US 12,442,009 B2
Sulfite tolerance in recombinant yeast host cells
Aaron Argyros, Lebanon, NH (US); Charles F. Rice, Plainfield, NH (US); Trisha Barrett, Bradford, VT (US); Michelle Oeser, Croydon, NH (US); and Janet Fisher, Norwich, VT (US)
Assigned to DANSTAR FERMENT AG, Zug (CH)
Appl. No. 16/472,742
Filed by Lallemand Hungary Liquidity Management LLC, Budapest (HU)
PCT Filed Dec. 22, 2017, PCT No. PCT/EP2017/084540
§ 371(c)(1), (2) Date Jun. 21, 2019,
PCT Pub. No. WO2018/115513, PCT Pub. Date Jun. 28, 2018.
Claims priority of provisional application 62/438,391, filed on Dec. 22, 2016.
Prior Publication US 2022/0090102 A1, Mar. 24, 2022
Int. Cl. C12N 1/20 (2006.01); C12N 1/18 (2006.01); C12N 15/81 (2006.01); C12R 1/865 (2006.01)
CPC C12N 15/81 (2013.01) [C12N 1/185 (2021.05); C12R 2001/865 (2021.05)] 14 Claims
 
1. A recombinant yeast host cell comprising:
(i) at least one first genetic modification selected from the group consisting of:
a heterologous nucleic acid molecule encoding a heterologous STL1 polypeptide, and
a heterologous nucleic acid molecule encoding a heterologous glucoamylase, wherein the heterologous glucoamylase comprises:
(i) the amino acid sequence of any one of SEQ ID NO: 13, 14, 15, 16, 17, 18, and 19,
(ii) a variant having at least 70% identity with the amino acid sequence of any one of SEQ ID NO: 13, 14, 15, 16, 17, 18, and 19, or
(iii) a fragment of (i) or (ii) having at least 70% identity with the amino acid sequence of any one of SEQ ID NO: 13, 14, 15, 16, 17, 18, and 19,
wherein the variant or the fragment has glucoamylase activity; and
(ii) at least one second genetic modification selected from the group consisting of:
a heterologous nucleic acid molecule encoding a heterologous transcription factor FZF1 polypeptide, wherein the heterologous transcription factor FZF1 polypeptide comprises:
(i) the amino acid sequence of any one of SEQ ID NO: 1 to 6, 21 and 22,
(ii) a variant having at least 70% identity with the amino acid sequence of any one of SEQ ID NO: 1 to 6, 21 and 22, or
(iii) a fragment of (i) or (ii) having at least 70% identity with the amino acid sequence of any one of SEQ ID NO: 1 to 6, 21 and 22,
wherein the variant or the fragment of the heterologous transcription factor FZF1 polypeptide favors the expression of a native SSU1 polypeptide, the heterologous SSU1 polypeptide, or both the native SSU1 polypeptide and the heterologous SSU1 polypeptide; and
a heterologous nucleic acid molecule encoding a heterologous SSU1 polypeptide, wherein the heterologous SSU1 polypeptide comprises:
(i) the amino acid sequence of any one of SEQ ID NO: 7 to 12, 23, 24, and 25,
(ii) a variant having at least 70% identity with the amino acid sequence of any one of SEQ ID NO: 7 to 12, 23, 24, and 25, or
(iii) a fragment of (i) or (ii) having at least 70% identity with the amino acid sequence of any one of SEQ ID NO: 7 to 12, 23, and 24,
wherein the variant or the fragment has sulfite efflux activity.