Enoate reductases are a promising class of biocatalysts which have been shown to reduce the carbon-carbon double bonds of cis,cis-muconic acid in vivo, generating adipic acid, an important precursor used in the synthesis of nylon-6,6. Bacillus coagulans (ERBC) is a well researched enoate reductase proven to work with several catechol ring cleavage products. Our research has shown that ERBC is capable of reducing carbon-carbon double bonds in a variety of molecules produced using the extradiol dioxygenase BphC. Since the native substrate of ERBC is unknown, studying its activity with a variety of similar substrates will be beneficial for evaluating the scope of its reactivity. Our research aims to catalogue viable substrates using UV-visible light spectroscopy and to characterize enzymatic products through high performance liquid chromatography (HPLC) analysis. Furthermore, optimizing these reaction conditions will permit high throughput product formation and isolation. Identifying substrates and subsequent enhancement of the catalytic activity of ERBC can enable the development of environmentally benign synthetic methods for the production of a variety of commodity chemicals. In the future, other enoate reductases will be studied to evaluate their potential as viable candidates for the adipic acid production pathway.
