CN-116471938-B - Baked and partially baked products with thermostable AMG variants from penicillium

Abstract

The present invention relates to a method for producing a baked or partially baked product comprising a first step of providing a dough comprising a mature thermostable variant of a parent glucoamylase having at least 70% identity with SEQ ID No.1, SEQ ID No. 6, SEQ ID No. 7 or SEQ ID No. 8 and a second step of baking or partially baking the dough to produce a baked or partially baked product, as well as a baked composition comprising said variant and use of said variant.

Inventors

• H. LUNDQVIST
• C. Valmin
• C. ANDERSON
• H. Sinica
• E. Ozkemlexi

Assignees

• 诺维信公司

Dates

Publication Date

20260505

Application Date

20211102

Priority Date

20201102

Claims (20)

1. 1. A method of producing a baked or partially baked product, the method comprising: a) Providing a dough comprising a mature thermostable variant of a parent glucoamylase having 100% identity to SEQ ID NO. 6, and B) Baking or partially baking the dough to produce a baked or partially baked product; wherein the mature thermostable variant has a thermostability improvement of at least 5 ℃ relative to its parent.
2. 2. A method according to claim 1, wherein the baked or partially baked product is a type of bread.
3. 3. The method of claim 2, wherein the bread is molded bread.
4. 4. The method of claim 2, wherein the bread is toast bread, open bread, buns, feno bread, ha Mam bread, samoli bread, long roll bread, bristled bread, hamburger buns, rolls, black bread, whole wheat bread, high-fat bread, bran bread, or flat bread.
5. 5. The method of claim 1, wherein the baked or partially baked product is a pastry.
6. 6. The method of claim 5, wherein the pastry is a tortilla, a cookie, or a cake.
7. 7. The method according to any one of claims 1-6, wherein the parent glucoamylase is from a species of penicillium.
8. 8. The method of claim 7, wherein the parent glucoamylase is from penicillium oxalicum, penicillium michaeli, penicillium rochanterium, or penicillium photospora.
9. 9. The method according to any one of claims 1-6, wherein the mature thermostable variant has a thermostability improvement of at least 6 ℃,7 ℃, or 8 ℃ relative to its parent.
10. 10. The method according to any one of claims 1-6, wherein the mature thermostable variant has a relative activity of at least 150 at 91 ℃ compared to its parent.
11. 11. The method according to claim 10, wherein the mature thermostable variant has a relative activity of at least 200 at 91 ℃ compared to its parent.
12. 12. The method according to claim 11, wherein the mature thermostable variant has a relative activity of at least 250 at 91 ℃ compared to its parent.
13. 13. The method according to claim 12, wherein the mature thermostable variant has a relative activity of at least 300 at 91 ℃ compared to its parent.
14. 14. The method according to any one of claims 1-6, wherein the baked product after final full baking has reduced initial firmness and/or increased initial elasticity when cooled to room temperature, packaged in a sealed container and stored at room temperature until analyzed, and/or reduced firmness increase and/or higher elasticity after 1, 7 or 14 days, compared to a control made without any glucoamylase.
15. 15. The method according to any one of claims 1-6, wherein the baked product after final full baking has at least the same sweetness as a control product made with twice the amount of mature glucoamylase whose amino acid sequence is shown in SEQ ID No. 10.
16. 16. The method according to any one of claims 1-6, wherein the mature thermostable variant glucoamylase is included in the dough in an amount of 0.01-1,000 mg enzyme protein per kg flour.
17. 17. The method according to claim 16, wherein the mature thermostable variant glucoamylase is included in the dough in an amount of 0.01-500 mg enzyme protein/kg flour.
18. 18. The method according to claim 17, wherein the mature thermostable variant glucoamylase is included in the dough in an amount of 0.1-100 mg enzyme protein/kg flour.
19. 19. The method of any one of claims 1-6, wherein the dough further comprises one or more additional enzymes selected from the group consisting of alpha-amylase, maltogenic amylase, beta-amylase, aminopeptidase, carboxypeptidase, catalase, cellulolytic enzyme, chitinase, cutinase, cyclodextrin glycosyltransferase, deoxyribonuclease, esterase, glucan 1, 4-alpha-maltotetraose hydrolase, glucosidase, galactanase, alpha-galactosidase, beta-galactosidase, alpha-glucosidase, beta-glucosidase, hemicellulolytic enzyme, invertase, laccase, lipase, mannanase, mannosidase, oxidase, pectinolytic enzyme, peptide glutaminase, phospholipase, phytase, proteolytic enzyme, ribonuclease, transglutaminase, and xylanase.
20. 20. The method of claim 19, wherein the alpha-amylase is a raw starch degrading alpha-amylase.

Description

Baked and partially baked products with thermostable AMG variants from penicillium Reference to sequence Listing The present application comprises a sequence listing in computer readable form, which is incorporated herein by reference. Technical Field The present invention relates to a method for producing a baked or partially baked (par-baked) product comprising a first step of providing a dough comprising a mature thermostable variant of a parent glucoamylase having at least 70% identity with SEQ ID No.1, SEQ ID No. 6, SEQ ID No. 7 or SEQ ID No. 8 and a second step of baking or partially baking the dough to produce a baked or partially baked product, as well as a baking composition comprising said variant and use of said variant. Background Sugar-containing baked products (breads, biscuits, etc.) worldwide are one of the most popular product categories (products segments). The amount of sugar in the formulation is typically 1% to 25% of the total weight of the flour. However, because of the rising market price of sugar, inadequate sugar supply in certain parts of the world, and health problems, there is a need for methods of producing baked products with reduced amounts of added sugar without sacrificing the quality of the baked product or even potentially improving the quality of the baked product. WO 2019/238423 (Novozymes a/S), denmark discloses methods of producing dough with reduced amounts of added sugar, which methods comprise adding a raw starch degrading alpha-amylase and a glucoamylase to the dough ingredients. Disclosure of Invention The inventors found that thermostable variants of certain glucoamylases exhibit greatly improved performance in the preservation or anti-aging of baked or partially baked products. Another improved property of thermostable variants is that they increase the sweetness or sweetness of the product, thereby reducing the amount of added sugar in traditional formulations. Thus, in a first aspect, the present invention relates to a method of producing a baked or partially baked product, the method comprising: a) Providing a dough comprising a mature thermostable variant of a parent glucoamylase having at least 70% identity with SEQ ID NO. 1, SEQ ID NO. 6, SEQ ID NO. 7 or SEQ ID NO. 8, and B) Baking or partially baking the dough to produce a baked or partially baked product. A second aspect of the invention relates to baking compositions comprising mature thermostable variants of the parent glucoamylase as defined in the first aspect. Other aspects of the invention relate to the use of the baked composition of the second aspect for replacing sugar in a method of producing a baked or partially baked product, increasing the sweetness of a baked or partially baked product, reducing the amount of sugar in a dough in a method of producing a baked or partially baked product and/or extending the shelf life of a baked or partially baked product in a method of producing a baked or partially baked product, and its use in a method as defined in the first aspect, whereby a final fully baked (bakeout) baked or partially baked product has a reduced initial firmness (firmness) and/or increased initial elasticity when cooled to room temperature, packaged in a sealed container and stored at room temperature until analyzed, and/or has a reduced firmness increase and/or higher elasticity after 1, 7 or 14 days, as compared to a control made without any glucoamylase. Preferably, the mature thermostable variant of a parent glucoamylase of the invention has at least 71% identity with SEQ ID No. 1, SEQ ID No.6, SEQ ID No.7 or SEQ ID No.8, e.g. at least 72%, e.g. at least 73%, e.g. at least 74%, e.g. at least 75%, e.g. at least 76%, e.g. at least 77%, e.g. at least 78%, e.g. at least 79%, e.g. at least 80%, e.g. at least 81%, e.g. at least 82%, e.g. at least 83%, e.g. at least 84%, e.g. at least 85%, e.g. at least 86%, e.g. at least 87%, e.g. at least 88%, e.g. at least 89%, e.g. at least 90%, e.g. at least 91%, e.g. at least 92%, e.g. at least 93%, e.g. at least 94%, e.g. at least 95%, e.g. at least 96%, e.g. at least 97%, e.g. at least 98%, e.g. at least 99% identity with SEQ ID No. 1, SEQ ID No. 1.6. Drawings FIG. 1 shows multiple alignments of amino acid sequences of the following mature proteins: Wild-type AMG (PoAMG) from Penicillium oxalicum (Penicillium oxalicum) according to SEQ ID NO. 1 PoAMG variant of SEQ ID NO. 2 denoted "AMG NL PoAMG variant of SEQ ID NO. 3 denoted "AMG anPAV498 PoAMG variant of SEQ ID NO. 4 denoted "AMG JPO001 PoAMG variant of SEQ ID NO. 5 denoted "AMG JPO124 PoAMG variant of SEQ ID NO. 6 denoted "AMG JPO172 Wild-type AMG (PoAMG) from Mikrill (Penicillium miczynskii) according to SEQ ID NO. 7 Wild-type AMG (PoAMG) from Penicillium rochanterium (Penicillium russellii) according to SEQ ID NO. 8 Wild-type AMG (PoAMG) from Penicillium light (Penicillium glabrum) of SEQ ID NO 9 Detailed Description Definition of the definition Sequence identity the degree of relatedness