Journal article
A lipase with broad temperature range from an alkaliphilic gamma-proteobacterium isolated in Greenland
Division of Microbiology and Risk Assessment, National Food Institute, Technical University of Denmark1
National Food Institute, Technical University of Denmark2
Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark3
Department of Chemical and Biochemical Engineering, Technical University of Denmark4
University of Copenhagen5
A gamma-proteobacterium related to the genera Alteromonadales and Pseudomonadales , isolated from a cold and alkaline environment in Greenland, has been shown to produce a lipase active between 5 ° C and 80 ° C, with optimal activity at 55 ° C and pH 8. PCR-based screening of genomic DNA from the isolated bacterium, followed by genome walking, resulted in two complete open reading frames, which were predicted to encode a lipase and its helper protein, a lipase foldase.
The amino acid sequence derived for the lipase showed resemblance to lipases from Pseudomonas , Rhodoferax, Aeromonas and Vibrio . The two genes were cloned into different expression systems in E. coli with or without a putative secretion sequence, but despite the fact that both recombinant lipase and lipase foldase were observed on SDS–PAGE, no recombinant lipase activity was detected.
Attempts to refold the recombinant lipase in vitro using a purified lipase foldase remained unsuccessful.
| Language: | English |
|---|---|
| Year: | 2010 |
| Pages: | 1091-1100 |
| ISSN: | 1479487x and 09593330 |
| Types: | Journal article |
| DOI: | 10.1080/09593331003770289 |
| ORCIDs: | 0000-0002-2796-7137 |
