Mechanism of the enzymatic synthesis of furan-containing compound

This research project stems from the desire to explore and eventually harness the enzymatic mechanism of 4-(hydroxymethyl)-2-furan-
carboxaldehyde-phosphate synthase (MfnB). The enzyme of interest, MfnB, is found most prominently in methanogens and has the ability to catalyze five or more separate chemical transformations in a single active site. This singular enzyme takes two molecules of glyceraldehyde-3-phosphate to create a furan-containing compound 4-(hydroxymethyl)-2-furan-carboxaldehyde-phosphate. The US. Department of Energy (DOE) has published a list of the “Top 10 + 4” bio-based chemicals, with furan-containing compounds highlighted for their high potential in the production of biofuels and biomaterial compounds(1). As one can imagine, industrial applications of MfnB might usher in a new era for the synthesis of furan compounds to be used in the generation of liquid fuel or other biomaterial via enzyme-catalyzed reactions. Despite the initial characterization of MfnB and the identification of Schiff base-forming lysine, the detail mechanism of MfnB remains speculative (2). Our team seeks to understand the details of catalytic mechanism through site-directed mutagenesis, comprehensive kinetic evaluation and structural study. Uncovering the molecular basis of this catalytic mystery will help us in applying the knowledge to enzyme engineering. The engineered catalytic machinery can then be used for generating building block compounds to meet the needs of industrial biofuel and other biomaterial products.

1. Werpy, T.; Petersen, G. Top Value Added Chemicals from Biomass Vol. I-Results of Screening for Potential Candidates from Sugars and Synthesis Gas, NREL/TP-510-35523; National Renewable Energy Laboratory: Golden, CO, 2004.

2. Wang, Y., Jones, M. K., Xu, H., Ray, W. K., and White, R. H. (2015) Mechanism of the Enzymatic Synthesis of 4-(Hydroxymethyl)-2-furancarboxaldehyde-phosphate (4-HFC-P) from Glyceraldehyde-3-phosphate catalyzed by 4-HFC-P synthase. Biochemistry 54, 2997–3008