araban and arabitol

Hypocrea jecorina (anamorph: Trichoderma reesei) can grow on plant arabinans by the aid of secreted arabinan-degrading enzymes. This growth on arabinan and its degradation product L-arabinose requires the operation of the aldose reductase XYL1 and the L-arabinitol dehydrogenase LAD1. Growth on arabinan and L-arabinose is also severely affected in a strain deficient in the general cellulase and hemicellulase regulator XYR1, but this impairment can be overcome by constitutive expression of the xyl1 encoding the aldose reductase. An inspection of the genome of H. jecorina reveals four genes capable of degrading arabinan, i.e., the alpha-L-arabinofuranosidase encoding genes abf1, abf2, and abf3 and also bxl1, which encodes a beta-xylosidase with a separate alpha-L-arabinofuranosidase domain and activity but no endo-arabinanase. Transcriptional analysis reveals that in the parent strain QM9414 the expression of all of these genes is induced by L-arabinose and to a lesser extent by L-arabinitol and absent on D-glucose. Induction by L-arabinitol, however, is strongly enhanced in a Deltalad1 strain lacking L-arabinitol dehydrogenase activity and severely impaired in an aldose reductase (Deltaxyl1) strain, suggesting a cross talk between L-arabinitol and the aldose reductase XYL1 in an alpha-L-arabinofuranosidase gene expression. Strains bearing a knockout in the cellulase regulator xyr1 do not show any induction of abf2 and bxl1, and this phenotype cannot be reverted by constitutive expression of xyl1. The loss of function of xyr1 has also a slight effect on the expression of abf1 and abf3. We conclude that the expression of the four alpha-L-arabinofuranosidases of H. jecorina for growth on arabinan requires an early pathway intermediate (L-arabinitol or L-arabinose), the first enzyme of the pathway XYL1, and in the case of abf2 and bxl1 also the function of the cellulase regulator XYR1.

Topics: Aldehyde Reductase; Arabinose; Base Sequence; Cellulase; Consensus Sequence; Gene Expression Regulation, Fungal; Glycoside Hydrolases; Hypocrea; Molecular Sequence Data; Phylogeny; Polysaccharides; Sugar Alcohol Dehydrogenases; Sugar Alcohols; Transcription, Genetic; Trichoderma

This paper describes two Aspergillus niger mutants (araA and araB) specifically disturbed in the regulation of the arabinanase system in response to the presence of L-arabinose. Expression of the three known L-arabinose-induced arabinanolytic genes, abfA, abfB and abnA, was substantially decreased or absent in the araA and araB strains compared to the wild-type when incubated in the presence of L-arabinose or L-arabitol. In addition, the intracellular activities of L-arabitol dehydrogenase and L-arabinose reductase, involved in L-arabinose catabolism, were decreased in the araA and araB strains. Finally, the data show that the gene encoding D-xylulose kinase, xkiA, is also under control of the arabinanolytic regulatory system. L-Arabitol, most likely the true inducer of the arabinanolytic and L-arabinose catabolic genes, accumulated to a high intracellular concentration in the araA and araB mutants. This indicates that the decrease of expression of the arabinanolytic genes was not due to lack of inducer accumulation. Therefore, it is proposed that the araA and araB mutations are localized in positive-acting components of the regulatory system involved in the expression of the arabinanase-encoding genes and the genes encoding the L-arabinose catabolic pathway.

Topics: Arabinose; Aspergillus niger; Fungal Proteins; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Glycoside Hydrolases; Mutation; Oxidoreductases; Polysaccharides; Sugar Alcohols; Xylans