Dadd M R, Sharp D C, Pettman A J, Knowles C J
Oxford Centre for Environmental Biotechnology, Center of Ecology and Hydrology, UK.
J Microbiol Methods. 2000 Jun;41(1):69-75. doi: 10.1016/s0167-7012(00)00138-x.
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.
在本研究中，利用红球菌LL100 - 21的全细胞悬浮液，采用中红外光谱法跟踪腈生物催化过程中的酶动力学。细菌在乙腈上生长以诱导两步酶促途径。乙腈通过腈水合酶生物转化为乙酰胺，随后通过酰胺酶生物转化为乙酸（羧酸根离子）。细菌也在苯甲腈上生长以诱导一步酶促途径。苯甲腈通过腈水解酶直接生物转化为苯甲酸（羧酸根离子）。使用硅探针通过React IR跟踪这些反应，得到了腈生物催化反应的出色定量和定性实时数据。该研究表明，这项新技术在生物催化中具有潜在的有用应用。
