fulltext.study @t Gmail

Improving the thermostability of lipase Lip2 from Yarrowia lipolytica

Paper ID Volume ID Publish Year Pages File Format Full-Text
23304 43431 2013 6 PDF Available
Title
Improving the thermostability of lipase Lip2 from Yarrowia lipolytica
Abstract

Yarrowia lipolytica lipase Lip2 (YlLip2) is a highly versatile biocatalyst. However, its practical use is often hampered by its low stability. Here three complementary protein engineering strategies were used to improve the thermostability of this enzyme. The first strategy was error-prone PCR based directed evolution, which resulted in a YlLip2 variant with a 2.5-fold longer half-life of thermal inactivation at 50 °C compared to the wild-type enzyme. The second strategy was semi-rational design using the so-called B-factor iterative test (B-FIT), which led to the discovery of two thermostable YlLip2 variants that showed a half-life of thermal inactivation 2-fold and 5-fold longer than that of the wild-type enzyme, respectively, at 50 °C. The third strategy was to use site-directed mutagenesis to combinatorially combine all three thermostabilizing mutations identified in the first two strategies, which improved the half-life of thermal inactivation of YlLip2 by 7-fold compared to that of the wild-type enzyme. Such engineered lipases provide not only new insights on the protein structure and function relationship but also potentially useful catalysts for practical applications.

► Three complementary strategies were used to improve the thermostability of a lipase. ► Directed evolution and the semi-rational design B-FIT method were compared side-by-side. ► Several thermostable lipase mutants were obtained and characterized. ► New insights on the lipase structure-function relationship were provided.

Keywords
Protein engineering; Directed evolution; Semi-rational design; B-FIT; Thermostability
First Page Preview
Improving the thermostability of lipase Lip2 from Yarrowia lipolytica
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
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 164, Issue 2, March 2013, Pages 248–253
Authors
, , ,
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