Production, partial purification and characterization of organic solvent tolerant lipase from Burkholderia multivorans V2 and its application for ester synthesis.
Burkholderia multivorans V2 (BMV2) isolated from soil was found to produce an extracellular solvent tolerant lipase (6.477 U/mL). This lipase exhibited maximum stability in n-hexane retaining about 97.8% activity for 24h. After performing statistical optimization of medium components for lipase production, a 2.2-fold (14 U/mL) enhancement in the lipase production was observed. The crude lipase from BMV2 was partially purified by ultrafiltration and gel permeation chromatography with 24.64-fold purification. The K(m) and V(max) values for partially purified BMV2 lipase were found to be 1.56 mM and 5.62 micromoles/mg min. The metal ions Ca(2+), Mg(2+) and Mn(2+) had stimulatory effect on lipase activity, whereas Cu(2+), Fe(2+) and Zn(2+) strongly inhibited the lipase activity. EDTA and PMSF at 10mM concentration strongly inhibited the lipase activity. Non-ionic and anionic surfactants stimulated the lipase activity. BMV2 lipase was proved to be efficient in synthesis of ethyl butyrate ester under non-aqueous environment.
Dandavate
Vrushali
V
BRD School of Biosciences, Sardar Patel Maidan, Sardar Patel University, Vallabh Vidyanagar, Gujarat, India. vdandavate@gmail.com
Jinjala
Jayesh
J
Keharia
Haresh
H
Madamwar
Datta
D
D001709:Biotechnology / Q000379:methods*; D019117:Burkholderia / Q000201:enzymology*; D002087:Butyrates / Q000737:chemistry; D002244:Carbon / Q000737:chemistry; D002625:Chemistry, Organic / Q000379:methods; D002845:Chromatography / Q000379:methods; D004952:Esters / Q000138:chemical synthesis* / Q000737:chemistry; D005374:Filtration; D007218:Industrial Microbiology / Q000379:methods; D007477:Ions; D007700:Kinetics; D008049:Lipase / Q000737:chemistry* / Q000302:isolation & purification*; D008670:Metals / Q000737:chemistry; D010802:Phylogeny; D012997:Solvents / Q000737:chemistry
D002087:Butyrates; D004952:Esters; D007477:Ions; D008670:Metals; D012997:Solvents; D002244:Carbon; D008049:Lipase; C045572:ethyl butyrate