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Molecular cloning and characterization of thermostable esterase and lipase from Geobacillus thermoleovorans YN isolated from desert soil in Egypt

Paper ID Volume ID Publish Year Pages File Format Full-Text
35864 45111 2007 11 PDF Available
Title
Molecular cloning and characterization of thermostable esterase and lipase from Geobacillus thermoleovorans YN isolated from desert soil in Egypt
Abstract

Genes encoding an esterase (EstA) and lipase (LipA) from Geobacillus thermoleovorans YN, a strain isolated from Egyptian desert soil, were cloned and the respective proteins were expressed in Escherichia coli and characterized. Whereas LipA was cloned directly by PCR amplification from genomic DNA, a genomic library composed of 3000 clones was screened on tributyrin agar plates to find EstA. An open reading frame of 744 bps encoding a polypeptide of 247 amino acid residues was identified as esterase due to its conserved GXSXG motif and its high similarity toward other carboxyl esterases. LipA (416 aa residues) is encoded by an ORF of 1251 bps and constitutes a pre-protein with a calculated molecular mass of 46 kDa including a signal sequence of 28 aa resulting in a mature lipase of 43 kDa. Both, LipA and EstA were sub-cloned and expressed under control of the temperature-inducible λ-promoter and purified by IMAC and gel filtration. The molecular mass of the purified EstA was 29 kDa. Both enzymes were most active at pH ∼9.5 and remarkably stable at pH 5 and 10.5. Temperature optima and stabilities (up to 70 °C) of both enzymes as well as their reaction kinetics and substrate spectra were determined.

Keywords
Esterase; Lipase; Geobacillus thermoleovorans; Thermophiles; Biocatalysis
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Molecular cloning and characterization of thermostable esterase and lipase from Geobacillus thermoleovorans YN isolated from desert soil in Egypt
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 42, Issue 7, July 2007, Pages 1090–1100
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us