Molecular cloning and characterization of a protease gene from antarctic psychrotrophic bacterium Pseudomonas sp. 3-37
| dc.contributor.author | Tham, Hui Mian | |
| dc.date.accessioned | 2014-10-27T07:24:10Z | |
| dc.date.available | 2014-10-27T07:24:10Z | |
| dc.date.issued | 2009 | |
| dc.description | Master | en_US |
| dc.description.abstract | Cold-active enzymes are usually characterized by a higher specific activity at low temperature when compared to their mesophilic counterparts and offer a high potential in numerous biotechnological applications. In this study, the gene encoding metalloprotease of the psychrotrophic bacterium Pseudomonas sp. 3-37 isolated from water samples collected at Odbert Island, Antarctica, was cloned and characterized. The bacterium could survive a wide range of temperature from 4 ºC to 30 ºC and produces a protease on Skim Milk Agar when cultured at 4 ºC. The metalloprotease structural gene was isolated from the Pseudomonas sp. 3-37 genomic library by using a homologous metalloprotease PCR fragment probe. Nucleotide sequence analysis revealed an open reading frame (ORF) of 1431 bp encoding a 477 amino acid residues with an estimated molecular mass of 53 kDa that exhibited 100% identity to extracellular alkaline metalloprotease of Pseudomonas fluorescens A506 (accession no AAP4489). Two other genes were identified flanking the metalloprotease gene. The 2 genes were putative Daminopeptidase gene and zinc protease transporter gene. The metalloprotease gene was expressed in a protease deficient strain of E. coli Tuner(DE3) and the functional activity of the recombinant metalloprotease was confirmed by casein zymography. This metalloprotease contains a zinc-binding motif (HExxH), repetitive glycine-rich nonapeptide sequence motif (GGxGxDxux) and contains no cysteine residue, these features are specifically found in serralysin subfamily. (accession no CAA07699.1) and zinc-protease transporter of Pseudomonas fluorescens CY091 , respectively. (Appendix H.3). | en_US |
| dc.identifier.uri | http://hdl.handle.net/123456789/168 | |
| dc.language.iso | en | en_US |
| dc.subject | Biological Science | en_US |
| dc.subject | Molecular cloning | en_US |
| dc.subject | Protease gene | en_US |
| dc.title | Molecular cloning and characterization of a protease gene from antarctic psychrotrophic bacterium Pseudomonas sp. 3-37 | en_US |
| dc.type | Thesis | en_US |
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