Journal Article

The effect of proline insertions on the thermostability of a barley α-glucosidase

E.H. Muslin, S.E. Clark and C.A. Henson

in Protein Engineering, Design and Selection

Volume 15, issue 1, pages 29-33
Published in print January 2002 | ISSN: 1741-0126
Published online January 2002 | e-ISSN: 1741-0134 | DOI: https://dx.doi.org/10.1093/protein/15.1.29
The effect of proline insertions on the thermostability of a barley α-glucosidase

Show Summary Details

Preview

The thermal stability of α-glucosidase is important because the conversion of starch to fermentable sugars during industrial production of ethanol (e.g. brewing, fuel ethanol production) typically takes place at temperatures of 65–73°C. In this study we investigate the thermostability of α-glucosidases from four plant species, compare their deduced amino acid sequences, and test the effect of substituting a proline for the residue present in the wild-type enzyme on the thermostability of α-glucosidase. The α-glucosidase from barley (Hordeum vulgare) was significantly less thermostable than the other three α-glucosidases. A comparison of the published deduced amino acid sequences of these four α-glucosidases revealed conserved proline residues in the three most thermostable α-glucosidases that were not found in the barley enzyme. Site-directed mutagenesis was done on recombinant barley α-glucosidase to create proteins with prolines at these conserved positions. The thermostability (T50) of one of these mutant enzymes, T340P, was 10°C higher than the non-mutated enzyme.

Keywords: Hordeum vulgare; maltase; starch degradation

Journal Article.  3226 words.  Illustrated.

Subjects: Proteins

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content. subscribe or purchase to access all content.