Enzymes have become important tools in several industries due to their ability to produce chirally pure and complex molecules with interesting biological properties. The NAD+-dependent LDH (lactate dehydrogenase) [bsLDH [Geobacillus stearothermophilus (formerly Bacillus stearothermophilus) LDH] from G. stearothermophilus and the NAD+-dependent FDH (formate dehydrogenase) [cmFDH (Candida methylica FDH)] enzyme from C. methylica are particularly crucial enzymes in the pharmaceutical industry and are related to each other in terms of NADH use and regeneration. LDH catalyses the interconversion of pyruvate (oxo acid) and lactate (α-hydroxy acid) using the NADH/NAD+ pair as a redox cofactor. Employing LDH to reduce other oxo acids can generate chirally pure α-hydroxy acids of use in the production of pharmaceuticals. One important use of FDH is to regenerate the relatively expensive NADH cofactor that is used by NAD+-dependent oxidoreductases such as LDH. Both LDH and FDH from organisms of interest were previously cloned and overproduced. Therefore they are available at a low cost. However, both of these enzymes show disadvantages in the large-scale production of chirally pure compounds. We have applied two routes of protein engineering studies to improve the properties of these two enzymes, namely DNA shuffling and site-directed mutagenesis. Altering the substrate specificity of bsLDH by DNA shuffling and changing the coenzyme specificity of cmFDH by site-directed mutagenesis are the most successful examples of our studies. The present paper will also include the details of these examples together with some other applications of protein engineering regarding these enzymes.
Skip Nav Destination
Article navigation
December 2007
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
Conference Article|
November 23 2007
Protein engineering applications of industrially exploitable enzymes: Geobacillus stearothermophilus LDH and Candida methylica FDH
N.G. Karagüler;
N.G. Karagüler
1
*Department of Molecular Biology and Genetics, Faculty of Science and Letters, Istanbul Technical University, 34469 Istanbul, Turkey
†Istanbul Technical University Molecular Biology–Biotechnology and Genetics Research Center (MOBGAM), 34469 Istanbul, Turkey
1To whom correspondence should be addressed (email karaguler@itu.edu.tr).
Search for other works by this author on:
R.B. Sessions;
R.B. Sessions
‡Department of Biochemistry, School of Medical Sciences, University Walk, Bristol BS8 TD, U.K.
Search for other works by this author on:
B. Binay;
B. Binay
*Department of Molecular Biology and Genetics, Faculty of Science and Letters, Istanbul Technical University, 34469 Istanbul, Turkey
†Istanbul Technical University Molecular Biology–Biotechnology and Genetics Research Center (MOBGAM), 34469 Istanbul, Turkey
Search for other works by this author on:
E.B. Ordu;
E.B. Ordu
*Department of Molecular Biology and Genetics, Faculty of Science and Letters, Istanbul Technical University, 34469 Istanbul, Turkey
†Istanbul Technical University Molecular Biology–Biotechnology and Genetics Research Center (MOBGAM), 34469 Istanbul, Turkey
Search for other works by this author on:
A.R. Clarke
A.R. Clarke
‡Department of Biochemistry, School of Medical Sciences, University Walk, Bristol BS8 TD, U.K.
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
June 19 2007
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© The Authors Journal compilation © 2007 Biochemical Society
2007
Biochem Soc Trans (2007) 35 (6): 1610–1615.
Article history
Received:
June 19 2007
Citation
N.G. Karagüler, R.B. Sessions, B. Binay, E.B. Ordu, A.R. Clarke; Protein engineering applications of industrially exploitable enzymes: Geobacillus stearothermophilus LDH and Candida methylica FDH. Biochem Soc Trans 1 December 2007; 35 (6): 1610–1615. doi: https://doi.org/10.1042/BST0351610
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.