Purification and characterization of a furfural reductase (FFR) from Escherichia coli strain LYO1 - An enzyme important in the detoxification of furfural during ethanol production

Tony Gutierrez, L O Ingram, J F Preston

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78 Citations (Scopus)

Abstract

Furfural, an inhibitor of ethanol production from hemicellulose acid hydrolysates, is reductively detoxified to furfuryl alcohol by the ethanologenic bacterium Escherichia coli strain LYO1. Furfural reductase was purified 106-fold from this bacterium to approximately 50% homogeneity. It has a native molecular mass of 135 kDa, determined by gel filtration, and subunit molecular mass of similar to 68kDa, determined by denaturing gel electropboresis, indicating the holoenzyme is a dimer of two similar if not identical subunits. The enzyme shows strong activity from pH 4 to 8 (optimum pH 7.0), relatively high temperature tolerance (50-55 degrees C), and an apparent K-m and V-max for furfural of 1.5 x 10(-4) M and 28.5 mu mol/min/mg of protein, respectively. It catalyzes the essentially irreversible reduction of furfural with NADPH, is specific for NADPH as. cofactor, and is relatively specific for the reduction of furfural and benzaldehyde; 2-acetylfuran, xylose, and glucose were not reduced, while acetaldehyde was reduced at a rate 25-fold lower than furfural. This is the first description of a furfural reductase which, based upon size and substrate specificity, appears to represent a new type of alcohol-aldehyde oxido-reductase. The conversion of relatively toxic furfural to less toxic furfuryl alcohol suggests a beneficial role for this enzyme in mitigating furfural toxicity encountered during ethanol production from lignocellulosic biomass. (c) 2005 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)154-164
Number of pages11
JournalJ BIOTECHNOL
Volume121
Issue number2
DOIs
Publication statusPublished - 2006

Keywords

  • SACCHAROMYCES-CEREVISIAE
  • STRICTLY ANAEROBIC BACTERIUM
  • Biotechnology & Applied Microbiology
  • INHIBITION
  • XYLOSE FERMENTATION
  • OXIDOREDUCTASES
  • GROWTH
  • ALCOHOL-DEHYDROGENASE
  • ACID
  • BATCH FERMENTATION
  • CONVERSION

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