Expression and purification of organic solvent stable lipase from soil metagenomic library.
Gene for organic solvent stable lipase was overexpressed from soil metagenomic library. The clone with maximum activity was selected, and enzyme was purified by gel-permeation chromatography with a molecular mass of approx. 40 kDa. The deduced aminoacid sequence indicated that the protein belongs to the lipase family I.3 and containing a C-terminal secretion signal for ABC dependent transport together with possible motifs for Ca(2+) binding sites. The enzyme expressed maximum activity at 30 °C and pH 7.0 and found to be stable in pH and temperature ranging from 6.0-9.0 and 20-60 °C, respectively. Furthermore, the enzyme was found highly resistant to many organic solvents, especially isopropanol, DMSO, methanol, xylene and hexane. The enzyme showed enhanced activity in the presence of divalent cations (Mg(2+), Mn(2+), Ca(2+), Hg(2+), Cu(2+)), whereas the presence of trivalent cation (Fe(3+)) inhibited the activity.
Department of Food Microbiology, Central Food Technological Research Institute, CSIR, Mysore, 570020, India. mahejibin@cftri.org
Mahejibin
M
Khan
Kottur
K
Jithesh
D004795:Enzyme Stability / Q000187:drug effects; D015723:Gene Library; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D056186:Metagenomics; D012987:Soil; D012997:Solvents / Q000494:pharmacology; D013696:Temperature
D012987:Soil; D012997:Solvents; D008049:Lipase