Biocatalytic production of l-fucose
AssigneeBASF SE
InventorsVaidotas Navickas, Michael Breuer, Michael Puhl, Melanie Weingarten, Kai-Uwe Baldenius, Wolfgang Siegel
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Abstract
The present invention relates to biocatalytic methods, comprising purely enzymatic, mixed enzymatic-fermentative and purely fermentative methods, for the direct single-step conversion of L-fucitol to L-fucose, in order to easily obtain L-fucose at high amounts and levels of purity. Suitable recombinant microorganisms and fungi are further disclosed and also the use thereof in said method for the single-step conversion to L-fucose.
Claims22
A method for producing L-fucose, comprising the following steps:
- (a) providing L-fucitol, a galactose oxidase of the enzyme class EC 1.1.3.9, a peroxidase and a catalase,
- (b) combining L-fucitol, the galactose oxidase, the peroxidase, and the catalase to form a mixture,
- (c) incubating the resulting mixture under conditions permitting the biocatalytic oxidation of L-fucitol to L-fucose, and
- (d) optionally isolating the synthesized L-fucose.
A method for producing L-fucose, comprising:
- adding L-fucitol to a culture medium of a recombinant microorganism or fungus expressing a transgene(s) in functional form, and
- culturing the mixture under conditions permitting the biocatalytic oxidation of L-fucitol to L-fucose,
- wherein the recombinant microorganism or fungus comprises a transgene which encodes a galactose oxidase of the enzyme class EC 1.1.3.9 from a fungus of the genus Fusarium, and further comprises a transgene selected from a peroxidase selected from a peroxidase of the group consisting of the enzyme class EC 1.11.1.7, and a catalase selected from a catalase of the group consisting of the enzyme class EC 1.11.1.6, wherein the recombinant microorganism or fungus is selected from the group consisting of Escherichia coli spp., Bacillus spp., Saccharomycetes, Kluyveromyces spp., Aspergillus spp., Trichoderma spp., and a fungus of the phylum Ascomycota.
A method for the biocatalytic conversion of L-fucitol to L-fucose, comprising the following steps:
- (a) providing a recombinant microorganism or fungus for the recombinant expression of a galactose oxidase, a peroxidase, and a catalase, wherein the microorganism comprises a transgene which encodes a galactose oxidase from a fungus of the genus Fusarium, a transgene which encodes a peroxidase, and a transgene with encodes a catalase, and wherein the recombinant microorganism or fungus is selected from the group consisting of:
- an Escherichia coli spp., a Bacillus spp., a Saccharomycetes, a Kluyverormyces spp., an Aspergillus spp., a Trichoderma spp., and a fungus of the phylum Ascomycota, or
- (b1) providing a first recombinant microorganism or fungus for the recombinant expression of a galactose oxidase, wherein the microorganism or fungus comprises a transgene encoding a galactose oxidase from a fungus of the genus Fusarium;
- (b2) providing a second recombinant microorganism or fungus for the recombinant expression of a peroxidase, wherein the microorganism or fungus comprises a transgene encoding a peroxidase selected from a peroxidase of the group consisting of the enzyme class EC 1.11.1.7;
- and
- (b3) providing a third recombinant microorganism or fungus for the recombinant expression of a catalase, wherein the microorganism or fungus comprises a transgene encoding a catalase selected from a catalase of the group consisting of the enzyme class EC 1.11.1.6;
- (c) culturing the recombinant microorganism and/or fungus under suitable conditions which permits the synthesis of the galactose oxidase, the peroxidase and the catalase,
- (d) optionally isolating the synthesized galactose oxidase and the peroxidase and/or the catalase;
- (e) combining L-fucitol, the galactose oxidase, the peroxidase, and the catalase to form a mixture;
- and
- (f) incubating the resulting mixture under conditions permitting the biocatalytic oxidation of L-fucitol to L-fucose.
Dependencies are inferred from the claim text.