WO2021218995A1 - 一种酶法减少烘焙产品中油脂使用量的方法 - Google Patents

一种酶法减少烘焙产品中油脂使用量的方法 Download PDF

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Publication number
WO2021218995A1
WO2021218995A1 PCT/CN2021/090509 CN2021090509W WO2021218995A1 WO 2021218995 A1 WO2021218995 A1 WO 2021218995A1 CN 2021090509 W CN2021090509 W CN 2021090509W WO 2021218995 A1 WO2021218995 A1 WO 2021218995A1
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Prior art keywords
dough
edible
fat
amylase
amount
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PCT/CN2021/090509
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English (en)
French (fr)
Inventor
徐清
王雅珍
李守宏
卢小明
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诺维信公司
徐清
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Application filed by 诺维信公司, 徐清 filed Critical 诺维信公司
Priority to US17/996,835 priority Critical patent/US20230320367A1/en
Priority to CA3179735A priority patent/CA3179735A1/en
Priority to CN202180026738.3A priority patent/CN115867141A/zh
Priority to AU2021264896A priority patent/AU2021264896A1/en
Priority to EP21797394.0A priority patent/EP4144220A4/en
Publication of WO2021218995A1 publication Critical patent/WO2021218995A1/zh

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Classifications

    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • A21D13/00—Finished or partly finished bakery products
    • A21D13/06—Products with modified nutritive value, e.g. with modified starch content
    • A21D13/068—Products with modified nutritive value, e.g. with modified starch content with modified fat content; Fat-free products
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • A21D8/00—Methods for preparing or baking dough
    • A21D8/02—Methods for preparing dough; Treating dough prior to baking
    • A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
    • A21D8/042—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with enzymes
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/14—Organic oxygen compounds
    • A21D2/16—Fatty acid esters
    • A21D2/165—Triglycerides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS OR COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings or cooking oils
    • A23D9/007—Other edible oils or fats, e.g. shortenings or cooking oils characterised by ingredients other than fatty acid triglycerides

Definitions

  • the present invention relates to a method for preparing a baked product, and more specifically, to a method for enzymatically reducing the amount of fat in the baked product without reducing or substantially reducing the quality of the baked product.
  • bakery industry has shown a trend of rapid development since the end of the 20th century. With the increase in residents’ acceptance of Western food and changes in consumption concepts, food consumption has further shown a trend of diversification, which will continue to drive the total consumption of bakery products. Continued growth. Due to differences in living habits and tastes, bread in European and American countries is usually light. Such bread can be used as a staple food or eaten early. It will not eat a lot of sugar and oil because it is a staple food. In contrast, Chinese bread is fragrant. It is sweet and has a relatively high fat content.
  • the fat content of different breads varies greatly, ranging from a few percent to tens of percent.
  • the fat content of shortbreads such as shortbread pie and mille bread is very high.
  • the fat content of toast bread is very high. Not low.
  • the addition of fat makes the bread sweeter and softer, the cost of fat is very high. It increases the calories and price of the bread, and the excessively high fat content is not conducive to cardiovascular health and weight control, reducing the amount of oil contained in the noodles or becoming China
  • maltose alpha-amylase can decompose amylopectin in flour to produce small molecular weight dextrin, prevent the interaction between starch and gluten and cause starch aging and retrogradation, thereby keeping baked products from being hard. Therefore, the shelf life is prolonged.
  • the inventor of the present application has discovered that when preparing baked products with high oil content such as toast, by adding an effective amount of maltose alpha-amylase to the dough, the desired edible oil can be prepared. The amount is significantly reduced, and the quality of the baked product made is not worse or substantially worse than the technical solution that does not use the enzyme and does not reduce the amount of edible fats and oils.
  • the present invention relates to a method for preparing a baked product from dough, which includes the steps of incorporating at least one maltose alpha-amylase into the dough and controlling the amount of edible fat added to the dough, and the baked product is made by baking ,
  • the amount of edible oil used in the dough can be reduced by at least 10% by weight compared to the amount of edible oil used in the dough under the same conditions but without the enzyme treatment Proportion.
  • the content of the edible oil and fat relative to the baked product by weight is at least 1% (w/w), for example, at least 2% (w/w), preferably At least 3% (w/w).
  • the baked product here is, for example, a "low-oil baked product", which means that the baked product prepared by the present invention has a lower fat content than the process of the present invention is not used.
  • the present invention also relates to a method for reducing the amount of edible fats and oils in baked products prepared from dough, which includes the step of incorporating at least one maltose alpha-amylase and edible fats into the dough, and the baked products are made by baking, wherein Due to the addition of maltose alpha-amylase, the amount of edible oil used in the dough can be reduced by at least 10% by weight compared to the amount of edible oil used under the same other conditions but without the enzyme treatment.
  • the content of the edible oil and fat relative to the baked product is at least 1% (w/w), for example, at least 2% (w/w), preferably at least 3% (w/w).
  • the bakery products here are, for example, "high-oil bakery products”.
  • the "high-oil bakery products” means that the oil content in this type of bakery products is relatively high compared to other bakery products with light taste, so it is worthwhile
  • the process of the present invention is used to reduce the amount of grease in it.
  • the present invention also relates to a method for preparing a baked product from the dough, which includes the step of incorporating cellulase, at least one maltose alpha-amylase and edible fats and oils into the dough, and the baked product is made by baking.
  • the content of the edible oil and fat relative to the baked product is at least 1% (w/w) by weight, such as at least 2% (w/w), preferably At least 3% (w/w).
  • the amount of edible fats and oils used in the dough is the same as that of other conditions but without the addition of maltose alpha-amylase.
  • the amount of edible fats and oils used during enzyme treatment is reduced by at least 15%, at least 20%, preferably at least 25%, at least 30%, at least 35%, or more preferably at least 40% by weight.
  • the dough further contains cellulase and/or phospholipase.
  • the dough further contains the following composition: flour, edible salt and edible sugar, and preferably may also contain edible flavor, yeast and/or vitamin C.
  • the baked product made is on the 4th day, or Take the 7th day or the 14th day as an example, the product hardness value is lower and/or the product elasticity value is higher.
  • the sensory evaluation of the baked product is not worse than that when the enzyme is not added to the dough and the amount of edible fat is not reduced. There is no substantial deterioration.
  • the sensory evaluation is a comprehensive evaluation of softness to the touch, crumb structure, softness of the mouthfeel, and mouthfeel moistness, and more preferably, the sensory evaluation It further includes the evaluation of olfactory aroma and taste aroma.
  • the sensory evaluation is a comprehensive evaluation of the softness of the mouthfeel, the moistness of the mouthfeel, and the mouthfeel of the cake.
  • the sensory evaluation further includes the cake Evaluation of cohesiveness.
  • the dough comprises flour selected from the group consisting of wheat, barley, rye, oats, corn, sorghum, rice, millet, and any mixtures thereof.
  • the present invention also relates to the use of maltose alpha-amylase or a combination of maltose alpha-amylase and cellulase to reduce the amount of edible fats and oils when preparing baked products with a shelf life of at least 4 days from dough.
  • the process of the present invention can be used for any kind of baked product prepared from dough, whether it is soft or crisp in nature, whether it is white, light or dark type.
  • the content of the edible oil and fat relative to the baked product is at least 1% (w/w) or at least 2% (w/w), preferably at least 3% (w /w), or at least 4% (w/w) relative to the baked product, or at least 5% (w/w) relative to the baked product, or at least 6% (w/w) relative to the baked product .
  • a typical example of the baked product is bread, such as soft bread, preferably buns, toasts, French buns, milk stick breads, and the like.
  • examples of baked products may also be crisp breads, such as croissants and shredded breads.
  • a typical example of a baked product is a cake, such as a sponge cake or a chiffon cake.
  • examples of baked products may also be Chinese snacks with high oil content such as moon cakes.
  • the use of edible fats and oils in the dough can be reduced by at least 15%, such as at least 20%, preferably at least 25%, at least 30%, at least 35%, or more preferably at least 40% .
  • the amount of edible oil used in the preparation of toast bread on the market generally accounts for about 10% by weight of the amount of flour used.
  • the amount of edible oil used can be reduced to about 6% by weight of the amount of flour used, that is to say, the amount of edible oil used in the dough can be reduced by 40%, but if only It is also completely feasible to reduce the amount of grease used by about 25%.
  • the amount of edible oil used in the preparation of the existing commercially available toast bread generally accounts for about 25% by weight of the amount of flour used.
  • the amount of edible oil used in the dough can be reduced by at least 15% by weight compared to the amount of edible oil used under the same conditions but without the enzyme treatment. %, for example at least 20%, preferably at least 25%, at least 30%, at least 35%, or more preferably at least 40%.
  • the present invention is compared with the case where the enzyme composition of maltose amylase or maltose alpha-amylase and cellulase is not added to the dough under the same conditions as other conditions, and the amount of edible fats and oils used is not reduced.
  • the preparation is The sensory evaluation of the finished bakery product has not deteriorated or has not substantially deteriorated.
  • the sensory evaluation is a comprehensive evaluation of softness to the touch, crumb structure, softness of the mouthfeel, and mouthfeel moistness. More preferably, the sensory evaluation also includes the evaluation of olfactory aroma and taste aroma. Evaluation.
  • the finished baked product has a shelf life of at least 4 days, or at least 7 days, or at least 14 days.
  • the baked product prepared by the present invention has an improved shelf life, and the shelf life is at least 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days after baking. 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, or 21 days.
  • the shelf life can be measured as follows: a baked product prepared using the process of the present invention, and compared with a control baked product (that is, when other conditions are the same but the enzyme is not added to the dough, and the amount of edible fat is not reduced). Use a texture analyzer to measure the hardness of the baked product and compare it with a control baked product stored under the same conditions. Improved shelf life is defined as a baked product that is not as hard (ie softer) compared to the control as measured by a texture analyzer.
  • Maltose alpha-amylase is an enzyme classified in EC 3.2.1.133.
  • the enzyme activity does not require the non-reducing end of the substrate.
  • the main enzyme activity leads to the degradation of amylopectin and amylose into maltose or longer Maltodextrin, which can hydrolyze amylose and amylopectin into ⁇ -configuration maltose, and also hydrolyze maltotriose and cyclodextrin.
  • the maltose alpha-amylase is selected from the following group:
  • a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3 or SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO : Composition of the amino acid sequence shown in 3;
  • polypeptide which is a polypeptide derived from (a) by substituting, deleting or adding one or several amino acids to the amino acid sequence in (a);
  • polypeptide which has at least 80% sequence identity with the amino acid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3.
  • the maltose alpha-amylase enzyme provided by the present invention is a polypeptide comprising the amino acid sequence shown in SEQ ID NO. 1, SEQ ID NO: 2 or SEQ ID NO: 3, and a modified polypeptide thereof Or its homologous polypeptide.
  • a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3 includes, for example, a polypeptide consisting of SEQ ID NO: 1, SEQ ID NO: A polypeptide composed of the amino acid sequence shown in SEQ ID NO: 2 or SEQ ID NO: 3, and a polypeptide composed of a signal peptide sequence added to the amino acid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3 , And a polypeptide composed of an amino acid sequence obtained by adding appropriate marker sequences to the N-terminus and/or C-terminus of the amino acid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3.
  • modified polypeptide in the present invention refers to the deletion, substitution, insertion or addition of one or several amino acids in the amino acid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3.
  • the obtained amino acid sequence and a protein with maltose alpha-amylase activity refers to the deletion, substitution, insertion or addition of one or several amino acids in the amino acid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3.
  • the amino acid modification in the modified polypeptide or its homologous polypeptide is a "conservative modification".
  • “conservative substitution” refers to the substitution of one or more amino acid residues with other amino acids that are chemically similar under conditions that do not substantially change the activity of the protein. Examples include the case where a certain hydrophobic residue is substituted with another hydrophobic residue, and the case where a certain polar residue is substituted with another polar residue having the same charge.
  • non-polar (hydrophobic) amino acids include alanine, valine, isoleucine, leucine, proline, tryptophan, phenylalanine, methionine, etc.
  • polar (neutral) amino acids include glycine, serine, threonine, tyrosine, glutamic acid, asparagine, and cysteine.
  • positively charged (basic) amino acids include arginine, histidine, and lysine.
  • negatively charged (acidic) amino acids include aspartic acid, glutamic acid, and the like.
  • the maltose alpha-amylase according to the present invention differs from the amino acids of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3 by no more than 10 amino acids, such as 1, 2, 3, 4, and 5. , 6, 7, 8, 9, or 10.
  • the "homologous polypeptide” in the present invention refers to the amino acid sequence that contains at least 80%, at least 85%, at least 90%, or at least the amino acid sequence represented by SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3 91%, at least preferably 92%, at least preferably 93%, at least preferably 94%, at least preferably 95%, at least preferably 96%, at least preferably 97%, at least preferably 98%, at least preferably 99%, more preferably 100% homology (Sequence identity) amino acid sequence.
  • the degree of sequence identity between two amino acid sequences uses, for example, the EMBOSS software package (EMBOSS: The European Molecular Biology Open Software Suite, Rice, etc., 2000, Trends Genet. 16: 276-277), preferably 3.0
  • the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453) executed in the Needle program of version .0 or higher.
  • the optional parameters used are gap open penalty 10, gap extension penalty 0.5 and EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.
  • the polypeptide of the present invention can be natural, synthetic, semi-synthetic, or recombinantly produced.
  • the polypeptide of the present invention can be produced by genetic engineering, by known peptide synthesis or by digesting the polypeptide of the present invention with an appropriate peptidase.
  • the polypeptide of the present invention can be encoded by a recombinant DNA sequence in a host cell to produce a polypeptide product according to a bioengineering method, or according to solid phase synthesis or liquid phase synthesis, for example, according to Steward and Young (Steward, JM and Young, JD , Solid Phase Peptide Synthesis, 2nd Ed., Pierce Chemical Company, Rockford, I11., (1984)) was synthesized using Applied Biosystem Synthesizer or PioneerTM Peptide Synthesizer according to solid-phase chemical technology.
  • the maltose alpha-amylase is derived from Bacillus, especially from Bacillus licheniformis, Bacillus stearothermophilus or Bacillus amyloliquefaciens.
  • the term "derived from” means that it can be produced or expressed in the original wild-type strain or in another strain, that is, the term covers the expression of wild-type, naturally occurring proteins, as well as any recombinant or genetic engineering The expression of chemical or synthetase in the host.
  • the maltose alpha-amylase can also be a maltose-producing alpha-amylase or a variant thereof as disclosed in, for example, WO 1999/043794; WO 2006/032281; or WO 2008/148845.
  • Suitable commercial maltose alpha-amylases include Novamyl, OPTICAKE 50BG, Novamyl Boost and Novamyl 3D (available from Novozymes A/S).
  • the "maltose alpha-amylase" added to the dough includes the maltose alpha-amylase shown in SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3. Or a modified polypeptide, or a homologous polypeptide, or a mixture thereof.
  • the "maltose alpha-amylase" added to the dough includes the maltose alpha-amylase shown in SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3.
  • the mixture of is, for example, a mixture comprising the maltose alpha-amylase shown in SEQ ID NO: 2 and/or SEQ ID NO: 3.
  • the "maltose alpha-amylase" added to the dough contains 10-95% (w/w) of the maltose alpha-amylase shown in SEQ ID NO: 2 and 90-5% (w/w) the maltose alpha-amylase shown in SEQ ID NO: 3, preferably comprising 20-95% (w/w) of the maltose alpha-amylase shown in SEQ ID NO: 2 and 80- 5% (w/w) of the maltose alpha-amylase shown in SEQ ID NO: 3, preferably comprising 30-95% (w/w) of the maltose alpha-amylase shown in SEQ ID NO: 2 and 70-5% (w/w) of the maltose alpha-amylase shown in SEQ ID NO: 3, preferably comprising 40-95% (w/w) of the maltose alpha-amylase shown in SEQ ID NO: 2 Enzyme and 60-5% (w/w) of the malto
  • the amount of maltose alpha-amylase is usually in the range of 1-1500 ppm, preferably 10-100 ppm per kilogram of flour. Calculated according to the activity of the enzyme, the added amount of malt alpha-amylase is preferably 10-1000MANU/kg flour, for example 50-500MANU/kg. In a preferred embodiment of the present invention, the enzyme activity corresponding to one ppm maltose amylase is about 2.7 MANU.
  • a MANU (Maltamylase Novo Unit) can be defined as the release of 1 ⁇ mol per minute at 37°C and pH 5.0 at a substrate concentration of 10 mg maltotriose (Sigma M 8378) per milliliter of 0.1M citrate buffer The amount of enzyme required for maltose.
  • cellulase as used herein should be understood as an enzyme composition or enzyme mixture comprising cellulase, in particular endoglucanase (EC 3.2.1.4).
  • the cellulase used according to the present invention is an enzyme composition comprising endoglucanase (EC 3.2.1.4).
  • the cellulase may include a sugar-binding module (CBM), which enhances the binding of the enzyme to the cellulose-containing fiber and increases the efficacy of the catalytically active part of the enzyme.
  • CBM is defined as a contiguous amino acid sequence in a sugar-active enzyme that has independent folds with sugar-binding activity.
  • CAZy Internet server or Tomme et al. (1995) in Enzymatic Degradation of Insoluble Polysaccharides (Saddler and Penner), Cellulose-binding domains: classification and properties [cellulose Binding domains: classification and characteristics]. Pages 142-163, American 25 Chemical Society [American Chemical Society No. 25], Washington.
  • Endoglucanase catalyzes 1,4- ⁇ -D-glycosidic bonds in cellulose, cellulose derivatives (such as carboxymethyl cellulose and hydroxyethyl cellulose), lichen starch, and mixing The internal hydrolysis of ⁇ -1,3 glucans such as cereal ⁇ -D-glucan or xyloglucan and other plant materials containing cellulosic parts.
  • the endoglucanase activity can be determined according to the procedure of Ghose, 1987, Pure and Appl. Chem. 59:257-268, for example, by using carboxymethyl cellulose (CMC) hydrolysis.
  • CMC carboxymethyl cellulose
  • the cellulase mixture may also include cellobiohydrolases (E.C.3.2.1.91) and/or ⁇ -glucosidases (E.C.3.2.1.21); especially cellobiohydrolases.
  • Cellobiohydrolase catalyzes the hydrolysis of 1,4- ⁇ -D-glycosidic bonds in cellulose, cellooligosaccharides, or any polymer containing ⁇ -1,4-linked glucose, thereby starting from the reducing end of the chain or The non-reducing end releases cellobiose.
  • cellobiohydrolases examples include CBH I and CBH II from Trichoderma reesei.
  • the cellulase or cellulase mixture may be derived from a Trichoderma strain, such as a Trichoderma reesei strain; or a Humicola strain, such as a Humicola insolens strain; or a Chrysosporium strain , Preferably a Chrysosporium lucknowense strain.
  • a Trichoderma strain such as a Trichoderma reesei strain
  • a Humicola strain such as a Humicola insolens strain
  • a Chrysosporium strain Preferably a Chrysosporium lucknowense strain.
  • the enzyme composition used in the method and/or use of the present invention may be the product of expressing one or more enzymes in a suitable host cell (for example, a fermentation product).
  • the cellulase composition is obtainable from (for example obtained from) Trichoderma, preferably from Trichoderma reesei.
  • Trichoderma reesei An example of a commercial cellulase product produced by Trichoderma reesei is Celluclast BG TM , available from Novozymes A/S.
  • Cellulase can be applied in an effective amount of addition, based on per kilogram of flour, for example, at least 0.5 ppm is added, such as in the range of 0.5-5000 ppm.
  • the use amount of cellulase can be in the range of 1-10 ppm. But it can be added as needed.
  • Phospholipase may have phospholipase A1, A2, B, C, D or lysophospholipase activity; it may or may not have lipase activity. It may be of animal origin, such as from pancreas, snake venom or bee venom, or it may be of microbial origin, such as from filamentous fungi, yeast or bacteria, such as Aspergillus or Fusarium, such as Aspergillus niger, Aspergillus oryzae or Fusarium oxysporum spore. The preferred lipase/phospholipase from Fusarium oxysporum is disclosed in WO 98/26057. Furthermore, the variants described in WO 00/32758 can be used.
  • Suitable phospholipase compositions are LIPOPAN F TM and LIPOPAN XTRA TM (available from Novozymes) or PANAMORE GOLDEN TM and PANAMORE SPRING TM (available from DSM).
  • a suitable commercial lipase preparation is, for example, LIPOPAN TM , such as LIPOPAN TM 50BG available from Novozymes.
  • additional enzymes such as amylase, alpha-amylase, beta-amylase, fungal alpha-amylase, carboxypeptidase, catalase, lipolytic enzyme, galactanase, alpha-galactanase Glycosidase, ⁇ -galactosidase, glucanase, glucoamylase, glucose oxidase, lipase, oxidase, peroxidase, phytase, polyphenol oxidase, protease and/or wood Glycanase can be used together with the enzymes involved in the process according to the present invention.
  • additional enzymes such as amylase, alpha-amylase, beta-amylase, fungal alpha-amylase, carboxypeptidase, catalase, lipolytic enzyme, galactanase, alpha-galactanase Glycosidase, ⁇ -galactosidase, glucan
  • the present invention also relates to methods for preparing dough or baked products. These methods include incorporating an effective amount of enzyme into the dough to control or reduce the amount of edible fat used in the preparation of baked products.
  • incorporación is defined herein as the addition of a material according to the present invention to the dough, to any ingredients of the dough to be made, and/or to any mixture of dough ingredients in the dough to be made.
  • the baking composition of the present invention can be added in any step of dough preparation, and can be added in one, two, or more steps.
  • the composition can be added to the kneaded and baked dough ingredients using methods well known in the art.
  • the term "effective amount” is defined herein as being sufficient to provide a measurable effect on at least one characteristic of interest of the dough and/or baked product according to the needs of the preparation of the baked product.
  • the term “dough” is defined herein as a mixture of flour and other ingredients that is hard enough to knead or roll.
  • the dough of the present invention may comprise flour derived from any grain, including wheat, barley, rye, oats, corn, sorghum, rice, millet, and any mixtures thereof.
  • the dough also contains the following composition: flour, yeast, edible salt, edible sugar, vitamin C and/or edible flavor.
  • the baked product can be made in an appropriate amount although there is no change or substantial change in the softness of the touch, crumb structure, softness of the mouthfeel and mouthfeel moistness.
  • margarine may contain edible flavors, the reduction in the amount of these flavors and edible fats and oils may have a certain impact on the odor, especially the odor after storage.
  • an appropriate amount can be used Add flavors to make up for it.
  • the dough can also contain other dough ingredients, such as protein, such as milk powder, gluten, and soy; eggs (whole eggs, egg yolks, or whites); oxidants, such as ascorbic acid, potassium bromate, potassium iodate, azodicarbonamide (ADA) or peroxide Ammonium sulfate; amino acids, such as L-cysteine; starch; and/or salts, such as sodium chloride, calcium acetate, sodium sulfate, calcium propionate, or calcium sulfate.
  • protein such as milk powder, gluten, and soy
  • eggs whole eggs, egg yolks, or whites
  • oxidants such as ascorbic acid, potassium bromate, potassium iodate, azodicarbonamide (ADA) or peroxide
  • Ammonium sulfate such as ascorbic acid, potassium bromate, potassium iodate, azodicarbonamide (ADA) or peroxide
  • Ammonium sulfate such as ascorbic acid, potassium bromate
  • the starch may be wheat starch, corn starch, maize starch, tapioca starch, cassava starch, potato starch; and/or sugar, such as sucrose, cane sugar, lactose or high fructose Corn syrup.
  • the edible fats and oils are, for example, natural fats or reprocessed fats, where the natural fats are, for example, vegetable fats and animal fats, such as vegetable oil or butter, the vegetable oils are, for example, rapeseed oil or soybean oil, and the reprocessed fats are, for example, Margarine, shortening, non-dairy cream, etc.
  • an emulsifier is not required; for some applications, an emulsifier may be required.
  • the emulsifier suitable for the present invention is preferably an emulsifier selected from the group consisting of: monoglyceride diacetyl tartrate (DATEM), sodium stearoyl lactylate (SSL), calcium stearoyl lactate (CSL), ethoxylate Alkylated monoglycerides and diglycerides (EMG), distilled monoglycerides (DMG), polysorbate (PS), and succinylated monoglycerides (SMG).
  • DATEM monoglyceride diacetyl tartrate
  • SSL sodium stearoyl lactylate
  • CSL calcium stearoyl lactate
  • EMG ethoxylate Alkylated monoglycerides and diglycerides
  • EMG distilled monoglycerides
  • PS polysorbate
  • the dough of the present invention can be fresh, frozen or partially baked (pre-baked).
  • the dough of the present invention is usually a dough that has been fermented or a dough that will undergo fermentation.
  • the dough can be fermented in various ways, for example by adding a chemical leavening agent (such as sodium bicarbonate) or by adding a leavening agent (leavened dough), but preferably by adding a suitable yeast culture such as Saccharomyces cerevisiae (baker's yeast)
  • a suitable yeast culture such as Saccharomyces cerevisiae (baker's yeast)
  • a culture such as a commercially available strain of Saccharomyces cerevisiae, is used to ferment the dough.
  • the present invention also relates to a baking fat for baking, which comprises edible fat, cellulase and at least one maltose alpha-amylase, preferably the fat also contains emulsifier and/or antioxidant.
  • a baking fat for baking which comprises edible fat, cellulase and at least one maltose alpha-amylase, preferably the fat also contains emulsifier and/or antioxidant.
  • the preparation process is the same as other conditions but not when the baking fat is added with the enzyme.
  • the amount of edible fat used in the edible fat can be reduced by at least 10% by weight.
  • the content of the edible fat relative to the baked product is at least 1% (w/w ), for example, at least 2% (w/w), preferably at least 3% (w/w).
  • the edible fat contained in the fat used for baking is vegetable oil, for example, rapeseed oil, soybean oil, tallow, butter, coconut oil or palm kernel oil.
  • the emulsifier is at least one of mono- and di-fatty acid glycerides, Span 60, polyglycerides, sucrose esters, and sodium stearoyl lactylate.
  • the present invention is particularly useful for preparing dough and baked products in an industrialized process, wherein the dough used for preparing baked products is mechanically prepared using automated or semi-automated equipment.
  • the process of preparing bread usually involves the following sequential steps: making dough (and an optional proofing step), pressing or dividing the dough, shaping or rolling, and proofing, these steps are well known in the art.
  • Example 1 Application of enzymes in reducing oil on toast
  • Toast bread is prepared by a direct dough fermentation process. All raw materials used are food grade. Among them, fungal amylase is Fungamyl 2500SG, xylanase is Pentopan Mono BG, glucose oxidase is Gluzyme Mono 10000BG, and cellulase is Celluclast BG is a commercial product of Novozymes, maltoamylase (a combination of SEQ ID NO: 2 and SEQ ID NO: 3), which is derived from Bacillus stearothermophilus.
  • the ingredient list in Table 1 and the test plan in Table 2 weigh the flour, edible salt, sucrose, yeast, calcium propionate, SSL, CSL, DMG into the noodle tank (vertical mixer, DIOSNA brand) Add enzymes, water and milk flavor, stir, and adjust the water temperature to keep the temperature of the mixed dough at 23°C. After adding margarine, continue to stir until the gluten is fully expanded.
  • the dough is divided into 450g/piece, the divided dough is initially formed and left for 10 minutes, and then further formed by the noodle forming machine 2 times, put into the mold, and then put into the proofing box, the proofing temperature is 35°C , The proofing humidity is 85%, proofing for 120min, the proofed dough is put into the oven for baking, the temperature of the oven (the upper fire is 185°C, the lower fire is 185°C), the baked toast is cooled and the product is made.
  • Hardness measurement method use a toast slicer to cut the toast bread (each bread slice is 1.2cm thick), and the cut bread slices are made into a set of two slices (thickness 2.4cm), use TA.XT Plus
  • the texture analyzer is used to determine the unit of gram. The higher the hardness value, the lower the softness of the bread and the worse the quality.
  • Method for measuring elasticity cut the toast with a toast slicer (each slice of bread has a thickness of 1.2 cm). Two slices of bread after cutting are made into a group (thickness of 2.4cm), and the measurement is carried out with the TA.XT Plus texture analyzer, with% as the unit. The higher the elasticity value, the better the quality of the bread.
  • the total sensory evaluation of each batch of bread 24 hours after it was made was that the average value of batch A was 5.0 points, the average value of batch B was 4.8 points, and the average value of batch B was 4.8 points.
  • the average value of C is 5.2 points, and the average value of batch D is 5.3 points;
  • the sensory evaluation of each batch of bread made at 336 hours (that is, the 14th day) after preparation is the average of batch A
  • the value is 5.0 points
  • the average value of batch B is 4.2 points
  • the average value of batch C is 5.4 points
  • the average value of batch D is 5.6 points.
  • the quality of the bread prepared by batch C on the 14th day was better than that of the bread prepared by batch A on the 7th day, and the texture of the bread prepared by batch D on the 14th day was significantly better than that of the batch.
  • Toast bread is prepared by direct dough fermentation process. All raw materials used are food grade. Among them, fungal amylase is Fungamyl 2500SG, xylanase is Pentopan Mono BG, glucose oxidase is Gluzyme Mono 10000BG, and cellulase is Celluclast BG is a commercial product of Novozymes. Maltose amylase (SEQ ID NO: 1 or SEQ ID NO: 2) is derived from Bacillus stearothermophilus.
  • the ingredient list in Table 4 and the test plan in Table 5 weigh the flour, edible salt, sucrose, yeast, calcium propionate, SSL, CSL, DMG into the noodle tank (vertical mixer, DIOSNA brand) ), add enzymes, water and milk flavors, stir, and adjust the water temperature to keep the temperature of the mixed dough at 23°C. After adding margarine, continue to stir until the gluten is fully expanded. Leave the beaten dough for 10 minutes. Then divide the dough into 450g/piece. The divided dough is preliminarily formed and allowed to stand for 10 minutes.
  • the proofing temperature is 35 °C, proofing humidity 85%, proofing for 120min, put the proofed dough into the oven for baking, the oven temperature (up to 185°C, down to 185°C), and the baked toast will be cooled to make the product.
  • the bread made by adding maltogenic amylase to the dough is on the 1st and 14th days Both show excellent softness and elasticity.
  • the sensory evaluation of the made bread is better, and there is a certain synergistic effect between maltogenic amylase and cellulase. .
  • the 14-day evaluation is sufficient for short-term breads such as toast bread.
  • the addition of maltogenic amylase to the dough can ensure that the overall evaluation of the key indicators of the toast bread does not deteriorate or change substantially. In the case of poor or even improved conditions, the amount of edible fats and oils used during preparation is greatly reduced.
  • Example 3 Application of enzymes in reducing oil in French bread
  • Toast bread is prepared by a direct dough fermentation process. All raw materials used are food grade. Among them, fungal amylase is Fungamyl 2500SG, xylanase is Pentopan Mono BG, glucose oxidase is Gluzyme Mono 10000BG, and cellulase is Celluclast BG is a commercial product of Novozymes, maltoamylase (a combination of SEQ ID NO: 2 and SEQ ID NO: 3), which is derived from Bacillus stearothermophilus.
  • ingredient table 8 weigh the flour, edible salt, sucrose, yeast, CSL and sodium dehydroacetate into the noodle tank (vertical mixer, DIOSNA brand), mix well, add eggs, Stir the syrup, water and enzymes, add shortening, beat the noodles until the gluten expands, and keep the temperature of the dough at 23°C.
  • noodle tank vertical mixer, DIOSNA brand
  • Roll the noodles divide the noodles into 25g pieces, put them in the mold, and put them in the proofing box, proofing temperature 35°C, proofing humidity 85%, proofing for 90 minutes, and then put the proofed dough into the oven for baking , Oven temperature (up to 200°C, down to 155°C), baking time 10min.
  • the baked French bread is packaged after cooling.
  • the batches of French bread are made 24 hours (i.e. day 1), 72 hours (day 3), 336 hours (day 14), 504 hours (day 21), and 672 hours ( Sensory evaluation on day 28):
  • the sponge cake is prepared by the whole egg beaten process, and all the raw materials used are food grade.
  • the oil reducing enzyme used is a combination of cellulase and maltose amylase
  • the cellulase is Celluclast BG, which is a commercial product of Novozymes
  • the maltose amylase SEQ ID NO: 2 and SEQ ID NO: 3
  • Baking percentage% (based on batter) Flour (Queen Low Gluten Flour) 25 Eggs (purchased from Deqingyuan) 28 Sugar powder 27 Vegetable oil (purchased from Arowana) 6 or 11 water 5 Whole milk powder (purchased from Nestle) 1 cooking salt 0.3 Baking powder (purchased from Puratos) 0.3 Calcium Propionate (Medaner) 0.4 Oil reducing enzyme 200ppm
  • Hardness measurement method cut the surface of the cake flat (the height of each cake is 45cm), and use the TA.XT Plus texture analyzer to measure it. The unit is gram. The higher the hardness value, the lower the softness of the finished cake. The worse the quality.
  • Cohesiveness measurement method The ratio of the energy (the integral of the positive peak area under the curve) of the two pressing processes in the texture test is used as the texture of the sample to test the cohesiveness. Cohesiveness reflects the taste quality of the cake. The higher the cohesion, the better the cohesion and the better the quality of the cake.
  • the total sensory evaluation of each batch of cakes made at 72 hours (that is, the 3rd day) after being made is that the average value of batch A is 5 points, the average value of batch B is 3 points, and the batch The average value of C is 5 points;
  • the sensory evaluation of each batch of cakes made at 168 hours (that is, the 7th day) after being made is, the average value of batch A is 5 points, and the average value of batch B
  • the value is 4 points, and the average value of batch C is 5.8 points;
  • the sensory evaluation of each batch of cakes made 360 hours after the preparation (that is, the 15th day) is, the average value of batch A is 5
  • the average value of batch B is 4 points, and the average value of batch C is 5.3 points.

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Abstract

一种减少烘焙产品中可食用油脂用量的方法，包括将至少一种麦芽糖α-淀粉酶和可食用油脂掺入面团，通过烘焙制成烘焙产品，所述面团中可食用油脂的使用量相对于未采用所述酶处理可至少减少10wt％；所述酶还可包括纤维素酶和/或磷脂酶。该方法不降低或实质性降低烘焙产品的品质，并具有至少4天的货架期。还涉及由上述酶与可食用油脂制得的烘焙油脂。

Description

一种酶法减少烘焙产品中油脂使用量的方法
涉及序列表
本申请含有计算机可读形式的序列表，其通过提述并入本文。
技术领域
本发明涉及一种烘焙产品的制备方法，更具体的说，涉及一种酶法减少烘焙产品中油脂使用量的方法，而不降低或者实质性降低烘焙产品的品质。
技术背景
中国的烘焙行业从20世纪末开始呈现出快速发展的趋势，随着居民对西方食品接受程度的提高以及消费观念的转变，食品消费进一步呈现多样化的趋势，也将继续推动着烘焙食品消费总量持续增长。由于生活习惯和口味的差异，欧美国家的面包通常比较清淡，这样的面包可以作为主食或早点吃，不会因为是主食而吃进大量的糖和油脂，相比之下，中国的面包又香又甜，油脂含量比较高。
不同面包的脂肪含量差别很大，从百分之几到百分之几十不等，例如起酥派、千层面包之类的起酥面包的油脂含量都非常高的，土司面包的油脂含量也不低。虽然油脂的添加让面包更香甜柔软，可是油脂的成本很高，它增加了面包的热量和价格，而且过高的油脂添加量不利于心脑血管健康和体重控制，降低面包含油量或成为中国面包发展的趋势和需求之一。
中国市场监管总局发文的《关于规范使用食品添加剂的指导意见》(市监食生〔2019〕53号)要求食品生产经营者生产加工食品应当积极推行减盐、减油、减糖行动。根据中国食品安全国家标准GB-28050，要想在预包装食品营养标签上宣称能够减少脂肪，就必须能够与消费者熟知的同一属类食品相比实现脂肪含量减少25％以上。
因此，仍有需求去改进现有含油量高的烘焙产品的制备工艺，去减少制备过程中的食用油脂的使用量，而不降低或实质性降低该烘焙产品的品质。
发明概述
尽管总所周知，麦芽糖α-淀粉酶能对面粉中的支链淀粉进行分解，产生小分子量的糊精，防止淀粉和面筋之间相互反应而导致淀粉老化回生，从而保持烘焙产品不变硬，从而延长保质期，但本申请的发明人已经发现，在制备土司面包等含油量较高的烘焙产品的时候，通过在面团中添加有效量的麦芽糖α-淀粉酶，能够实现制备所需可食用油脂用量的显著减少，而且制成的烘焙产品的品质，相对于不使用该酶也没有减少可食用油脂用量的技术方案相比，其没有变差或者实质性变差。特别的是，当在面团添加了纤维素酶，形成了含麦芽糖α-淀粉酶和纤维素酶的酶组合物时，由于存在麦芽糖α-淀粉酶与纤维素酶的协同作用，在显著减少制备过程时的可食用油脂含量时制成的烘焙产品的品质更优。
鉴于此，本发明涉及一种由面团制备烘焙产品的方法，包括将至少一种麦芽糖α-淀粉酶掺入该面团以及控制在面团中添加的可食用油脂用量的步骤，通过烘焙制成烘焙产品，其中，由于麦芽糖α-淀粉酶的添加，所述面团中可食用油脂的使用量相对于其他条件相同但未采用所述酶处理时的可食用油脂用量，按重量计可至少减少10％的比例。优选的是在该制成的低油烘焙产品中，所述食用油脂的含量相对于该烘焙产品按重量计至少是1％(w/w)，例如是至少2％(w/w)、优选至少是3％(w/w)。此处的烘焙产品例如是“低油烘焙产品”，该“低油烘焙产品”是指，相对于未使用本发明工艺而言，本发明制备的烘焙产品中的油脂含量较低。
本发明还涉及一种减少由面团制备的烘焙产品中的食用油脂用量的方法，包括将至少一种麦芽糖α-淀粉酶和可食用油脂掺入该面团的步骤，通过烘焙制成烘焙产品，其中，由于麦芽糖α-淀粉酶的添加，所述面团中可食用油脂的使用量相对于其他条件相同但未采用所述酶处理时的可食用油脂用量，按重量计可至少减少10％的比例。优选的是，在该制成的烘焙产品中，所述食用油脂的含量相对于该烘焙产品至少是1％(w/w)，例如是至少2％(w/w)、优选至少是3％(w/w)。此处的烘焙产品例如是“高油烘焙产品”，该“高油烘焙产品”是 指，该类烘焙产品中的油脂含量相对于口感清淡的其他烘焙产品而言，含油量偏高，因而值得采用本发明工艺去降低其中的油脂用量。
本发明还涉及一种由面团制备烘焙产品的方法，包括将纤维素酶和至少一种麦芽糖α-淀粉酶以及可食用油脂掺入该面团中的步骤，通过烘焙制成烘焙产品。优选的是，在该制成的烘焙产品中，所述可食用油脂的含量相对于该烘焙产品按重量计至少是1％(w/w)，例如是至少2％(w/w)、优选至少是3％(w/w)。
在本发明的优选的实施方式中，由于麦芽糖α-淀粉酶的添加，相对于未麦芽糖α-淀粉酶时，所述面团中可食用油脂的使用量，相对于其他条件相同但未采用所述酶处理时的可食用油脂用量，按重量计其比例至少减少15％、至少减少20％、优选至少减少25％、至少减少30％、至少减少35％，或更优选至少减少40％。
在本发明的优选的实施方式中，其中，在所述面团中还进一步包含纤维素酶和/或磷脂酶。
在本发明的优选的实施方式中，其中，在所述面团中还含有以下组成的组：面粉、食用盐和食用糖，优选的还可以含有食用香精、酵母和/或维生素C。
在本发明的优选的实施方式中，其中，其他条件相同但与未在面团添加所述酶、也没有减少可食用油脂使用量时相比，其制成的烘焙产品，以第4天、或第7天、或第14天为例，其产品硬度值更低和/或其产品弹性值更高。
在本发明的优选的实施方式中，其中，其他条件相同但与未在面团添加所述酶、也没有减少可食用油脂使用量时相比，其制成的烘焙产品的感官评价没有变差或者没有实质性变差。
在本发明的优选的实施方式中，其中，对面包来说，所述感官评价是触感柔软度、面包瓤结构、、口感柔软度和口感湿润度的综合评价，更优选的是，该感官评价进一步含对嗅觉香气和味觉香味的评价。
在本发明的优选的实施方式中，其中，对蛋糕来说，所述感官评价是对蛋糕的口感柔软度、口感湿润度和化口性的综合评价，优选，所述感官评价还含有对蛋糕的粘聚性的评价。
根据前述权利要求中任一项所述的方法，其中该面团包含选自由以下组成的组的面粉：小麦、大麦、黑麦、燕麦、玉米、高粱、稻、粟、以及其任何混合物。
本发明还涉及麦芽糖α-淀粉酶或麦芽糖α-淀粉酶和纤维素酶的组合在由面团制备具有至少4天货架期的烘焙产品时减少可食用油脂用量中的用途。
发明详述
烘焙产品
本发明的工艺可用于从面团制备的任何一种烘焙产品，无论是柔软的或脆的性质，无论是白色、浅色或深色类型。有本发明工艺制成的烘焙产品中，所述可食用油脂的含量相对于该烘焙产品至少是1％(w/w)或者至少是2％(w/w)，优选至少是3％(w/w)，或者相对于该烘焙产品至少是4％(w/w)，或者相对于该烘焙产品至少是5％(w/w)，或者相对于该烘焙产品至少6％(w/w)。尽管在该烘焙产品中仍然存在该一定含量的可食用油脂，但正是由于采用了本发明的工艺，使得制成的烘焙产品在感官评价没有变差或者没有实质性变差的情况下，相对于不才采用本发明工艺时相比，已经显著了减少了可食用油脂的含量。
在本发明的优选的实施方式中，烘焙产品的典型实例是面包，例如是软式面包，优选是小圆面包、土司面包、法式小面包和奶棒面包等。在本发明的优选的实施方式中，烘焙产品的实例还可以是起酥面包，例如牛角面包和手撕面包等。在本发明的优选的实施方式中，烘焙产品的典型实例是蛋糕，例如是海绵蛋糕或者是戚风蛋糕。在本发明的优选的实施方式中，烘焙产品的实例还可以是月饼等含油量较高的中式点心等。
在本发明的优选的实施方式中，由于采用了本发明工艺，相对于其他条件相同但采未用麦芽糖α-淀粉酶或麦芽糖α-淀粉酶和纤维素酶的酶组合物处理时，按重量计，所述面团中可食用油脂的使用量可减少的比例是至少是15％、例如至少20％、优选至少减少25％、至少减少30％、至少减少35％，或更优选至少减少40％。
在本发明的优选的实施方式中，在制备土司面包的时候，由于一般现有市售的土司面包在制备时，可食用油脂的使用量一般占面粉使用量按重量计10％ 左右，由于采用了本发明工艺，可食用油脂的使用量可减少到面粉使用量按重量计的6％左右，也就是说，所述面团中可食用油脂的使用量可减少的比例是40％,但若仅仅减少油脂使用量25％左右，也是完全可行的。
在本发明的优选的实施方式中，在制备手撕面包的时候，由于一般现有市售的土司面包在制备时，可食用油脂的使用量一般占面粉使用量按重量计25％左右，由于采用了本发明工艺，在制备过程中，所述面团中可食用油脂的使用量，相对于其他条件相同但未采用所述酶处理时的可食用油脂用量，按重量计其比例可至少减少15％，例如至少减少20％、优选至少减少25％、至少减少30％、至少减少35％，或更优选至少减少40％。
在本发明的优选的实施方式中，其中，与其他条件相同但未在面团添加麦芽糖淀粉酶或麦芽糖α-淀粉酶和纤维素酶的酶组合物、也没有减少可食用油脂使用量时相比，其制成的烘焙产品，以第4天、第7天、或第14天为例，其产品硬度值更低和/或其产品弹性值更高。
在本发明的优选的实施方式中，与其他条件相同但未在面团添加麦芽糖淀粉酶麦芽糖α-淀粉酶和纤维素酶的酶组合物、也没有减少可食用油脂使用量时相比，其制成的烘焙产品的感官评价没有变差或者没有实质性变差。优选的是，对面包来说，所述感官评价是触感柔软度、面包瓤结构、口感柔软度和口感湿润度的综合评价，更优选的是，该感官评价还含对嗅觉香气和味觉香味的评价。
在本发明的优选的实施方式中，尽管显著降低了可食用油脂的用量，但是，制成的所述烘焙产品具有至少4天、或至少7天、或至少14天的货架期。
在本发明的优选实施例中，本发明制备的烘焙产品具有改善的保质期，其保质期为烘焙后至少4天、5天、6天、7天、8天、9天、10天、11天、12天、13天、14天、15天、16天、17天、18天、19天、20天或21天。
保质期可以按如下测量：使用本发明工艺制备的烘焙产品，并与对照烘焙产品(即以其他条件相同但未在面团添加所述酶、也没有减少可食用油脂使用量时相比)进行比较。使用质构分析仪来测量烘焙产品的硬度，并且与在相同条件下储存的对照烘焙产品行比较。改善的保质期被定义为如通过质构分析仪测量的与对照相比不那么硬(即较软)的烘焙产品。
麦芽糖α-淀粉酶
麦芽糖α-淀粉酶是在EC3.2.1.133中进行分类的酶，所述酶活性不需要底物的非还原末端，主要的酶活性导致支链淀粉和直链淀粉降解为麦芽糖或较长的麦芽糊精，它能够将直链淀粉和支链淀粉水解为α构型的麦芽糖，而且还能水解麦芽三糖和环糊精。
在本发明优选的实施方式中，麦芽糖α-淀粉酶选自下组：
(a)多肽，其包含SEQ ID NO:1、SEQ ID NO:2和/或SEQ ID NO:3所示的氨基酸序列或由SEQ ID NO:1、SEQ ID NO:2和/或SEQ ID NO:3所示的氨基酸序列组成；
(b)多肽，其是在(a)中的氨基酸序列经过取代、缺失或添加一个或几个氨基酸由(a)衍生的多肽；
(c)多肽，其与SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3所示的氨基酸序列具有至少80％的序列同一性。
在本发明的优选实施方式中，本发明提供的麦芽糖α-淀粉酶酶是包含SEQ ID NO.1、SEQ ID NO:2或SEQ ID NO:3所示的氨基酸序列的多肽，以及其修饰多肽或其同源多肽。
在本发明的优选实施方式中，“包含SEQ ID NO：1、SEQ ID NO:2或SEQ ID NO:3所示的氨基酸序列的多肽”包括，例如，由SEQ ID NO：1、SEQ ID NO:2或SEQ ID NO:3所示的氨基酸序列组成的多肽，及在由SEQ ID NO：1、SEQ ID NO:2或SEQ ID NO:3所示的氨基酸序列中添加信号肽序列构成的多肽，以及由在SEQ IDNO：1、SEQ ID NO:2或SEQ ID NO:3所示的氨基酸序列的N末端和/或C末端添加适当标记序列而得的氨基酸序列构成的多肽。
本发明中的“修饰多肽”指的是，包含在SEQ ID NO：1、SEQ ID NO:2或SEQ ID NO:3所示的氨基酸序列中缺失、取代、插入或添加一个或几个氨基酸而得到的氨基酸序列，并具有麦芽糖α-淀粉酶活性的蛋白质。
在本发明优选的实施方式中，所述的修饰多肽或其同源多肽中的对氨基酸的修饰是“保守性修饰”。例如“保守性取代”指的是，在不实质性改变蛋白质活性的条件下，将一个或多个氨基酸残基以化学上类似的其它氨基酸取代。可举出，将某疏水性残基以其它疏水性残基取代的情形，将某极性残基以具有 相同电荷的其它极性残基取代的情形。
可进行这种保守性取代的功能类似的氨基酸，在各氨基酸相应的技术领域都是公知的。具体而言，作为非极性(疏水性)氨基酸，可以举出，丙氨酸、缬氨酸、异亮氨酸、亮氨酸、脯氨酸、色氨酸、苯丙氨酸、蛋氨酸等。作为极性(中性)氨基酸，可以举出，甘氨酸、丝氨酸、苏氨酸、酪氨酸、谷氨酸、天冬酰胺、半胱氨酸等。作为带有正电荷(碱性)的氨基酸，可以举出，精氨酸、组氨酸、赖氨酸等。此外，作为负电荷(酸性)氨基酸，可以举出，天冬氨酸、谷氨酸等。
在一个方面，根据本发明的麦芽糖α-淀粉酶与SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:3的氨基酸相差不超过10个氨基酸，例如1、2、3、4、5、6、7、8、9或10个。
本发明中的“同源多肽”指的是，包含和SEQ ID NO：1、SEQ ID NO:2或SEQ ID NO:3表示的氨基酸序列具有至少80％，至少85％，至少90％，至少91％，至少优选92％，至少优选93％，至少优选94％，至少优选95％，至少优选96％，至少优选97％，至少优选98％，至少优选99％，更优选100％同源性(序列同一性)的氨基酸序列。
就本发明而言，两个氨基酸序列之间的序列同一性程度使用如EMBOSS软件包(EMBOSS:The European Molecular Biology Open Software Suite,Rice等,2000,Trends Genet.16:276-277)，优选3.0.0版或更高版本的Needle程序中所执行的Needleman-Wunsch算法(Needleman和Wunsch,1970,J.Mol.Biol.48:443-453)来测定。使用的可选参数为缺口开放罚分(gap open penalty)10，缺口延伸罚分(gap extension penalty)0.5和EBLOSUM62(BLOSUM62的EMBOSS版)取代矩阵。使用Needle标记为“最高同一性(longest identity)”的输出结果(使用-nobrief选项获得)作为同一性百分比，并计算如下：
(同样的残基×100)/(比对长度－比对中缺口的总数)
本发明多肽可以是天然的、合成的、半合成的、或者重组产生。本发明的多肽可通过基因工程、通过已知的肽合成或通过用适当的肽酶消化本发明之多肽来生成。优选的是，本发明多肽可按生物工程方法由宿主细胞中的重组DNA序列编码产生多肽产物，也可按照固相合成或液相合成，例如可按照Steward和Young(Steward，J.M.和Young，J.D.，Solid Phase Peptide Synthesis，2nd Ed.， Pierce Chemical Company，Rockford，I11.，(1984))描述的方法用Applied Biosystem合成仪或PioneerTM肽合成仪按固相化学技术合成。
在本发明的优选实施方式中，所述的麦芽糖α-淀粉酶源自芽孢杆菌属，特别是源自地衣芽孢杆菌、嗜热脂肪芽孢杆菌或解淀粉芽孢杆菌。所述术语“源自”是指它可以在该原始野生型菌株、在另一种菌株中产生或表达，即该术语涵盖了野生型、自然发生的蛋白质的表达，以及任一重组、基因工程化或合成酶在宿主中的表达。
所述的麦芽糖α-淀粉酶还可以是在如例如WO 1999/043794；WO 2006/032281；或WO 2008/148845中所披露的产麦芽糖α-淀粉酶或其酶的变体。
适合的商业化麦芽糖α-淀粉酶包括Novamyl、OPTICAKE 50BG、Novamyl Boost和Novamyl 3D(可获得自诺维信公司(Novozymes A/S))。
在本发明优选的实施例中，添加至面团中的“麦芽糖α-淀粉酶”是包含SEQ ID NO:1、SEQ ID NO:2、或SEQ ID NO:3所示的麦芽糖α-淀粉酶，或其修饰多肽，或其同源多肽、或其混合物。
在本发明优选的实施例中，添加至面团中的“麦芽糖α-淀粉酶”是包含SEQ ID NO:1、SEQ ID NO:2和/或SEQ ID NO:3所示的麦芽糖α-淀粉酶的混合物，例如是包含SEQ ID NO:2和/或SEQ ID NO:3所示的麦芽糖α-淀粉酶的混合物。
在本发明优选的实施例中，添加至面团中的“麦芽糖α-淀粉酶”包含10-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和90-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶，优选的是包含20-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和80-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶，优选的是包含30-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和70-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶，优选的是包含40-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和60-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶，优选的是包含50-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和50-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶，优选的是包含60-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和40-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶，优选的是包含70-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和30-5％(w/w)的SEQ ID NO:3所 示的麦芽糖α-淀粉酶，优选的是包含80-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和20-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶，优选的是包含90-95％(w/w)的SEQ ID NO:2所示的麦芽糖α-淀粉酶和10-5％(w/w)的SEQ ID NO:3所示的麦芽糖α-淀粉酶。
麦芽糖α-淀粉酶的量通常在每千克面粉1-1500ppm、优选10-100ppm的范围内。按酶的活性计算，麦芽α-淀粉酶的添加量优选为10-1000MANU/kg面粉，例如50-500MANU/kg。在本发明优选的实施例中，一个ppm麦芽糖淀粉酶对应的酶活约为2.7个MANU。一个MANU(麦芽淀粉酶Novo单位)可以定义为在每毫升0.1M柠檬酸盐缓冲液10mg麦芽三糖(Sigma M 8378)底物浓度下，在37℃、pH 5.0下30分钟内每分钟释放1μmol麦芽糖所需的酶量。
纤维素酶
如本文使用的术语“纤维素酶”应理解为包含纤维素酶的酶组合物或酶混合物，特别是内切葡聚糖酶(EC 3.2.1.4)。
在一个实施例中，根据本发明使用的纤维素酶是包含内切葡聚糖酶(EC 3.2.1.4)的酶组合物。
纤维素酶可以包含糖-结合模块(CBM)，其增强酶与含纤维素纤维的结合，并增加酶催化活性部分的功效。CBM定义为糖-活性酶中的相连的(contiguous)氨基酸序列，该酶具有糖-结合活性的独立折叠。要了解CBM的进一步信息，可以参见CAZy互联网服务器或Tomme等人(1995)于Enzymatic Degradation of Insoluble Polysaccharides[不溶性多糖的酶降解](Saddler和Penner编)，Cellulose-binding domains:classification and properties[纤维素结合结构域：分类和特性].第142-163页，American 25 Chemical Society[美国化学学会第25期]，华盛顿。
内切葡聚糖酶(E.C.3.2.1.4)催化纤维素、纤维素衍生物(例如羧甲基纤维素和羟乙基纤维素)、地衣淀粉中1,4-β-D-糖苷键、混合β-1,3葡聚糖如谷类β-D-葡聚糖或木葡聚糖和包含纤维素部分的其他植物材料中的β-1,4键的内水解。
内切葡聚糖酶活性可以根据Ghose,1987,Pure and Appl.Chem.[纯粹与应用化学]59:257-268的程序，例如，通过使用羧甲基纤维素(CMC)水解来进 行确定。
除内切葡聚糖酶外，纤维素酶混合物还可以包括纤维二糖水解酶(E.C.3.2.1.91)和/或β-葡糖苷酶(E.C.3.2.1.21)；特别是纤维二糖水解酶。
纤维二糖水解酶催化纤维素、纤维寡糖或任何含有β-1,4-连接的葡萄糖的聚合物中的1,4-β-D-糖苷键的水解，从而从该链的还原端或非还原端释放纤维二糖。
纤维二糖水解酶的实例包括来自里氏木霉的CBH I和CBH II。
在一些实施例中，该纤维素酶或纤维素酶混合物可以衍生自木霉属菌株，例如里氏木霉菌株；或腐质霉属菌株，例如特异腐质霉菌株；或金孢子菌属菌株，优选地卢克诺文思金孢子菌(Chrysosporium lucknowense)菌株。
在一些实施例中，用于在本发明的方法和/或用途中使用的酶组合物可以是在合适的宿主细胞(例如发酵产物)中表达一种或多种酶的产物。
优选地，纤维素酶组合物可以获得自(例如从以下获得)木霉属，优选获得自里氏木霉。
由里氏木霉产生的商业纤维素酶产物的实例是Celluclast BG TM，可获得自诺维信公司(Novozymes A/S)。
纤维素酶可以以有效量的添加量施用，基于每千克面粉，例如至少添加0.5ppm，例如是0.5-5000ppm的范围内,优选的是，纤维素酶的使用量可以是1-10ppm的范围，但可根据需要来添加。
磷脂酶
磷脂酶可以具有磷脂酶A1、A2、B、C、D或溶血磷脂酶活性；它可能有也可能没有脂肪酶活性。它可以具有动物来源，例如来自胰腺、蛇毒或蜂毒，或它可以具有微生物来源，例如来自丝状真菌、酵母或细菌，例如曲霉属或镰孢属，例如黑曲霉、米曲霉或尖孢镰孢。来自尖孢镰孢的优选的脂肪酶/磷脂酶披露于WO 98/26057中。而且，可以使用描述于WO 00/32758中的变体。
适合的磷脂酶组合物是LIPOPAN F TM和LIPOPAN XTRA TM(可获得自诺维信公司)或PANAMORE GOLDEN TM和PANAMORE SPRING TM(可获得自DSM公司)。
适合的商业化脂肪酶制剂是例如LIPOPAN TM，例如可获得自诺维信公司的LIPOPAN TM50BG。
另外的酶
任选地，另外的酶如淀粉酶、α-淀粉酶、β-淀粉酶、真菌α-淀粉酶、羧肽酶、过氧化氢酶、脂肪分解酶、半乳聚糖酶、α-半乳糖苷酶、β-半乳糖苷酶、葡聚糖酶、葡糖淀粉酶、葡糖氧化酶、脂肪酶、氧化酶、过氧化物酶、植酸酶、多酚氧化酶、蛋白酶和/或木聚糖酶可以与根据本发明工艺涉及的酶一起使用。
面团
本发明还涉及用于制备面团或烘焙产品的方法，这些方法包括向该面团中掺入有效量的酶，以控制或减少烘焙产品在制备过程的食用油脂的使用量。
短语“掺入”在本文中定义为将根据本发明涉及的物料添加到面团中、添加到待制作面团的任何成分中、和/或添加到待制作面团中面团成分的任何混合物中。
换言之，本发明的烘焙组合物可以在面团制备的任何步骤中添加，并且可以在一个、两个、或多个步骤中添加。可以使用本领域熟知的方法将组合物添加到揉捏和烘焙的面团成分中。
术语“有效量”在本文中定义为根据烘焙产品制备的需要，其足以对面团和/或烘焙产品的至少一种感兴趣的特性提供可测量的影响。
术语“面团”在本文中定义为面粉和其他成分的混合物，其足够硬以揉捏或滚压。
本发明的面团可以包含衍生自任何谷粒的面粉，包括小麦、大麦、黑麦、燕麦、玉米、高粱、稻、粟、以及其任何混合物。
在本发明优选的实施例中，在所述面团中还含有以下组成的组：面粉、酵母、食用盐、食用糖、维生素C和/或食用香精。优选的是，考虑到可食用油脂使用量的显著降低，制成的烘焙产品尽管在触感柔软度、面包瓤结构、口感柔软度和口感湿润度方面均没有发生改变或实质性改变，但可以适量添加食用 香精，以弥补由于可食用油脂使用量的大量减少带来的嗅觉香气和/或味觉香味的部分改变。特别是，由于人造奶油中可含有食用香精，这部分食用香精和可食用油脂的使用量的减少可能对气味特别是储存后的气味有一定影响，在本发明优选的实施例中，可以通过适量添加食用香精进行弥补。
面团还可以包含其他的面团成分，例如蛋白质，如奶粉、面筋和大豆；蛋(全蛋、蛋黄或蛋白)；氧化剂，如抗坏血酸、溴酸钾、碘酸钾、偶氮二甲酰胺(ADA)或过硫酸铵；氨基酸，如L-半胱氨酸；淀粉；和/或盐，如氯化钠、乙酸钙、硫酸钠、丙酸钙或硫酸钙。
淀粉可以是小麦淀粉、玉米淀粉、玉蜀黍淀粉、树薯(tapioca)淀粉、木薯(cassava)淀粉、马铃薯淀粉；和/或糖，如蔗糖(sucrose)、甘蔗糖(cane sugar)、乳糖或高果糖玉米糖浆。
在本发明中，可食用油脂例如是天然油脂或再加工油脂，其中，天然油脂例如是植物油脂和动物油脂，例如是植物油或者黄油，植物油例如是菜籽油或大豆油，再加工油脂例如是人造奶油、起酥油、植脂奶油等。
在本发明的一些实施例中，对于一些应用，不需要乳化剂；对于一些应用，可能需要乳化剂。适用于本发明的乳化剂优选为选自由以下组成的组的乳化剂：甘油单酯的二乙酰酒石酸酯(DATEM)、硬脂酰乳酸钠(SSL)、硬脂酰乳酸钙(CSL)、乙氧基化甘油单酯和甘油二酯(EMG)、蒸馏的甘油单酯(DMG)、聚山梨醇酯(PS)、和琥珀酰化的甘油单酯(SMG)。
本发明的面团可以是新鲜的、冷冻的或部分烘焙的(预烘焙的)。
本发明的面团通常是经发酵的面团或将要经受发酵的面团。该面团可以各种方式来发酵，例如通过添加化学发酵剂(例如碳酸氢钠)或通过添加发酵剂(发酵面团)，但优选的是通过添加适合的酵母培养物如酿酒酵母(面包酵母)的培养物(例如可商购的酿酒酵母菌株)来发酵该面团。
本发明还涉及一种用于烘焙的烘焙油脂，其包括可食用油脂、纤维素酶和至少一种麦芽糖α-淀粉酶，优选该油脂还含有乳化剂和/或抗氧化剂。当使用该烘焙油脂由面团制备烘焙产品时，由于所述纤维素酶和至少一种麦芽糖α-淀粉酶的添加，与其他条件相同但未在该烘焙油脂添加所述酶时相比，制备过程中可食用油脂的使用量按重量计可减少至少10％的比例，优选的是，在 该制成的烘焙产品中，所述可食用油脂的含量相对于烘焙产品至少是1％(w/w)、例如是至少2％(w/w)，优选是至少是3％(w/w)。
在本发明的一些实施例中，所述用于烘焙的油脂中包含的可食用油脂是植物油，例如是菜籽油、大豆油、牛油、奶油、椰子油或棕榈仁油。优选的，在该用于烘焙的油脂中，所述乳化剂为单双脂肪酸甘油酯、司盘60、聚甘油酯、蔗糖酯、硬脂酰乳酸钠中的至少一种。
工业工艺
本发明特别有用于在工业化工艺中制备面团和烘焙产品，其中用来制备烘焙产品的面团是使用自动化或半自动化的设备以机械方式制备的。
制备面包的工艺通常涉及以下顺序步骤：制作面团(与任选的醒发步骤)，对该面团进行压片或分割、成形或滚压、以及醒发，这些步骤在本领域中是熟知的。
如果使用任选的醒发步骤，优选地添加更多面粉并且可以添加碱来中和产生的或将要在第二醒发步骤期间产生的酸。在根据本发明的工业烘焙生产工艺中，使用自动化或半自动化的设备来进行这些步骤中的一个或多个。
本文中描述并要求保护的发明不限于本文中公开的具体实施方案的范围，这些实施方案仅旨在说明本发明的几个方面。
实施例1：酶在吐司面包减油中的应用
采用直接面团发酵工艺来制备吐司面包，所有使用的原料均为食品级，其中，真菌淀粉酶是Fungamyl 2500SG，木聚糖酶是Pentopan Mono BG，葡萄糖氧化酶是Gluzyme Mono 10000BG，纤维素酶是Celluclast BG，均是诺维信公司的商业化产品，麦芽糖淀粉酶(SEQ ID NO:2和SEQ ID NO:3的复配)，其源自于嗜热脂肪芽孢杆菌。
根据表1的配料表和表2的试验方案，将面粉、食用盐、蔗糖、酵母、丙酸钙、SSL、CSL、DMG称好放入面缸中(立式搅拌和面机，DIOSNA牌)中，加入酶和水及牛奶香精，搅拌，并调节水温使混合后面团的温度保持在23℃，加入人造黄油后，继续搅拌，至面筋充分扩展，将打好后的面团静置10min， 然后进行面团按450g/个进行分割，分割后的面团进行初步成形后静置10min，之后通过压面成形机进一步成形2次，放入模具，然后将其放入醒发箱，醒发温度35℃，醒发湿度85％，醒发120min，醒发好的面团放入烤箱进行焙烤，烤箱温度(上火185℃，下火185℃)，烘焙后的土司面包放冷却后制成产品。
表1：原料配方
Figure PCTCN2021090509-appb-000001
表2：试验方案
批次 油脂用量 麦芽糖淀粉酶 纤维素酶
A 10％ 0ppm 0ppm
B 6％ 0ppm 0ppm
C 6％ 60ppm 0ppm
D 6％ 55ppm 5ppm
参数表征
硬度的测定方法：将吐司面包使用吐司切片机进行切割(每个面包片厚度是1.2cm)，切后的面包片每两片为一组(厚度为2.4cm)，使用TA.XT Plus质构仪进行测定，以克为单位，硬度值越高，代表制成的面包面包柔软度越低，品质越差。
弹性的测定方法：将吐司面包使用吐司切片机进行切割(每个面包片厚度是1.2cm)。切后的面包片每两片为一组(厚度为2.4cm)，使用TA.XT Plus质构仪进行测定，以％为单位。弹性值越高，代表制成的面包品质越好。
感官评价：由5名熟练的测试面包师对上述各批次制备的土司面包的触感柔软度、面包瓤结构、嗅觉香气、口感柔软度、口感湿润度和味觉香味分别进行打分(对批次A制成的面包计5.0分，作为基准，分值不低于4.0分表示没有实质性变差)，取其平均值，进行综合评价，平均值的分值越高，代表制成的面包品质越好。
制成的各批次面包在制成之后的24小时(即第1天)时的总感官评价分别为，批次A的平均值是5.0分，批次B的平均值是4.8分，批次C的平均值是5.2分，批次D的平均值是5.3分；制成的各批次面包在制成之后的336小时(即第14天)时的感官评价分别为，批次A的平均值是5.0分，批次B的平均值是4.2分，批次C的平均值是5.4分，批次D的平均值是5.6分。
由此表明，在土司面包的制备过程，尽管大幅度的减少了食用油脂的用量(减少了40％的用量)，大幅度节约了面包的制备成本和减少了面包中的油脂含量，但是由于麦芽糖淀粉酶的添加，制成的面包的感观评价总体上相对于参照方案没有变差，甚至还有一定程度的改善，其中，在面团中添加麦芽糖淀粉酶和纤维素酶时，制成的面包的感观评价更佳，麦芽糖淀粉酶和纤维素酶存在某种协同作用。
此外，由下表3可知，与作为对照的批次A的产品相比，当仅仅将制备过程中的食用油脂的用量由基于面粉重量计的10％减少到6％时，制成的土司面包的弹性值相对较好，但硬度值显著偏高。但是，在同样低水平的油脂用量 的情况下，通过在面团中添加麦芽糖淀粉酶制成的面包则表现出非常出色的柔软度和弹性。特别的，在面团中添加麦芽糖淀粉酶和纤维素酶时，制成的面包的硬度值最低。所有涉及酶法制备的土司面包，随着面包存放时间的增加，其表现出在柔软度和弹性方面的优势更加明显。例如，在由批次C制备的面包的第14天的品质优于由批次A制备的面包的第7天的水平，由批次D制备的面包在14天的质构明显优于批次A制备的面包的第7天的水平，特别是在提高柔软度的同时还保留了非常好的弹性。
表3：各批次产品的硬度和弹性
Figure PCTCN2021090509-appb-000002
由此说明，在含油量较高的土司面包的制备过程中，通过在面团中添加麦芽糖淀粉酶，优选复配纤维素酶，是能够在确保制成的土司面包的关键指标总体评价不变差或实质性变差的情况下，大幅度减少制备时的可食用油脂的用量，这不仅能大大减少烘焙产品的制备成本，而且由于在制备过程中能够显著降低可食用油脂的用量，因而制成的烘焙类产品也更符合健康食品的概念。
实施例2：酶在吐司面包减油中的应用
采用直接面团发酵工艺来制备吐司面包，所有使用的原料均为食品级，其中，真菌淀粉酶是Fungamyl 2500SG，木聚糖酶是Pentopan Mono BG，葡萄糖氧化酶是Gluzyme Mono 10000BG，纤维素酶是Celluclast BG，均是诺维信公司的商业化产品，麦芽糖淀粉酶(SEQ ID NO:1或SEQ ID NO:2)，均源自于嗜热脂肪芽孢杆菌。
据以表4的配料表和表5的试验方案，将面粉、食用盐、蔗糖、酵母、丙酸钙、SSL、CSL、DMG称好放入面缸中(立式搅拌和面机，DIOSNA牌)中， 加入酶和水及牛奶香精，搅拌，并调节水温使混合后面团的温度保持在23℃，加入人造黄油后，继续搅拌，至面筋充分扩展，将打好后的面团静置10min，然后进行面团按450g/个进行分割，分割后的面团进行初步成形后静置10min，之后通过压面成形机进一步成形2次，放入模具，然后将其放入醒发箱，醒发温度35℃，醒发湿度85％，醒发120min，醒发好的面团放入烤箱进行焙烤，烤箱温度(上火185℃，下火185℃)，烘焙后的土司面包放冷却后制成产品。
表4：原料配方
Figure PCTCN2021090509-appb-000003
表5：试验方案：
Figure PCTCN2021090509-appb-000004
Figure PCTCN2021090509-appb-000005
根据实施例1中的参数表征方法，对制成的各批次面包在制成之后的24小时(即第1天)、96小时(第4天)、168小时(第7天)、336小时(第14天)进行感官评价，如表6所表示：
表6：各批次面包的感官评价
批次 第1天 第4天 第7天 第14天
A 5.0 5.0 5.0 5.0
B 5.1 5.4 5.5 5.6
C 5.3 5.4 5.5 5.6
D 5.2 5.2 5.4 5.3
由此表明，在土司面包的制备过程，尽管大幅度的减少了可食用油脂的用量(减少了40％的用量)，大幅度节约了面包的制备成本和面包中的油脂含量，但是由于麦芽糖淀粉酶(SEQ ID NO:1或SEQ ID NO:2)的添加，制成的面包的感观评价总体上相对于参照方案没有变差，甚至还有一定程度的改善。
此外，由下表7可知，与作为对照的批次A的产品相比，在低水平的油脂用量的情况下，通过在面团中添加麦芽糖淀粉酶制成的面包在第1天和第14天均则表现出非常出色的柔软度和弹性，其中，在面团中添加麦芽糖淀粉酶和纤维素酶时，制成的面包的感观评价更佳，麦芽糖淀粉酶和纤维素酶存在某种协同作用。跨度14天的评价对土司面包之类的短保面包来说已经足够说明，通过在面团中添加麦芽糖淀粉酶，是能够在确保制成的土司面包的关键指标总体评价不变差或实质性变差、甚至还能改善的情况下，大幅度减少制备时的食 用油脂的用量。
表7各批次产品的硬度和弹性
Figure PCTCN2021090509-appb-000006
实施例3：酶在法式小面包减油中的应用
采用直接面团发酵工艺来制备吐司面包，所有使用的原料均为食品级，其中，真菌淀粉酶是Fungamyl 2500SG，木聚糖酶是Pentopan Mono BG，葡萄糖氧化酶是Gluzyme Mono 10000BG，纤维素酶是Celluclast BG，均是诺维信公司的商业化产品，麦芽糖淀粉酶(SEQ ID NO:2和SEQ ID NO:3的复配)，其源自于嗜热脂肪芽孢杆菌。
根据下述配料表8，将面粉、食用盐、蔗糖、酵母、CSL和脱氢醋酸钠物料称好放入面缸中(立式搅拌和面机，DIOSNA牌)中，混合均匀，加入鸡蛋，糖浆和水及酶，搅拌，加入起酥油，打面至面筋扩展，控制面团的温度保持在23℃。分割面团200g/个后进行压面30次，至面筋完全。卷面，将面分割成25g/个，放入模具，然后将其放入醒发箱，醒发温度35℃，醒发湿度85％，醒发90min，醒发好的面团放入烤箱进行焙烤，烤箱温度(上火200℃，下火155℃)，烘焙时间10min。烘焙后的法式小面包放冷却后包装。
表8：原料配方
Figure PCTCN2021090509-appb-000007
Figure PCTCN2021090509-appb-000008
表9：试验方案
Figure PCTCN2021090509-appb-000009
感官评价：由5名熟练的测试面包师对上述各批次制备的法式小面包的触感柔软度、触感弹性、面包瓤结构、嗅觉香气、口感柔软度、口感湿润度和味觉香味分别进行打分(对批次A制成的小面包计5.0分，作为基准)，取其平均值，进行综合评价，平均值的分值越高，代表制成的小面包品质越好。
制成的各批次法式小面包在制成之后的24小时(即第1天)、72小时(第3天)、336小时(第14天)、504小时(第21天)和672小时(第28天)时进行感官评价：
表10：各批次面包的感官评价
  批次A 批次B 批次C
第1天 5.0 4.7 4.9
第3天 5.0 4.7 5.0
第14天 5.0 4.8 5.1
第21天 5.0 4.9 5.1
第28天 5.0 4.7 5.3
如上表10所示，在法式小面包的制备过程中，尽管大幅度的减少了起酥油的用量(减少了36％的用量)，节约了面包的制备成本，但是通过添加在面团中添加麦芽糖淀粉酶与纤维素酶的复合减油酶制成的面包的感观评价没有变差，甚至还有一定程度的改善。跨度28天的评价对法式小面包之类的中保面包来说已经足够说明，在含油量较高的法式小面包的制备过程中，通过在面团中添加麦芽糖淀粉酶，优选复配纤维素酶，是能够在确保制成的土司面包的关键指标总体评价不变差或实质性变差的情况下，大幅度减少制备时的可食用油脂的用量，这不仅能大大减少烘焙产品的制备成本，而且由于在制备过程中能够显著降低可食用油脂的用量，因而制成的烘焙类产品也更符合健康食品的概念。
实施例4：酶在海绵蛋糕减油中的应用
采用全蛋打法工艺来制备海绵蛋糕，所有使用的原料均为食品级。
根据表11的配料表和表12的试验方案，将丙酸钙、泡打粉、全脂奶粉、面粉和酶混合过筛后备用。搅拌缸(立式搅拌缸，DIOSNA牌)中加入鸡蛋、食用糖、食用盐和酶搅打2分钟；加入除油外所有液体原料和粉末原料，搅打1分钟，刮缸，继续搅拌2分钟；加入植物油，搅打1分钟，刮缸，然后将面糊按300g/份倒入模具，放入烤箱进行焙烤，烤箱温度(上火180℃，下火170℃)，烘焙后的蛋糕放冷却后制成产品。其中，使用的减油酶是纤维素酶和麦芽糖淀粉酶的复配，所述的纤维素酶是Celluclast BG，是诺维信公司的商业化产品，麦芽糖淀粉酶(SEQ ID NO:2和SEQ ID NO:3的复配)，源自于嗜热脂 肪芽孢杆菌。
表11：原料配方
配方 烘焙百分比％(基于面糊)
面粉(王后低筋粉) 25
鸡蛋(购自德青源) 28
砂糖粉 27
植物油(购自金龙鱼) 6或11
水 5
全脂奶粉(购自雀巢) 1
食用盐 0.3
泡打粉(购自焙乐道) 0.3
丙酸钙(蜜丹儿) 0.4
减油酶 200ppm
表12：试验方案
批次 油脂用量 减油酶
A 11％ 0ppm
B 6％ 0ppm
C 6％ 200ppm
参数表征
硬度的测定方法：将蛋糕表面切割平整(每个蛋糕高度是45cm)，使用TA.XT Plus质构仪进行测定，以克为单位，硬度值越高，代表制成的蛋糕柔软度越低，品质越差。
粘聚性的测定方法：在质构测试中两次下压过程能量(曲线下正峰面积积分)的比值，作为样品的质构测试粘聚性。粘聚性反应蛋糕的口感品质，粘聚 性越高，代表制成的蛋糕粘聚性越好，品质越好。
感官评价：由5名熟练的测试烘焙师对上述各批次制备的蛋糕的口感柔软度、口感粘聚性、口感湿润度和化口性分别进行打分(对批次A制成的蛋糕计5.0分，作为基准，0.5分的差距表示有实质性的品质差异)，取其平均值，进行综合评价，平均值的分值越高，代表制成的蛋糕品质越好。
制成的各批次蛋糕在制成之后的72小时(即第3天)时的总感官评价分别为，批次A的平均值是5分，批次B的平均值是3分，批次C的平均值是5分；制成的各批次蛋糕在制成之后的168小时(即第7天)时的感官评价分别为，批次A的平均值是5分，批次B的平均值是4分，批次C的平均值是5.8分；制成的各批次蛋糕在制成之后的360小时(即第15天)时的感官评价分别为，批次A的平均值是5分，批次B的平均值是4分，批次C的平均值是5.3分。
由此表明，在蛋糕的制备过程，尽管大幅度的减少了食用油脂的用量(减少了约45％的用量)，大幅度节约了蛋糕的制备成本和减少了蛋糕中的油脂含量，但是由于减油酶的添加，制成的蛋糕的感观评价总体上相对于参照方案没有变差，甚至还有一定程度的改善，制成的蛋糕的感观评价更佳。
表13：各批次产品的感官评价
Figure PCTCN2021090509-appb-000010
此外，由下表14可知，与作为对照的批次A的产品相比，当仅仅将制备过程中的食用油脂的用量由11％减少到6％时，即由批次B制成的蛋糕的粘聚性值相对较好，但硬度值显著偏高。但是，在同样低水平的油脂用量的情况下，通过在蛋糕配方中添加减油酶制成的蛋糕则表现出非常出色的柔软度和粘聚性，而且涉及酶法制备的蛋糕，随着蛋糕存放时间的增加，其表现出在柔软度 和粘聚性方面的优势更加明显。例如，由批次C制备的蛋糕在15天的质构明显优于批次A制备的蛋糕的第7天的水平，特别是在提高柔软度的同时还提高了粘聚性。
表14：各批次产品的硬度和粘聚性
Figure PCTCN2021090509-appb-000011
由此说明，在含油量较高的蛋糕的制备过程中，通过在配方中添加减油酶，是能够在确保制成的蛋糕的关键指标总体评价不变差或实质性变差的情况下，大幅度减少制备时的可食用油脂的用量，这不仅能大大减少烘焙产品的制备成本，而且由于在制备过程中能够显著降低可食用油脂的用量，因而制成的烘焙类产品也更符合健康食品的概念。

Claims (17)

  1. 一种由面团制备烘焙产品的方法，包括将至少一种麦芽糖α-淀粉酶掺入该面团以及控制在面团中添加的可食用油脂用量的步骤，通过烘焙制成烘焙产品，其中，由于麦芽糖α-淀粉酶的添加，所述面团中可食用油脂的使用量相对于其他条件相同但未采用所述酶处理时的可食用油脂用量，按重量计可至少减少10％的比例。
  2. 一种减少由面团制备的烘焙产品中的可食用油脂用量的方法，包括将至少一种麦芽糖α-淀粉酶和可食用油脂掺入该面团的步骤，通过烘焙制成烘焙产品，其中，由于麦芽糖α-淀粉酶的添加，所述面团中可食用油脂的使用量相对于其他条件相同但未采用所述酶处理时的可食用油脂用量，按重量计可至少减少10％的比例。
  3. 一种由面团制备烘焙产品的方法，包括将纤维素酶和至少一种麦芽糖α-淀粉酶以及可食用油脂掺入该面团的步骤，通过烘焙制成烘焙产品。
  4. 根据上述权利要求任一项中所述的方法，该制成的烘焙产品中，所述可食用油脂的含量相对于该烘焙产品按重量计至少是1％(w/w)、至少是2％(w/w)，或者至少是3％(w/w)。
  5. 根据上述权利要求任一项中所述的方法，由于所述酶的添加，相对于其他条件相同但未添加所述酶时，所述面团中可食用油脂的使用量，按重量计可减少的比例至少是15％、至少是20％、优选至少减少25％、至少减少30％、至少减少35％，或更优选至少减少40％。
  6. 根据上述权利要求任一项中所述的方法，其中，在所述面团中还进一步包含纤维素酶和/或磷脂酶。
  7. 根据前述权利要求中任一项所述的方法，其中所述的烘焙产品是面包，蛋糕或中式糕点，例如是软式面包或者是起酥面包，例如是土司面包、法式小面包或者小圆面包，或者例如是海绵蛋糕，或者是戚风蛋糕。
  8. 根据前述权利要求中任一项所述的方法，其中，所述烘焙产品具有至少4天、或至少7天、或至少14天的货架期，或者，与其他条件相同但未在面团添加所述酶、也没有减少可食用油脂使用量时相比，其制成的烘焙产品，以第4天、或第7天、或第14天为例，其产品硬度值更低和/或其产品弹性值更高。
  9. 根据前述权利要求中任一项所述的方法，其中，在所述面团中，相对于每一千克面粉，麦芽糖淀粉酶的添加用量是是10-1000MANU，优选是50-500MANU。
  10. 根据前述权利要求中任一项所述的方法，其中，所述的可食用油脂是黄油、人造黄油、植物油、人造奶油和/或起酥油。
  11. 根据前述权利要求中任一项所述的方法，其中，在所述面团中还含有以下组成的组：面粉、食用盐和食用糖，优选还含有食用香精、酵母和/或维生素C。
  12. 根据前述权利要求中任一项所述的方法，其中，与其他条件相同但未在面团添加所述酶、也没有减少可食用油脂使用量时相比，其制成的烘焙产品的感官评价没有变差或者没有实质性变差。
  13. 根据前述权利要求中任一项所述的方法，其中，对面包来说，所述感官评价是触感柔软度、面包瓤结构、、口感柔软度和口感湿润度的综合评价，优选的是，所述感官评价还含对嗅觉香气和味觉香味的评价。
  14. 根据前述权利要求中任一项所述的方法，其中，对蛋糕来说，所述感官评价是对蛋糕的口感柔软度、口感湿润度和化口性的综合评价，优选的是，所述感官评价还含有对蛋糕的粘聚性的评价。
  15. 麦芽糖α-淀粉酶和/或麦芽糖α-淀粉酶和纤维素酶的组合在由面团制备具有至少4天货架期的烘焙产品时减少可食用油脂用量中的用途，包括将所述酶和和可食用油脂掺入该面团和通过烘焙制成烘焙产品的步骤，其中，由于所述酶的添加，与其他条件相同但未在面团添加所述酶时相比，制备过程中可食用油脂的使用量按重量计可减少至少10％的比例，优选的是，在该制成的烘焙产品中，所述可食用油脂的含量相对于烘焙产品至少是1％(w/w)、例如是至少2％(w/w)，优选是至少是3％(w/w)。
  16. 一种烘焙油脂，其包括可食用油脂、纤维素酶和至少一种麦芽糖α-淀粉酶，优选该油脂还含有乳化剂和/或抗氧化剂。
  17. 权利要求16所述的烘焙油脂，其中，当使用该烘焙油脂由面团制备烘焙产品时，由于所述纤维素酶和至少一种麦芽糖α-淀粉酶的添加，与其他条件相同但未在该烘焙油脂添加所述酶时相比，制备过程中可食用油脂的使用量按重量计可减少至少10％的比例，优选的是，在该制成的烘焙产品中， 所述可食用油脂的含量相对于烘焙产品至少是1％(w/w)、例如是至少2％(w/w)，优选是至少是3％(w/w)。
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