Materials and methods for producing alkaloids

A host cell able to produce more of at least one loline alkaloid or precursor thereof, than does a control cell, as a result of the host cell being transformed or modified to comprise at least the following polynucleotides:

• i) a polynucleotide encoding a polypeptide comprising the sequence of any one of SEQ ID NO:1, 12, 13 and 61 or a variant thereof with at least 70% identity to any one of SEQ ID NO:1, 12, 13 and 61 with activity of a gamma-class pyridoxal phosphate (PLP) enzyme,
• ii) a polynucleotide encoding a polypeptide comprising the sequence of any one of SEQ ID NO:2, 14, 15 and 62 or a variant thereof with at least 70% identity to any one of SEQ ID NO:2, 14, 15 and 62 with activity of an alpha-class PLP enzyme,
• iii) a polynucleotide encoding a polypeptide comprising the sequence of any one of SEQ ID NO:3, 16, 17 and 63 or a variant thereof with at least 70% identity to of any one of SEQ ID NO:3, 16, 17 and 63 with monooxygenase activity, and
• iv) a polynucleotide encoding a polypeptide comprising the sequence of any one of SEQ ID NO:5, 20, 21 and 64 or a variant thereof with at least 70% identity to of any one of SEQ ID NO:5, 20, 21 and 64 with activity of an alpha-class PLP enzyme;
• and
• wherein the host cell is tolerant of endogenous (3-amino-3-carboxypropyl)proline (ACPP) production;
• wherein the genome of the untransformed (wild type) host cell does not prior to transformation with the polynucleotides of i)-iv) comprise one or more LOL genes selected from lolC, lolD, lolF and lolT;
• and
• wherein the host cell is a fungal host cell.