Abstract
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.
Original language | English |
---|---|
Journal | Environmental Technology (United Kingdom) |
Volume | 31 |
Issue number | 10 |
Pages (from-to) | 1091-1100 |
ISSN | 0959-3330 |
DOIs | |
Publication status | Published - 2010 |
Keywords
- Lipase-specific foldase
- Pseudomonas
- Lipase