|
|
Publication
|
| Research Title |
Cloning and expression of pyridoxal kinase from Geobacillus sp.H6a. |
| Date of Distribution |
5 October 2012 |
| Conference |
| Title of the Conference |
International Conference on Microbial Taxonomy, Basic and Applied Microbiology |
| Organiser |
Department of Microbiology, Faculty of Science Khon Kaen University |
| Conference Place |
Kosa Hotel |
| Province/State |
Khon Kaen, Thailand |
| Conference Date |
4 October 2012 |
| To |
6 October 2012 |
| Proceeding Paper |
| Volume |
1 |
| Issue |
1 |
| Page |
99 |
| Editors/edition/publisher |
|
| Abstract |
Vitamin B6 is an important coenzyme for metabolism of protein, carbohydrate, lipid, nervous system, hormone and heme synthesis. In all organisms, the pyridoxal (PL) kinase is in the salvage pathway of vitamin B6 that this enzyme is coded by pdxK gene. In the presence of Zn2+ and ATP, PL kinase catalyzes the phosphorylation of PL, pyridoxine (PN), and pyridoxamine (PM) to form pyridoxine 5-phosphate (PNP), pyridoxal 5-phosphate (PLP), and pyridoxamine 5-phosphate (PMP), respectively. In this study, we cloned and expressed pdxK gene from Geobacillus sp. H6a (Gh), isolated from hot springs at the North of Thailand. Using touchdown PCR, the GhpdxK gene had a length of 810 base pairs (bp) encoding a protein of 270 amino acids (aa). The GhpdxK gene was inserted into pET28a (+) expression vector at restriction site of NdeI and BamHI and then transformed into E.coli BL21 (DE3). The transformant was induced with 1 mM IPTG at 25, 30 and 37C for 0, 1, 3 and 6 hours. The recombinant GhPL kinase with His-tag had molecular weight 37 kDa and was expressed as soluble protein and inclusion bodies. The soluble rGhPL kinase showed the enzyme activity. The optimum condition of solubilization and refolding of rGhPL kinase inclusion bodies will be further studied.
Keywords: pyridoxal kinase; cloning; expression; Geobacillus sp.H6a; inclusion body protein
|
| Author |
|
| Peer Review Status |
มีผู้ประเมินอิสระ |
| Level of Conference |
นานาชาติ |
| Type of Proceeding |
Full paper |
| Type of Presentation |
Oral |
| Part of thesis |
true |
| Presentation awarding |
false |
| Attach file |
|
| Citation |
0
|
|
|
|
|
|
|
|
|
Journal Publication
ไทย