institute of Bioinformatics and Biotechnology), Savitribai Phule Pune UniversityAbstract The recombinant Yleh from a tropical marine yeast Yarrowia lipolytica NCIM 3589 exhibited a high epoxide hydrolase activity of 9.34 ± 1.80 µmol min-1 mg-1 protein towards 1,2-epoxyoctane (EO), at pH 8.0 and 30 °C. The reaction product was identified as 1,2-Octanediol (OD) by GC-MS using EO and H2O18 as substrate, affirming the functionality of Yleh as an epoxide hydrolase. For EO, the K m, V max, and k cat/K m values were 0.43 ± 0.017 mM, 0.042 ± 0.003 mM min-1, and 467.17 ± 39.43 mM-1 min-1, respectively. To optimize the reaction conditions for conversion of racemic EO by Yleh catalyst to enantiopure (R)-1,2-octanediol, initially, Response Surface Methodology was employed. Under optimized reaction conditions of 15 mM EO, 150 µg purified Yleh at 30 °C a maximal diol production of 7.11 mM was attained in a short span of 65 min with a yield of 47.4%. Green technology using deep eutectic solvents for the hydrophobic substrate (EO) were tested as co-solvents in Yleh catalyzed EO hydrolysis. Choline chloride-Glycerol, produced 9.08 mM OD with an increased OD yield of 60.5%. Thus, results showed that deep eutectic solvents could be a promising solvent for Yleh-catalyzed reactions making Yleh a potential biocatalyst for the biosynthesis of enantiopure synthons.https://doi.org/10.1186/s13568-023-01584-1Deep Eutectic solventsOptimizationRecombinant epoxide hydrolaseResponse surface methodologyYarrowia lipolytica |
| spellingShingle | Vijaya P. Godase V. Ravi Kumar Ameeta Ravi Kumar Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol AMB Express Deep Eutectic solvents Optimization Recombinant epoxide hydrolase Response surface methodology Yarrowia lipolytica |
| title | Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol |
| title_full | Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol |
| title_fullStr | Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol |
| title_full_unstemmed | Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol |
| title_short | Potential of Y. lipolytica epoxide hydrolase for efficient production of enantiopure (R)-1,2-octanediol |
| title_sort | potential of y lipolytica epoxide hydrolase for efficient production of enantiopure r 1 2 octanediol |
| topic | Deep Eutectic solvents Optimization Recombinant epoxide hydrolase Response surface methodology Yarrowia lipolytica |
| url | https://doi.org/10.1186/s13568-023-01584-1 |
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