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 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.
Dadd
M R
MR
Oxford Centre for Environmental Biotechnology, Center of Ecology and Hydrology, UK. mrdad@ceh.ac.uk
Sharp
D C
DC
Pettman
A J
AJ
Knowles
C J
CJ
D000097:Acetonitriles / Q000378:metabolism; D003470:Culture Media; D009570:Nitriles / Q000378:metabolism*; D012240:Rhodococcus / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared
D000097:Acetonitriles; D003470:Culture Media; D009570:Nitriles; C014356:benzonitrile; C032159:acetonitrile