GLUTATHIONE REDUCTASE

• US 20190330600A1
• Filed: 12/14/2017
• Published: 10/31/2019
• Est. Priority Date: 12/22/2016
• Status: Abandoned Application

• Current Assignee: DSM IP Assets BV (DSM-Firmenich AG)
• Original Assignee: DSM IP Assets BV (DSM-Firmenich AG)
• Inventors: VAN DEN BERG, Marco Alexander, JAMALZADEH, Elaheh, RUINARD, Samuel Adrianus Maria
• Application Number:
  US16/472,099
• Publication Number:
  US 20190330600A1
• Time in Patent Office: Days
• CPC Class Codes: C12N 9/0051 acting on a sulfur group of donors (1.8)

Abstract

The present invention relates to a polypeptide having glutathione reductase activity selected from the group consisting of: a) polypeptide having an amino acid sequence comprising the mature polypeptide sequence of SEQ ID NO: 1 or a 2) a polypeptide comprising an amino acid sequence that has at least 50% sequence identity with the mature polypeptide sequence of SEQ ID NO: 1.

Claims

1. A polypeptide having glutathione reductase activity selected from the group consisting of:

a) a polypeptide having an amino acid sequence comprising the mature polypeptide sequence of SEQ ID NO;

1;

b) a polypeptide comprising an amino acid sequence that has at least 50% sequence identity with the mature polypeptide sequence of SEQ ID NO;

1;

c) a polypeptide encoded by a nucleic acid comprising a sequence that hybridizes under medium stringency conditions to the complementary strand of the mature polypeptide encoding sequence of SEQ ID NO;

2; and

d) a polypeptide comprising an amino acid sequence encoded by a nucleic acid that has at least 50% sequence identity to the mature polypeptide coding sequence of SEQ ID NO;

2.

2. A polypeptide according to claim 1, which is derivable from Chaetomium thermophilum.

3. A nucleic acid encoding a polypeptide having glutathione reductase activity according to claim 1, which comprises a sequence that at least 50% sequence identity to the mature polypeptide encoding sequence of SEQ ID NO:

2.

4. An expression vector comprising a nucleic acid according to claim 3, operably linked to one or more control sequences that direct expression of the glutathione reductase in a host cell.

5. A recombinant host cell comprising a polypeptide according to claim 1, a nucleic acid encoding said polypeptide or an expression vector comprising said nucleic acid.

6. Recombinant host cell according to claim 5, which is a recombinant yeast cell, optionally a recombinant Saccharomyces cerevisiae cell.

7. A method for preparation of a polypeptide according to claim 1, which method comprises:

cultivating a suitable host cell in a suitable fermentation medium under conditions that allow for production of the polypeptide; and

, optionally,

recovering the polypeptide.

8. A product comprising a polypeptide according to claim 1, for reduction of oxidized thiols.

9. A method for enzymatic reduction of cystine to cysteine, which method comprises:

contacting cystine with a reduction solution comprising a polypeptide according to claim 1, a cofactor and a mediator; and

, optionally, recovering the cysteine.

10. Method according to claim 9, wherein contacting the cystine with the reduction solution is carried out at a temperature within a range of 25°

C. to 75°

C.

11. Method according to claim 10, wherein the mediator is glutathione.

12. Method according to claim 9, wherein the cofactor is nicotinamide adenine dinucleotide phosphate (NADPH), preferably in oxidized form (NADP+) or nicotinamide adenine dinucleotide (NADH), preferably in oxidized form (NAD+).

13. Method according to claim 9, wherein the reduction solution has a pH of at least 6.

14. A composition comprising cysteine and a polypeptide according to claim 1.

15. A composition according to claim 14, further comprising one or more selected from gluconic acid, formic acid, glucose, formate, nicotinamide adenine dinucleotide phosphate, nicotinamide adenine dinucleotide, glutathione, acetone and acetaldehyde.