B. Ngamsom
Development of a high throughput screening tool for biotransformations utilising a thermophilic L-aminoacylase enzyme
Ngamsom, B.; Hickey, A. M.; Greenway, G. M.; Littlechild, J. A.; Watts, P.; Wiles, C.
Authors
A. M. Hickey
G. M. Greenway
J. A. Littlechild
P. Watts
C. Wiles
Abstract
Micro-reactors containing a monolith-immobilised thermophilic l-aminoacylase, from Thermococcus litoralis, have been developed for use in biotransformation reactions and a study has been carried out to investigate the stereospecificity and stability of the immobilised enzyme. The potential to use the developed micro-reactors as a tool for rapid screening of enzyme specificity was demonstrated, confirming that the l-aminoacylase showed a similar substrate specificity to that previously reported of the free enzyme. From this baseline, the technique was employed as a tool to evaluate potential unreported substrates with N-benzoyl- (l-threonine, l-leucine and l-arginine) and N-acetyl- (d,l-serine, d,l-leucine, l-tyrosine and l-lysine) protecting groups. The order of preferred substrates was found to be Phe > Thr > Leu > Arg for N-benzoyl substrates and Phe ≫ Ser > Leu > Met > Tyr > Trp for N-acetyl substrates. It was found that by using the micro-reactor a significantly smaller quantity of enzyme and substrates was required. It was shown that the micro-reactors were still operational in the presence of selected organic solvents, such as ethanol, methanol, acetone, dimethylformamide (DMF) and dimethylsulfoxide (DMSO). The results indicated that a combination of a small amount of an appropriate solvent (5% DMSO) and a higher reaction temperature could be employed in biotransformations where substrate solubility was an issue. © 2009 Elsevier B.V. All rights reserved.
Citation
Ngamsom, B., Hickey, A. M., Greenway, G. M., Littlechild, J. A., Watts, P., & Wiles, C. (2010). Development of a high throughput screening tool for biotransformations utilising a thermophilic L-aminoacylase enzyme. Journal of Molecular Catalysis B: Enzymatic, 63(1-2), 81-86. https://doi.org/10.1016/j.molcatb.2009.12.013
Journal Article Type | Article |
---|---|
Acceptance Date | Dec 14, 2009 |
Online Publication Date | Dec 24, 2009 |
Publication Date | 2010-04 |
Journal | JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC |
Print ISSN | 1381-1177 |
Publisher | Elsevier |
Peer Reviewed | Peer Reviewed |
Volume | 63 |
Issue | 1-2 |
Pages | 81-86 |
DOI | https://doi.org/10.1016/j.molcatb.2009.12.013 |
Keywords | Process Chemistry and Technology; Biochemistry; Bioengineering; Catalysis |
Public URL | https://hull-repository.worktribe.com/output/391490 |
Publisher URL | https://www.sciencedirect.com/science/article/pii/S1381117709003294?via%3Dihub |
You might also like
Electro-catalytic reactions
(2009)
Thesis
Pollution reduction with processed waste materials
(2013)
Thesis
Downloadable Citations
About Repository@Hull
Administrator e-mail: repository@hull.ac.uk
This application uses the following open-source libraries:
SheetJS Community Edition
Apache License Version 2.0 (http://www.apache.org/licenses/)
PDF.js
Apache License Version 2.0 (http://www.apache.org/licenses/)
Font Awesome
SIL OFL 1.1 (http://scripts.sil.org/OFL)
MIT License (http://opensource.org/licenses/mit-license.html)
CC BY 3.0 ( http://creativecommons.org/licenses/by/3.0/)
Powered by Worktribe © 2024
Advanced Search