Method for producing selenoneine

Patent metadata

Field	Value
Publication number	US-11629341-B2
Application number	US-202117319362-A
Country	US
Kind code	B2
Filing date	May 13, 2021
Priority date	Aug 7, 2015
Publication date	Apr 18, 2023
Grant date	Apr 18, 2023

Abstract

The purpose of the present invention is to provide a method for producing selenoneine that allows production of selenoneine at higher yields as compared with a conventional technology, and, therefore, enables selenoneine production on an industrial scale. This purpose can be achieved by a method for producing selenoneine, comprising the step of applying histidine and a selenium compound to a transformant that has a gene encoding an enzyme of (1) below introduced therein and that can overexpress the introduced gene, to obtain selenoneine.(1) An enzyme that catalyzes a reaction in which hercynylselenocysteine is produced from histidine and selenocysteine in the presence of S-adenosylmethionine and iron (II).

First claim

The invention claimed is:

1. A method for producing selenoneine, comprising the step of;
contacting histidine and selenous acid with a transformant by using as a host organism Aspergillus that has a gene encoding an enzyme of (1) introduced therein as foreign gene and that can overexpress the introduced gene to obtain selenoneine,
wherein the enzyme of (1) catalyzes a reaction in which hercynylselenocysteine of formula is produced from histidine and selenocysteine in the presence of S-adenosylmethionine and iron (II),
wherein the gene encoding the enzyme of (1) has the nucleotide sequence of SEQ ID NO: 1, or the enzyme of (1) has the amino acid sequence of SEQ ID NO: 4, and
wherein formula [I] is:

wherein the host organism further has a second ne encoding an enzyme of (2) expressed therein,
wherein the enzyme of (2) catalyzes a reaction in which selenoneine is produced from hercynylselenocysteine of formula [I] using pyridoxal 5′-phosphate as a coenzyme, and
wherein the second gene encoding the enzyme of (2) has the nucleotide sequence of SEQ ID NO: 2 or 3, or the enzyme of (2) has the amino acid sequence of SEO ID NO: 5 or 6.

2. The method according to claim 1 , wherein the transformant further expresses at least one enzyme selected from the group consisting of selenic acid reductase, selenocysteine lyase, and serine dehydratase.

3. The method according to claim 1 , wherein the transformant is produced by using as a host organism Aspergillus selected from the group consisting of Aspergillus sojae, Aspergillus oryzae, Aspergillus niger, Aspergillus tamarii, Aspergillus awarmori, Aspergillus usamii, Aspergillus kawachii , and Aspergillus saitoi.

4. The method according to claim 1 , wherein the transformant is a transformant in which the expression of the gene encoding the enzyme of (1) is enhanced to increase the amount of selenoneine as compared to the host organism.

5. The method according to claim 1 , wherein the transformant is a transformant in which the expression of the gene encoding the enzyme of (1) is enhanced so that the amount of selenoneine produced when the transformant is cultured in a selenocystine-containing medium suitable for the growth of the host organism at 30° C. for 5 days is not less than 10 μg per gram of wet cell mass.

6. The method according to claim 1 , wherein the enzyme of (2) is AsEgtC.

7. The method according to claim 1 , wherein the second gene encoding the enzyme of (2) has the nucleotide sequence of SEQ ID NO: 2, or the enzyme of (2) has the amino acid sequence of SEQ ID NO: 5.

8. The method according to claim 1 , wherein the second gene encoding the enzyme of (2) has the nucleotide sequence of SEQ ID NO: 3, or the enzyme of (2) has the amino acid sequence of SEQ ID NO: 6.

9. The method according to claim 1 , wherein the microorganism is Aspergillus sojae.

10. The method according to claim 6 , wherein microorganism is Aspergillus sojae.

11. The method according to claim 7 , wherein the gene is obtained from the microorganism Aspergillus sojae.

12. The method according to claim 8 , wherein the microorganism is Aspergillus sojae.

Assignees

• Kikkoman Corp

Inventors

• Ichikawa Keiichi
• Shinohara Yasutomo
• Hara Seiichi
• Kurosawa Keiko
• Yamashita Yumiko
• Yamashita Michiaki

Classifications

• C12N15/09

  Recombinant DNA-technology · CPC title

• C12R2001/66

  Aspergillus · CPC title

• C12R2001/69

  Aspergillus oryzae · CPC title

• C12N9/88

  Lyases (4.) · CPC title

• C12Y403/01017

  L-Serine ammonia-lyase (4.3.1.17) · CPC title