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Kinetic and thermodynamic characterization of lipase produced by Cellulomonas flavigena UNP3
Author(s) -
Prajapati Vimal,
Patel Honey,
Trivedi Ujjval,
Patel Kamlesh
Publication year - 2014
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201300065
Subject(s) - lipase , hydrolysis , chemistry , ammonium sulfate precipitation , chromatography , activation energy , precipitation , ammonium , enzyme , triacylglycerol lipase , nuclear chemistry , biochemistry , organic chemistry , size exclusion chromatography , physics , meteorology
Lipase of Cellulomonas flavigena UNP3 was purified by two‐step purification process comprising ammonium sulfate precipitation followed by gel permeation chromatography (GPC). The recovery of lipase after GPC was found to be 1.70% with 20.98‐fold increase in specific activity. The molecular weight of lipase protein was found to be 45.2 kDa by SDS–PAGE. Activation energy for p ‐nitrophenol palmitate (pNPP) hydrolysis was 26.45 kJ mol −1 , while temperature quotient ( Q 10 ) was found to be 1.64. The enzyme was found to be stable over wide pH range and thermally stable at 30–40 °C up to 60 min of incubation while exhibited maximum activity at 30 °C with pH 7.0. V max , K m , and K cat for pNPP were found to be 666.71 U ml −1 , 1.33 mM (pNPP) and 433 min −1 , respectively. Activation energy for irreversible inactivation E a(d) of lipase was 64.32 kJ mol −1 . Thermodynamic parameters of irreversible inactivation of lipase and pNPP hydrolysis were also determined.