A Broad Temperature Active Lipase Purified From a Psychrotrophic Bacterium of Sikkim Himalaya With Potential Application in Detergent Formulation

Frontiers in Bioengineering and Biotechnology
Anil KumarRakshak Kumar

Abstract

Bacterial lipases with activity spanning over a broad temperature and substrate range have several industrial applications. An efficient enzyme-producing bacterium Chryseobacterium polytrichastri ERMR1:04, previously reported from Sikkim Himalaya, was explored for purification and characterization of cold-adapted lipase. Optimum lipase production was observed in 1% (v/v) rice bran oil, pH 7 at 20°C. Size exclusion and hydrophobic interaction chromatography purified the enzyme up to 21.3-fold predicting it to be a hexameric protein of 250 kDa, with 39.8 kDa monomeric unit. MALDI-TOF-MS analysis of the purified lipase showed maximum similarity with alpha/beta hydrolase (lipase superfamily). Biochemical characterization of the purified enzyme revealed optimum pH (8.0), temperature (37°C) and activity over a temperature range of 5-65°C. The tested metals (except Cu2+ and Fe2+) enhanced the enzyme activity and it was tolerant to 5% (v/v) methanol and isopropanol. The Km and Vmax values were determined as 0.104 mM and 3.58 U/mg, respectively for p-nitrophenyl palmitate. Bioinformatics analysis also supported in vitro findings by predicting enzyme's broad temperature and substrate specificity. The compatibility of the purified lipase ...Continue Reading

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Citations

Jan 6, 2021·Journal, Genetic Engineering & Biotechnology·Shimaa E HelalMahmoud M Amer
Feb 6, 2021·Applied Biochemistry and Biotechnology·Nutan MhetrasDigambar Gokhale

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Methods Mentioned

BETA
genome-based predictions
lipase assay
MDS
size exclusion chromatography
gel filtration

Software Mentioned

Pymol
PRODRG
COACH
ProSAweb
PSI
ProtParam
MASCOT
NK
ProSA
Autodock Vina

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