Biosynthesis of isoprenoids and precursors thereof
AssigneeGinkgo Bioworks, Inc.
InventorsGuillaume Beaudoin, Elena Brevnova, Alkiviadis Orfefs Chatzivasileiou, Alexandra Exner, Annapuma Kamineni, Matthew McMahon, Joshua Trueheart
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Abstract
Described in this application are proteins and host cells involved in methods of producing isoprenoid precursors and/or isoprenoids.
Claims145
A host cell for producing an isoprenoid precursor or isoprenoid, wherein the host cell comprises a heterologous polynucleotide encoding a lanosterol synthase with reduced activity as compared to a wild-type lanosterol synthase, wherein the host cell is capable of producing more of an isoprenoid or isoprenoid precursor as compared to a control host cell that does not comprise the heterologous polynucleotide.
A host cell that comprises a heterologous polynucleotide encoding a lanosterol synthase, wherein the lanosterol synthase comprises a sequence that is at least 90% identical to SEQ ID NO:
- 3, 83-87, 89-92, 94-95, 99, 100-102, 118-120, 316-319, 321-326, 329, or 331.
A host cell that comprises a heterologous polynucleotide encoding a lanosterol synthase, wherein the lanosterol synthase comprises relative to SEQ ID NO:
- 1:
- a) R33Q, R193C, D289G, N295I, S296T, N620S, and Y736F;
- b) K47E, L92I, T360S, S372P, T444M, and R578P;
- c) D50G, K66R, N94S, G417S, E617V, and F726L;
- d) N14Y, N132S, Y145C, R193H, I286F, L316R, F432I, E442V, T444S, I479S, K631R, and T655A;
- e) E287G, K329N, E617V, and F726V;
- f) E231V, A407V, Q423L, A529T, and Y564C;
- g) V248F, D371V, and G702D;
- h) G122C, H249L, and K738M;
- or
- i) K85N, G158S, S515L, P526T, and Q619L, and a truncation resulting in a deletion of the residue corresponding to Q742 in SEQ ID NO:
- 1.
A host cell that comprises a heterologous polynucleotide encoding a lanosterol synthase, wherein the heterologous polynucleotide comprises a sequence that is at least 90% identical to SEQ ID NO:
- 4, 62-66, 68-71, 73-74, 78, 80-82, 103-109, 111-117, 328, or 330.
A host cell for producing an isoprenoid precursor or isoprenoid, wherein the host cell comprises:
- (a) a heterologous polynucleotide encoding a squalene epoxidase with reduced activity as compared to a wild-type squalene epoxidase;
- or
- (b) a heterologous polynucleotide that reduces squalene epoxidase activity, wherein the host cell is capable of producing more of an isoprenoid or isoprenoid precursor as compared to a control host cell that does not comprise the heterologous polynucleotide of (a) and/or (b).
A host cell comprising:
- (a) one or more enzymes in the yeast mevalonate pathway;
- and
- (b) a heterologous polynucleotide encoding a lanosterol synthase with reduced activity as compared to a wild-type lanosterol synthase;
- and/or
- (c) a heterologous polynucleotide encoding a squalene epoxidase with reduced activity as compared to a wild-type squalene epoxidase;
- or
- (d) a heterologous polynucleotide that reduces squalene epoxidase activity.
A host cell comprising:
- (a) one or more enzymes in the Archaea I mevalonate pathway;
- and
- (b) a heterologous polynucleotide encoding a lanosterol synthase with reduced activity as compared to a wild-type lanosterol synthase;
- or
- (c) a heterologous polynucleotide that reduces lanosterol synthase activity;
- and/or
- (d) a heterologous polynucleotide encoding a squalene epoxidase with reduced activity as compared to a wild-type squalene epoxidase;
- or
- (e) a heterologous polynucleotide that reduces squalene epoxidase activity.
A host cell comprising:
- (a) one or more enzymes in the Archaea II mevalonate pathway;
- and
- (b) a heterologous polynucleotide encoding a lanosterol synthase with reduced activity as compared to a wild-type lanosterol synthase;
- or
- (c) a heterologous polynucleotide that reduces lanosterol synthase activity;
- and/or
- (d) a heterologous polynucleotide encoding a squalene epoxidase with reduced activity as compared to a wild-type squalene epoxidase;
- or
- (e) a heterologous polynucleotide that reduces squalene epoxidase activity.
A host cell comprising:
- (a) one or more enzymes in the MEP pathway;
- and
- (b) a heterologous polynucleotide encoding a lanosterol synthase with reduced activity as compared to a wild-type lanosterol synthase;
- or
- (c) a heterologous polynucleotide that reduces lanosterol synthase activity;
- and/or
- (d) a heterologous polynucleotide encoding a squalene epoxidase with reduced activity as compared to a wild-type squalene epoxidase;
- or
- (e) a heterologous polynucleotide that reduces squalene epoxidase activity.
A method of producing an isoprenoid precursor or isoprenoid comprising culturing a host that comprises:
- (a) a heterologous polynucleotide encoding a lanosterol synthase with reduced activity as compared to a wild-type lanosterol synthase;
- and/or
- (b) a heterologous polynucleotide encoding a squalene epoxidase with reduced activity as compared to a wild-type squalene epoxidase;
- or
- (c) a heterologous polynucleotide that reduces squalene epoxidase activity, wherein the host cell is capable of producing more of an isoprenoid or isoprenoid precursor as compared to a control host cell that does not comprise one or more of (a)-(c).
Dependencies are inferred from the claim text.