madrid2i

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In one aspect there is provided a method for producing a recombinant enzyme capable of hydrolysing chlorophyll or a chlorophyll derivative, comprising a step of intracellular expression of the recombinant enzyme in a eukaryotic host cell.

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CLAIMS 1. A method for producing a recombinant enzyme capable of hydrolysing chlorophyll or a chlorophyll derivative, comprising a step of intracellular expression of the recombinant enzyme in a eukaryotic host cell. 2. A method according to claim 1 , wherein the host cell is a fungal cell. 3. A method according to claim 2, wherein the host cell is derived from Trichoderma spp. 4. A method according to any preceding claim, wherein the recombinant enzyme is a chlorophyllase. 5. A method according to any preceding claim, wherein the enzyme comprises one or more amino acid sequences selected from GHSXGG (SEQ ID NO:36), DPVXG (SEQ ID NO:37) and YGHXD (SEQ ID NO:38). 6. A method according to any preceding claim, wherein the recombinant enzyme is derived from Arabidopsis thaliana, Brassica oleracea, Ricinus communis, Ginkgo biloba, Populus trichocarpa, Vitis vinifera, Phyllostachys heterocycla, Sorghum bicolor, Glycine max, Pachira macrocarpa, Triticum aestivum, Citrus sinensis, Zea mays, Chenopodium album, Picea sitchensis or Chlamydomonas reinhardtii. 7. A method according to any preceding claim, wherein the recombinant enzyme comprises a polypeptide sequence as defined in any one of SEQ ID NO:s 1 to 18, or a functional fragment or variant thereof having at least 75% sequence identity to any one of SEQ ID NO:s 1 to 18 over at least 50 amino acid residues. 8. A method according to any preceding claim, wherein the recombinant enzyme is encoded by a nucleic acid sequence as defined in any one of SEQ ID NO:s 19 to 35, or a functional fragment or variant thereof having at least 75% sequence identity to any of SEQ ID NO:s 19 to 35 over at least 50 nucleotide residues. 9. A method according to any preceding claim, further comprising lysing the host cells and recovering the recombinant enzyme from the lysate. 10. A method according to claim 8, wherein the host cells are exposed to a detergent, an organic acid, homogenization and/or a heat treatment. 11. A eukaryotic expression vector, comprising a nucleic acid sequence encoding an enzyme capable of hydrolysing chlorophyll or a chlorophyll derivative, operably linked to one or more control sequences suitable for directing intracellular expression of the enzyme in a eukaryotic host cell. 12. An expression vector according to claim 10, comprising a nucleic acid sequence as defined in any one of SEQ ID NO:s 19 to 35, or a functional fragment or variant thereof having at least 75% sequence identity to any of SEQ ID NO:s 19 to 35 over at least 50 nucleotide residues. 13. A eukaryotic host cell comprising an expression vector as defined in claim 10 or claim 11. 14. A host cell according to claim 12, which is a fungal cell. 15. A recombinant enzyme capable of hydrolyzing chlorophyll or a chlorophyll derivative, which is obtainable by a method as defined in any of claims 1 to 9. 16. A recombinant enzyme according to claim 14, wherein the recombinant enzyme comprises a polypeptide sequence as defined in any one of SEQ ID NO:s 1 to 18, or a functional fragment or variant thereof having at least 75% sequence identity to any one of SEQ ID NO:s 1 to 18 over at least 50 amino acid residues.

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