Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp.
The 3-HBDH gene from Pseudomonas sp. was amplified, cloned, and sequenced. The deduced amino acid sequence was highly matched with 3-HBDH sequences in DDBJ/GenBank/EMBL. But Pseudomonas sp. 3-HBDH was lack of the C-terminal region found in mammalian enzymes containing a lipid-binding domain that is...
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| Language: | en |
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Universiti Malaysia Sabah
2003
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| Online Access: | http://eprints.utm.my/6922/1/NikAzmiNikMahmod2003_CloningAndExpressionOf3-hydroxybutyrateDehydrogenase.pdf http://eprints.utm.my/6922/ |
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| author | Nik Mahmod, Nik Azmi Mohd Illias, Rosli Syed Muhammad, Syed Annuar Fauad H. D., Habib Abdullah Fikri S., Wan Nor Azlinda Lau, S. Y. |
| author_facet | Nik Mahmod, Nik Azmi Mohd Illias, Rosli Syed Muhammad, Syed Annuar Fauad H. D., Habib Abdullah Fikri S., Wan Nor Azlinda Lau, S. Y. |
| author_sort | Nik Mahmod, Nik Azmi |
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| collection | Institutional Repository |
| content_provider | Universiti Teknologi Malaysia |
| content_source | UTM Institutional Repository |
| continent | Asia |
| country | Malaysia |
| description | The 3-HBDH gene from Pseudomonas sp. was amplified, cloned, and sequenced. The deduced amino acid sequence was highly matched with 3-HBDH sequences in DDBJ/GenBank/EMBL. But Pseudomonas sp. 3-HBDH was lack of the C-terminal region found in mammalian enzymes containing a lipid-binding domain that is important for the 3-HBDH's activity. The E-coli XL1 blue cells transformed with the resultant plasmid, pBDH-1 showed 3-HBDH's activity. The E. coli XL1 Blue cells transformed with resultant plasmid, pBDH-1 showed 3-HBDH's activity. The nucleotide sequence of recombinant pBDH-1 indicate functional oligomers composed of subunits of 225 amino acid with a calculated Mr of 26, 855 Da and proved on SDS-PAGE. Ammonium sulfate farctionation resulted in low yield of the enzyme but its activity was comparably high with other 3-HBDHs. The recombinant enzyme showed a broad range of stability with respect to pH and temperature. |
| format | Article |
| id | my.utm.eprints-6922 |
| institution | Universiti Teknologi Malaysia |
| language | en |
| publishDate | 2003 |
| publisher | Universiti Malaysia Sabah |
| record_format | eprints |
| spelling | my.utm.eprints-69222010-06-01T15:48:23Z http://eprints.utm.my/6922/ Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. Nik Mahmod, Nik Azmi Mohd Illias, Rosli Syed Muhammad, Syed Annuar Fauad H. D., Habib Abdullah Fikri S., Wan Nor Azlinda Lau, S. Y. T Technology (General) The 3-HBDH gene from Pseudomonas sp. was amplified, cloned, and sequenced. The deduced amino acid sequence was highly matched with 3-HBDH sequences in DDBJ/GenBank/EMBL. But Pseudomonas sp. 3-HBDH was lack of the C-terminal region found in mammalian enzymes containing a lipid-binding domain that is important for the 3-HBDH's activity. The E-coli XL1 blue cells transformed with the resultant plasmid, pBDH-1 showed 3-HBDH's activity. The E. coli XL1 Blue cells transformed with resultant plasmid, pBDH-1 showed 3-HBDH's activity. The nucleotide sequence of recombinant pBDH-1 indicate functional oligomers composed of subunits of 225 amino acid with a calculated Mr of 26, 855 Da and proved on SDS-PAGE. Ammonium sulfate farctionation resulted in low yield of the enzyme but its activity was comparably high with other 3-HBDHs. The recombinant enzyme showed a broad range of stability with respect to pH and temperature. Universiti Malaysia Sabah 2003 Article PeerReviewed application/pdf en http://eprints.utm.my/6922/1/NikAzmiNikMahmod2003_CloningAndExpressionOf3-hydroxybutyrateDehydrogenase.pdf Nik Mahmod, Nik Azmi and Mohd Illias, Rosli and Syed Muhammad, Syed Annuar Fauad and H. D., Habib Abdullah Fikri and S., Wan Nor Azlinda and Lau, S. Y. (2003) Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. Proceedings of International Conference On Chemical and Bioprocess Engineering . pp. 262-265. |
| spellingShingle | T Technology (General) Nik Mahmod, Nik Azmi Mohd Illias, Rosli Syed Muhammad, Syed Annuar Fauad H. D., Habib Abdullah Fikri S., Wan Nor Azlinda Lau, S. Y. Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. |
| title | Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. |
| title_full | Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. |
| title_fullStr | Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. |
| title_full_unstemmed | Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. |
| title_short | Cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated Pseudomonas sp. |
| title_sort | cloning and expression of 3-hydroxybutyrate dehydrogenase gene from locally isolated pseudomonas sp. |
| topic | T Technology (General) |
| url | http://eprints.utm.my/6922/1/NikAzmiNikMahmod2003_CloningAndExpressionOf3-hydroxybutyrateDehydrogenase.pdf http://eprints.utm.my/6922/ |
| url_provider | http://eprints.utm.my/ |