Real-time monitoring of nitrile biotransformations by mid-infrared spectroscopy

In this study mid-infrared spectroscopy was used to follow the enzyme kinetics involved in nitrile biocatalysis using whole cell suspensions of the bacterium Rhodococcus rhodochrous LL100-21. The bacteria were grown on acetonitrile to induce a two-step enzymatic pathway. Acetonitrile was biotransfor...

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Bibliographic Details
Published in:Journal of microbiological methods 2000-06, Vol.41 (1), p.69-75
Main Authors: Dadd, Michael R., Sharp, David C.A., Pettman, Alan J., Knowles, Christopher J.
Format: Article
Language:English
Subjects:
Acetonitriles - metabolism
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biocatalysis
Bioconversions. Hemisynthesis
Biological and medical sciences
Biotechnology
Culture Media
Fundamental and applied biological sciences. Psychology
infrared spectroscopy
Methods. Procedures. Technologies
Microbiology
Mid-infrared spectroscopy
Nitrile hydrolysis
Nitriles - metabolism
On-line analysis
React IR
Rhodococcus
Rhodococcus - metabolism
Rhodococcus rhodochrous
Spectroscopy, Fourier Transform Infrared
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Summary:In this study mid-infrared spectroscopy was used to follow the enzyme kinetics involved in nitrile biocatalysis using whole cell suspensions of the bacterium Rhodococcus rhodochrous LL100-21. The bacteria were grown on acetonitrile to induce a two-step enzymatic pathway. Acetonitrile was biotransformed to acetamide by a nitrile hydratase enzyme and subsequently to acetic acid (carboxylate ion) by an amidase enzyme. The bacteria were also grown on benzonitrile to induce a one-step enzymatic pathway. Benzonitrile was biotransformed directly to benzoic acid (carboxylate ion) by a nitrilase enzyme. These reactions were followed by React IR using a silicon probe and gave excellent quantitative and qualitative real-time data of both nitrile biocatalytic reactions. This study has shown that this novel technique has potentially useful applications in biocatalysis.
ISSN:0167-7012
1872-8359
DOI:10.1016/S0167-7012(00)00138-X