Engineering Fungi for the Utilisation of L-Arabinose
AssigneeValtion Teknillinen Tutkimuskeskus
InventorsJohn Londesborough, Merja Penttila, Peter Richard
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Abstract
A fungal microorganism can be engineered by means of genetic engineering to utilise L-arabinose. The genes of the L-arabinose pathway, which were unknown, i.e. L-arabinitol 4-dehydrogenase and L-xylulose reductase, were identified. These genes, together with the known genes of the L-arabinose pathway, form a functional pathway. This pathway can be introduced to a fungus, which is completely or partially lacking this pathway.
Claims11
A method for producing ethanol, lactic acid or xylitol using fungal biofermentation of L-arabinose, which comprises
- (a) providing a first fungus that is unable to use or is inefficient in metabolising L-arabinose;
- (b) providing a nucleic acid that
- (1) encodes a fungal enzyme selected from the group consisting of LA4D and LXR wherein said LA4D converts L-arabinitol and NAD to L-xylulose and NADH and wherein said LXR converts xylitol and NADP to L-xylulose and NADPH;
- (2) is contained within the genomic DNA of a second fungus that is able to efficiently metabolise L-arabinose;
- and
- (3) is amplifiable from said genomic DNA by PCR, using the primers of SEQ ID NOS:13 and 14 when said nucleic acid encodes LA4D and the primers of SEQ ID NOS:15 and 16 when said nucleic acid encodes LXR;
- (c) expressing said nucleic acid in said first fungus;
- and
- (d) performing biofermentation by growing said first fungus in the presence of L-arabinose to produce ethanol, lactic acid or xylitol.
A method for producing ethanol, lactic acid or xylitol using fungal biofermentation of L-arabinose, which comprises
- (a) providing a fungus that is unable to use or is inefficient in metabolising L-arabinose;
- (b) providing a nucleic acid that encodes a protein sequence selected from the group consisting of SEQ ID NOs:17 and 18;
- (c) expressing said nucleic acid in said fungus;
- and
- (d) performing biofermentation by growing said fungus in the presence of L-arabinose to produce ethanol, lactic acid or xylitol.
A method of modifying the L-arabinose metabolic pathway of a fungus to allow metabolic conversion of L-arabinose to ethanol, lactic acid or xylitol, which comprise transferring to said fungus an LA4D-encoding nucleic acid from Penicillium chrysogenum or Aspergillus niger, an LXR-encoding nucleic acid from Erwinia uredovora or Aspergillus niger, or both.
A method of producing a useful product from biomass containing L-arabinose, which comprises:
- (a) providing a genetically modified fungus which is transformed with a DNA encoding an enzyme selected from the group consisting of L-arabinitol 4-dehydrogenase (LA4D) and L-xylulose reductase (LXR), each of the fungal L-arabinose pathway, wherein said fungus has an increased ability to utilise L-arabinose;
- and
- (b) fermenting said fungus under conditions suitable to produce said useful product,
- wherein the DNA is selected from the group consisting of nucleic acids that encode SEQ ID NOs:17 and 18.
Dependencies are inferred from the claim text.