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EP2126024A2 - Compositions de nettoyage comprenant une transglucosidase - Google Patents

Compositions de nettoyage comprenant une transglucosidase

Info

Publication number
EP2126024A2
EP2126024A2 EP08727093A EP08727093A EP2126024A2 EP 2126024 A2 EP2126024 A2 EP 2126024A2 EP 08727093 A EP08727093 A EP 08727093A EP 08727093 A EP08727093 A EP 08727093A EP 2126024 A2 EP2126024 A2 EP 2126024A2
Authority
EP
European Patent Office
Prior art keywords
enzyme
transglucosidase
cleaning
natural gum
composition
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP08727093A
Other languages
German (de)
English (en)
Inventor
Hugh C. Mcdonald
Ayrookaran J. Poulose
Jayarama K. Shetty
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Danisco US Inc
Original Assignee
Danisco US Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Danisco US Inc filed Critical Danisco US Inc
Publication of EP2126024A2 publication Critical patent/EP2126024A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/20—Organic compounds containing oxygen
    • C11D3/22—Carbohydrates or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
    • C11D3/38636—Preparations containing enzymes, e.g. protease or amylase containing enzymes other than protease, amylase, lipase, cellulase, oxidase or reductase
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/20—Organic compounds containing oxygen
    • C11D3/22—Carbohydrates or derivatives thereof
    • C11D3/222—Natural or synthetic polysaccharides, e.g. cellulose, starch, gum, alginic acid or cyclodextrin
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1048—Glycosyltransferases (2.4)
    • C12N9/1051—Hexosyltransferases (2.4.1)
    • C12N9/107—1,4-Alpha-glucan branching enzyme (2.4.1.18)
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06L—DRY-CLEANING, WASHING OR BLEACHING FIBRES, FILAMENTS, THREADS, YARNS, FABRICS, FEATHERS OR MADE-UP FIBROUS GOODS; BLEACHING LEATHER OR FURS
    • D06L1/00—Dry-cleaning or washing fibres, filaments, threads, yarns, fabrics, feathers or made-up fibrous goods
    • D06L1/12—Dry-cleaning or washing fibres, filaments, threads, yarns, fabrics, feathers or made-up fibrous goods using aqueous solvents
    • D06L1/14—De-sizing
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06L—DRY-CLEANING, WASHING OR BLEACHING FIBRES, FILAMENTS, THREADS, YARNS, FABRICS, FEATHERS OR MADE-UP FIBROUS GOODS; BLEACHING LEATHER OR FURS
    • D06L4/00—Bleaching fibres, filaments, threads, yarns, fabrics, feathers or made-up fibrous goods; Bleaching leather or furs
    • D06L4/40—Bleaching fibres, filaments, threads, yarns, fabrics, feathers or made-up fibrous goods; Bleaching leather or furs using enzymes

Definitions

  • the present invention provides compositions comprising a transglucosidase enzyme and a natural gum polysaccharide, wherein the natural gum polysaccharide is a substrate for the transglucosidase enzyme.
  • the present invention also provides methods for using a transglucosidase enzyme to degrade natural gum polysaccharide.
  • the compositions and methods find use in cleaning applications.
  • Detergent and other cleaning compositions often include a complex combination of active ingredients.
  • many cleaning products contain a surfactant system, enzymes for cleaning, bleaching agents, builders, suds suppressors, soil-suspending agents, soil-release agents, optical brighteners, softening agents, dispersants, dye transfer inhibition compounds, abrasives, bactericides, and perfumes.
  • the present invention provides compositions comprising a transglucosidase enzyme and a natural gum polysaccharide, wherein the natural gum polysaccharide is a substrate for the transglucosidase enzyme.
  • the present invention also provides methods for using a transglucosidase enzyme to degrade natural gum polysaccharide.
  • the compositions and methods find use in cleaning applications.
  • the compositions comprise natural gum polysaccharides such as xanthan gums.
  • the transglucosidase enzyme has an activity defined as EC 2.4.1.24, according to IUBMB nomenclature.
  • the transglucosidase enzyme has an amino acid sequence that is at least about 90% identical to an Aspergillus transglucosidase enzyme.
  • the composition further comprises at least one surfactant.
  • the natural gum polysaccharide is present as a stain on an object. In some particularly preferred embodiments, the object is a fabric.
  • the present invention also provides methods comprising combining a transglucosidase enzyme with at least one natural gum polysaccharide to degrade the natural gum polysaccharide.
  • the transglucosidase enzyme has an activity defined as EC 2.4.1.24, according to IUBMB nomenclature.
  • the natural gum polysaccharide is a xanthan gum.
  • the present invention also provides cleaning methods comprising: contacting an object soiled with a natural gum polysaccharide with a cleaning composition comprising a transglucosidase enzyme; and maintaining the object and cleaning composition under conditions sufficient to effect degradation of the natural gum polysaccharide and thereby clean the object.
  • the object is soiled with a food that contains at least one natural gum polysaccharide.
  • the natural gum polysaccharide comprises a xanthan gum.
  • the object is selected from fabrics and hard surfaces.
  • the transglucosidase enzyme has an amino acid sequence that is at least about 90% identical to an Aspergillus transglucosidase enzyme.
  • the cleaning composition further comprises at least one surfactant.
  • the cleaning composition further comprises at least one enzyme selected from proteases, amylases, cellulases, lipases, cutinases, mannanases, pectinases, pectate lyases, and oxido-reductases, for the degradation of other stain components.
  • the cleaning composition has a pH of about pH 5.0 to about pH 1 1.5.
  • the cleaning composition comprises the transglucosidase enzyme at a concentration in the range of about 0.01 ppm to about 100 ppm.
  • the present invention provides compositions comprising: a transglucosidase enzyme; and at least one natural gum polysaccharide; wherein the natural gum polysaccharide is a substrate for the transglucosidase enzyme.
  • the natural gum polysaccharide comprises a xanthan and/or guar gum.
  • the natural gum polysaccharide is present as a stain on an object (e.g., a fabric), while in other embodiments, it is free in solution.
  • the transglucosidase enzyme comprises an amino acid sequence that is at least about 70% identical to, at least about 80% identical to, at least about 85% identical to, at least about 90% identical to, at least about 95% identical to, or at least about 98% identical to an Aspergillus transglucosidase.
  • the compositions further comprise at least one surfactant and/or other cleaning agent.
  • the present invention also provides methods that include combining a transglucosidase enzyme with at least one natural gum polysaccharide to degrade the natural gum polysaccharide.
  • the methods comprise: contacting an object soiled with a natural gum polysaccharide with a cleaning composition comprising a transglucosidase enzyme; and maintaining the object and cleaning composition together under conditions sufficient to effect degradation of the natural gum polysaccharide and thereby clean the object.
  • the object ⁇ e.g., a fabric
  • the object is soiled with a food that contains at least one natural gum polysaccharide ⁇ e.g., a food that contains at least one xanthan and/or guar gum).
  • the cleaning composition further comprises at least one surfactant, other cleaning agents, and/or at least one additional enzyme ⁇ e.g., a protease, amylase, cellulase, lipase, cutinase, oxido-reductase, or the like), for the degradation of other stain components.
  • the cleaning compositions have a pH in the range of about pH 5.0 to about pH 1 1.5, and the transglucosidase enzyme is present in the cleaning composition at a concentration in the range of about 0.01 ppm to about m.
  • the methods provided herein result in more efficient removal of stains that contain natural gum polysaccharides than equivalent methods that do not employ a transglucosidase enzyme.
  • the present invention provides an isolated enzyme containing an Aspergillus transglucosidase produced by a Trichoderm ⁇ reesei host cell.
  • the present invention also provides cleaning compositions ⁇ e.g., laundry detergents), containing the enzyme are provided, as well as well as a method of using the enzyme for cleaning an object ⁇ e.g., a fabric).
  • Figure 1 provides a vector map of pTrex3(AGL51M ).
  • Figure 2 provides the nucleotide sequence of an expression cassette of pTrex3(AGL51M) (SEQ ID NO: 1).
  • Figure 3 provides a graph showing the enzymatic activity of transglucosidase on 0.2% xanthan, as measured by a reducing sugar assay.
  • Figure 4 presents graphs showing the cleaning performance of Trip-TG on guar soiled microswatches (top panel) and salad dressing soiled microswatches (bottom panel) in 50 mM Hepes buffer (pH 7.4) and in AATCC HDL Detergent (pH 7.4).
  • Figure 5 provides a graph of a dose response experiment, showing that Trip-TG is active on salad dressing soiled microswatches at a concentration of 1 to 5 ppm in AATCC HDL.
  • Figure 6 provides a graph showing Trip-TG cleaning activity on salad dressing microswatches in heavy duty solid detergent (HDD).
  • HDD heavy duty solid detergent
  • Figure 7 provides a graph showing Trip-TG cleaning activity on marmalade stain in a Tergotometer assay using 0.15% AATCC HDL.
  • the present invention provides compositions comprising a transglucosidase enzyme and a natural gum polysaccharide, wherein the natural gum polysaccharide is a substrate for the transglucosidase enzyme.
  • the present invention also provides methods for using a transglucosidase enzyme to degrade natural gum polysaccharide.
  • the compositions and methods find use in cleaning applications.
  • nucleic acids are written left to right in 5' to 3' orientation; amino acid sequences are written left to right in amino to carboxy orientation, respectively.
  • the headings provided herein are not limitations of the various aspects or embodiments of the invention that can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the Specification as a whole.
  • recombinant refers to a polynucleotide or polypeptide that does not naturally occur in a host cell. In some embodiments, recombinant molecules contain two or more naturally-occurring sequences that are linked together in a way that does not occur naturally.
  • a recombinant cell contains a recombinant polynucleotide or polypeptide.
  • heterologous refers to elements that are not normally associated with each other. For example, if a host cell produces a heterologous protein, that protein is not normally produced in that host cell.
  • a promoter that is operably linked to a heterologous coding sequence is a promoter that is operably linked to a coding sequence that it is not usually operably linked to in a wild-type host cell.
  • homologous with reference to a polynucleotide or protein, refers to a polynucleotide or protein that occurs naturally in a host cell.
  • a “signal sequence” is a sequence of amino acids present at the N-terminal portion of a protein which facilitates the secretion of the mature form of the protein from the cell. The definition of a signal sequence is a functional one. The mature form of the extracellular protein lacks the signal sequence, which is cleaved off during the secretion process.
  • a “coding sequence” is a DNA segment that encodes a polypeptide.
  • nucleic acid encompasses DNA, RNA, single stranded or double stranded and chemical modifications thereof.
  • the terms “nucleic acid” and “polynucleotide” are used interchangeably herein, in context.
  • a "vector” refers to a polynucleotide designed to introduce nucleic acids into one or more host cells. Suitable vectors autonomously replicate in different host cells and include: cloning vectors, expression vectors, shuttle vectors, plasmids, phage particles, cassettes and the like.
  • An "expression vector” as used herein means a DNA construct comprising a protein- coding region that is operably linked to a suitable control sequence that is capable of effecting expression of the protein in a suitable host cell.
  • control sequences include a promoter to effect transcription, an optional operator sequence to control transcription to produce mRNA, a sequence encoding suitable ribosome binding sites on the mRNA, and enhancers and other sequences which control termination of transcription and translation.
  • a "promoter” is a regulatory sequence that initiates transcription of a downstream nucleic acid.
  • the term “operably linked” refers to an arrangement of elements that allows them to be functionally related.
  • a promoter is operably linked to a coding sequence if it controls the transcription of the sequence.
  • selectable marker refers to a protein capable of expression in a host that allows for ease of selection of those hosts containing an introduced nucleic acid or vector. Examples of selectable markers include, but are not limited to, antimicrobials (e.g., hygromycin, bleomycin, or chloramphenicol) and/or genes that confer a metabolic advantage, such as a nutritional advantage on the host cell.
  • derived encompasses the terms "originated from,” “obtained,” “obtainable from,” and “isolated from” and refers to the source of a sequence and/or protein.
  • a "non-pathogenic" organism is an organism that is not pathogenic to human and/or other animals.
  • the terms “recovered, “isolated,” and “separated,” as used herein refer to a protein, cell, nucleic acid or amino acid that is removed from at least one component with which it is naturally associated.
  • the terms “transformed,” “stably transformed,” and “transgenic” used in reference to a cell means the cell has a non-native (e.g., heterologous) nucleic acid sequence integrated into its genome or as an episomal plasmid that is maintained through multiple generations.
  • the term "expression” refers to the process by which a polypeptide is produced based on the nucleic acid sequence of a gene. The process includes both transcription and translation.
  • the term “introduced” in the context of inserting a nucleic acid sequence into a cell refers to "transfection,” “transformation,” or “transduction,” and includes reference to the incorporation of a nucleic acid sequence into a eukaryotic or prokaryotic cell wherein the nucleic acid sequence may be incorporated into the genome of the cell (e.g., chromosome, plasmid, plastid, or mitochondrial DNA), converted into an autonomous replicon, or transiently expressed (e.g., transfected mRNA).
  • hybridization refers to the process by which a strand of nucleic acid joins with a complementary strand through base pairing as known in the art.
  • a nucleic acid is considered to be "selectively hybridizable" to a reference nucleic acid sequence if the two sequences specifically hybridize to one another under moderate to high stringency hybridization and wash conditions.
  • Moderate and high stringency hybridization conditions are known to those of skill in the art.
  • One example of high stringency conditions include hybridization at about
  • cleaning composition and "cleaning formulation” refer to a composition that finds use in the removal of undesired compounds (e.g., a "stain") from items to be cleaned, such as fabric, dishes, contact lenses, other solid substrates, hair (shampoos), skin (soaps and creams), teeth (mouthwashes, toothpastes), etc.
  • undesired compounds e.g., a "stain”
  • the term encompasses any materials/compounds selected for the particular type of cleaning composition desired in the form of the product (e.g., liquid, gel, granule, or spray composition), as long as the composition is compatible with the subject enzyme in the composition.
  • the specific selection of cleaning composition materials are readily made by considering the surface, item and/or fabric to be cleaned, and the desired form of the composition for the cleaning conditions during use.
  • detergent compositions e.g., liquid, gel, and/or solid laundry detergents and fine fabric detergents
  • hard surface cleaning formulations such as for glass, wood, ceramic and metal counter tops and windows
  • carpet cleaners oven cleaners
  • fabric fresheners fabric softeners
  • textile and laundry pre-spotters as well as dish detergents
  • cleaning composition includes, unless otherwise indicated, granular, tablet or powder-form all-purpose or heavy-duty washing agents, especially cleaning detergents; liquid, gel or paste-form all-purpose washing agents, especially the so-called heavy- duty liquid (HDL) types; liquid fine-fabric detergents; hand dishwashing agents or light duty dishwashing agents, including those of the high- foaming type; machine dishwashing agents, including the various tablet, granular, liquid and rinse-aid types for household and institutional use; liquid cleaning and disinfecting agents, including antibacterial hand-wash types, cleaning bars, mouthwashes, denture cleaners, car or carpet shampoos, bathroom cleaners; hair shampoos and hair-rinses; shower gels and foam baths; metal cleaners; as well as cleaning auxiliaries such as bleach additives and "stain-stick,” pre-treat, or laundry additive types.
  • HDL heavy-duty liquid
  • washing agents including those of the high- foaming type
  • machine dishwashing agents including the various tablet, granular, liquid and rinse-aid types for household and
  • detergent composition and “detergent formulation” are used in reference to compositions that are formulated for use in a wash medium for the cleaning of soiled objects.
  • the term is used in reference to laundering fabrics and/or garments (e.g., “laundry detergents”).
  • laundry detergents e.g., "laundry detergents”
  • the term refers to other detergents, such as those used to clean dishes, cutlery, etc. (e.g., "dishwashing detergents”). It is not intended that the present invention be limited to any particular detergent formulation or composition.
  • a detergent composition may also contain surfactants, transferase(s), hydrolytic enzymes, oxido reductases, builders, bleaching agents, bleach activators, bluing agents and fluorescent dyes, caking inhibitors, masking agents, enzyme activators, antioxidants, and/or solubilizers, etc.
  • "enhanced performance" in a cleaning composition is defined as increasing cleaning (e.g., removal and/or decolorization) of stains (particularly those comprising natural gum polysaccharide-related stains such as chocolate cream, salad dressing, guar, etc.), as determined by usual evaluation after a standard wash cycle.
  • hard surface cleaning composition refers to detergent compositions for cleaning hard surfaces such as floors, walls, tile, bath and kitchen fixtures, and the like. Such compositions are provided in any form, including but not limited to solids, liquids, emulsions, etc.
  • dishwashing composition refers to all forms for compositions for cleaning dishes, including but not limited to gel, granular and liquid forms.
  • fabric cleaning composition refers to all forms of detergent compositions for cleaning fabrics, including but not limited to, gel, granular, liquid and bar forms.
  • textile refers to woven fabrics, as well as staple fibers and filaments suitable for conversion to or use as yarns, woven, knit, and non-woven fabrics. The term encompasses yarns made from natural, as well as synthetic (e.g., manufactured) fibers.
  • textile materials is a general term for fibers, yarn intermediates, yarn, fabrics, and products made from fabrics (e.g., garments and other articles).
  • fabric encompasses any textile material. Thus, it is intended that the term encompass garments, as well as fabrics, yarns, fibers, non-woven materials, natural materials, synthetic materials, and any other textile material.
  • an effective amount of transglucosidase refers to the quantity of transglucosidase enzyme necessary to achieve the enzymatic activity required in the specific application (e.g., cleaning composition, etc.). Such effective amounts are readily ascertained by one of ordinary skill in the art and are based on many factors, such as the particular enzyme variant used, the cleaning application, the specific composition of the cleaning composition, and whether a liquid or dry (e.g., granular, bar) composition is required, and the like.
  • transglucosidase refers to an enzyme that transfers an ⁇ -D-glucosyl residue in a 1,4- ⁇ -D-glucan to the primary hydroxy group of glucose, free or combined in a 1 ,4- ⁇ -D-glucan.
  • the transglucosidase described herein has an activity described as EC 2.4.1.24, according to IUBMB enzyme nomenclature.
  • the systematic name for the transglucosidase described herein is l,4- ⁇ -D-glucan:l,4- ⁇ -D-glucan(D-glucose) 6- ⁇ -D-glucosyltransferase. This enzyme may be referred to as ⁇ -glucosidase in certain publications.
  • soiled object refers to an object (e.g., a fabric or dish), that is soiled (e.g., "stained"), with a second composition.
  • soiled object are dirty fabrics, such as dirty clothing, linens, and fabrics stained with foodstuffs containing a natural gum polysaccharide.
  • the stain exhibits a visible color, while in other embodiments, it does not exhibit a visible color.
  • natural gum polysaccharide refers to a non-starch polysaccharide of natural origin that is capable of causing a large viscosity increase in solution at low concentration.
  • Such polysaccharides are commonly employed in the food industry and are used as thickening agents, gelling agents, emulsifiers and stabilizers in many foodstuffs (e.g., sauces, creams, dairy products, ice creams, mousses, milkshakes, salad dressings, etc.).
  • Guar gum (food additive E412), an edible thickening agent extracted from the leguminous guar bean shrub, and xanthan gum (food additive E415), a polysaccharide that is produced by fermentation of glucose or sucrose (e.g., by Xanthomonas campestris), are examples of natural gum polysaccharides.
  • Non-starch food polysaccharide degrading enzyme refers to an enzyme that degrades non-starch food polysaccharides.
  • Exemplary enzymes include, but are not limited to, hemicellulase, mannanase, pectinase, xylanase, ⁇ -galactosidase and ⁇ -galactosidase.
  • working pH refers to the pH of a detergent during its use.
  • the working pH of a laundry detergent is the pH of the detergent when it is used to wash fabrics in a washing machine and/or during hand washing of laundry.
  • the working pH of a dishwashing detergent is the pH of that detergent as it is being used in a dishwasher and/or during hand washing of dishes.
  • detergents that are in concentrated or solid form are diluted or dissolved before the pH of that detergent is at its working pH.
  • working concentration refers to the concentration of an enzyme in a detergent during its use.
  • the working concentration of an enzyme in a laundry detergent is the concentration of that enzyme when the laundry detergent is used to wash fabrics in a washing machine and/or during hand washing of laundry.
  • the working concentration of an enzyme in a dishwashing detergent is the concentration of that enzyme in the detergent as it is being used in a dishwasher and/or during hand washing of dishes.
  • detergents that are in concentrated or solid form are diluted or dissolved before the concentration of an enzyme in a detergent is at its working concentration.
  • the present invention provides compositions comprising a transglucosidase enzyme and a natural gum polysaccharide, wherein the natural gum polysaccharide is a substrate for the transglucosidase enzyme.
  • the present invention also provides methods for using a transglucosidase enzyme to degrade natural gum polysaccharide.
  • the compositions and methods find use in cleaning applications. Indeed the present invention provides various compositions containing transglucosidase enzymes, as well as methods of using the compositions.
  • the present invention provides compositions comprising transglucosidase.
  • the composition comprises at least one transglucosidase enzyme, and a natural gum polysaccharide, where the natural gum polysaccharide is a substrate for the transglucosidase enzyme.
  • the transglucosidase enzyme generally has an activity defined as EC 2.4.1.24, according to IUBMB enzyme nomenclature, which activity transfers glucosyl residues in certain glucans to the primary hydroxy group of glucose.
  • the enzyme may also have an activity that degrades natural gum polysaccharide (e.g., xanthan, and galactomannan-containing polysaccharides such as guar gum or lima bean gum), by clipping off sugar side chains or cleaving internal bonds to break the polysaccharide backbone.
  • Any suitable transglucosidase enzyme finds use in the present invention (See e.g., Pazur et al, Carbohydr.
  • the transglucosidase enzyme that find use in the present invention are commercially available (e.g., including but not limited to enzymes obtained from Megazyme, Wicklow, Ireland; or Danisco US Inc., Genencor Division, Palo Alto, CA).
  • the enzyme is an Aspergillus niger transglucosidase produced in Trichoderma reesei cells.
  • the transglucosidase is a wild type fungal transglucosidase (e.g., including but not limited to a fungal transglucosidase having an amino acid sequence deposited in NCBI's GENBANK® database as accession numbers: D45356 (GID:2645159; Aspergillus niger), BAD06006.1 (GID:4031328; Aspergillus awamori), BAA08125.1 ⁇ GlO: ⁇ 054565; Aspergillus oryzae), XPJ)Ol 210809.1 (GID: 1 15492363; Aspergillus terreus), XP_001271891.1 (GID: 121707620; Aspergillus clavatus), XPJ)01266999.1 (GID: 1 19500484; Neosartorya fischeri), XP 75181 1.1 (GID:70993928; Aspergillus fumigatus), X
  • the enzyme is generally present in the composition at a concentration in the range of about 0.01 ppm (parts per million, w/v) to about 100 ppm ⁇ e.g., about 0.01 ppm to about 0.05 ppm, about 0.05 ppm to about 0.1 ppm, about 0.1 ppm to about 0.5 ppm, about 0.5 ppm to about 1 ppm, about 1 ppm to about 5 ppm, about 5 ppm to about 10 ppm, or about 1 Oppm to about 1 OOppm).
  • the composition is a cleaning composition.
  • the composition further comprises at least one surfactant, cleaning agents, and/or other fabric care agents described in greater detail below.
  • the natural gum polysaccharide is in an aqueous solution, while in other embodiments it is affixed to an object ⁇ e.g., as a stain that is present on the surface of an object such as a fabric or hard surface).
  • the natural gum polysaccharide is in dried form. Since natural gum polysaccharides are commonly employed in a variety of foodstuffs, in some embodiments, the object is soiled with a foodstuff that contains a natural gum polysaccharide.
  • Exemplary foodstuffs that contain a natural gum polysaccharide include, but are not limited to sauces, creams, dairy products, ice creams, mousses, milkshakes, salad dressings, fruit juice beverages, canned fruit, jelly, soy sauce, oyster sauce, packaged meats, cheese, and bakery products.
  • the natural gum polysaccharide is present in the foodstuff at a concentration of about 0.1% to about 1.5%, about 0.1% to about 0.5%, about 0.5% to about 1.0%, about 1.0%, or about 1.5%.
  • the transglucosidase is produced using conventional methods.
  • the transglucosidase enzyme is produced by expressing a fusion protein containing a signal sequence operably linked to the transglucosidase enzyme in a T. reesei host cell.
  • the transglucosidase enzyme is secreted into culture medium, from which it is harvested.
  • Any signal sequence finds use in suitable fusion proteins, such that protein secretion from the Trichoderma host cell is facilitated.
  • the signal sequence is endogenous, while in other embodiments it is non- endogenous to the Trichoderma host cell and, in some embodiments, it is a signal sequence of a protein that is known to be highly secreted from a Trichoderma sp. host cell.
  • Such signal sequence include, but are not limited to: the signal sequence of cellobiohydrolase I, cellobiohydrolase II, endoglucanases I, endoglucanases II, endoglucanases III, ⁇ -amylase, aspartyl proteases, glucoamylase, mannanase, glycosidase and barley endopeptidase B ⁇ See e.g., Saarelainen, Appl. Environ. Microbiol., 63: 4938-4940 [1997]).
  • the transglucosidase is secreted using its own signal sequence (i.e., the AGLl, AGL2 or AGL3 signal sequences).
  • the transglucosidase is produced using a nucleic acid that comprises: a signal sequence-encoding nucleic acid operably linked to an transglucosidase- encoding nucleic acid, where translation of the nucleic acid produces a fusion protein comprising an transglucosidase portion having an N-terminal signal sequence for secretion of the transglucosidase portion from a Trichoderma host cell.
  • the fusion protein further contains, in addition to a signal sequence, a "carrier protein" that is a portion of a protein that is endogenous to and highly secreted by the T. reesei sp. host cell.
  • carrier proteins include, but are not limited to those of T. reesei mannanase I (Man5A, or MANI), T. reesei cellobiohydrolase II (Cel ⁇ A, or CBHII) (See e.g., Paloheimo et ah, Appl. Environ. Microbiol., 69: 7073-7082 [2003]) or T.
  • the carrier protein is a truncated T. reesei CBHl protein that includes the CBHl core region and part of the CBHl linker region.
  • a nucleic acid encoding a fusion protein containing, from amino- terminus to carboxy-terminus, a signal sequence, a carrier protein and a subject phytase in operable linkage are employed.
  • the coding sequence of the transglucosidase is codon optimized for expression of the transglucosidase in the host cell used.
  • nucleic acids in addition to a coding sequence, the nucleic acid further contains other elements that are necessary for expression of the transglucosidase enzyme in the host cell.
  • the nucleic acid contains a promoter for transcription of the coding sequence, and a transcriptional terminator.
  • Exemplary promoters that find use in T. reesei include, but are not limited to the T. reesei cbhl, cbh2, egll, egl2, eg5, xlnl and xln2 promoters, or a hybrid or truncated version thereof.
  • the promoter is a T. reesei cbhl promoter.
  • Suitable terminators include the T. reesei cbhl, cbh2, egll, egl2, eg5, xlnl and xln2 terminators, and many others, including, for example, the terminators from A. niger or A.
  • awamori glucoamylase genes as known to those of skill in the art, as well as Aspergillus nidulans anthranilate synthase genes, Aspergillus oryzae TAKA amylase genes, or A. nidulans trpC (Punt et al., Gene 56: 1 17-124 [1987]).
  • the promoter and/or terminator are native to the Trichoderma sp. host cell, while in other embodiments, they are non-endogenous to the Trichoderma sp. host cell. [077] In some embodiments in which a T.
  • the cell is genetically modified to reduce expression of secreted proteins that are endogenous to the cell.
  • the cell comprises one or more native genes, particularly genes that encode secreted proteins, that have been deleted or inactivated.
  • one or more protease-encoding genes e.g., an aspartyl protease-encoding gene; See, Berka et al, Gene 86: 153-162 [1990]; and US Pat. No. 6,509,171
  • cellulase-encoding genes are deleted or inactivated.
  • the Trichoderma sp. host cell is a T.
  • the above-described nucleic acid is present in the nuclear genome of the Trichoderma sp. host cell, while in other embodiments, it is present in a plasmid that replicates in the Trichoderma host cell,.
  • Any suitable method for introducing nucleic acids into Trichoderma host cells finds use in the present invention (e.g., electroporation, nuclear microinjection, transduction, transfection, lipofection-mediated and DEAE-dextrin mediated transfection, incubation with calcium phosphate DNA precipitate, high velocity bombardment with DNA-coated microprojectiles, and protoplast fusion).
  • electroporation, nuclear microinjection, transduction, transfection, lipofection-mediated and DEAE-dextrin mediated transfection, incubation with calcium phosphate DNA precipitate, high velocity bombardment with DNA-coated microprojectiles, and protoplast fusion e.g., Campbell et al, Curr. Genet., 16:53-56 [1989]; WO 05/001036; US Pat. No. 6,022,725; US Pat. No. 6,103,490; US Pat No. 6,268,328; and published U.S. Patent Appln Ser.
  • the preparation of Trichoderma for transformation includes the preparation of protoplasts from fungal mycelia. (See e.g.,, Campbell et al, supra).
  • the mycelia are obtained from germinated vegetative spores. [079]
  • the transglucosidase enzyme is recovered using any suitable, convenient method ⁇ e.g., by precipitation, centrifugation, affinity, filtration or any other method known in the art.
  • transglucosidase is used without purification from the other components of the culture medium.
  • the culture medium is simply concentrated and then used without further purification of the protein from the components of the growth medium, while in other embodiments, it is used without any further modification or treatment.
  • the present invention also provides methods for degrading a natural gum polysaccharides.
  • the methods include combining (e.g., mixing) a transglucosidase enzyme with a natural gum polysaccharide under conditions such that the natural gum polysaccharide is degraded.
  • Conditions suitable for the activity of the transglucosidase enzymes e.g., the temperature range, pH range, and other reaction components suitable for the activity of a transglucosidase enzyme) are known in the art.
  • the present invention also provides cleaning methods. These methods generally include: contacting an object soiled with at least one natural gum polysaccharide with a cleaning composition comprising a transglucosidase enzyme; and maintaining the object and cleaning composition together under conditions sufficient to effect degradation of the natural gum polysaccharide and thereby clean the object.
  • the cleaning composition employed in these methods is a fabric cleaning composition (e.g., a laundry detergent), a surface cleaning composition, a dish cleaning composition, an automatic dishwasher detergent composition, or the like.
  • a fabric cleaning composition e.g., a laundry detergent
  • a surface cleaning composition e.g., a dish cleaning composition
  • an automatic dishwasher detergent composition e.g., a dishwasher sprayed-on-dioxide-sprayed-onitride, a dish cleaning composition, or the like.
  • Various formulations for cleaning compositions find use in the present invention, including, but not limited to those described in great detail in WOOOOl 826, incorporated by reference herein.
  • the subject cleaning composition e.g., laundry detergent
  • the subject cleaning composition comprises from about 1% to about 80% (e.g., about 5% to about 50% (by weight)) of at least one surfactant (e.g., non-ionic surfactant, cationic surfactant, anionic surfactant, and/or a zwitterionic surfactant, or any mixture thereof, such as a mixture of anionic and nonionic surfactants).
  • at least one surfactant e.g., non-ionic surfactant, cationic surfactant, anionic surfactant, and/or a zwitterionic surfactant, or any mixture thereof, such as a mixture of anionic and nonionic surfactants.
  • Exemplary surfactants include, but are not limited to: alkyl benzene sulfonate (ABS), including linear alkyl benzene sulfonate and linear alkyl sodium sulfonate, alkyl phenoxy polyethoxy ethanol (e.g., nonyl phenoxy ethoxylate or nonyl phenol), diethanolamine, triethanolamine and monoethanolamine.
  • ABS alkyl benzene sulfonate
  • alkyl phenoxy polyethoxy ethanol e.g., nonyl phenoxy ethoxylate or nonyl phenol
  • diethanolamine e.g., nonyl phenoxy ethoxylate or nonyl phenol
  • triethanolamine e.g., triethanolamine
  • Exemplary surfactants that find use in various detergents, particularly laundry detergents are described in U.S. Patent Nos. 3,664,961, 3,919,678, 4,222,905, and 4,239,659
  • the subject cleaning composition is solid (e.g., powder or tablet form), liquid, or a gel.
  • the composition further comprises at least one buffer (e.g. , sodium carbonate, or sodium bicarbonate), detergent builder, bleach, bleach activator, enzyme, enzyme stabilizing agent, suds booster, suppresser, anti-tarnish agent, anti-corrosion agent, soil suspending agent, soil release agent, germicide, pH adjusting agent, non-builder alkalinity source, chelating agent, organic or inorganic filler, solvent, hydrotrope, optical brightener, dye and/or perfume.
  • the cleaning composition is combined with a detergent before use as a laundry additive.
  • the cleaning compositions further contain at least one non-starch food polysaccharide degrading enzyme (e.g., hemicellulase, mannanase, pectinase, xylanase, or pectate lyase) and, optionally, one or more other enzymes, (e.g., proteases, such as a subtilisin protease and/or SSI protein; lipase, amylase, cellulase, cutinase, lipase, oxidoreductase, etc.), for the removal of other stains.
  • non-starch food polysaccharide degrading enzyme e.g., hemicellulase, mannanase, pectinase, xylanase, or pectate lyase
  • other enzymes e.g., proteases, such as a subtilisin protease and/or SSI protein
  • compositions provided herein A wide variety of other ingredients useful in detergent cleaning compositions find use in the compositions provided herein, including, but not limited to other active ingredients, carriers, hydrotropes, processing aids, dyes or pigments, solvents for liquid formulations, etc.
  • suds boosters such as the C] 0 -Ci 6 alkolamides are incorporated into the compositions, typically at about 1% to about 10% of the composition by weight.
  • detergent compositions comprise water and other solvents as carriers.
  • Low molecular weight primary or secondary alcohols exemplified by methanol, ethanol, propanol, and isopropanol find use as such carriers.
  • monohydric alcohols are preferred for solubilizing surfactants, but polyols, such as those containing from about 2 to about 6 carbon atoms and from about 2 to about 6 hydroxy groups (e.g., 1,3- propanediol, ethylene glycol, glycerine, and 1 ,2-propanediol) also find use.
  • the compositions comprise from about 5% to about 90%, typically from about 10% to about 50% of such carriers.
  • the detergent compositions provided herein are formulated such that during use in aqueous cleaning operations, the wash water has a pH between about 5.0 and about 11.5. Finished products, thus, are typically formulated at this range. Techniques for controlling pH at recommended usage levels include the use of buffers, alkalis, acids, etc., and are well known to those skilled in the art.
  • the cleaning compositions are automatic dishwashing detergents that have working pHs in the range of about pH 9.0 to about pH 1 1.5, about pH 9.0 to about pH 9.5, about pH 9.5 to about pH 10.0, about pH 10.0 to about pH 10.5, about pH 10.5 to about pH 11.0, or about pH 1 1.0 to about pH 1 1.5.
  • the cleaning compositions are liquid laundry detergents that have working pHs in the range of about pH 7.5 to about pH 8.5, about pH 7.5 to about pH 8.0, or about pH 8.0 to about pH 8.5.
  • the cleaning compositions are solid laundry detergents that have working pHs in the range of about pH 9.5 to about pH 10.5, about pH 9.5 to about pH 10.0, or about pH 10.0 to about pH 10.5.
  • Various bleaching compounds such as the percarbonates, perborates and the like, also find use in the various compositions provided herein, typically at levels from about 1% to about 15% by weight.
  • compositions also contain bleach activators such as tetraacetyl ethylenediamine, nonanoyloxybenzene sulfonate, and the like, which are also known in the art. Usage levels typically range from about 1% to about 10% by weight.
  • Various soil release agents especially of the anionic oligoester type, various chelating agents, especially the aminophosphonates and ethylenediaminedisuccinates, various clay soil removal agents, especially ethoxylated tetraethylene pentamine, various dispersing agents, especially polyacrylates and polyasparatates, various brighteners, especially anionic brighteners, various suds suppressors, especially silicones and secondary alcohols, various fabric softeners, especially smectite clays, and the like, also find use in various compositions, at levels ranging from about 1% to about 35% by weight. Standard formularies and published patents contain multiple, detailed descriptions of such conventional materials that find use in compositions provided by the present invention.
  • enzyme stabilizers also find use in the cleaning compositions provided herein.
  • Such stabilizers include, but are not limited to propylene glycol (preferably from about 1% to about 10%), sodium formate (preferably from about 0.1% to about 1%) and calcium formate (preferably from about 0.1% to about 1%).
  • the hard surface cleaning compositions and/or fabric cleaning compositions are formulated to include various builders at levels from about 5% to about 50% by weight. Typical builders include, but are not limited to 1-10 micron zeolites, polycarboxylates such as citrate and oxydisuccinates, layered silicates, phosphates, and the like. Other conventional builders are listed in standard formularies, and find use in the present invention.
  • cleaning methods provided by the present invention are more effective at removal of certain stains (e.g., stains from foodstuffs containing natural gum polysaccharides), than equivalent methods that do not employ a transglucosidase enzyme. In some preferred embodiments, in comparison to an otherwise equivalent method that does not contain a transglucosidase enzyme, the cleaning compositions of the present invention are more effective at stain removal.
  • the subject method removes and/or discolors at least about 20%, at least about 40%, at least about 60%, at least about 80% or, in some embodiments, at least about 90% more stain than an equivalent method that does not employ a transglucosidase enzyme.
  • a nucleic acid encoding the mature transglucosidase enzyme of A. niger was amplified from the genomic DNA of A. niger by PCR and cloned into the vector pTrex3 to make pTrex3(AGL51M ).
  • pTrex3(AGL51M ) is illustrated in Figure 1.
  • the transglucosidase protein encoded by this vector was operably linked to the CBHl signal sequence to facilitate its secretion into the growth medium.
  • the transglucosidase coding sequence was flanked by the T. reesei cbhl promoter and terminator.
  • the nucleotide sequence of the expression cassette of pTrex3(AGL51M ) vector is set forth in Figure 2.
  • the 7.57 kb Xba ⁇ -Xba ⁇ fragment of the plasmid pTrex3(AGL5 IM ) was purified by agarose gel electrophoresis and used to transform the spores of T. reesei by electroporation.
  • the electroporation parameters were as follows: voltage - 16 kV/cm, capacitance - 25 ⁇ F , resistance - 50 ⁇ . Electroporation was carried out using a suspension of freshly harvested T. reesei spores suspended in ice-cold 1.2 M sorbitol.
  • the spores were incubated overnight in a rotary shaker (3O 0 C, 200 rpm) in a medium containing 1 M sorbitol, 0.3% glucose, 0.3% Bacto peptone and 0.15% yeast extract.
  • the germinating spores were plated on a selective medium containing acetamide as a sole source of nitrogen (acetamide 0.6 g/1; cesium chloride 1.68 g/1; glucose 20 g/1; potassium dihydrogen phosphate 15 g/1; magnesium sulfate heptahydrate 0.6 g/1; calcium chloride dihydrate 0.6 g/1; iron (II) sulfate 5mg/l; zinc sulfate 1.4 mg/1; cobalt (II) chloride lmg/1; manganese (II) sulfate 1.6 mg/1; agar 20 g/1; pH 4,25).
  • acetamide 0.6 g/1; cesium chloride 1.68 g/1; glucose 20 g/1; potassium dihydrogen phosphate 15 g/1; magnesium sulfate heptahydrate 0.6 g/1; calcium chloride dihydrate 0.6 g/1; iron (II) sulfate 5mg/l; zinc sul
  • Transformant colonies appeared in about 1 week. Individual transformants were transferred onto fresh acetamide selective plates and allowed to grow for 3-4 days.
  • Isolates showing stable growth on selective medium were used to inoculate 5 ml of lactose defined medium ⁇ See e.g., WO 2005/001036, p. 60) in 20x175 mm test tubes. The tubes were fixed in a rotary shaker at about 45° angle and shaken at 200 rpm and 28 0 C for 4-5 days. Electrophoretic analysis of the culture supernatants demonstrated the presence of a new protein band of approximately expected molecular weight. [0100] The production of transglucosidase activity by the transformants was also confirmed using an enzymatic assay.
  • the assay was carried out in 100 mM sodium acetate buffer, pH 4.5, containing 4 mM para-nitrophenyl- ⁇ -glucoside and 1 mg/ml BSA. After 30 min incubation at 3O 0 C the reaction was terminated by the addition of an equal volume 1 M sodium carbonate and OD 4 o 5 was recorded. Typically, transformants expressed 1-2U transglucosidase activity (expressed as micromoles of para-nitrophenol liberated per min) per ml of culture broth. In untransformed controls, the activity was below detection limit.
  • EXAMPLE 2 Transglucosidase Degrades Xanthan Gum [0101] Hydrolytic activity by enzymes on xanthan gum was measured by the reducing sugar assay using the PAHBAH (para-hydroxybenzoic acid hydrazide) reagent, as known in the art ⁇ See, Lever et al, Anal. Biochem., 47: 273 [1972]).
  • PAHBAH para-hydroxybenzoic acid hydrazide
  • Xanthan gum (CAS 111 38-66-2) was purchased from Sigma Chemicals, St. Louis MO and dissolved in 50 mM sodium acetate buffer pH 6.0 at a concentration of 0.2%.
  • AATCC standard heavy duty liquid detergent AATCC HDL 2003 without brightner, Test Fabrics, Inc.
  • the assay was performed as follows in a 24 well microplate (COSTAR 3526, Corning Incorporated, Corning, N. Y.): one ml of buffer was added to well 1, one ml of buffer plus enzyme was added to well 2, one ml of buffer and substrate to well 3, and one ml of buffer, plus substrate and enzyme was added to well 4. For statistical purposes, each well may be set up 2 to 4 times.
  • Enzymes to be tested are usually diluted in reaction buffer from 1 to 10 to 1 to 1000. After all reagents were added, a plastic cover was place over the microplate and the cover and plate intersection was wrapped tightly with several layers of Paraf ⁇ lm (Pechiney Plastic Packing, Menasha, WI) to prevent evaporation. The reaction plate was incubated for 1 to 16 hr, at 37 0 C on a shaker rotating at 100 rpm. [0103] Reducing sugar activity was measured using an Eppendorf Mastercycler Gradient (Eppendorf Scientific, Westbury, NY) thermal cycler and 0.2 ml disposable PCR (polymerase chain reaction) strip tubes and caps purchased from VWR International, West Chester, PA.
  • Reducing sugar reagent was prepared as follows: to 10 ml of 2% sodium hydroxide in distilled water, add 0.15 g of sodium potassium tartrate tetrahydrate (Rochelle Salt, Sigma Chemical Co.) and 0.15 g of parahydroxybenzoic acid hydrazide (H-9882, Sigma Chemical Co.). The solution (called "PAHBAH reagent”) was swirled to solubilize all ingredients and put on ice in the dark until used. This reagent was made fresh daily. Immediately before sample analysis, 0.160 ml of PAHBAH reagent was added to each tube of a PCR strip followed by 5 to 20 ul of enzyme samples and controls.
  • FIG. 3 shows that transglucosidases produced in Trichoderma (Trip-TG) and in Aspergillus (Mega-TG) showed significant reducing sugar activity on xanthan gum in 50 mM acetate buffer plus AATCC heavy duty liquid detergent at pH 6.0.
  • microplate was covered with a plastic lid and aluminum foil and incubated at 37 0 C with lOOrpm gentle rotation for 4-16 hr.
  • the plates were removed from the shaker and the detergent solution was removed by aspiration.
  • Each microplate well was washed three (3) times with 1.5 ml of Dulbecco's PBS pH 7.3 and three (3) times with 1.5 ml of distilled water.
  • Each disk was removed from its well and dried overnight between sheets of paper towels and not exposed to direct light. Disks were inspected visually and then analyzed with a Minolta Reflectometer CR-200 calibrated on a standard white tile. The average L values with standard deviations were calculated.
  • FIG. 4 The graphs provided in Figure 4 show that Trip-TG (diluted 1/50 in 50 mM Hepes buffer) removes soil from both salad dressing stains and guar-pigment stains in 50 mM Hepes buffer pH 7.4 and in 50 mM Hepes buffer pH 7.4 plus 0.15% AATCC HDL.
  • Figure 5 shows the results obtained in a Trip-TG dose response microswatch experiment. Trip-TG removed salad dressing soils at 1 ppm in 0.15% AATCC heavy duty liquid..
  • Figure 6 shows that Trip-TG removed salad dressing soil in a microswatch cleaning experiment in 0.1% North American AATCC- 1993 HDD standard without brightner.
  • This detergent contained 18% linear alkyl sulfonates, 25% Zeolite A, 18% soda ash, 0.5% sodium silicate, 22% sodium sulfate, 10% moisture, and a 6.3% copolymer or other additives. [0110] This experiment showed transglucosidase soil removal activity in both heavy duty liquids (HDL) and in heavy duty solids (HDD).
  • HDL heavy duty liquids
  • HDD heavy duty solids
  • Percent soil release was calculated by standard methods after analysis of each stain by reflectometer.
  • Figure 7 shows that transglucosidase significantly cleaned marmalade stain compared to a no enzyme control or a control with an unrelated protein, bovine serum albumin (BSA-50, Fraction V, Immunoglobulin and Protease Free) obtained from Rockland Immunochemicals, Gilbertsville, PA.
  • BSA-50 bovine serum albumin

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Abstract

L'invention concerne des compositions comprenant une enzyme transglucosidase et un polysaccharide de gomme naturelle qui sert de substrat à la transglucosidase. L'invention concerne également des procédés d'utilisation de la transglucosidase pour dégrader un polysaccharide de gomme naturelle. Dans certains modes de réalisation préférés, les compositions et les procédés sont utilisés dans des applications de nettoyage.
EP08727093A 2007-03-22 2008-03-21 Compositions de nettoyage comprenant une transglucosidase Withdrawn EP2126024A2 (fr)

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WO2010115664A2 (fr) 2009-03-30 2010-10-14 Unilever Nv Composition de conditionnement de tissus
RU2013122800A (ru) 2010-10-20 2014-11-27 ДАНИСКО ЮЭс ИНК. Термостабильная целлюлаза из trichoderma
AR090971A1 (es) 2012-05-07 2014-12-17 Novozymes As Polipeptidos que tienen actividad de degradacion de xantano y polinucleotidos que los codifican
US9988657B2 (en) 2014-02-27 2018-06-05 E I Du Pont De Nemours And Company Enzymatic hydrolysis of disaccharides and oligosaccharides using alpha-glucosidase enzymes
CN113834757B (zh) * 2021-07-30 2024-02-23 广东美味鲜调味食品有限公司 一种对蚝油原料的耐高温淀粉酶微量残留的快速检测方法
AU2022354202A1 (en) 2021-09-30 2024-03-14 International N&H Denmark Aps Method for reducing sugar in food stuff
EP4604740A1 (fr) 2022-10-17 2025-08-27 International N&H Denmark ApS Procédé d'amélioration de l'arôme dans un produit alimentaire à base de plante
WO2024206631A1 (fr) 2023-03-29 2024-10-03 International N&H Denmark Aps Procédés de modification de texture dans un produit alimentaire par l'intermédiaire d'alpha-glucane de préférence produit in situ
WO2025198996A1 (fr) 2024-03-19 2025-09-25 Danisco Us Inc. Procédé d'amélioration du goût dans un produit alimentaire

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EP0896998A1 (fr) * 1997-08-14 1999-02-17 The Procter & Gamble Company Compositions détergentes pour le linge contenant une enzyme dégradant la gomme de polysaccharide
US6486112B1 (en) * 1997-08-14 2002-11-26 The Procter & Gamble Company Laundry detergent compositions comprising a saccharide gum degrading enzyme
WO1999055817A1 (fr) * 1998-04-29 1999-11-04 The Procter & Gamble Company Detergent a lessive et/ou compositions d'entretien des tissus comprenant la transferase a
JP2002513563A (ja) * 1998-05-01 2002-05-14 ザ、プロクター、エンド、ギャンブル、カンパニー 修飾トランスフェラーゼを含有した洗濯洗剤および／または布帛ケア組成物
US6410498B1 (en) * 1999-04-30 2002-06-25 Procter & Gamble Company Laundry detergent and/or fabric care compositions comprising a modified transferase
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