Method and means for manufacturing terpene indole alkaloids

An expression vector comprising

• (a) a nucleic acid encoding a first enzyme which is a precondylocarpine acetate synthase comprising the amino acid sequence of SEQ ID NO:
• 1 or a functional variant or homologue thereof comprising an amino acid sequence having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 1;
• (b) a nucleic acid encoding a second enzyme which is (i) a dehydroprecondylocarpine acetate synthase comprising the amino acid sequence of SEQ ID NO:
• 2 or a functional variant or homologue thereof comprising an amino acid sequence having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 2, or (ii) a dehydroprecondylocarpine acetate synthase comprising the amino acid sequence of SEQ ID NO:
• 66 or a functional variant or homologue thereof having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 66;
• (c) a nucleic acid encoding a third enzyme which is a catharanthine synthase comprising the amino acid sequence of SEQ ID NO:
• 3 or a functional variant or homologue thereof comprising an amino acid sequence having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 3;
• (d) a nucleic acid encoding a fourth enzyme which is a tabersonian synthase comprising the amino acid sequence of SEQ ID NO:
• 4 or a functional variant or homologue thereof having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 4;
• or
• (e) a combination of (a), (b), (c) or (d).

A host cell comprising

• (a) a nucleic acid encoding a first enzyme which is a precondylocarpine acetate synthase comprising the amino acid sequence of SEQ ID NO:
• 1 or a functional variant or homologue thereof comprising an amino acid sequence having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 1;
• (b) a nucleic acid encoding a second enzyme which is (i) a dehydroprecondylocarpine acetate synthase comprising the amino acid sequence of SEQ ID NO:
• 2 or a functional variant or homologue thereof comprising an amino acid sequence having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 2, or (ii) a dehydroprecondylocarpine acetate synthase comprising the amino acid sequence of SEQ ID NO:
• 66 or a functional variant or homologue thereof having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 66;
• (c) a nucleic acid encoding a third enzyme which is a catharanthine synthase comprising the amino acid sequence of SEQ ID NO:
• 3 or a functional variant or homologue thereof comprising an amino acid sequence having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 3;
• (d) a nucleic acid encoding a fourth enzyme which is a tabersonian synthase comprising the amino acid sequence of SEQ ID NO:
• 4 or a functional variant or homologue thereof having at least 90% sequence identity with and over a full length of SEQ ID NO:
• 4;
• or
• a combination of (a), (b), (c) or (d).