Heterologous protease expression for improving alcoholic fermentation

Patent metadata

Field	Value
Publication number	US-2020165592-A1
Application number	US-201816482633-A
Country	US
Kind code	A1
Filing date	Feb 1, 2018
Priority date	Feb 2, 2017
Publication date	May 28, 2020
Grant date	—

Abstract

The present disclosure relates to proteases for improving alcoholic fermentation. The proteases are expressed from a recombinant host cell. The present disclosure also provides a population of recombinant host cells expressing an heterologous protease that can be used in combination with recombinant host cells expressing an heterologous glucoamylase and/or an heterologous glycerol reduction system.

First claim

1 . A first recombinant yeast host cell comprising a first genetic modification allowing expression of an heterologous protease, wherein the heterologous protease is:
a) a polypeptide having the amino acid sequence of SEQ ID NO: 2, 6, 8, 10, 12, 14, 30, 36, 38, 40, 42, 52 or 92; b) a variant having at least 70% identity to the polypeptide of a) and exhibiting proteolytic activity; or c) a fragment having at least 70% identity to the polypeptide of a) or the variant of b) and exhibiting proteolytic activity.

2 . The first recombinant yeast host cell of claim 1 , wherein the heterologous protease is the polypeptide having the amino acid sequence of SEQ ID NO: 2, 14, 40 or 52.

3 .- 8 . (canceled)

9 . The first recombinant yeast host cell of claim 1 , having a second genetic modification allowing expression of an heterologous glucoamylase.

10 . (canceled)

11 . The first recombinant yeast host cell of claim 1 having a third genetic modification for reducing production of one or more native enzymes that function to produce glycerol or regulate glycerol synthesis.

12 .- 13 . (canceled)

14 . The first recombinant yeast host cell of claim 1 having a fourth genetic modification for reducing production of one or more native enzymes that function to catabolize formate.

15 . (canceled)

16 . The first recombinant yeast host cell of claim 1 being from a genus that is the genus Saccharomyces.

17 . The first recombinant yeast host cell of claim 16 being from a species of the genus Saccharomyces that is the species Saccharomyces cerevisiae.

18 . A cellular population comprising:
a first recombinant yeast host cell comprising the first genetic modification defined in claim 1 ; and a second recombinant yeast host cell comprising a second, a third and/or a fourth genetic modification wherein:
the second genetic modification allows the expression of an heterologous glucoamylase;
the third genetic modification is for reducing the production of one or more native enzymes that function to produce glycerol or regulate glycerol synthesis; and
the fourth genetic modification is for reducing the production of one or more native enzymes that function to catabolize formate.

19 .- 26 . (canceled)

27 . A process for promoting ethanolic fermentation, the process comprising fermenting a medium with
(a) the first recombinant yeast host cell defined in claim 1 , or with (b) a cellular population comprising: a first recombinant yeast host cell comprising the first genetic modification defined in claim 1 ; and a second recombinant yeast host cell comprising a second, a third and/or a fourth genetic modification wherein:
the second genetic modification allows the expression of an heterologous glucoamylase;
the third genetic modification is for reducing the production of one or more native enzymes that function to produce glycerol or regulate glycerol synthesis; and
the fourth genetic modification is for reducing the production of one or more native enzymes that function to catabolize formate.

28 . The process of claim 27 , wherein the medium comprises raw starch.

29 . The process of claim 27 , wherein the medium is derived from corn.

30 . The process of claim 27 , wherein the medium is derived from barley.

31 . The process of claim 30 , wherein the barley is malted barley.

32 .- 36 . (canceled)

37 . A composition comprising the heterologous protease of claim 1 .

38 . The composition of claim 37 being obtainable from the first recombinant yeast host cell of claim 1 .

39 . (canceled)

40 . The composition of claim 37 further comprising a medium.

41 . The composition of claim 40 , wherein the medium comprises raw starch.

42 . The composition of claim 40 , wherein the medium is derived from corn.

43 . The composition of claim 40 , wherein the medium is derived from barley.

44 . The composition of claim 43 , wherein the barley is malted barley.

Assignees

• Lallemand Hungary Liquidity Man Llc

Inventors

• Barrett Trisha
• Rice Charles F
• Argyros Aaron

Classifications

• C12N9/60 Primary

  from yeast · CPC title

• C12P7/06

  Ethanol, i.e. non-beverage · CPC title

• C12N9/50 Primary

  Proteinases {, e.g. Endopeptidases (3.4.21-3.4.25)} · CPC title

• C12N1/18

  Baker's yeast; Brewer's yeast · CPC title

• Y02E50/10

  Biofuels, e.g. bio-diesel · CPC title