Bidirectional multi-enzymatic scaffolds for biosynthesizing cannabinoids

A host cell capable of producing one or more cannabinoids, said host cell comprising:

• (a) a first exogenous nucleic acid encoding a first polypeptide having CBGA synthase activity and comprising a first heterologous interaction domain,
• (b) a second exogenous nucleic acid encoding a second polypeptide having olivetolic acid cyclase activity and comprising a second heterologous interaction domain,
• (c) a third exogenous nucleic acid encoding a third polypeptide having olivetol synthase activity and comprising a third heterologous interaction domain,
• (d) a fourth exogenous nucleic acid encoding a fourth polypeptide having trans-2-enoyl-CoA reductase activity and comprising a fourth heterologous interaction domain,
• (e) a fifth exogenous nucleic acid encoding a fifth polypeptide having enoyl-CoA hydratase activity and comprising a fifth heterologous interaction domain,
• (f) a sixth exogenous nucleic acid encoding a sixth polypeptide having 3-hydroxybutyryl-CoA dehydrogenase activity and comprising a sixth heterologous interaction domain,
• (g) a seventh exogenous nucleic acid encoding a seventh polypeptide having acetyl-CoA acetyltransferase activity and comprising a seventh heterologous interaction domain,
• (h) an eighth exogenous nucleic acid encoding an eighth polypeptide having ATP citrate lyase activity and comprising an eighth heterologous interaction domain,
• (i) a ninth exogenous nucleic acid encoding a ninth polypeptide having geranyl pyrophosphate synthase activity and comprising a ninth heterologous interaction domain,
• (j) a tenth exogenous nucleic acid encoding a tenth polypeptide having isopentyl-diphosphate isomerase activity and comprising a tenth heterologous interaction domain,
• (k) an eleventh exogenous nucleic acid encoding an eleventh polypeptide having diphospho-mevalonate decarboxylase activity and comprising an eleventh heterologous interaction domain,
• (l) a twelfth exogenous nucleic acid encoding a twelfth polypeptide having phosphomevalonate kinase activity and comprising a twelfth heterologous interaction domain,
• (m) a thirteenth exogenous nucleic acid encoding a thirteenth polypeptide having mevalonate kinase activity and comprising a thirteenth heterologous interaction domain,
• (n) a fourteenth exogenous nucleic acid encoding a fourteenth polypeptide having HMG-CoA reductase activity and comprising a fourteenth heterologous interaction domain,
• (o) a fifteenth exogenous nucleic acid encoding a fifteenth polypeptide having HMG-CoA synthase activity and comprising a fifteenth heterologous interaction domain, and
• (p) a sixteenth exogenous nucleic acid encoding a polypeptide scaffold comprising a peptide ligand for each of said first to fifteenth heterologous interaction domains,
• wherein each of said first to fifteenth heterologous interaction domains is different,
• wherein each peptide ligand for each of said first to fifteenth heterologous interaction domains is different,
• wherein said polypeptide scaffold comprises, in an order extending in a first direction away from said peptide ligand for said first heterologous interaction domain, (1) said peptide ligand for said second heterologous interaction domain, (2) said peptide ligand for said third heterologous interaction domain, (3) said peptide ligand for said fourth heterologous interaction domain, (4) said peptide ligand for said fifth heterologous interaction domain, (5) said peptide ligand for said sixth heterologous interaction domain, (6) said peptide ligand for said seventh heterologous interaction domain, and (7) said peptide ligand for said eighth heterologous interaction domain, and
• wherein said polypeptide scaffold comprises, in an order extending in the other direction away from said peptide ligand for said first heterologous interaction domain, (1) said peptide ligand for said ninth heterologous interaction domain, (2) said peptide ligand for said tenth heterologous interaction domain, (3) said peptide ligand for said eleventh heterologous interaction domain, (4) said peptide ligand for said twelfth heterologous interaction domain, (5) said peptide ligand for said thirteenth heterologous interaction domain, (6) said peptide ligand for said fourteenth heterologous interaction domain, (7) said peptide ligand for said fifteenth heterologous interaction domain, (8) said peptide ligand for said seventh heterologous interaction domain, and (9) said peptide ligand for said eighth heterologous interaction domain.