Granted Patent B2

US 12,644,103 · App. 18/008,939 · Granted Jun 2, 2026

Inventors: Bernhard Neumayer (Vienna, AT); Elisabeth Streit (Vienna, AT); Barbara Weber (Vienna, AT); Gudrun Vogtentanz (Krems, AT)

Assignee: DSM AUSTRIA GMBH

C12N9/0008A23K20/189A23L29/06C12Y197/00

Abstract

Claims (116)

1 . A method of converting 3-keto-DON into 3-epi-DON, the method comprising contacting one or more polypeptide(s) comprising or consisting of: (a) SEQ ID NO: 1, or (b) a sequence having a sequence identity of at least 72.0% to SEQ ID NO: 1, with 3-keto DON, wherein the polypeptide comprises one or more of the following polypeptide sequences:

(i)

(SEQ ID NO. 31)

YRKLGNSG;

(ii)

(SEQ ID NO. 32)

LGTMTFG;

(iii)

(SEQ ID NO. 33)

AGGNFX 1 DTAX 2 VYS, wherein X 1  is I or L and X 2  is

N or D;

(iv)

(SEQ ID NO. 34)

ETLRFLDD;

(v)

(SEQ ID NO. 35)

GKIX 3 YYGFSN, wherein X 3  is A or G;

(vi)

(SEQ ID NO. 36)

RDIEHEX 4 VPA, wherein X 4  = I or V;

(vii)

(SEQ ID NO. 37)

GLLPWSPLGGGWL;

(viii)

(SEQ ID NO. 38)

GATRLGENP;

(ix)

(SEQ ID NO. 39)

AQVALAW;

and/or

(x) 

(SEQ ID NO. 40)

PAVX 5 SVILGART, wherein X 5  = T or A.

2 . The method of claim 1 , wherein the polypeptide comprises 2, 3, 4, 5, 6, 7, 8, or 9 of polypeptide sequences (i)-(x).

3 . The method of claim 1 , wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON and into the product DON with a ratio of the activity of the polypeptide for DON to the activity of the polypeptide for 3-epi-DON (DON: 3-epi-DON) between 0.045 and 0.26.

4 . The method of claim 1 , wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, wherein between 79.0% and 96.0%, between 80.0% and 95.0%, or between 81.0% and 94.0% of the total product is 3-epi-DON.

5 . The method of claim 1 , wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON and additionally into the product DON amounting to 100% of total product, wherein between 3.9% and 21.5%, between 4.0% and 21.0%, or between 5.0% and 19.0% of the total product is DON.

6 . The method of claim 1 , wherein the polypeptide is capable of converting the substrate 3-keto-DON into the products 3-epi-DON and DON with a ratio of between 24.5:1 to 3.7:1, 24:1 to 4:1, 20:1 to 4:1, 18:1 to 4:1, 19:1 to 5:1, or 18:1 to 6:1.

7 . A method of reducing the content of DON in a composition comprising DON or of reducing the toxicity of a composition comprising DON by converting DON into 3-epi-DON, the method comprising

a) contacting the composition with an enzyme capable of converting DON into 3-keto DON; and

b) subsequently or concurrently contacting the composition with one or more polypeptide(s) comprising or consisting of SEQ ID NO: 1, or a sequence having a sequence identity of at least at least 72.0% to SEQ ID NO: 1; wherein the polypeptide comprises one or more of the following polypeptide sequences:

(i)

(SEQ ID NO. 31)

YRKLGNSG;

(ii)

(SEQ ID NO. 32)

LGTMTFG;

(iii)

(SEQ ID NO. 33)

AGGNFX 1 DTAX 2 VYS, wherein X 1  is I or L and X 2  is

N or D;

(iv)

(SEQ ID NO. 34)

ETLRFLDD;

(v)

(SEQ ID NO. 35)

GKIX 3 YYGFSN, wherein X 3  is A or G;

(vi)

(SEQ ID NO. 36)

RDIEHEX 4 VPA, wherein X 4  = I or V;

(vii)

(SEQ ID NO. 37)

GLLPWSPLGGGWL;

(viii)

(SEQ ID NO. 38)

GATRLGENP;

(ix)

(SEQ ID NO. 39)

AQVALAW;

and/or

(x) 

(SEQ ID NO. 40)

PAVX 5 SVILGART, wherein X 5  = T or A.

8 . The method of claim 1 wherein the polypeptide comprises at least 330 or at least 340 amino acids, with respect to SEQ ID NO: 1.

9 . The method of claim 8 wherein the polypeptide comprises or consists of 341, 342, or 343 amino acids, with respect to SEQ ID NO: 1.

10 . The method of claim 2 wherein the polypeptide comprises all of the polypeptide sequences (i)-(x).

11 . A method of converting 3-keto-DON into 3-epi-DON, the method comprising contacting one or more polypeptide(s) with 3-keto DON, wherein the one or more polypeptide(s) comprise or consist of: (a) SEQ ID NO: 1, or (b) a sequence having a sequence identity of at least 72.0% to SEQ ID NO: 1, and at least 95.0% sequence identity to any one of SEQ ID NOs: 1-28.

12 . The method of claim 11 wherein the one or more polypeptide(s) comprise or consist of: (b) a sequence having a sequence identity of at least 72.0% to SEQ ID NO: 1, and at least 97.0% sequence identity to any one of SEQ ID NOs: 1-28.

13 . The method of claim 12 wherein the one or more polypeptide(s) comprise or consist of: (b) a sequence having a sequence identity of at least 72.0% to SEQ ID NO: 1, and at least 99.0% sequence identity to any one of SEQ ID NOs: 1-28.

14 . The method of claim 13 wherein the one or more polypeptide(s) comprise or consist of any one of SEQ ID NOs: 1-28.

15 . The method of claim 11 , wherein the polypeptide comprises one or more of the following polypeptide sequences:

(i)

(SEQ ID NO. 31)

YRKLGNSG;

(ii)

(SEQ ID NO. 32)

LGTMTFG;

(iii)

(SEQ ID NO. 33)

AGGNFX 1 DTAX 2 VYS, wherein X 1  is I or L and X 2  is

N or D;

(iv)

(SEQ ID NO. 34)

ETLRFLDD;

(v)

(SEQ ID NO. 35)

GKIX 3 YYGFSN, wherein X 3  is A or G;

(vi)

(SEQ ID NO. 36)

RDIEHEX 4 VPA, wherein X 4  = I or V;

(vii)

(SEQ ID NO. 37)

GLLPWSPLGGGWL;

(viii)

(SEQ ID NO. 38)

GATRLGENP;

(ix)

(SEQ ID NO. 39)

AQVALAW;

and/or

(x) 

(SEQ ID NO. 40)

PAVX 5 SVILGART, wherein X 5  = T or A.

16 . The method of claim 15 wherein the polypeptide comprises all of the polypeptide sequences (i)-(x).

17 . The method of claim 11 wherein the polypeptide comprises at least 330 or at least 340 amino acids, with respect to SEQ ID NO: 1.

18 . The method of claim 17 wherein the polypeptide comprises or consists of 341, 342, or 343 amino acids, with respect to SEQ ID NO: 1.

Assignments (1)

Priority Claims (1)

Continuity (1)

References Cited (29)