Thursday, April 22, 2010 - 4:00 PM
11-06

Furaldehyde metabolism of Cupriavidus basilensis HMF14: Opportunities for feedstock detoxification and chemicals production

Harald J. Ruijssenaars1, Nick J.P. Wierckx1, Frank W. Koopman2, and Han De Winde2. (1) BIRD Engineering, Westfrankelandsedijk 1, B-Basic, Schiedam, 3115HG, Netherlands, (2) Dept of Biotechnology, Delft University of Technology, Julianalaan 67, Delft, 2628BC, Netherlands

The toxic fermentation inhibitors in lignocellulosic hydrolysates, like the furaldehydes 5-hydroxymethylfurfural (HMF) and furfural, put serious constraints on the efficient production of second-generation biofuels and –chemicals. Recently, we isolated a novel HMF and furfural metabolizing bacterium, Cupriavidus basilensis HMF14, that also metabolizes inhibitors such as acetate, formate and aromatic compounds. Remarkably, this bacterium does not utilize the most abundant sugars in lignocellulosic hydrolysates: glucose, xylose and arabinose. The furaldehyde catabolic pathways of this strain were elucidated using a transposon mutagenesis approach. Degradation of furfural was found to occur via CoA ligation of furoic acid, eventually leading to the formation of 2-ketoglutarate, whereas HMF was degraded via 2,5-furandicarboxylic acid (FDCA). Various approaches were taken to employ the metabolic capacities of C. basilensis HMF14. One application is the biological detoxification of hydrolysates: when incubated with wheat straw hydrolysate, C. basilensis HMF14 completely removed furfural, HMF, acetate and formate while the sugar fraction (80 g/l) was left intact. In another approach, the furaldehyde metabolic pathways of C. basilensis HMF were introduced into a heterologous host, resulting in a strain that removes the furaldehydes during fermentation. Finally, the enzyme of the HMF metabolic pathway involved in FDCA formation was utilized to construct a Pseudomonas putida-based whole-cell biocatalyst for the efficient production of this high-potential biobased platform chemical.

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