Journal
BIOTECHNOLOGY LETTERS
Volume 32, Issue 4, Pages 521-526Publisher
SPRINGER
DOI: 10.1007/s10529-009-0189-9
Keywords
Burkholderia cepacia; Lipase; Over-expression; Red recombinase; T7 RNA polymerase
Categories
Funding
- National High Technology Research and Development Program of China [2006AA020203, 2007AA05Z417, 2007AA100703, 2009AA03Z232]
- Program for New Century Excellent Talents in University [NCET-07-0336]
Ask authors/readers for more resources
Red recombinase system of the lambda phage is widely used for recombination of short linear DNA fragments and genome. Using this system, we obtained T7 RNA polymerase (RNAP) substitution mutants in Burkholderia cepacia. To test the expression abilities of the T7 mutants, four different lipase expression vectors were transformed and the lipase activity of these recombinants was evaluated. Our results suggest that 500 nt homology between the unit and the genome is sufficient to generate mutations and this strategy enables the rapid establishment of mutant strains with efficiencies of 85%. After expression and purification, the highest purified lipase activity obtained was 3,990 U/l, nearly triple that of the wild-type organism.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available