Biofuel production

Patent metadata

Field	Value
Publication number	US-9243256-B2
Application number	US-201013640902-A
Country	US
Kind code	B2
Filing date	Apr 12, 2010
Priority date	Apr 12, 2010
Publication date	Jan 26, 2016
Grant date	Jan 26, 2016

Abstract

The invention relates to the combination of the TLG1 glucoamylase from Thermomyces lanuginoses with: (i) the SFA1 alpha-amylase from Saccharomycopsis fibuligera ; and/or (ii) the LKA alpha-amylase. The enzyme combinations may be expressed in a host cell (e.g. in Saccharomyces cerevisiae ) or provided as an enzyme composition. Methods for making the enzyme combinations of the invention are provided. The invention also relates to a yeast strain which exhibits amylolytic activity and promising fermentative abilities. Processes for producing a fermentation product (in particular alcohol) from starch-containing material are described.

First claim

The invention claimed is:

1. A host cell which expresses a Thermomyces lanuginosus glucoamylase 1 (TLG1) protein and at least one of a Saccharomycopsis fibuligera alpha-amylase 1 (SFA1) protein and a Lypomyces kononenkoae alpha-amylase 1 (LKA1) protein, wherein:
the sequence of the TLG1 protein comprises a sequence which has at least 70% identity to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which has at least 70% identity to SEQ ID NO: 3;
the sequence of the LKA1 protein comprises a sequence which has at least 70% identity to SEQ ID NO: 2, and
the host cell is a yeast;
and wherein the proteins which are expressed by the host cell are capable of converting an uncooked grain starch-containing material to a fermentation product.

2. The host cell according to claim 1 , wherein:
the sequence of the TLG1 protein comprises a sequence which has at least 80% identity to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which has at least 80% identity to SEQ ID NO: 3; and/or
the sequence of the LKA1 protein comprises a sequence which has at least 80% identity to SEQ ID NO: 2.

3. The host cell according to claim 1 , wherein:
the sequence of the TLG1 protein comprises a sequence which has at least 90% identity to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which has at least 90% identity to SEQ ID NO: 3; and/or
the sequence of the LKA1 protein comprises a sequence which has at least 90% identity to SEQ ID NO: 2.

4. The host cell according to claim 1 , wherein:
the sequence of the TLG1 protein comprises a sequence which is identical to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which is identical to SEQ ID NO: 3; and/or
the sequence of the LKA1 protein comprises a sequence which is identical to SEQ ID NO: 2.

5. The host cell according to claim 1 , which expresses the Thermomyces lanuginosus TLG1 protein and the Saccharomycopsis fibuligera SFA1 protein.

6. The host cell according to claim 1 , which expresses the Thermomyces lanuginosus TLG1 protein and the Lypomyces kononenkoae LKA1 protein.

7. The host cell according to claim 1 , which expresses the Thermomyces lanuginosus TLG1 protein, the Saccharomycopsis fibuligera SFA1 protein and the Lypomyces kononenkoae LKA1 protein.

8. The host cell according to claim 1 , wherein the fermentation product is selected from the group consisting of an alcohol, organic acid, ketone, amino acid, gas, enzyme and vitamin.

9. The host cell according to claim 8 , wherein the alcohol is selected from the group consisting of ethanol, methanol and butanol.

10. A process for producing a fermentation product, the process comprising the steps of using a host cell which expresses a Thermomyces lanuginosus glucoamylase 1 (TLG1) protein and at least one of a Saccharomycopsis fibuhgera alpha-amylase 1 (SFA1) protein and a Lypomyces kononenkoae alpha-amylase 1 (LKA1) protein to convert grain starch in a starch-containing material to the fermentation product, wherein:
the sequence of the TLG1 protein comprises a sequence which has at least 70% identity to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which has at least 70% identity to SEQ ID NO: 3;
the sequence of the LKA1 protein comprises a sequence which has at least 70% identity to SEQ ID NO: 2;
the host cell is a yeast; and
wherein the starch-containing material is uncooked grain starch.

11. The process according to claim 10 , wherein:
the sequence of the TLG1 protein comprises a sequence which has at least 80% identity to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which has at least 80% identity to SEQ ID NO: 3; and/or
the sequence of the LKA1 protein comprises a sequence which has at least 80% identity to SEQ ID NO: 2.

12. The process according to claim 10 , wherein:
the sequence of the TLG1 protein comprises a sequence which has at least 90% identity to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which has at least 90% identity to SEQ ID NO: 3; and/or
the sequence of the LKA1 protein comprises a sequence which has at least 90% identity to SEQ ID NO: 2.

13. The process according to claim 10 , wherein:
the sequence of the TLG1 protein comprises a sequence which is identical to SEQ ID NO: 1;
the sequence of the SFA1 protein comprises a sequence which is identical to SEQ ID NO: 3; and/or
the sequence of the LKA1 protein comprises a sequence which is identical to SEQ ID NO: 2.

14. The process according to claim 10 , wherein the host cell expresses the Thermomyces lanuginosus TLG1 protein and the Saccharomycopsis fibuligera SFA1 protein.

15. The process according to claim 10 , wherein the host cell expresses the Thermomyces lanuginosus TLG1 protein and the Lypomyces kononenkoae LKA1 protein.

16. The process according to claim 10 , wherein the host cell expresses the Thermomyces lanuginosus TLG1 protein, the Saccharomycopsis fibuligera SFA1 protein and the Lypomyces kononenkoae LKA1 protein.

17. The process according to claim 10 , wherein the fermentation product is selected from the group consisting of an alcohol, organic acid, ketone, amino acid, gas, enzyme and vitamin.

18. The process according to claim 17 , wherein the alcohol is selected from the group consisting of ethanol, methanol and butanol.

Assignees

• Van Zyl Willem Heber
• Jooste Tania
• Gorgens Johann Ferdinand
• Saayman Maryna
• Favaro Lorenzo
• Basaglia Marina
• Casella Sergio
• Univ Stellenbosch
• Univ Padova

Inventors

• Van Zyl Willem Heber
• Jooste Tania
• Gorgens Johann Ferdinand
• Saayman Maryna
• Favaro Lorenzo
• Basaglia Marina
• Casella Sergio

Classifications

• C12N9/242 Primary

  Fungal source · CPC title

• C12N9/2428

  Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase · CPC title

• C12N9/2402

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title

• Y02E50/17

  Cross-Sectional Technologies · mapped topic

• C12P7/06

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