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

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Title Purification and biochemical characterization of an extracellular lipase from Pseudomonas fluorescens MTCC 2421.
Not Available
 
Creator Paulraj,R
Chakraborty,Kajal
 
Subject Lipase
P. fluorescens MTCC 2421
sorbitol
Ca2+
fatty acids
glycerol trioleate
 
Description Not Available
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.
Not Available
 
Date 2021-09-15T09:54:28Z
2021-09-15T09:54:28Z
2009
 
Type Article
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/62521
 
Language English
 
Relation Not Available
 
Publisher American Chemical Society