Biosynthesis of mogrosides
AssigneeGinkgo Bioworks, Inc.
InventorsGuillaume Beaudoin, Alexandra Exner, Annapurna Kamineni, Matthew McMahon, Joshua Trueheart
Life of this patent
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Abstract
Described in this application are proteins and host cells involved in methods of producing mogrol precursors, mogrol, and/or mogrosides.
Claims107
A host cell for producing mogrol, one or more mogrol precursors, and/or one or more mogrosides, 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:
- (a) one or more mogrol precursors selected from the group consisting of:
- squalene, 2-3-oxidosqualene, 2,3,22,23-dioxidosqualene, cucurbitadienol, 24, 25-expoxycucurbitadienol, 11-hydroxycucurbitadienol, 11-hydroxy-24,25-epoxycucurbitadienol, 11-hydroxycucurbitadienol, 11-oxo-cucurbitadienol, and 24,25-dihydroxycucurbitadienol;
- (b) mogrol;
- and/or
- (c) one or more mogrosides.
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 method of producing mogrol, one or more mogrol precursors, and/or one or more mogrosides comprising culturing a host cell that comprises a heterologous polynucleotide encoding a lanosterol synthase, wherein the lanosterol synthase comprises an amino acid substitution or deletion relative to SEQ ID NO:
- 1 at one or more residues corresponding to position 14, 33, 47, 50, 66, 80, 83, 85, 92, 94, 107, 122, 132, 145, 158, 170, 172, 184, 193, 197, 198, 212, 213, 227, 228, 231, 235, 248, 249, 260, 282, 286, 287, 289, 295, 296, 309, 314, 316, 329, 344, 360, 370, 371, 372, 398, 407, 414, 417, 423, 432, 437, 442, 444, 452, 474, 479, 491, 498, 515, 526, 529, 536, 544, 552, 559, 560, 564, 578, 586, 608, 610, 617, 619, 620, 631, 638, 650, 655, 660, 679, 686, 702, 710, 726, 736, 738, and/or 742 in SEQ ID NO:
- 1 and wherein the host cell is capable of producing:
- (a) one or more mogrol precursors selected from the group consisting of:
- squalene, 2-3-oxidosqualene, 2,3,22,23-dioxidosqualene, cucurbitadienol, 24, 25-expoxycucurbitadienol, 11-hydroxycucurbitadienol, 11-hydroxy-24,25-epoxycucurbitadienol, 11-hydroxycucurbitadienol, 11-oxo-cucurbitadienol, and 24,25-dihydroxycucurbitadienol;
- (b) mogrol;
- and/or
- (c) one or more mogrosides.
A method of producing mogrol, one or more mogrol precursors, and/or one or more mogrosides comprising culturing a host cell that comprises a heterologous polynucleotide encoding a lanosterol synthase, wherein the lanosterol synthase comprises an amino acid substitution or deletion relative to SEQ ID NO:
- 313 at one or more residues corresponding to position 64, 120, 121, 136, 226, 268, 275, 281, 300, 322, 333, 438, 502, 604, 619, 628, 656, 693, 726, 727, 728, 729, 730, and/or 731.
Dependencies are inferred from the claim text.