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Cloning and characterisation of a new 2-deoxy-d-ribose-5-phosphate aldolase from Rhodococcus erythropolis

Paper ID Volume ID Publish Year Pages File Format Full-Text
23799 43473 2012 7 PDF Available
Title
Cloning and characterisation of a new 2-deoxy-d-ribose-5-phosphate aldolase from Rhodococcus erythropolis
Abstract

A new 2-deoxy-d-ribose-5-phoshate aldolase (DERA) gene was cloned from Rhodococcus erythropolis strain DSM 311, recombinantly expressed in Escherichia coli, and purified via affinity chromatography which yielded a homo-dimeric enzyme of 44.3 kDa as apparent by size exclusion chromatography. To characterise the enzyme, investigations about pH and temperature tolerance, stability, as well as analyses on resistance to organic solvents and acetaldehyde were performed. In addition, kinetic constants of the new DERARE were compared to respective values of the DERA from E. coli (DERAEC). Stability of DERARE turned out to be a crucial factor: The pH for optimal DERARE activity was determined to be 7.0, whereas the highest stability was achieved at pH 9.0 with a half-life of approximately 20 days. The optimal temperature for DERARE activity was 65 °C, but coupled with a rather low stability (half-life of 2 min). The highest stability was achieved at 25 °C. The new enzyme exhibits high resistance to organic solvents and acetaldehyde with a half-life being 2.5× higher compared to DERAEC under the exposure of 300 mM acetaldehyde. Hence it has the potential as a new promising biocatalyst with applications in organic synthesis.

► New acetaldehyde depending aldolase identified from Rhodococcus erythropolis. ► Functional expression in E. coli and purification of enzyme was established. ► Enhanced solvent tolerance was observed. ► Increased stability towards substrate acetaldehyde was demonstrated.

Keywords
2-Deoxy-d-ribose-5-phosphate aldolase; Aldol reaction; Enzyme stability; Organic solvent tolerance; Rhodococcus erythropolis
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Cloning and characterisation of a new 2-deoxy-d-ribose-5-phosphate aldolase from Rhodococcus erythropolis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 161, Issue 2, 15 October 2012, Pages 174–180
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