A lipase with broad solvent stability from Burkholderia cepacia RQ3: isolation, characteristics and application for chiral resolution of 1-phenylethanol.
Using both polar and low polar organic solvents (DMSO and toluene) as screening stress, a solvent-stable bacterium Burkholderia cepacia RQ3 was newly isolated. An organic solvent-stable lipase from strain RQ3 was purified in a single step with 50.1% recovery by hydrophobic chromatography. The purified lipase was homogenous on SDS-PAGE and had an apparent molecular mass of 33 kDa. The gene of lipase RQ3 with an open reading frame of 1095 bp encoding 364-amino acid residues was cloned. The optimal pH and temperature for lipase activity were 9.0 and 40 °C. The lipase was stable in a wide pH range of 6.0-10.0 and at temperature below 50 °C. Strikingly, all the tested hydrophilic and hydrophobic organic solvents significantly extended the half-life of lipase RQ3 compared with that in a solvent-free system, which indicated that lipase RQ3 showed a broad solvent tolerance to various organic solvents. The lipase demonstrated excellent enantioselective transesterification toward the (S)-1-phenylethanol with a theoretical conversion yield of 50% and ee p of 99.9%, which made it an exploitable biocatalyst for organic synthesis and pharmaceutical industries.
Chengjia
C
Xie
Bin
B
Wu
Song
S
Qin
Bingfang
B
He
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D001592:Benzyl Alcohols / Q000737:chemistry*; D016956:Burkholderia cepacia / Q000201:enzymology*; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification
D001426:Bacterial Proteins; D001592:Benzyl Alcohols; C002017:methylphenyl carbinol; D008049:Lipase