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US 2023/0332188 A1Published application
Improved fermenting organism for ethanol production
AssigneesNovozymes A/S, Microbiogen Pty. Ltd.
InventorsRoberto Nobuyuki Maeda, Philip John Livingstone Bell
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Abstract
The present invention relates to processes for producing ethanol comprising saccharifying cellulosic or starch-containing material and fermenting the saccharified material with a fermenting microorganism to produce ethanol. The fermenting organism is Saccharomyces cerevisiae strain MBG5151 (deposited under Accession No. Y-67971 at the Agricultural Research Service Culture Collection (NRRL), Illinois 61604 U.S.A.), Saccharomyces cerevisiae strain MBG5248 (deposited under Accession No. Y-68015 at the Agricultural Research Service Culture Collection (NRRL), Illinois 61604 U.S.A.) or a fermenting organism that has properties that the same or about the same as that of Saccharomyces cerevisiae MBG5151 or MBG5248.
Claims20
1:
- A method of producing a fermentation product from a cellulosic-containing and/or starch-containing material, the method comprising:
- (a) saccharifying the cellulosic-containing or starch-containing material;
- and
- (b) fermenting the saccharified material of step (a) with a fermenting organism under suitable conditions to produce the fermentation product;
- wherein the fermenting organism is:
- (1) a recombinant strain of Saccharomyces cerevisiae deposited under the Budapest Treaty at the Agricultural Research Service Patent Culture Collection (NRRL) having deposit accession no. NRRL Y-67971 (Saccharomyces cerevisiae strain MBG5151), or a derivative thereof (e.g., expressing a heterologous polypeptide such as a glucoamylase and/or alpha-amylase) or a fermenting organism having properties that are about the same as that of Saccharomyces cerevisiae MBG5151;
- or
- (2) a recombinant strain of Saccharomyces cerevisiae deposited under the Budapest Treaty at the Agricultural Research Service Patent Culture Collection (NRRL) having deposit accession no. NRRL Y-68015 (Saccharomyces cerevisiae strain MBG5248), or a derivative thereof (e.g., expressing a heterologous polypeptide such as a glucoamylase and/or alpha-amylase) or a fermenting organism having properties that are about the same as that of Saccharomyces cerevisiae M BG5248.
5 more elements
2:
- A recombinant Saccharomyces yeast strain selected from:
- Saccharomyces cerevisiae strain deposited under the Budapest Treaty at the Agricultural Research Service Patent Culture Collection (NRRL) having deposit accession no. NRRL Y-67971 (Saccharomyces cerevisiae strain MBG5151), or a derivative thereof (e.g., expressing a heterologous polypeptide such as a glucoamylase and/or alpha-amylase) or a fermenting organism having properties that are about the same as that of Saccharomyces cerevisiae MBG5151;
- and
- Saccharomyces cerevisiae strain deposited under the Budapest Treaty at the Agricultural Research Service Patent Culture Collection (NRRL) having deposit accession no. NRRL Y-68015 (Saccharomyces cerevisiae strain MBG5248), or a derivative thereof (e.g., expressing a heterologous polypeptide such as a glucoamylase and/or alpha-amylase) or a fermenting organism having properties that are about the same as that of Saccharomyces cerevisiae MBG5248.
1 more element
3:
- The recombinant Saccharomyces cerevisiae strain of claim 2, wherein the strain has one or more of the following properties:
- higher ethanol fermentation kinetics compared to Saccharomyces cerevisiae CIBTS1260 at 1 g DWC/L, 32° C., pH 5.5;
- higher xylose consumption compared to Saccharomyces cerevisiae CIBTS1260 after 48 hours fermentation at 1 g DWC/L, 35° C., pH 5.5;
- higher glucose consumption compared to Saccharomyces cerevisiae CIBTS1260 after 48 hours fermentation at 1 g DWC/L, 35° C., pH 5.5.
1 more element
4:
- The recombinant Saccharomyces cerevisiae strain of claim 2, wherein the strain is capable of higher ethanol yield compared to Saccharomyces cerevisiae CIBTS1260 at 1 g DWC/L, 32° C., pH 5.5 between 10 to 30 hours of fermentation.
5:
- The recombinant Saccharomyces cerevisiae strain of claim 2, wherein the strain is capable of greater than 95% xylose consumption by 48 hours fermentation under the process conditions of 1 g DCW/L, 35° C., pH 5.5.
6:
- The recombinant Saccharomyces cerevisiae strain of claim 2, wherein the strain is capable of greater than 95% glucose consumption by 24 hours fermentation under the process conditions of 1 g DCW/L, 35° C., pH 5.5.
7:
- The recombinant Saccharomyces cerevisiae of claim 2, wherein the strain is capable of providing more than 30 g/L ethanol after 48 hours fermentation under the process conditions of 1 g DCW/L, 35° C., pH 5.5.
8:
- The recombinant Saccharomyces cerevisiae of claim 2, comprising a heterologous gene encoding a xylose isomerase.
9:
- The recombinant Saccharomyces cerevisiae of claim 2, comprising a heterologous gene encoding a pentose transporter.
10:
- The recombinant Saccharomyces cerevisiae of claim 2, wherein the pentose transporter gene is a GFX gene.
11:
- The recombinant Saccharomyces cerevisiae of claim 2, comprising a heterologous gene encoding a xylulokinase (XKS).
12:
- The recombinant Saccharomyces cerevisiae of claim 2, comprising a heterologous gene encoding a ribulose 5 phosphate 3-epimerase (RPE1), a heterologous gene encoding a ribulose 5 phosphate isomerase (RKI1), or a heterologous gene encoding a transketolase (TKL1) and a heterologous gene encoding a transaldolase (TAL1).
13:
- A method of producing a derivative of Saccharomyces cerevisiae strain MBG5151 (deposited under Accession No. NRRL Y-67971 at the Agricultural Research Service Patent Culture Collection (NRRL)) or Saccharomyces cerevisiae strain MBG5248 (deposited under Accession No. NRRL Y-68015 at the Agricultural Research Service Patent Culture Collection (NRRL)) which exhibits the defining characteristics of Saccharomyces cerevisiae strain MBG5151 or Saccharomyces cerevisiae strain MBG5248, respectively, the method comprising:
- (a) providing:
- (i) a first yeast strain;
- and
- (ii) a second yeast strain, wherein the second yeast strain is Saccharomyces cerevisiae strain MBG5151, Saccharomyces cerevisiae strain MBG5248, or a derivative thereof;
- (b) culturing the first yeast strain and the second yeast strain under conditions which permit combining of DNA between the first and second yeast strains;
- (c) screening or selecting for a derivative of Saccharomyces cerevisiae strain MBG5151 or Saccharomyces cerevisiae strain MBG5248.
4 more elements
14:
- The method of claim 13, wherein step (c) comprises screening or selecting for a hybrid strain which exhibits one or more defining characteristic of Saccharomyces cerevisiae strain MBG5151 or Saccharomyces cerevisiae strain MBG5248.
15:
- The method of claim 13, comprising the further step of:
- (d) repeating steps (a) and (b) with the screened or selected strain from step (c) as the first and/or second strain, until a derivative is obtained which exhibits the defining characteristics of Saccharomyces cerevisiae strain MBG5151 or Saccharomyces cerevisiae strain MBG5248.
16:
- The method of claim 13, wherein the culturing step (b) comprises:
- (i) sporulating the first yeast strain and the second yeast strain;
- (ii) hybridizing germinated spores produced by the first yeast strain with germinated spores produced by the second yeast strain.
17:
- A method of producing a recombinant derivative of Saccharomyces cerevisiae strain MBG5151 (deposited under Accession No. NRRL Y-67971 at the Agricultural Research Service Patent Culture Collection (NRRL)) or Saccharomyces cerevisiae strain MBG5248 (deposited under Accession No. NRRL Y-68015 at the Agricultural Research Service Patent Culture Collection (NRRL)), the method comprising:
- (a) transforming Saccharomyces cerevisiae strain MBG5151 (or a derivative of Saccharomyces cerevisiae strain MBG5151) or Saccharomyces cerevisiae strain MBG5248 (or a derivative of Saccharomyces cerevisiae strain MBG5248) with one or more expression vectors;
- and
- (b) isolating the transformed strain.
1 more element
18:
- A Saccharomyces cerevisiae strain produced by the method of claim 17.
19:
- A method of producing ethanol, comprising incubating a Saccharomyces cerevisiae strain of claim 2 with a substrate comprising a fermentable sugar under conditions which permit fermentation of the fermentable sugar to produce ethanol.
20:
- A composition comprising a Saccharomyces cerevisiae strain of claim 2, and one or more naturally occurring and/or non-naturally occurring components.