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Purification and biochemical characterization of an extracellular lipase from Pseudomonas fluorescens MTCC 2421.

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Relation http://eprints.cmfri.org.in/10784/
 
Title Purification and biochemical characterization of an extracellular lipase from Pseudomonas fluorescens MTCC 2421.
 
Creator Chakraborty, Kajal
Paulraj, R
 
Subject Bioactive compounds
Biochemistry
 
Description An extracellular lipase produced by Pseudomonas fluorescens MTCC 2421 was purified 184.37-fold
with a specific activity of 424.04 LU/mg after anion exchange and gel exclusion chromatography. The
enzyme is a homomeric protein with an apparent molecular mass of 65.3 kDa. The lipase exhibited
hydrolytic resistance toward triglycerides with longer fatty acyl chain length containing unsaturation as
evident from the lowerVmax (0.23 mM/mg/min) of the lipase toward glycerol trioleate (C18:1n9) compared
with the fatty acid triglycerides having short to medium carbon chain lengths (C18:0-12:0, Vmax 0.32-
0.51 mM/mg/min). This indicates a preferential specificity of the lipase toward cleaving shorter carbon
chain length fatty acid triglycerides. The lipase exhibited optimum activity at 40 �C and pH 8.0,
respectively. A combination of Ca2+ and sorbitol induced a synergistic effect on the thermostability of
lipase with a significantly high residual activity (100%) after 30 min at 40 �C, as compared to 90.6% after
incubation with Ca2+ alone. The lipase activity was inhibited by Cu2+ and Fe2+ (42 and 48%,
respectively) at 10 mM. The enzyme lost 31% of its initial activity by 0.001 mM EDTA and 42% by
0.1 mM EDTA. Significant reduction in lipase activity was apparent by 2-mercaptoethanol and
phenylmethanesulfonyl fluoride at diluted concentration (0.001 mM), thereby indicating an important
role of sulfhydryl groups in the catalytic mechanism.
 
Publisher American Chemical Society
 
Date 2009
 
Type Article
PeerReviewed
 
Format text
 
Language en
 
Identifier http://eprints.cmfri.org.in/10784/1/Purification_%20biochemical%20characterization_2009_57_Kajal.pdf
Chakraborty, Kajal and Paulraj, R (2009) Purification and biochemical characterization of an extracellular lipase from Pseudomonas fluorescens MTCC 2421. Journal of Agricultural and Food Chemistry, 57 (9). pp. 3859-3866.