Transformed fungus having enhanced ergothioneine productivity and method for producing ergothioneine

Patent metadata

Field	Value
Publication number	US-10760107-B2
Application number	US-201515533215-A
Country	US
Kind code	B2
Filing date	Dec 25, 2015
Priority date	Jan 30, 2015
Publication date	Sep 1, 2020
Grant date	Sep 1, 2020

Abstract

The purpose of the present invention is to provide an organism having an ergothioneine productivity that is capable of easily producing ergothioneine within a short period of time at a high yield, as compared with a conventional technology, and, therefore, enables ergothioneine production on an industrial scale. This purpose can be achieved by a transformed fungus into which a gene encoding enzyme (1) or genes encoding enzymes (1) and (2) have been inserted and in which the inserted gene(s) are overexpressed. (1) an enzyme catalyzing a reaction of synthesizing hercynyl cysteine sulfoxide from histidine and cysteine in the presence of S-adenosyl methionine, iron (II) and oxygen. (2) An enzyme catalyzing a reaction of synthesizing ergothioneine from hercynyl cysteine sulfoxide using pyridoxal 5′-phosphate as a coenzyme.

First claim

The invention claimed is:

1. A transformed filamentous fungus comprising a gene encoding an enzyme (1) or genes encoding enzymes (1) and (2) inserted therein and can overexpress the inserted gene or genes,
wherein the gene encoding the enzyme (1) is a gene having the base sequence set forth in SEQ ID NO: 1, 23 or 33, or the enzyme (1) is an enzyme having the amino acid sequence set forth in SEQ ID NO: 4, 26 or 34, and
wherein the gene encoding the enzyme (2) is a gene having the base sequence set forth in SEQ ID NO: 2, 3, 24 or 25, or the enzyme (2) is the enzyme having the amino acid sequence set forth in SEQ ID NO: 5, 6, 27 or 28, wherein the enzyme (1) catalyzes the reaction in which hercynyl cysteine sulfoxide represented by the following formula (I)

is produced from histidine and cysteine in the presence of S-adenosylmethionine, iron (II), and oxygen; and wherein the enzyme (2) catalyzes the reaction in which ergothioneine is produced from hercynyl cysteine sulfoxide using pyridoxal 5′-phosphate as a coenzyme.

2. The transformed filamentous fungus of claim 1 , wherein the filamentous fungus is a microorganism of the genus Aspergillus.

3. The transformed filamentous fungus of claim 1 , wherein the filamentous fungus is a fungus of the genus Aspergillus selected from the group consisting of Aspergillus sojae, Aspergillus oryzae, Aspergillus niger, Aspergillus tamarii, Aspergillus awamori, Aspergillus usamii, Aspergillus kawachii , and Aspergillus saitoi.

4. The transformed filamentous fungus of claim 1 , wherein the transformed filamentous fungus is a transformed filamentous fungus in which the expression of the gene encoding the enzyme (1) or the genes encoding the enzymes (1) and (2) is enhanced such that the amount of ergothioneine is increased compared to a host filamentous fungus.

5. The transformed filamentous fungus of claim 1 , wherein the transformed filamentous fungus is a transfmmed filamentous fungus in which the expression of the genes encoding the enzymes (1) and (2) is enhanced such that the amount of ergothioneine is increased compared to transformed filamentous fungi in which the expression of the gene encoding the enzyme (1) is enhanced.

6. The transformed filamentous fungus of claim 1 , wherein the transformed filamentous fungus is a transformed filamentous fungus in which the expression of the gene encoding the enzyme (1) or the genes encoding the enzymes (1) and (2) is enhanced such that when the transformed filamentous fungi is cultured at 30° C. for 3 days in a culture medium suitable for the growth of the host filamentous fungus, the amount of ergothioneine is 10.0 mg or more per 1 g of dry cell mass.

7. A method for producing ergothioneine, comprising the step of applying histidine and cysteine to the transfoiined filamentous fungus according to claim 1 to obtain ergothioneine.

8. A method for producing a high-purity-ergothioneine-containing composition, comprising the step of obtaining an ergothioneine-containing composition having purity of 5% or higher from a culture obtained by culturing a transformed filamentous fungus according to claim 1 with the use of a culture medium containing histidine and cysteine.

9. The transformed filamentous fungus of claim 1 , wherein the enzyme (1) and/or the enzyme (2) is overexpressed under the control of a high expression promoter selected from the group consisting of a promoter region of translation elongation factor TEF1 gene (tef1), a promoter region of α-amylase gene (amy), and a promoter region of alkaline protease gene (alp).

Assignees

• Kikkoman Corp

Inventors

• Hara Seiichi
• Kurosawa Keiko
• Ichikawa Keiichi

Classifications

• C12N9/1007 Primary

  Methyltransferases (general) (2.1.1.) · CPC title

• C12R2001/665

  Aspergillus awamori · CPC title

• C12R2001/69

  Aspergillus oryzae · CPC title

• C12R2001/66

  Aspergillus · CPC title

• C12R2001/685

  Aspergillus niger · CPC title