Alpha-glucosidase, compositions and methods

Patent metadata

Field	Value
Publication number	US-10294466-B2
Application number	US-201615553134-A
Country	US
Kind code	B2
Filing date	Feb 25, 2016
Priority date	Feb 25, 2015
Publication date	May 21, 2019
Grant date	May 21, 2019

Abstract

The present disclosure relates to polypeptides having alpha-glucosidase activity isolated, derived or derivable from Rasamsonia or engineered polypeptides having alpha-glucosidase activity isolated, derived or derivable from Rasamsonia homologs. The present disclosure also pertains to polynucleotides encoding the polypeptides, nucleic acid constructs, vectors, host cells and mutant cells comprising the polynucleotides. The disclosure further pertains to compositions comprising such polypeptides, methods of producing the polypeptides and compositions, as well as methods for using such polypeptides and compositions for industrial applications.

First claim

What is claimed is:

1. A non-native polypeptide comprising an amino acid sequence that is at least 97% identical to, but not 100% identical to, the amino acid sequence of SEQ ID NO:4 or 8, wherein the non-native polypeptide has alpha-glucosidase activity.

2. The non-native polypeptide of claim 1 , wherein the non-native polypeptide comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO:4 or 8.

3. The non-native polypeptide of claim 2 , wherein the non-native polypeptide comprises an amino acid sequence that is at least about 98% 99% identical to the amino acid sequence of SEQ ID NO:4 or 8.

4. A composition comprising the non-native polypeptide of any one of claims 1 - 3 and one or more of an alpha-amylase, a beta-amylase, a glucoamylase, a pullulanase, an isoamylase, a different alpha-glucosidase, and/or a cyclodextrin glycosyltransferase.

5. The non-native polypeptide of claim 1 , wherein the non-native polypeptide comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO:4.

6. The non-native polypeptide of claim 5 , wherein the non-native polypeptide comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO:4.

7. A polynucleotide encoding the non-native polypeptide of any one of claims 1 - 3 , 5 , or 6 .

8. The polynucleotide of claim 7 , comprising at least one transcriptional or translational regulatory sequence that allows the polynucleotide sequence to be expressed by a host cell.

9. The polynucleotide of claim 8 , wherein the polynucleotide comprises a transcriptional regulatory sequence that is a promoter sequence.

10. A host cell comprising the polynucleotide of claim 9 .

11. The host cell of claim 10 , which is a Trichoderma or Aspergillus host cell.

12. The host cell of claim 10 , which is an E. coli, Bacillus, Streptomyces , or Pseudomonas cell.

13. The host cell of claim 10 , which is a yeast cell.

14. A method of making a polypeptide, the method comprising cultivating the host cell of claim 10 , wherein said polypeptide having alpha-glucosidase activity is produced by the host cell.

15. The method of claim 14 , further comprising a step of recovering, enriching and/or purifying the polypeptide.

16. A polynucleotide comprising a nucleotide sequence that encodes a polypeptide having alpha-glucosidase activity, wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO:4 or 8, and wherein the nucleotide sequence is
(i) a complementary DNA (cDNA) sequence, and/or
(ii) operably linked to at least one transcriptional or translational regulatory sequence that is heterologous to the nucleotide sequence.

17. The polynucleotide of claim 16 , wherein the polypeptide comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO:4 or 8.

18. The polynucleotide of claim 17 , wherein the polypeptide comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:4 or 8.

19. The polynucleotide of claim 16 , wherein the polypeptide comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO:4.

20. The polynucleotide of claim 19 , wherein the polypeptide comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO:4.

21. The polynucleotide of any one of claims 16 - 20 , wherein the nucleotide sequence is operably linked to at least one transcriptional or translational regulatory sequence that is heterologous to the nucleotide sequence.

22. The polynucleotide of claim 21 , wherein the nucleotide sequence is operably linked to a transcriptional regulatory sequence that is a promoter sequence.

23. A host cell comprising the polynucleotide of claim 22 .

24. A method of making a polypeptide, the method comprising cultivating the host cell of claim 23 , wherein said polypeptide having alpha-glucosidase activity is produced by the host cell.

Assignees

• Danisco Us Inc

Inventors

• Yu Zheyong
• Zhang Zhenghong
• Ge Jing
• Qian Zhen
• Liu Guoqing
• Xie Zhiyong
• Tang Zhongmei

Classifications

• C12N1/20

  Bacteria; Culture media therefor · CPC title

• C12N1/14

  Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor · CPC title

• Y02E50/16

  Cross-Sectional Technologies · mapped topic

• C12P19/14

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12P7/10

  substrate containing cellulosic material · CPC title