IP Library › Granted Patent US 12,203,108
Granted Patent B2
US 12,203,108 · App. 16/635,474 · Granted Jan 21, 2025

Psicose-6-phosphate phosphatase, composition for producing psicose including said enzyme, method for producing psicose using said enzyme

Inventors: Byung-Sam Son (Ansan-Si, KR); Hyun Kug Cho (Suwon-Si, KR); Sung Jae Yang (Suwon-Si, KR); Seong Bo Kim (Seongnam-Si, KR); Seung Hwan Kim (Seoul, KR); Hyun June Park (Suwon-Si, KR)
Assignee: CJ CHEILJEDANG CORPORATION
C12N9/16C12N9/1051C12N9/1205C12N9/2414C12N9/2428C12N9/2457C12N9/246C12N9/90C12P19/02C12Y204/01001C12Y204/01007C12Y207/01002C12Y301/03025C12Y501/03C12Y503/01009C12Y504/02002
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Quick Facts
Patent No.
US 12,203,108
App. No.
16/635,474
Granted
Jan 21, 2025
Kind
B2
Abstract

The present application relates to a psicose-6-phosphate phosphatase comprising motif A and motif B, a composition for producing D-psicose comprising the enzyme, and a method for producing D-psicose using the enzyme.

Claims (62)

1. A method for producing D-psicose comprising contacting a psicose-6-phosphate phosphatase, a microorganism expressing the psicose-6-phosphate phosphatase or a culture of the microorganism expressing the psicose-6-phosphate phosphatase with D-psicose-6-phosphate to convert the D-psicose-6-phosphate to D-psicose;

wherein the psicose-6-phosphate phosphatase comprises:

i) a motif A comprising at least one sequence selected from the group consisting of SEQ ID NOS: 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, and 91;

ii) a motif B comprising a sequence having at least 90% sequence identity to at least one sequence selected from the group consisting of SEQ ID NOS: 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102,103, 104, 105, 106, 107, and 108; and

iii) A remainder sequence comprising remainder having at least 85% sequence identity to the amino acids sequence other than motif A and/or motif B in any sequence selected from the group consisting of SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20.

2. The method according to claim 1 , further comprising, prior to the conversion of D-psicose-6-phosphate to D-psicose, contacting a D-fructose-6-phosphate-3-epimerase, a microorganism expressing the D-fructose-6-phosphate-3-epimerase, or a culture of the microorganism expressing the D-fructose-6-phosphate-3-epimerase with D-fructose-6-phosphate to convert the D-fructose-6-phosphate to D-psicose-6-phosphate.

3. The method according to claim 1 , further comprising, prior to the conversion of D-fructose-6-phosphate to D-psicose-6-phosphate, contacting a D-glucose-6-phosphate-isomerase, a microorganism expressing the D-glucose-6-phosphate-isomerase, or a culture of the microorganism expressing the D-glucose-6-phosphate-isomerase with D-glucose-6-phosphate to convert the D-glucose-6-phosphate to D-fructose-6-phosphate.

4. The method according to claim 3 , further comprising, prior to the conversion of D-glucose-6-phosphate to D-fructose-6-phosphate, contacting a phosphoglucomutase, a microorganism expressing the phosphoglucomutase, or a culture of the microorganism expressing the phosphoglucomutase with D-glucose-1-phosphate to convert the D-glucose-1-phosphate to D-glucose-6-phosphate.

5. The method according to claim 4 , further comprising, prior to the conversion of D-glucose-6-phosphate to D-fructose-6-phosphate, contacting a glucokinase, a microorganism expressing the glucokinase, or a culture of the microorganism expressing the glucokinase and a phosphate with glucose to convert the glucose to D-glucose-6-phosphate.

6. The method according to claim 5 , further comprising, prior to the conversion of D-glucose-1-phosphate to D-glucose-6-phosphate, contacting (a) an α-glucan phosphorylase, a starch phosphorylase, a maltodextrin phosphorylase or a sucrose phosphorylase, (b) a microorganism expressing the α-glucan phosphorylase, the starch phosphorylase, the maltodextrin phosphorylase or the sucrose phosphorylase, or (c) a culture of the microorganism expressing the α-glucan phosphorylase, the starch phosphorylase, the maltodextrin phosphorylase or the sucrose phosphorylase, and a phosphate with starch, maltodextrin, sucrose or a combination thereof to convert the starch, maltodextrin, sucrose or combination thereof to D-glucose-1-phosphate.

7. The method according to claim 5 , further comprising, prior to the conversion of glucose to D-glucose-6-phosphate, contacting an α-amylase, a pullulanase, a glucoamylase, a sucrase or an isoamylase, (b) a microorganism expressing the α-amylase, pullulanase, glucoamylase, sucrase or isoamylase, or (c) a culture of the microorganism expressing the α-amylase, the pullulanase, the glucoamylase, the sucrase or the isoamylase with starch, maltodextrin, sucrose or a combination thereof to convert the starch, maltodextrin, sucrose or combination thereof to glucose.

8. A method for producing D-psicose comprising contacting (a) enzymes of (i) a psicose-6-phosphate phosphatase, (ii) a D-fructose-6-phosphate-3-epimerase, (iii) a D-glucose-6-phosphate-isomerase, (iv) a phosphoglucomutase or a glucokinase and (v) one or more selected from the group consisting of an α-glucan phosphorylase, a starch phosphorylase, a maltodextrin phosphorylase, a sucrose phosphorylase, an α-amylase, a pullulanase, an isoamylase, a glucoamylase and a sucrase, (b) microorganisms expressing the enzymes listed in (a), or (c) a culture of microorganisms expressing the enzymes listed in (a) with (i) one or more selected from the group consisting of starch, maltodextrin, and sucrose, and (ii) a phosphate wherein the psicose-6-phosphate phosphatase comprises:

i) a motif A comprising at least one sequence selected from the group consisting of SEQ ID NOS: 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, and 91;

ii) a motif B comprising a sequence having at least 90% sequence identity to at least one sequence selected from the group consisting of SEQ ID NOS: 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, and 108; and

iii) A remainder sequence comprising remainder having at least 85% sequence identity to the amino acids sequence other than motif A and/or motif B in any sequence selected from the group consisting of SEQ ID NOS: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20.

9. The method according to claim 1 , wherein the contact reaction is carried out at a pH of 5.0 to 9.0, at a temperature of 40° C. to 80° C. and/or for 2 hours to 24 hours.

10. The method of producing D-psicose according to claim 1 , wherein the psicose-6-phosphate phosphatase consists of the amino acid sequence set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

11. The method of producing D-psicose according to claim 1 , wherein the psicose-6-phosphate phosphatase comprises an amino acid sequence comprising:

i. the motif A comprising a sequence of SEQ ID No: 79, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 92, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 1;

ii. the motif A comprising a sequence of SEQ ID No: 79, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 93, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 2;

iii. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 94, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 3;

iv. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 95, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 4;

V. the motif A comprising a sequence of SEQ ID No: 81, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 96, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 5;

vi. the motif A comprising a sequence of SEQ ID No: 82, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 97, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 6;

vii. the motif A comprising a sequence of SEQ ID No: 83, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 98, and the amino acids sequence comprising the remainder having at least 85% sequence identity to sequence other than motif A and/or motif B in SEQ ID NO: 7;

viii. the motif A comprising a sequence of SEQ ID No: 84, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 99, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 8;

ix. the motif A comprising a sequence of SEQ ID No: 85, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 100, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 9;

x. the motif A comprising a sequence of SEQ ID No: 86, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 101, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 10;

xi. the motif A comprising a sequence of SEQ ID No: 87, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 102, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 11;

xii. the motif A comprising a sequence of SEQ ID No: 88, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 103, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 12;

xiii. the motif A comprising a sequence of SEQ ID No: 89, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 104, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 13;

xiv. the motif A comprising a sequence of SEQ ID No: 90, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 105, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 14;

xv. the motif A comprising a sequence of SEQ ID No: 91, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 106, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 15;

xvi. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 107, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 16;

xvii. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 107, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 17;

xviii. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 94, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 18;

xix. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 98, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 19; or

xx. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 108, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 20.

12. The method of producing D-psicose according to claim 8 , wherein the psicose-6-phosphate phosphatase comprises an amino acid sequence comprising:

i. the motif A comprising a sequence of SEQ ID No: 79, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 92, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 1;

ii. the motif A comprising a sequence of SEQ ID No: 79, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 93, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 2;

iii. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 94, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 3;

iv. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 95, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 4;

V. the motif A comprising a sequence of SEQ ID No: 81, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 96, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 5;

vi. the motif A comprising a sequence of SEQ ID No: 82, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 97, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 6;

vii. the motif A comprising a sequence of SEQ ID No: 83, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 98, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 7;

viii. the motif A comprising a sequence of SEQ ID No: 84, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 99, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 8;

ix. the motif A comprising a sequence of SEQ ID No: 85, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 100, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 9;

x. the motif A comprising a sequence of SEQ ID No: 86, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 101, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 10;

xi. the motif A comprising a sequence of SEQ ID No: 87, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 102, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 11;

xii. the motif A comprising a sequence of SEQ ID No: 88, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 103, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 12;

xiii. the motif A comprising a sequence of SEQ ID No: 89, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 104, and the sequence comprising remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 13;

xiv. the motif A comprising a sequence of SEQ ID No: 90, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 105, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 14;

xv. the motif A comprising a sequence of SEQ ID No: 91, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 106, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 15;

xvi. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 107, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 16;

xvii. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 107, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 17;

xviii. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 94, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 18;

xix. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 98, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 19; or

xx. the motif A comprising a sequence of SEQ ID No: 80, the motif B comprising a sequence having at least 90% sequence identity to SEQ ID NO: 108, and the sequence comprising the remainder having at least 85% sequence identity to amino acids sequence other than motif A and/or motif B in SEQ ID NO: 20.

13. The method according to claim 1 , wherein the psicose-6-phosphate phosphatase is encoded by a nucleotide sequence set forth as SEQ ID NO: 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40.

14. The method according to claim 8 , wherein the psicose-6-phosphate phosphatase consists of the amino acid sequence set forth as SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

15. The method according to claim 8 , wherein the psicose-6-phosphate phosphatase is encoded by a nucleotide sequence set forth as SEQ ID NO: 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39 or 40.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 9, 2020
From: SON, BYUNG-SAM; CHO, HYUN KUG; YANG, SUNG JAE; KIM, SEONG BO; KIM, SEUNG HWAN; PARK, HYUN JUNE
To: CJ CHEILJEDANG CORPORATION
Reel/Frame 052121/0973 →
Priority Claims (1)
KR 10-2017-0097334 · Jul 31, 2017 · national
Continuity (1)
Related Publication 20210355461A1 · Nov 18, 2021
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