Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3
A recombinant A3 lipase from Pseudomonas sp. strain AMS 3 was previously expressed in E. coli. In present study, recombinant A3 lipase was successfully purified through Nickel-Sepharose Fast Flow affinity chromatography. This purification recovered 58.47% of yield with the purification fold 3.69. So...
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| Format: | Project Paper Report |
| Language: | English |
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2015
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| Online Access: | http://psasir.upm.edu.my/id/eprint/78207/1/FBSB%202015%2056%20-%20IR.pdf |
| _version_ | 1796980441732874240 |
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| author | Leong, Mun Hong |
| author_facet | Leong, Mun Hong |
| author_sort | Leong, Mun Hong |
| collection | UPM |
| description | A recombinant A3 lipase from Pseudomonas sp. strain AMS 3 was previously expressed in E. coli. In present study, recombinant A3 lipase was successfully purified through Nickel-Sepharose Fast Flow affinity chromatography. This purification recovered 58.47% of yield with the purification fold 3.69. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to determine the molecular weight of purified A3 lipase. A3 lipase was screened for crystal under crystallization condition of Crystal Screen and Crystal Screen 2 from Hampton Research in MRC 2 well crystallization plate (Swissci) with the aid of crystallization robot, Oryx 8 for crystallization hits from the screening. The best A3 lipase crystal being observed in the formulation of 0.5 M ammonium sulfate, 0.1 M HEPES pH 7.5 and 30% v/v (+/-)-2-methyl-2,4-pentanediol. 3D structure of A3 lipase was predicted from RaptorX and analyzed by YASARA software. The predicted 3D structure of A3 lipase contained catalytic triad covered with 2 lid subunits, 2 metal ions binding site and glutathione-s-transferase located at the N-terminal. |
| first_indexed | 2024-03-06T10:23:03Z |
| format | Project Paper Report |
| id | upm.eprints-78207 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T10:23:03Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | upm.eprints-782072020-06-26T01:38:37Z http://psasir.upm.edu.my/id/eprint/78207/ Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3 Leong, Mun Hong A recombinant A3 lipase from Pseudomonas sp. strain AMS 3 was previously expressed in E. coli. In present study, recombinant A3 lipase was successfully purified through Nickel-Sepharose Fast Flow affinity chromatography. This purification recovered 58.47% of yield with the purification fold 3.69. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to determine the molecular weight of purified A3 lipase. A3 lipase was screened for crystal under crystallization condition of Crystal Screen and Crystal Screen 2 from Hampton Research in MRC 2 well crystallization plate (Swissci) with the aid of crystallization robot, Oryx 8 for crystallization hits from the screening. The best A3 lipase crystal being observed in the formulation of 0.5 M ammonium sulfate, 0.1 M HEPES pH 7.5 and 30% v/v (+/-)-2-methyl-2,4-pentanediol. 3D structure of A3 lipase was predicted from RaptorX and analyzed by YASARA software. The predicted 3D structure of A3 lipase contained catalytic triad covered with 2 lid subunits, 2 metal ions binding site and glutathione-s-transferase located at the N-terminal. 2015-06 Project Paper Report NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/78207/1/FBSB%202015%2056%20-%20IR.pdf Leong, Mun Hong (2015) Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3. [Project Paper Report] |
| spellingShingle | Leong, Mun Hong Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3 |
| title | Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3 |
| title_full | Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3 |
| title_fullStr | Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3 |
| title_full_unstemmed | Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3 |
| title_short | Purification, crystallization and molecular modeling of recombinant A3 lipase from an antartic Pseudomonas sp. strain AMS3 |
| title_sort | purification crystallization and molecular modeling of recombinant a3 lipase from an antartic pseudomonas sp strain ams3 |
| url | http://psasir.upm.edu.my/id/eprint/78207/1/FBSB%202015%2056%20-%20IR.pdf |
| work_keys_str_mv | AT leongmunhong purificationcrystallizationandmolecularmodelingofrecombinanta3lipasefromanantarticpseudomonasspstrainams3 |