WO2002019828A1 - Composition de pate de cuisson comprenant une enzyme encapsulee dans un lipide - Google Patents

Composition de pate de cuisson comprenant une enzyme encapsulee dans un lipide Download PDF

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Publication number
WO2002019828A1
WO2002019828A1 PCT/DK2001/000583 DK0100583W WO0219828A1 WO 2002019828 A1 WO2002019828 A1 WO 2002019828A1 DK 0100583 W DK0100583 W DK 0100583W WO 0219828 A1 WO0219828 A1 WO 0219828A1
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WIPO (PCT)
Prior art keywords
enzyme
dough
lipid
composition according
flour
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PCT/DK2001/000583
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English (en)
Inventor
Claus Crone Fuglsang
Thomas Hønger CALLISEN
Gitte Budolfsen
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Novozymes A/S
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Priority to AU2001283817A priority Critical patent/AU2001283817A1/en
Publication of WO2002019828A1 publication Critical patent/WO2002019828A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01032—Xylan endo-1,3-beta-xylosidase (3.2.1.32), i.e. endo-1-3-beta-xylanase
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • 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
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10—Cereal-derived products
    • A23L7/104—Fermentation of farinaceous cereal or cereal material; Addition of enzymes or microorganisms
    • A23L7/107—Addition or treatment with enzymes not combined with fermentation with microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • A23P10/35—Encapsulation of particles, e.g. foodstuff additives with oils, lipids, monoglycerides or diglycerides

Definitions

  • the present invention relates to a dough composition
  • a dough composition comprising one or more lipid-encapsulated or lipid-coated enzyme (s), methods for preparing said dough composition, a use of' one or more lipid-encapsulated or lipid-coated enzyme (s) in said dough composition, a method for improving one or more properties of a dough, a method for preparing a baked product, and a dough and/or a baked product produced thereby.
  • the strength of a dough is an important aspect of baking for both small-scale and large-scale applications.
  • a strong dough has a greater tolerance of mixing time, proofing time, and mechanical vibrations during dough transport, whereas a weak dough is less tolerant to these treatments.
  • a strong dough with superior rheological and handling properties results from flour containing a strong gluten network.
  • Flour with a low protein content or a poor gluten quality results in a weak dough .
  • Dough "conditioners” are well known in the baking industry.
  • the addition of conditioners to bread dough has resulted in improved machinability of the dough and improved texture, volume, flavour, and freshness (anti-staling) of the bread.
  • Nonspecific oxidants such as iodates, peroxides, ascorbic acid, potassium bromate and azodicarbona ide have a gluten strengthening effect . It has been suggested that these conditioners induce the formation of interprotein bonds which strengthen the gluten and thereby the dough.
  • the use of several of the currently available chemical oxidising agents has been met with consumer resistance or is not permitted by regulatory agencies .
  • the use of enzymes as dough conditioners has been considered as an alternative to the chemical conditioners.
  • a number of enzymes have been used recently as dough and/or bread improving agents, in particular enzymes that act on components present in large amounts in the dough. Examples of such enzymes are found within the groups of amylases, proteases, glucose oxidases, trans-glutaminases and (hemi) cellulases, including pentosanases .
  • EP-A1-0669082 discloses an aqueous bread improver composition comprising at least one water-soluble bread-improving enzyme and optionally a lecithin.
  • the lecithin component if any, is added in form of an emulsion in water.
  • WO 98/38869 discloses an edible composition comprising at least two components being encapsulated or coated by a fatty substance.
  • the fatty substances disclosed are optionally esterified mono-, di- and triglycerides and waxes. Lecithins may also be used.
  • WO 00/01793 discloses an enzyme-containing granular composition comprising an enzyme-containing core and a protective layer or coating.
  • WO 99/27907 discloses a composition containing an active principle encapsulated in multilamellar vesicles comprising at least one surfactant .
  • WO 97/16076 discloses a particulate enzyme-containing preparation suitable for e.g. the production of an animal feed composition.
  • WO 92/12645 discloses the use of an enzyme-containing T- granulate, which is coated with a coating agent comprising a high melting fat or wax, as a component of a mixture which is well-suited as a fodder.
  • WO 89/08694 discloses an enzyme containing granulate with a coating comprising a mono- or diglyceride of a fatty acid.
  • WO 90/09440 discloses an enzyme containing granulate having two coatings and an enzyme.
  • the present invention relates to a dough composition
  • a dough composition comprising:
  • (a) provides, at a temperature of less than 25° C, a barrier, which inhibits release of said enzyme (s) to the surrounding dough, and
  • the present invention also relates to a method for preparing a dough composition, comprising:
  • the present invention also relates to a use of one or more lipid-encapsulated or lipid-coated enzyme (s) in a dough composition, wherein said lipid substance undergoes a phase transition in the temperature range from 25° C to 60° C.
  • the present invention also relates to a method for improving one or more properties of a dough, comprising adding one or more lipid-encapsulated or lipid-coated enzyme (s) to a dough mixture before baking.
  • the present invention also relates to a method for preparing a baked product .
  • the present invention also relates to a dough product and to a baked product .
  • the present invention relates to a dough composition
  • a dough composition comprising one or more lipid-encapsulated or lipid-coated enzyme (s) which improve one or more properties of the dough or the baked product obtained from the dough relative to a dough or a baked product in which an enzyme is not incorporated.
  • the enzyme (s) is/are preferably present in the dough encapsulated in mono-, oligo-, or multi-lamellar vesicles or in the form of coated solid particles or granules. By encapsulating or coating the enzyme (s) a controlled release of said enzymes can be obtained, so that the enzyme (s) are not released to the other dough components until a selected point in time.
  • selected point in time is meant a point in time, in which the effect of the active enzyme is desired. This means that release of the enzyme is inhibited during the initial processing of the dough, at which stage where it would cause undesired effects such as increased stickiness of the dough, reduced machinability, increased softness of the dough, and increased tightness of the dough. Rather it will be released during leavening of the dough, i.e. when the temperature of the dough has increased from usually room temperature to a temperature in the range 25-60°C, preferably to a temperature in the range 35-50°C.
  • improved property is defined herein as any property of a dough and/or a product obtained from the dough, particularly a baked product, which is improved by the action of an enzyme relative to a dough or product in which said enzyme is not incorporated.
  • the improved property may include, but is not limited to, increased strength of the dough, increased elasticity of the dough, increased stability of the dough, reduced stickiness of the dough, improved extensibility of the dough, improved machinability of the dough, increased volume of the baked product, improved crumb structure of the baked product, improved softness of the baked product, improved flavor of the baked product, and/or improved antistaling of the baked product.
  • the use of an enzyme may result in an increased strength, stability, and/or reduced stickiness of the dough, resulting in improved machinability, as well as in an increased volume and improved crumb structure and softness of the baked product .
  • the effect on the dough may be particularly advantageous when a poor quality flour is used.
  • Improved machinability is of particular importance in connection with dough that is to be processed industrially.
  • the improved property may be determined by comparison of a dough and/or a baked product prepared with and without addition of an enzyme in accordance with the methods of the present invention.
  • Organoleptic qualities may be evaluated using procedures well established in the baking industry, and may include, for example, the use of a panel of trained taste- testers .
  • the term "increased strength of the dough” is defined herein as the property of a dough that has generally more elastic properties and increased resistance towards overmixing.
  • the term “increased elasticity of the dough” is defined herein as the property of a dough which has a higher tendency to regain its original shape after being subjected to a certain physical strain.
  • the term "increased stability of the dough” is defined herein as the property of a dough that is less susceptible to mechanical abuse thus better maintaining its shape and volume.
  • reduced stickiness of the dough is defined herein as the property of a dough that has less tendency to adhere to surfaces, e.g., in the dough production machinery, and is either evaluated empirically by the skilled test baker or measured by the use of a texture analyzer (e.g., TA-XT2 Texture Analyser available from Stable Micro Systems, England) as known in the art .
  • a texture analyzer e.g., TA-XT2 Texture Analyser available from Stable Micro Systems, England
  • improved extensibility of the dough is defined herein as the property of a dough that can be subjected to increased strain or stretching without rupture.
  • improved machinability of the dough is defined herein as the property of a dough that is generally less sticky and/or more firm and/or more elastic .
  • increased volume of the baked product is measured as the specific volume of a given loaf of bread (volume/weight) determined typically by the traditional rape seed displacement method.
  • improved crumb structure of the baked product is defined herein as the property of a baked product with finer and/or thinner cell walls in the crumb and/or more uniform/homogenous distribution of cells in the crumb and is usually evaluated empirically by the skilled test baker.
  • improved softness of the baked product is the opposite of "firmness” and is defined herein as the property of a baked product that is more easily compressed and is evaluated either empirically by the skilled test baker or measured by the use of a texture analyzer (e.g., TA-XT2) as known in the art .
  • TA-XT2 a texture analyzer
  • improved flavor of the baked product is evaluated as mentioned above by a trained test panel .
  • improved antistaling of the baked product is defined herein as the properties of a baked product that have a reduced rate of deterioration of quality parameters, e.g., softness and/or elasticity, during storage.
  • the term "dough” is defined herein as a mixture of flour and other ingredients firm enough to knead or roll.
  • the dough may be fresh, frozen, pre-bared, or pre-baked.
  • the preparation of frozen dough is described by Kulp and Lorenz in Frozen and
  • baked product is defined herein as any product prepared from a dough, either of a soft or a crisp character.
  • Examples of baked products whether of a white, light or dark type, which may be advantageously produced by the present invention are bread (in particular white, whole-meal or rye bread) , typically in the form of loaves or rolls, French baguette-type bread, pasta, pita bread, tortillas, tacos, cakes, pancakes, biscuits, cookies, pie crusts, steamed bread, and crisp bread, and the like.
  • bread in particular white, whole-meal or rye bread
  • French baguette-type bread pasta, pita bread, tortillas, tacos, cakes, pancakes, biscuits, cookies, pie crusts, steamed bread, and crisp bread, and the like.
  • the enzyme (s) may be any enzyme which provides an improved property to a dough and/or to a baked product obtained from the dough.
  • Enzymes to be used according to the invention are preferably selected among carbohydrases, proteases, oxidases, lipases, and trans-glutaminases .
  • the source of an enzyme is not critical for improving one or more properties of a dough and/or a baked product. Accordingly, the enzyme (s) may be obtained from any source such as a plant, microorganism, or animal.
  • the enzyme (s) is/are preferably obtained, e.g., from a microbial source, such as a bacterium or a fungus, e.g., a filamentous fungus or a yeast .
  • the enzyme (s) is/are obtained from a bacterial source.
  • the enzyme (s) may be obtained from an Acetobacter, Acinetobacter, Agrobacterium, Alcaligenes, Arthrobacter, Az ⁇ tobacter, Bacillus, Comamonas, Clostridium, Gluconobacter , Halobacterium, Mycobacterium, Rhizobium, Salmonella, Serratia, Streptomyces, Escherichia, Pseudomonas, Wolinella, or methylotrophic bacterium strain.
  • the enzyme (s) is/are obtained from an Aceto-bacter aceti , Alcaligenes faecalis, Arthrobacter oxidans, Azotobacter vinelandii, Bacillus alkalophilus , Bacillus amyloliquefaciens, Bacillus ani tratum, Bacillus brevis, Bacillus circulans, Bacillus coagulans, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus stearothermophilus , Bacillus subtilis , Bacillus thuringiensis , Comamonas testosteroni , Clostridium tyrobutyricum, Gluconobacter dioxyaceticus,
  • Gluconobacter liquefaciens Gluconobacter suboxydans, Halobacterium cutirubrum, Mycobacterium convolutum, Rhizobium melioti , Salmonella typhimurium, Serratia marcescens,
  • the enzyme (s) is/are obtained from a fungal source.
  • the enzyme (s) may be obtained from a yeast strain such as a Candida, Kluyveromyces , Pichia, Saccharomyces, Schizosaccharomyces, or Yarrowia strain; or from a filamentous fungal strain such as an Acremonium, Aspergillus, Aureobasidium, Chrysosporium, Cryptococcus , Filibasidium, Fusarium, Humicola, Magnaporthe, Monilia, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces , Penicillium, Phanerochaete , Piromyces ,
  • a yeast strain such as a Candida, Kluyveromyces , Pichia, Saccharomyces, Schizosaccharomyces, or Yarrowia strain
  • a filamentous fungal strain such as an Acremonium, Aspergillus, Aureobasidium, Chrysosporium,
  • the enzyme (s) is/are obtained from a Saccharomyces carlsbergensis, Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii , Saccharomyces kluyveri , Saccharomyces norbensis, or Saccharomyces oviformis strain.
  • the enzyme (s) is obtained from an Aspergillus aculeatus, Aspergillus awamori , Aspergillus foetidus , Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Chrysosporium lignorum, Fusarium bactridioides , Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusarium graminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi, Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusarium sambucinum, Fusarium sarcochroum, Fusarium sulphureum, Fusarium toruloseum, Fusarium trichothecioides, Fu
  • Trichoderma rolfsii Sporotrichum thermophile, Trichoderma ci trinoviride, Trichoderma hamatum, Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma polysporum, Trichoderma reesei, Trichoderma saturnisporum, or Trichoderma vi ide strain.
  • enzyme variants are included within the meaning of the term "enzyme” .
  • examples of such enzyme variants are disclosed, e.g., in EP 251,446 (Genencor) , WO 91/00345 (Novo Nordisk) , EP 525,610 (Solvay) and WO 94/02618 (Gist-Brocades NV) .
  • the enzyme classification employed in the present specification and claims is in accordance with Recommendations (1992) of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology, Academic Press, Inc., 1992.
  • enzymes which may appropriately be used according to the invention include oxidoreductases (EC 1.-.-.-), transferases (EC 2.-.-.-), hydrolases (EC 3.-.-.-), lyases (EC 4.-.-.-), isomerases (EC 5.-.-.-) and ligases (EC 6.-.-.-) .
  • Preferred oxidoreductases in the context of the invention are peroxidases (EC 1.11.1) such as haloperoxidase, laccases (EC 1.10.3.2) and glucose oxidases (EC 1.1.3.4), while preferred transferases are transferases in any of the following sub-classes: a) Transferases transferring one-carbon groups (EC 2.1); b) Transferases transferring aldehyde or ketone residues (EC 2.2); acyltransferases (EC 2.3); c) Glycosyltransferases (EC 2.4); d) Transferases transferring alkyl or aryl groups, other than methyl groups (EC 2.5); and e) Transferases transferring nitrogenous groups (EC 2.6).
  • peroxidases EC 1.11.1
  • preferred transferases are transferases in any of the following sub-classes: a) Transferases transferring one-carbon groups (EC 2.1); b) Transferases transferring aldehyde or
  • a most preferred type of transferase in the context of the invention is a transglutaminase (protein-glutamine . ⁇ - glutamyltransferase; EC 2.3.2.13). Further examples of suitable transglutaminases are described in WO 96/06931 (Novo Nordisk A/S) .
  • Preferred hydrolases in the context of the invention are: Carboxylic ester hydrolases (EC 3.1.1.-) such as lipases (EC 3.1.1.3); phytases (EC 3.1.3.-), e.g. 3-phytases (EC 3.1.3.8) and 6-phytases (EC 3.1.3.26); glycosidases (EC 3.2, which fall within a group denoted herein as "carbohydrases”), such as . -amylases
  • carbohydrate chains e.g. starches
  • five- and six-membered ring structures i.e. glycosidases, EC 3.2
  • enzymes capable of isomerizing carbohydrates e.g. six- membered ring structures such as D-glucose to five-membered ring structures such as D-fructose.
  • Carbohydrases of relevance include the following (EC numbers in parentheses): ⁇ -amylases (3.2.1.1), ⁇ -amylases (3.2.1.1), ⁇ -amylases (3.2.1.1), ⁇ -amylases (3.2.1.1), ⁇ -amylases (3.2.1.1), ⁇ -amylases (3.2.1.1), ⁇ -amylases (3.2.1.1), ⁇ -amylases (3.2.1.1), ⁇ -amylases
  • glucan 1, 4- ⁇ -glucosidases (3.2.1.3), cellulases (3.2.1.4), endo-1,3 (4) - ⁇ -glucanases (3.2.1.6), endo-1,4-.- xylanases (3.2.1.8), dextranases (3.2.1.11), chitinases
  • oxidoreductases examples include GluzymeTM (enzyme available from Novo Nordisk A/S) .
  • proteases examples include KannaseTM, EverlaseTM, EsperaseTM, AlcalaseTM, NeutraseTM,
  • Bio-FeedTM Pro and Clear-LensTM Pro are available from Novo
  • proteases include MaxataseTM, MaxacalTM, MaxapemTM, OpticleanTM and PurafectTM (available from Genencor International Inc. or Gist-Brocades) .
  • Lipolase Ultra Lipolase Ultra
  • Lipozyme Palatase
  • NovozymTM 435 LipozymTM 435
  • LecitaseTM All available from Novo Nordisk A/S.
  • Other commercially available lipases include LumafastTM
  • LipomaxTM Ps. pseudoalcaligenes lipase from Gist- Brocades/Genencor Int. Inc.
  • Bacillus sp . lipase from Solvay enzymes
  • carbohydrases examples include Alpha-GalTM, Bio-FeedTM Alpha, Bio-FeedTM Beta, Bio- FeedTM Plus, Bio-FeedTM Plus, NovozymeTM 188, CelluclastTM CellusoftTM, CeremylTM, CitrozymTM, DenimaxTM, DezymeTM DextrozymeTM, FinizymTM, FungamylTM, GamanaseTM, GlucanexTM LactozymTM, MaltogenaseTM, PentopanTM, PectinexTM, PromozymeTM
  • Combinations of enzymes may be obtained by fermenting two or more enzymes simultaneously in the same fermentation broth.
  • combinations of enzymes may be obtained by fermenting the enzymes separately and using mixtures of different fermentation broths.
  • Carbohydrases such as amylases and pentosanases, may beneficially be added to flour in order to improve one or more properties thereof.
  • Flour has varying content of amylases leading to differences in the baking quality. Addition of amylases can be necessary in order to standardise the flour.
  • Amylases and pentosanases generally provide sugar for the yeast fermentation, improve the bread volume, retard retrogradation, and decrease the staling rate and stickiness that result from pentosan gums .
  • Certain maltogenic amylases can be used for prolonging the shelf life of bread for two or more days without causing gumminess in the product.
  • These enzymes selectively modify the gelatinised starch by cleaving from the non-reducing end of the starch molecules to produce low molecular weight sugars and dextrins .
  • the starch is modified in such a way that retrogradation is less likely to occur.
  • the produced low- molecular-weight sugars improve the water retention capacity of the baked goods without creating the intermediate-length dextrins that result in gumminess in the finished product.
  • Fungal ⁇ -amylases may be used to improve the bread volume and to provide a good and uniform structure of the bread crumb.
  • Said ⁇ -amylases are endoenzymes that produce maltose, dextrins and glucose. Cereal and some bacterial .-amylases are inactivated at temperatures above the gelatinisation temperature of starch, and therefore when added to a wheat dough result in a low bread volume and a sticky bread interior.
  • Fungal amylases, such as FungamylTM have the advantage of being thermolabile and are inactivated just below the gelatinisation temperature.
  • Enzyme preparations containing a number of pentosanase and hemi-cellulase activities can improve the handling and stability of the dough, and improves the freshness, the crumb structure and the volume of the bread.
  • Pentosanases can be used in combination with or as an alternative to emulsifiers.
  • the one or more lipid-encapsulated or lipid-coated enzymes is/are selected among ⁇ -amylase and hemicellulase .
  • said hemicellulase is a pentasanase, such as a xylanase.
  • the xylanase is preferably of microbial origin, e.g., derived from a bacterium or fungus, such as a strain of Aspergillus, in particular of Aspergillus aculeatus, Aspergillus niger (cf. WO 91/19782), Aspergillus awamori (WO 91/18977), or Aspergillus tubigensis (WO 92/01793), from a strain of Trichoderma, e.g., Trichoderma reesei , or from a strain of Humicola, e.g., Humicola insolens (WO 92/17573 , the contents of which is hereby incorporated by reference) .
  • a bacterium or fungus such as a strain of Aspergillus, in particular of Aspergillus aculeatus, Aspergillus niger (cf. WO 91/19782), Aspergillus awamori
  • Proteases may also be used according to the invention, as these enzymes may be useful for gluten weakening in particular when using hard wheat flour.
  • Further useful enzymes comprise oxidases, which are useful for improving dough consistency.
  • the oxidase is an aldose oxidase, a glucose oxidase, a pyranose oxidase, a lipoxygenase or an L-amino acid oxidase.
  • the enzyme is a lipase, which is useful for the modification of lipids present in the dough or dough constituents so as to soften the crumb.
  • the enzyme (s) has/have a pH optimum in the range of about 3 to about 10. In a more preferred embodiment, the enzyme (s) has/have a pH optimum in the range of about 4.5 to about 8.5. In another preferred embodiment, the enzyme (s) has/have a temperature optimum in the range of about 5°C to about 100°C. In a more preferred embodiment, the enzyme (s) has/have a temperature optimum in the range of about 25°C to about 75°C.
  • combinations of enzymes may also be used to improve one or more properties of the dough and/or baked product obtained from the dough.
  • the combination comprises a xylanase in combination with an amylase, or combinations of a xylanase and an amylase with a glucose-oxidase or a lipase.
  • the enzyme (s) is/are used in an amount sufficient to provide the desired effect, i.e., the improved properties in question.
  • the dosage of the enzyme (s) to be used in the present invention should be adapted to the nature and composition of the dough in question as well as to the nature of the enzyme (s) to be used.
  • the enzyme (s) is/are typically added in an amount corresponding to 0.01-100 mg enzyme protein per kg of flour, preferably 0.1-25 mg enzyme protein per kg of flour, more preferably 0.1-5 mg enzyme protein per kg of flour.
  • the appropriate dosage of a given enzyme for exerting a desirable improvement of dough and/or baked products will depend on the enzyme and the enzyme substrate in question.
  • the skilled person may determine a suitable enzyme unit dosage on the basis of methods known in the art .
  • the enzyme (s) is/are encapsulated or coated by a lipid substance, wherein said lipid substance a) provides, at a temperature of less than 25° C, a barrier, which inhibits release of said enzyme (s) to the surrounding dough, and b) undergoes a phase transition in the temperature range from 25° C to 60° C.
  • at least 75% by weight of said lipid substance undergoes a phase transition within a temperature interval of less than 20° C, comprised in the range 25-60°C.
  • Said phase transition of the lipid substance is preferably from a lamellar to a non-lamellar phase. In a particular embodiment said phase transition is melting.
  • a lipid substance wherein at least 85 % by weight of said lipid substance undergoes a phase transition within a temperature interval of less than 15°C. In another preferred embodiment of the invention at least 90 % by weight of said lipid substance undergoes a phase transition within a temperature interval of less than 12 °C. In a most preferred embodiment at least 95% by weight of said lipid substance undergoes a phase transition within a temperature interval of less than 10°C.
  • an important property of an enzyme entity is the size and/or the size distribution of the entity as the application of an enzyme entity involves mixing the entities with other particulate products, particularly flour.
  • Use of an enzyme entity of a proper size in such compositions may provide a more homogeneous distribution of the enzyme entity in the composition and a less tendency of the enzyme entities separating from the other composition components. If the enzyme entities do not possess the proper size distribution compared to the composition in which they are used the enzyme entity may concentrate in specific parts or layers of the composition. Also the average particle size of the entities should not be too large, since they would then tend not to remain intact during the mixing process.
  • the term particle size as used herein is to be understood as the diameter of the particle measured in its longest dimension.
  • the lipid- encapsulated or lipid-coated enzyme (s) is/are provided in the form of particles, wherein at least 95 % by weight thereof have a particle size in the range 10-200 ⁇ m. In a further embodiment of the invention the above particle size is in the range 20-150 ⁇ m.
  • the enzyme (s) and/or additional enzymes to be used according to the present invention may be in any form suitable for the use in question, e.g., in the form of a dispersion of vesicle encapsulated enzyme (s) or in the form of a dry powder of coated agglomerated or granulated particles, in particular a non-dusting granulate.
  • the enzyme (s) are encapsulated they are preferably used in the form of vesicles, i.e. the enzyme (s) are encapsulated by a multilamellar bilayer of lipid substance.
  • said vesicles are comprised of one or more lipid substances.
  • any lipid substance undergoing a phase transition in the above range to release the enzyme (s) and suitable for preparing vesicles may be employed.
  • said lipid substance comprises a phosphoglyceride, such as phosphatidylethanolamine (PE) , phosphatidylcholine (PC) or derivatives thereof.
  • derivatives thereof in the present context, is meant esters, ethers, anhydrides, salts etc. of the phosphoglycerides in question.
  • said phosphoglyceride is selected among egg phosphatidylethanolamine, transesterified phosphatidylethanolamine, N-methyl-di-oleoyl phosphatidylethanolamine , and dimyristoyl phosphaphosphatidylcholine .
  • the above phosphoglyceride in admixture with one or more components selected among saturated or mono- or polyunsaturated, linear or branched C6-C22 fatty acids and edible, amphiphilic polymers, preferably of sugar or cellulose origin.
  • one of the above phosphoglycerides is used in admixture with a C12-C22 fatty acid, such as myristic acid.
  • lipid composition determines the stability and the release profile of the entrapped enzyme (s) .
  • the above listed lipids provide onset temperatures for release between 25 and 60° C, in particular between 35 and 50 °C.
  • vesicles may be prepared by the following process : (i) mixing a buffered enzyme containing liquid with a lipid substance to produce a dispersion, (ii) forming at least one lipid bilayer around enzyme by repeatedly cooling and heating under agitation the dispersion, (iii) adding a dispersion of the enzyme containing vesicles obtained in step (i) and (ii) to a dough mixture comprising flour and any additional, conventional dough ingredients.
  • the enzyme containing liquid of step (i) is preferably an aqueous enzyme solution of dispersion and the lipid substance is preferably added in dry form under vigorous mixing.
  • the cooling and heating/mixing procedure of step (ii) is suitably repeated about 5 to 15 times.
  • the cooling is preferably rapidly by placing the dispersion e.g. in a bath of ethanol and solid carbon dioxide (dry ice) . While heating it is important that the temperature does not exceed the onset temperature for release of the specific system. If needed, the entrapped volume may be further increased by extrusion or ultrasonic treatment of the system as described in Hope et al., 1985 (Biochemica et Biophysica Acta 812: 55-65).
  • the encapsulated enzyme (s) is in the form of vesicles
  • said vesicles are preferably added to the dough as a buffered dispersion in water.
  • Buffers for use in the preparation of said enzyme vesicles are any conventionally used buffer known in the art, such as a phosphate buffer.
  • the encapsulated or coated enzyme (s) for use in the dough composition according to the invention may also be in the form of solid coated particles comprising an enzyme-containing core and a coating of a lipid substance.
  • Said particles may be prepared by any granulation technique known in the art .
  • Known formulation technologies include: - spray-dried products, wherein a liquid enzyme-containing solution is atomised in a spray drying tower to form small droplets which during its way down the drying tower dries up to form an enzyme- containing particulate material. Very small particles can be produced this way (Michael S. Showell (editor); Powdered detergents; Surfactant Science Series; 1998; vol. 71; page 140-142; Marcel Dekker) .
  • Extruded or pelletised products wherein an enzyme-containing paste is pressed to pellets or extruded under pressure through a small opening and cut into particles which are subsequently dried.
  • Such particles usually have a considerable size because the material of which the extrusion opening is made (usually a plate with bore holes) sets a limit on the allowable pressure drop over the extrusion opening, and since very high extrusion pressures, when using a small opening, would " " increase heat generation in the enzyme paste, which is harmful to the enzyme.
  • the granules are produced by spray-drying, since said technique provides very small particles, which is favourable in terms of mixing with flour.
  • the granules are produced via a mixer granulation technique as disclosed above.
  • a lipid coating is applied on said cores.
  • the coating is preferably applied in a conventional fluid-bed coating process, wherein the lipid-coating material is sprayed in liquid form onto fluidised enzyme particles.
  • suitable coating materials comprise the following: high melting fats such as glycerol esters (mono-, di- or triesters or mixtures thereof) , - lipids such as phosphoglycerides, waxes isolated from a natural source, such as Carnauba wax, Candelilla wax and bees wax. Other natural waxes or derivatives thereof are waxes derived from animals or plants, e.g. of marine origin.
  • the degree of hydrogenation may be optimised by the skilled person to obtain suitable phase transition properties of the hydrogenated lipis substance.
  • An example of hydrogenated palm oil is commercially available, e.g.
  • fatty acid alcohols such as linear long chain fatty acid alcohols such as NAFOL 1822 (C18, 20, 22) from Condea Chemie GmbH, Germany, having a melting point between 55- 60° C,
  • Mono-glycerides and/or di-glycerides such as glyceryl stearate.
  • An example of this is Di odan PM from Danisco Ingredients, Denmark.
  • Fatty acids such as hydrogenated, linear, long-chained fatty acids.
  • Paraffins i.e. solid hydrocarbons, Micro-crystalline wax.
  • Enzyme particles or granules typically also comprise auxiliary compounds such as: a) Edible fillers such as fillers conventionally used in the field of granulation, e.g. water soluble and/or insoluble inorganic salts such as finely ground alkali or alkaline earth sulphate, alkali carbonate and/or alkali halide, clays such as kaolin (e.g. Speswhite, English China Clay), bentonites, talcs, zeolites, and/or silicates. b) Edible binders such as binders conventionally used in the field of granulation, e.g.
  • binders with a high melting point or no melting point at all and of a non-waxy nature e.g. polyvinyl pyrrolidone, dextrins, polyvinylalkohol , cellulose derivatives, for example hydroxypropyl cellulose, methyl cellulose or CMC.
  • a suitable binder is a carbohydrate binder such as Glucidex 21D available from Roquette Freres, France .
  • Edible fibre materials such as fibres conventionally used in the field of granulation. Pure or 'impure cellulose in fibrous form can be sawdust, pure fibrous cellulose, cotton, or other forms of pure or impure fibrous cellulose. Also, filter aids based on fibrous cellulose can be used.
  • CEPOTM and ARBOCELTM are on the market, e.g. CEPOTM and ARBOCELTM.
  • Cepo S/20 the approximate maximum fibre length is 500 ⁇ m
  • the approximate average fibre length is 160 ⁇ m
  • the approximate maximum fibre width is 50 ⁇ m
  • the approximate average fibre width is 30 ⁇ m.
  • CEPO SS/200 cellulose has an approximate maximum fibre length of 150 ⁇ m, an approximate average fibre length of 50 ⁇ m, an approximate maximum fibre width of 45 ⁇ m and an approximate average fibre width of 25 ⁇ m.
  • a preferred fibrous cellulose is Arbocel BFC200.
  • Edible enzyme stabilising or protective agents such as conventionally used in the field of granulation. Stabilising or protective agents may fall into several categories: alkaline or neutral materials, reducing agents, antioxidants and/or salts of first transition series metal ions. Each of these may be used in conjunction with other protective agents of the same or different categories.
  • alkaline protective agents are alkali metal silicates, -carbonates or bicarbonates, which provide a chemical scavenging effect by actively neutralising e.g. oxidants.
  • antioxidants are methionine, butylated hydroxytoluene (BHT) or butylated hydroxyanisole (BHA) or ascorbic acid or salts thereof.
  • Edible sugars may act as process aid.
  • the additional enzyme may be of any origin, including mammalian and plant, and preferably of microbial (bacterial, yeast or fungal) origin and may be obtained by techniques conventionally used in the art.
  • the additional enzyme may be a cyclodextrin glucanotransferase, a peptidase, in particular, an exopeptidase (useful in flavour enhancement) , a phospholipase (useful for the modification of lipids present in the dough or dough constituents so as to soften the dough and improve gas retention in the dough) , a cellulase, a protein disulfide isomerase, e.g., a protein disulfide isomerase as disclosed in WO 95/00636, a glycosyltransferase, a peroxidase (useful for improving the dough consistency) , or a laccase.
  • a cyclodextrin glucanotransferase a peptidase, in particular, an exopeptidase (useful in flavour enhancement) , a phospholipase (useful for the modification of lipids present in the dough or dough constituents so as to soften the
  • a conventionally used baking agent may also be incorporated into the dough.
  • the baking agent may include proteins, such as milk powder (to provide crust colour) , gluten (to improve the gas retention power of weak flours) , and soy (to provide additional nutrients and improve water binding) ; eggs (either whole eggs, egg yolks or egg whites) ; fat such as granulated fat or shortening (to soften the dough and improve the texture of the bread) ; an emulsifier (to improve dough extensibility and, to some extent, the consistency of the resulting bread); an antioxidant, e.g., ascorbic acid, an oxidant such as potassium bromate, potassium iodate, azodicarbon amide (ADA) or ammonium persulfate (to strengthen the gluten structure); an amino acid, e.g., L- cysteine (to improve mixing properties) ; a sugar; a salt, e.g., sodium chloride, calcium acetate, sodium sulfate, and soy (to provide
  • Suitable emulsifiers are mono- or diglycerides, diacetyl tartaric acid esters of mono- or diglycerides, sugar esters of fatty acids, polyglycerol esters of fatty acids, lactic acid esters of monoglycerides, acetic acid esters of monoglycerides, polyoxyethylene stearates, phospholipids, and lecithin.
  • the dough and/or baked product prepared by a method of the present invention may be based on wheat meal or flour, optionally in combination with other types of meal or flour such as corn meal, corn flour, rye meal, rye flour, oat meal, oat flour, soy meal, soy flour, sorghum meal, sorghum flour, potato meal, or potato flour.
  • the handling of the dough and/or baking may be performed in any suitable manner for the dough and/or baked product in question, typically including the steps of kneading the dough, subjecting the dough to one or more proofing treatments, and baking the product under suitable conditions, i.e., at a suitable temperature and for a sufficient period of time.
  • the dough may be prepared by using conventional methods such as a normal straight dough process, a sour dough process, an overnight dough method, a low-temperature and long-time fermentation method, a frozen dough method, the Chorleywood Bread process, or the Sponge and Dough process.
  • the dough of the invention is generally a leavened dough or a dough to be subjected to leavening.
  • the dough may be leavened in various ways such as by adding sodium bicarbonate or the like, or by adding a leaven (fermenting dough) , but it is preferable that the dough be leavened by adding a suitable yeast culture, such as a culture of Saccharomyces cerevisiae (baker's yeast). Any of the commercially available Saccharomyces cerevisiae strains may be employed.
  • the present invention also relates to methods for preparing a baked product, comprising baking a dough obtained by a method of the present invention to produce a baked product . The baking of the dough to produce a baked product may be performed using methods well known in the art.
  • the present invention also relates to doughs and baked products, respectively, produced by the methods of the present invention.
  • a controlled release system according to the present invention in which a xylanase (PentopanTM Mono) has been encapsulated in a lipid matrix using the same method as described in Example 1 above was tested in a micro scale baking assay using a normal straight dough procedure and 12 g of flour for each dough. The obtained results were compared to a regular PentopanTM Mono baking granulate.
  • the flour was incubated in a heating cabinet (28 °C) for two days before dough preparation. Also water temperature was adjusted to obtain a dough temperature of 27 °C.
  • Yeast, salt, and sugar were added as a water solution with the respective concentrations of 0.67 g/ml for yeast, 0.072 g/ml for sugar, and 0.072 g/ml for salt.
  • the encapsulated enzyme, PentopanTM Mono was added as a vesicle dispersion in water. The ingredients were then combined and the dough was mixed on the Micro Mixer NSI-33R for 3.5 minutes.
  • the dough was then moulded and sheeted and incubated in the heating cabinet at 28°C for 10 minutes. Moulding and sheeting was repeated and the dough was placed in a 37 ml pan, which was then incubated in a heating cabinet at 32 °C and 85% relative humidity for 45 minutes.
  • the dough was baked at 230°C for 17 minutes and volume was evaluated after cooling of the bread.
  • FXU Fungal Xylanase Units
  • FAU Fungal Amylase Units
  • stickiness and softness are evaluated by comparing all the dough samples to the reference dough no. 1 (100 FXU and 10 FAU per kg flour) .
  • the dough samples are scored on a ten point scale, in which the reference sample no. 1 gets a score of 5.
  • a bread with 300 FXU encapsulated PentopanTM Mono furthermore gives bread with a better volume than bread with 300 FXU PentopanTM Mono as a baking granulate (BG) .
  • the lipid used for encapsulation of the xylanase was included as a control.
  • the lipid makes the dough slightly stickier than the reference sample (100 FXU as BG) , which means that the reduced stickiness of the encapsulated xylanase
  • control lipid gives a volume increase compared to the reference sample (100 FXU as BG) .

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Abstract

L'invention concerne une composition comportant i) une quantité efficace d'une ou plusieurs enzyme(s) encapsulée(s) recouvertes d'une substance lipidique, dans laquelle ladite substance lipidique a) procure, à une température inférieure à 25 °C, une protection qui empêche la diffusion de ladite/desdites enzyme(s) à la pâte de cuisson qui l'entoure, et b) subit une transition de phase lorsque la température est comprise entre 25 °C et 60 °C afin de diffuser ladite/lesdites enzyme(s), et ii) la farine ou tout ingrédient classique de pâte de cuisson supplémentaire à des procédés de préparation de ladite composition de pâte de cuisson, à l'utilisation d'une ou plusieurs enzyme(s) encapsulée(s) de lipide ou d'enzyme(s) recouverte(s) dans une composition de pâte de cuisson, à un procédé d'amélioration d'une ou plusieurs propriété(s) d'une pâte de cuisson, à un procédé de fabrication d'un produit cuit et à un pâte de cuisson et/ou un produit cuit ainsi réalisé.
PCT/DK2001/000583 2000-09-08 2001-09-07 Composition de pate de cuisson comprenant une enzyme encapsulee dans un lipide WO2002019828A1 (fr)

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WO2004023879A1 (fr) * 2002-09-11 2004-03-25 Puratos Naamloze Vennootschap Utilisation d'enzymes de famille 8 possedant une activite xylanolytique en boulangerie
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EP1586240A1 (fr) * 2004-03-31 2005-10-19 LESAFFRE et Cie Ameliorant de panification
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