Journal
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
Volume 51, Issue -, Pages 23-31Publisher
WILEY
DOI: 10.1042/BA20070186
Keywords
alkaline lipase; Burkholderia cepacia lipase; CD; cystic fibrosis; hydrophobic interaction chromatography; sodium alginate
Funding
- Department of Science and Technology (Government of India) Core Group Grant on Applied Biocatalysis
- Department of Biotechnology (Government of India)
- AICTE (All India Council for Technical Education)
- ICMR (Indian Council of Medical Research)
Ask authors/readers for more resources
A Burkholderia cepacia (bacteria) strain, A.TC.C. 25609, which had been isolated from the bronchial washings of a cystic fibrosis patient, was used to produce lipase. The presence of sodium alginate at an optimal concentration of 8 mg. ml(-1) in the growth medium nearly doubled the production of extracellular lipase activity. The enzyme could be purified with 38-fold purification and 96 % activity recovery using a two-step purification protocol. The molecular mass of the purified lipase determined by SDS/PAGE was shown to be 28 kDa. The pH optimum of the purified enzyme was 9 and it was stable up to 12 h at pH 9 and 10. The enzyme has a temperature optimum of 40 C and its half-life (t(1/2)) values were 54 and 46 min at 50 and 60 degrees C respectively. The lipase was found to be stable in the presence of the detergents Tween 20 and Triton X-100. The secondary-structure analysis of lipase by CD spectroscopy showed 52% a-helix, 7.7% beta-sheet, 12.6% beta-turn and 27.8% random structure. The lipase was cloned and overexpressed in Escherichia coli. The gene sequence of the cloned lipase was determined and compared with other lipases.
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