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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

B. Ngamsom

A. M. Hickey

G. M. Greenway G.M.Greenway@hull.ac.uk

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.

Journal Article Type Article
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
APA6 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
DOI https://doi.org/10.1016/j.molcatb.2009.12.013
Keywords Process Chemistry and Technology; Biochemistry; Bioengineering; Catalysis
Publisher URL https://www.sciencedirect.com/science/article/pii/S1381117709003294?via%3Dihub
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