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Applied Biochemistry and Biotechnology

, Volume 179, Issue 3, pp 474–484 | Cite as

Semi-Rational Design of Geobacillus stearothermophilus L-Lactate Dehydrogenase to Access Various Chiral α-Hydroxy Acids

  • Aşkın Sevinç Aslan
  • William R. Birmingham
  • Nevin Gül Karagüler
  • Nicholas J. Turner
  • Barış BinayEmail author
Article

Abstract

Chiral α-hydroxy acids (AHAs) are rapidly becoming important synthetic building blocks, in particular for the production of pharmaceuticals and other fine chemicals. Chiral compounds of a variety of functionalities are now often derived using enzymes, and L-lactate dehydrogenase from the thermophilic organism Geobacillus stearothermophilus (bsLDH) has the potential to be employed for the industrial synthesis of chiral α-hydroxy acids. Despite the thorough characterization of this enzyme, generation of variants with high activity on non-natural substrates has remained difficult and therefore limits the use of bsLDH in industry. Here, we present the engineering of bsLDH using semi-rational design as a method of focusing screening in a small and smart library for novel biocatalysts. In this study, six mutant libraries were designed in an effort to expand the substrate range of bsLDH. The eight variants identified as having enhanced activity toward the selected α-keto acids belonged to the same library, which targeted two positions simultaneously. These new variants now may be useful biocatalysts for chiral synthesis of α-hydroxy acids.

Keywords

Chiral α-hydroxy acids (AHAs) Geobacillus stearothermophilus L-lactate dehydrogenase (bsLDH) Protein engineering Modeling 

Notes

Acknowledgments

This work was supported by a grant from the TUBİTAK (grant number 213S099) and special thanks to COST Action (grant number CM1303).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Faculty of Science and Letters, Department of Molecular Biology and GeneticsIstanbul Technical UniversityIstanbulTurkey
  2. 2.Molecular Biology-Biotechnology and Genetics Research Center (MOBGAM)Istanbul Technical UniversityIstanbulTurkey
  3. 3.School of Chemistry, Manchester Institute of BiotechnologyUniversity of ManchesterManchesterUK
  4. 4.Molecular Biology and GeneticsIstanbul AREL UniversityIstanbulTurkey

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