Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4
The exogenous lipolytic activities of Kocuria sp. have been recognized earlier but the genus further contains many more unexplored strains. In this study, the extracellular lipase activity of Kocuria flava Y4 (GenBank accession no. MT773277), isolated from Dioscorea villosa during our previous study...
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| Format: | Article |
| Language: | English |
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Wiley
2022-01-01
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| Series: | International Journal of Analytical Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2022/6403090 |
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| author | Adyasa Barik Sudip Kumar Sen Geetanjali Rajhans Sangeeta Raut |
| author_facet | Adyasa Barik Sudip Kumar Sen Geetanjali Rajhans Sangeeta Raut |
| author_sort | Adyasa Barik |
| collection | DOAJ |
| description | The exogenous lipolytic activities of Kocuria sp. have been recognized earlier but the genus further contains many more unexplored strains. In this study, the extracellular lipase activity of Kocuria flava Y4 (GenBank accession no. MT773277), isolated from Dioscorea villosa during our previous study, was regulated by different physicochemical parameters, such as pH, temperature, shaking speed, and incubation time. For efficient immobilization of the extracellular lipase, 4% sodium alginate, 50 mL of 25 nM CaCl2.2H2O solution, and 15 min. Hardening time of gel beads in calcium chloride was used. For the first time, K. flava Y4 lipase was purified using ammonium sulphate precipitation followed by dialysis and DEAE-Sepharose anion exchange chromatography with Sepharose-6B gel filtration chromatography, yielding ∼15-fold purified lipase with a final yield of 96 U/mL. The SDS-PAGE of purified lipase displayed a single strong band, indicating a monomeric protein of 45 kDa. At a temperature of 35°C and pH 8, the purified lipase showed maximum hydrolytic activity. Using p-nitrophenyl acetate (p-NPA) as the hydrolysis substrate, the values of Km and Vmax derived from the Lineweaver–Burk plot were 4.625 mM and 125 mol/min−1mg−1, respectively. |
| format | Article |
| id | doaj-art-24732fa6d6fa4298bc78e45f2507fdfe |
| institution | Kabale University |
| issn | 1687-8779 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Analytical Chemistry |
| spelling | doaj-art-24732fa6d6fa4298bc78e45f2507fdfe2025-02-03T06:05:32ZengWileyInternational Journal of Analytical Chemistry1687-87792022-01-01202210.1155/2022/6403090Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4Adyasa Barik0Sudip Kumar Sen1Geetanjali Rajhans2Sangeeta Raut3Centre for BiotecnologyBiostadt India LimitedCentre for BiotecnologyCentre for BiotecnologyThe exogenous lipolytic activities of Kocuria sp. have been recognized earlier but the genus further contains many more unexplored strains. In this study, the extracellular lipase activity of Kocuria flava Y4 (GenBank accession no. MT773277), isolated from Dioscorea villosa during our previous study, was regulated by different physicochemical parameters, such as pH, temperature, shaking speed, and incubation time. For efficient immobilization of the extracellular lipase, 4% sodium alginate, 50 mL of 25 nM CaCl2.2H2O solution, and 15 min. Hardening time of gel beads in calcium chloride was used. For the first time, K. flava Y4 lipase was purified using ammonium sulphate precipitation followed by dialysis and DEAE-Sepharose anion exchange chromatography with Sepharose-6B gel filtration chromatography, yielding ∼15-fold purified lipase with a final yield of 96 U/mL. The SDS-PAGE of purified lipase displayed a single strong band, indicating a monomeric protein of 45 kDa. At a temperature of 35°C and pH 8, the purified lipase showed maximum hydrolytic activity. Using p-nitrophenyl acetate (p-NPA) as the hydrolysis substrate, the values of Km and Vmax derived from the Lineweaver–Burk plot were 4.625 mM and 125 mol/min−1mg−1, respectively.http://dx.doi.org/10.1155/2022/6403090 |
| spellingShingle | Adyasa Barik Sudip Kumar Sen Geetanjali Rajhans Sangeeta Raut Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4 International Journal of Analytical Chemistry |
| title | Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4 |
| title_full | Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4 |
| title_fullStr | Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4 |
| title_full_unstemmed | Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4 |
| title_short | Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4 |
| title_sort | purification and optimization of extracellular lipase from a novel strain kocuria flava y4 |
| url | http://dx.doi.org/10.1155/2022/6403090 |
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