Low-dusting granules

Abstract

The present invention provides layered particles comprising a core and a matrix coating surrounding the core. The matrix comprises a bioactive and a non-volatile liquid and can be applied using a fluidized bed sprayer (fluidized bed coating). Upon subjecting the particles to mechanical stress, the particles exhibit reduced release of bioactive dust, such as enzyme dust.

Description

[0001] This application is a divisional application of the invention patent application filed on October 15, 2018, with application number 201880066602.3 and invention title "Low-Powdered Particles". Technical Field

[0002] This invention relates to layered particles comprising a core and a matrix layer surrounding the core. The matrix comprises a bioactive substance and a non-volatile liquid, and can be applied using a fluidized bed coating machine. After subjecting these particles to mechanical stress, they exhibit reduced release of bioactive dust, such as enzyme dust. Background Technology

[0003] One of the main challenges in developing enzyme formulations is avoiding exposure to enzyme dust, which is known to cause irritation or allergic reactions. Since the 1970s, several breakthroughs have been made in this field, to the point that potential dust exposure only occurs when enzyme products undergo destructive processes due to mechanical or spillage processes, including shear stress, impact stress, or compressive stress as described by Meesters in Agglomeration of Enzymes, Micro-organisms and Flavours, Handbook of Power Technology, Volume 11, Granulation, edited by AD Salman et al., 2007.

[0004] Traditionally, bioactive substances (such as enzymes) are formulated as liquids or solids. Liquid formulations have an inherent advantage in inhibiting enzyme dust formation, while dry solid compositions are generally far superior for other properties (such as the stability of the enzyme activity) because bioactive substances (such as enzymes) can be very effectively separated from other components, and dry compositions typically do not provide a medium in which the active substance is degraded. Although several solid formulations known in the art effectively limit the formation of active dust, in reality, active dust can still be released from the particles, for example, as measured by the well-known Heubach method or washing method, and this release increases when the solid formulation breaks due to destructive stress during processing.

[0005] In this invention, we combine the advantages of both liquids and solids with the result of lower exposure to active dust (even after subjecting particles containing bioactive substances to destructive stress).

[0006] WO 2004 / 058933 describes the manufacture of mechanically robust granules by impregnating (absorbing) a plasticizer onto a coating / surface containing a plasticizable polymer. The method described in the patent requires additional, complex, and difficult steps in the granulation process and can only ensure plasticization of the granule surface or coating. Such plasticized coatings / surfaces can reduce the tendency of granules to release dust under physical stress / impact, provided the granule coating / surface remains intact; however, if the granule coating / surface is damaged / cut (e.g., by closing a valve or through some grinding process), dust emissions are not reduced, as these processes are commonly used, for example, in the production of detergent powders.

[0007] WO 02 / 28991 describes particles containing an active ingredient contained in a viscoelastic liquid matrix, wherein the viscoelastic parameters η' (elastic parameter) and η' (viscosity parameter), measured in a cone-plate rheometer at 25°C and a sinusoidal frequency ω of 1 Hz, are within a range of 10. 3 Pa to 10 14 The processing of such materials is inherently more difficult than that of materials without viscoelastic properties, meaning that commonly used granulation methods, such as high-shear granulation described in US 4,106,991 and US 4,661,452, and fluidized bed methods described in US 5,324,649, cannot be used to produce particles preferably ranging from 200 micrometers to 2000 micrometers. Viscoelastic properties will prevent the operation of high-shear granulation processes because the viscoelastic liquid matrix does not fracture upon impact, but only deforms, resulting in virtually no particles within the desired size range. In fluidized bed methods, viscoelastic properties induce severe agglomeration, which also leads to very low yields of particles with the desired size. Summary of the Invention

[0008] In a first aspect, the present invention provides layered particles comprising a core surrounded by a matrix layer (matrix coating), wherein the matrix layer comprises a bioactive substance and 1%-50% non-volatile liquid.

[0009] In one embodiment, the bioactive substance is an enzyme.

[0010] In one embodiment, the particles contain an additional (salt) coating.

[0011] In one embodiment, a matrix layer surrounding the core is applied in a fluidized bed sprayer.

[0012] The particles exhibit reduced dust release when exposed to mechanical stress and can be used as an ingredient in (powder) detergents.

[0013] Other aspects and embodiments of the invention will be apparent from the description and examples. Detailed Implementation

[0014] We have found that it is possible to prepare solid particles / particles containing a certain amount of non-volatile liquid and bioactive substances (e.g., enzymes), wherein the amount of active dust (bioactive substances) released by particles subjected to shear stress is lower than that released by reference particles. The release of active dust before and after the application of shear stress was quantified by active dust analysis, as described in the Examples section. This analysis incorporates a pre-analysis step in which the particles are compressed or even crushed, and the coating is broken, thus providing a more well-defined picture of the robustness of the particles against shear stress.

[0015] The reduction in the release of reactive dust after applying shear stress is surprising, since methods known in the art only involve reducing the release of reactive dust from undisturbed, typically coated particles containing bioactive substances.

[0016] Therefore, one object of the present invention is to provide a new and safer way of using and handling bioactive substances (e.g., enzymes and / or microorganisms). It is generally desirable to isolate bioactive substances from their surrounding environment until the moment they are used in an application. This is achieved by incorporating the active substances into discrete particles. Incorporating the active substances into particles can also be used to reduce the amount of potentially harmful bioactive substance dust that can be generated by the bioactive substances. The present invention relates to such improved particles.

[0017] The particles / granules of the present invention comprise a bioactive substance dispersed in a wet solid matrix, wherein the solid matrix is ​​wetted with a non-volatile liquid (e.g., glycerol). According to the "Test Methods" (see Examples), such wetted but still solid particles exhibit lower release of active dust compared to unwetted reference particles.

[0018] US 4,106,991 explores the possibility of using waxy components with melting points above 30°C to achieve this plastic behavior to some extent. In this invention, we use a non-volatile liquid to create wet solid particles that will be strong and plastic enough to withstand destructive stress.

[0019] The present invention includes a fluidized bed spraying process for applying a wet solid matrix onto core particles to achieve a final product exhibiting low dust characteristics.

[0020] definition

[0021] As used in the context of this invention, the term "liquid" should be understood to refer to the properties of a material. A liquid material is defined as a material that, when a deformation force, i.e., strain, is applied to it, it acquires a certain amount of stress, i.e., the force per unit area, as long as the material deforms. However, once deformation ceases, the stress level immediately decreases to a steady-state level, which is always precisely zero. A liquid cannot maintain or sustain internal persistent stress. The properties of a liquid are understood to be the properties of the liquid itself applied to the coating granulation process (not the properties of the generated particles / coating).

[0022] As used in the context of this invention, the term "viscoelastic" should be understood to refer to the properties of the liquid. A liquid (fluid) is viscoelastic if the time span from the occurrence of deformation to the stress in the material reaching a precise zero value is sufficiently long. Viscoelastic liquids can be described using a simple model containing two parameters, η'(ω) and η'(ω), which can be readily measured in a cone-plate rheometer (e.g., a Bohlin rheometer) at different sinusoidal frequencies ω. η'(ω) can be interpreted as the elastic parameter of the viscoelastic fluid. This definition is recognized in the art, for example, in Bird RB, Armstrong RC, Hassager O. "Dynamics of polymeric liquids", Volume 1: Fluid mechanics, John Wiley and Sons, Chapter 6, especially Example 6.1.2.1, p 281, 1977; and in WO 02 / 28991.

[0023] The term "viscosity" as used in the context of this invention should be understood to refer to the properties of such liquids. Unless otherwise stated, viscosity μ is given as dynamic viscosity (e.g., in Pascal-seconds (Pa·s)), which is measured at 25°C using 1 s⁻¹. -1 The shear rate is measured in a cone-plate rheometer (e.g., a Bohlin rheometer). For Newtonian (or near-Newtonian) liquids, viscosity can be measured at other shear rates.

[0024] Surface tension, denoted by γ, is defined as the energy required to increase the surface area of ​​a liquid per unit area, and is typically measured in mN / m (Kirk-Othmer Encyclopedia of Chemical Technology, "Surfactants"). The surface tension of a liquid in air can be measured by those skilled in the art and is expressed as γ. LV .

[0025] The wettability of a liquid to a solid is determined by the contact angle (θ) between the solid and the liquid.

[0026] As used in the context of this invention, the term "multiple particles" refers to the number of particles required to determine the particle size distribution with reasonable precision, but at least 50 particles (randomly sampled), such as at least 100, 500, or 1000 particles. Typically, multiple particles are one gram or several grams of particles. The particle size distribution can be measured using laser diffraction, optical digital imaging, or sieving analysis.

[0027] Fluidized bed spraying process

[0028] The layered particles of the present invention can be prepared during a fluidized bed spraying process. The fluidized bed spraying process according to the present invention is a process for producing layered products, wherein an enzyme is coated as a layer surrounding a pre-formed (inert) core particle, wherein an enzyme-containing solution is atomized, typically in a fluidized bed apparatus. The pre-formed core particle is fluidized in an airflow, and the enzyme-containing solution adheres to the core particle. Particles with a desired size can be obtained by using core particles of a desired size. This type of product is described, for example, in WO 97 / 23606.

[0029] Drying is preferably carried out at a product temperature ranging from 25°C to 90°C. After drying, these cores preferably contain 0.1% w / w to 10% w / w of water.

[0030] Non-volatile liquids

[0031] The non-volatile liquid mixed with the bioactive substance in the matrix layer according to the present invention is a non-volatile liquid chemical compound or a mixture of non-volatile liquid chemical compounds, each having the properties specified below.

[0032] At 25°C, the vapor pressure of the non-volatile liquid is less than 1 kPa, and when measured in a cone-plate rheometer (such as a Bohlin rheometer) using a sinusoidal frequency ω at 1 Hz at 25°C, it has an elastic parameter η' that is less than 0.1 kPa.

[0033] In a preferred embodiment, the vapor pressure of the non-volatile liquid is less than 0.5 kPa at 25°C. In another preferred embodiment, the elastic parameter η' is less than 10 Pa; for example, less than 1 Pa.

[0034] To minimize liquid loss from the particle-containing bioactive material during production and use, a low vapor pressure is preferred. For better dispersion of the active material in the liquid, a low elasticity parameter η' is preferred.

[0035] Preferably, the liquid has a melting point of 25°C (for components with a melting range, this means at least 50% of the component is in a liquid state) or lower, more preferably lower than 20°C, even more preferably lower than 10°C, and most preferably lower than 5°C. The liquid is advantageously selected to be in a liquid state under the conditions of use of the particles.

[0036] Preferably, the non-volatile liquid is water-soluble. The bioactive component is typically added to the granulation process as a solution or dispersion in water, and / or water is used as a granulation aid during the granulation process. Therefore, the non-volatile component can be conveniently added to the matrix by dissolving it in these aqueous liquids. At 25°C, the solubility of the non-volatile liquid in water should be at least 1% by weight (i.e., 1 g dissolved in 99 g of water), more preferably at least 10%, even more preferably at least 25%, and most preferably at least 50%.

[0037] In one embodiment, the non-volatile liquid has a surface tension of at least 30 mN / m at 20°C (or the melting point of a liquid with a higher melting point). Preferably, the surface tension is at least 40 mN / m; and more preferably, at least 50 mN / m. High surface tension is advantageous because it improves the binding effect of the liquid. This is important for balancing the plastic behavior of a wet solid matrix with sufficient yield strength, thereby preventing particle disintegration under shear stress and reducing the release of reactive dust.

[0038] In one embodiment, the dynamic viscosity of the non-volatile liquid is at least 0.001 Pa·s, more preferably at least 0.01 Pa·s, and most preferably at least 0.1 Pa·s; at 25°C, at a shear rate of 1 s⁻¹ -1 Measurements were taken in a cone-plate rheometer. High viscosity requires the liquid to have stronger binding properties, providing improved particle strength, thereby resulting in reduced release of reactive dust.

[0039] In one embodiment, the non-volatile liquid is capable of wetting a mixture of other components of the matrix. Advantageously, the non-volatile liquid's ability to wet the mixture of other components facilitates its diffusion and distribution within the matrix. This means that the contact angle of droplets placed on a perfect surface of the mixture of other components within the matrix should be less than 180 degrees, more preferably less than 135 degrees, and most preferably less than 90 degrees.

[0040] In a preferred embodiment, the non-volatile liquid used according to the present invention is a polyol, such as an alcohol having many hydroxyl groups, such as glycerol, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, tripropylene glycol, polyethylene glycol, and polar low molecular weight organic compounds.

[0041] The most preferred are glycerol, triethylene glycol, propylene glycol, and polyethylene glycol (PEG) with an average molecular weight of about 1000 or less.

[0042] As described above, the non-volatile liquid can be a mixture of two or more compounds / liquids, each of which exhibits the characteristics of the non-volatile liquid of the present invention.

[0043] The non-volatile liquid used in this invention can be a material that is anhydrous or contains at least a very small amount of water. The water may be bound to the components of the liquid, or the liquid may contain water absorbed from a humid environment. Therefore, the amount of water in the liquid depends on the components of the liquid, the hygroscopicity of the components, and the humidity of the surrounding environment. Typically, water is used as a processing aid, for example, to carry active ingredients from water droplets to the particle surface in a fluidized bed coated product. Typically, most of the water is removed during processing / drying.

[0044] bioactive substances

[0045] In the context of this invention, a bioactive substance is a compound or microorganism that exhibits biological activity, such as catalyzing biochemical reactions or carrying out biological processes.

[0046] Preferred examples of bioactive substances are enzymes and microorganisms, such as bacterial spores.

[0047] enzymes

[0048] Bioactive substances can be one or more enzymes, such as proteases, lipases, keratinases, amylases, glycoses, cellulases, pectinases, mannanases, arabinases, galactanases, xylanases, DNases, hydrolases, oxidases (e.g., laccase), and / or peroxidases.

[0049] The enzyme can be a naturally occurring bacterial or fungal enzyme, or it can be a variant derived from one or more naturally occurring enzymes through gene recombination and / or by substitution, deletion, or insertion of one or more amino acids. This includes chemically modified mutants or protein-engineered mutants.

[0050] Preferably, the particle contains at least one enzyme in an amount of more than 0.5% w / w and less than 50% w / w active enzyme protein; more preferably, in an amount of more than 0.6% w / w and less than 40% w / w active enzyme protein; even more preferably, in an amount of more than 0.75% w / w and less than 30% w / w active enzyme protein; and most preferably, in an amount of more than 1% w / w and less than 25% w / w active enzyme protein.

[0051] CellulaseSuitable cellulases include those of bacterial or fungal origin. This includes chemically modified mutants or protein-engineered mutants. Suitable cellulases include those from the genera *Bacillus*, *Pseudomonas*, *Pyrophyllus*, *Fusarium*, *Clostridium*, and *Cladosporium*, such as the fungal cellulases produced by *Pyrophyllus*, *Thermophyllus*, and *Fusarium* as disclosed in US 4,435,307, US 5,648,263, US 5,691,178, US 5,776,757, and WO 89 / 09259.

[0052] Particularly suitable cellulases are alkaline or neutral cellulases that offer color-care benefits. Examples of such cellulases are those described in EP 0 495 257, EP 0 531 372, WO 96 / 11262, WO 96 / 29397, and WO 98 / 08940. Other examples are cellulase variants, such as those described in WO 94 / 07998, EP 0 531 315, US 5,457,046, US 5,686,593, US 5,763,254, WO 95 / 24471, WO 98 / 12307, and PCT / DK 98 / 00299.

[0053] Commercially available cellulases include Celluzyme™, Carezyme™ and Cellluclean™ (Novozymes), Clazinase™ and Puradax HA™ (Genetronics), and KAC-500(B)™ (Kao Corporation).

[0054] protease Suitable proteases include those of bacterial, fungal, plant, viral, or animal origin, such as those of plant or microbial origin. Microbial origin is preferred. This includes chemically modified mutants or protein-engineered mutants. It can be an alkaline protease, such as a serine protease or a metalloproteinase. Serine proteases can be, for example, from the S1 family (such as trypsin) or the S8 family (such as subtilisin). Metalloproteinases can be, for example, thermophilic bacterial proteases from family M4 or other metalloproteinases, such as those from the M5, M7, or M8 families.

[0055] The term "subtilisinase" refers to the serine protease subgroup according to Siezen et al., Protein Engineering, 4 (1991) 719-737 and Siezen et al., Protein Science, 6 (1997) 501-523. Serine proteases are a subgroup of proteases characterized by the presence of a serine residue at its active site that forms a covalent adduct with the substrate. Subtilisins can be divided into six subfamilies: the subtilisin family, the thermophilic protease family, the proteinase K family, the lanoxanthipeptidase family, the Kexin family, and the Pyrolysin family.

[0056] Examples of subtilisinases are those derived from the genus Bacillus, such as *Bacillus lentus*, *Bacillus alkalophilus*, *Bacillus subtilis*, *Bacillus amyloliquefaciens*, *Bacillus pumilus*, and *Bacillus giganteus* as described in US 7262042 and WO 09 / 021867; and *Lentus*, *Novo*, *Carlsberg*, *Bacillus licheniformis*, *BPN'*, *309*, *147*, and *168* as described in WO 89 / 06279, and *PD138* as described in (WO 93 / 18140). Other useful proteases may be those described in WO 92 / 175177, WO 01 / 016285, WO 02 / 026024, and WO 02 / 016547. Examples of trypsin-like proteases are trypsin (e.g., from pigs or cattle) and Fusarium proteases (described in WO 89 / 06270, WO 94 / 25583 and WO 05 / 040372), as well as chymotrypsin derived from Cellumonas (described in WO 05 / 052161 and WO 05 / 052146).

[0057] Further preferred proteases are alkaline proteases from Bacillus tarda DSM 5483 (as described in, for example, WO95 / 23221) and their variants (described in WO 92 / 21760, WO 95 / 23221, EP 1921147 and EP 1921148).

[0058] Examples of metalloproteinases are, for example, neutral metalloproteinases described in WO 07 / 044993 (Genencor Int.), such as those derived from Bacillus amyloliquefaciens.

[0059] Examples of useful proteases are the variants described in the following: WO 92 / 19729, WO 96 / 034946, WO98 / 20115, WO 98 / 20116, WO 99 / 011768, WO 01 / 44452, WO 03 / 006602, WO 04 / 03186, WO 04 / 041979, WO 07 / 006305, WO 11 / 036263, WO 11 / 036264, especially with a variant that has a substitution in one or more of the following positions: 3, 4, 9, 15, 27, 36, 57, 68, 76, 87, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 106, 118, 120, 123, 128, 129, 130, 160, 167, 170, 194, 195, 199, 205, 206, 217, 218, 222, 224, 232, 235, 236, 245, 248, 252 and 274, numbered using BPN'. More preferably, the Bacillus subtilis enzyme variant may contain the following mutations: S3T, V4I, S9R, A15T, K27R, *36D, V68A, N76D, N87S,R, *97E, A98S, S99G,D,A, S99AD, S101G,M,R S103A, V104I,Y,N, S106A, G118V,R, H120D,N, N123S, S128L, P129Q, S130A, G160D, Y167A, R170S, A194P, G195E, V199M, V205I, L217D, N218D, M222S, A232V, K235L, Q236H, Q245R, N252K, T274A (numbered using BPN').

[0060] Suitable commercially available proteases include those sold under the following trade names: Alcalase®, Duralase Tm Durazym Tm Relase®, Relase® Ultra, Savinase®, Savinase® Ultra, Primease®, Polarzyme®, Kannase®, Liquanase®, Liquanase® Ultra, Ovozyme®, Coronase®, Coronase® Ultra, Neutrase®, Everlase®, and Esperase® (Novozymes), sold under the following trade names: Maxatase®, Maxacal®, Maxapem®, Purafect®, Purafect Prime®, PreferenzTm , Purafect MA®, Purafect Ox®, Purafect OxP®, Puramax®, Properase®, Effectenz Tm FN2®, FN3®, FN4®, Excellase®, Opticlean®, Optimase®, and Excellenz P1000 (Danisco / DuPont), Axapem™ (Gist-Brocases NV), BLAP (sequence shown in Figure 29 of US 5352604) and its variants (Henkel AG), as well as KAP (Alkaliophilic Bacillus subtilis protease) from Kao Corporation.

[0061] Lipase and keratinase Suitable lipases and keratins include those of bacterial or fungal origin. This includes chemically modified or protein-engineered mutant enzymes. Examples include lipases from the genus *Thermophilic*, such as those from *Thermophilic Hypotherium latifolium* (formerly named *Pythium latifolium*) as described in EP258068 and EP305216; cutinases from the genus *Pythium*, such as *Pythium salivarium* (WO 96 / 13580); lipases from strains of the genus *Pseudomonas* (some of which are now renamed *Burkholderia*), such as *Alcaligenes* or *Alcaligenes-like* (EP 218272), *Pseudomonas cepacia* (EP 331376), *Pseudomonas* strain SD705 (WO 95 / 06720 & WO 96 / 27002), *Pseudomonas wisconsinensis* (WO 96 / 12012); GDSL-type *Streptomyces* lipase (WO 10 / 065455); and cutinases from *Bacillus oryzae* (WO 10 / 065455). 10 / 107560); cutinase from Pseudomonas mendoza (US 5,389,536); lipase from Thermobifida fusca (WO 11 / 084412); lipase from Bacillus stearothermophilus (WO 11 / 084417); lipase from Bacillus subtilis (WO 11 / 084599); and lipase from Streptomyces griseus (WO 11 / 150157) and Streptomyces pristinaespiralis (WO 12 / 137147).

[0062] Other examples are lipase variants, such as those described in EP 407225, WO 92 / 05249, WO 94 / 01541, WO 94 / 25578, WO 95 / 14783, WO 95 / 30744, WO 95 / 35381, WO 95 / 22615, WO 96 / 00292, WO 97 / 04079, WO 97 / 07202, WO 00 / 34450, WO 00 / 60063, WO 01 / 92502, WO 07 / 87508, and WO 09 / 109500.

[0063] Preferred commercially available lipase products include Lipolase™, Lipex™; Lipolex™ and Lipoclean™ (Novozymes), Lumafast (Genencor), and Lipomax (Gist-Brocades).

[0064] Other examples are lipases sometimes called acyltransferases or perhydrolases, such as an acyltransferase homologous to Candida antarctica lipase A (WO 10 / 111143), an acyltransferase from Mycobacterium smegmatis (WO 05 / 56782), a perhydrolase from the CE 7 family (WO 09 / 67279), and variants of Mycobacterium smegmatis perhydrolases (particularly the S54V variant used in the commercial product Gentle Power Bleach from Huntsman Textile Effects Pte Ltd) (WO 10 / 100028).

[0065] amylase: Suitable amylases are α-amylases or glucosylamylases and can be of bacterial or fungal origin. This includes chemically modified mutants or protein-engineered mutants. Amylases include, for example, α-amylases obtained from specific strains of Bacillus, such as Bacillus licheniformis (described in more detail in GB 1,296,839).

[0066] Suitable amylases include the amylase having SEQ ID NO: 3 in WO 95 / 10603 or a variant thereof having 90% sequence identity with SEQ ID NO: 3. Preferred variants are described in WO 94 / 02597, WO 94 / 18314, WO 97 / 43424 and SEQ ID NO: 4 in WO 99 / 019467, for example, variants having substitutions at one or more of the following positions: 15, 23, 105, 106, 124, 128, 133, 154, 156, 178, 179, 181, 188, 190, 197, 201, 202, 207, 208, 209, 211, 243, 264, 304, 305, 391, 408 and 444.

[0067] Suitable amylases include the amylase having SEQ ID NO: 6 in WO 02 / 010355 or a variant thereof having 90% sequence identity with SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those with deletions at positions 181 and 182 and substitutions at position 193. Other suitable amylases are hybrid α-amylases comprising residues 1-33 of the α-amylase derived from Bacillus amyloliquefaciens shown in SEQ ID NO: 6 of WO 2006 / 066594 and residues 36-483 of the Bacillus licheniformis α-amylase shown in SEQ ID NO: 4 of WO 2006 / 066594, or variants thereof having 90% sequence identity. Preferred variants of this hybrid α-amylase are those having substitutions, deletions, or insertions at one or more of the following positions: G48, T49, G107, H156, A181, N190, M197, I201, A209, and Q264. The most preferred variant of the hybrid α-amylase derived from *Bacillus amyloliquefaciens* as shown in SEQ ID NO: 6 of WO 2006 / 066594, containing residues 1-33 and residues 36-483 of SEQ ID NO: 4, is those having the following substitutions:

[0068] M197T;

[0069] H156Y + A181T + N190F + A209V + Q264S; or

[0070] G48A+T49I+G107A+H156Y+A181T+N190F+I201F+A209V+Q264S.

[0071] Another suitable amylase is the amylase having SEQ ID NO: 6 in WO 99 / 019467 or a variant thereof having 90% sequence identity with SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having substitutions, deletions, or insertions in one or more of the following positions: R181, G182, H183, G184, N195, I206, E212, E216, and K269. Particularly preferred amylases are those having deletions in positions R181 and G182, or positions H183 and G184.

[0072] Other amylases that can be used are those of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 2, or SEQ ID NO: 7 with WO 96 / 023873, or variants thereof having 90% sequence identity with SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 7. Preferred variants of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 7 are those with substitutions, deletions, or insertions at one or more of the following positions: 140, 181, 182, 183, 184, 195, 206, 212, 243, 260, 269, 304, and 476. More preferred variants are those with deletions at positions 181 and 182 or positions 183 and 184. The most preferred amylase variants of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7 are those with deletions in positions 183 and 184 and substitutions in one or more of positions 140, 195, 206, 243, 260, 304 and 476.

[0073] Other amylases that may be used are amylases having SEQ ID NO: 2 of WO 08 / 153815, SEQ ID NO: 10 of WO 01 / 66712, or variants thereof having 90% sequence identity with SEQ ID NO: 2 of WO 08 / 153815 or 90% sequence identity with SEQ ID NO: 10 of WO 01 / 66712. Preferred variants of SEQ ID NO: 10 of WO 01 / 66712 are those having substitutions, deletions, or insertions at one or more of the following positions: 176, 177, 178, 179, 190, 201, 207, 211, and 264.

[0074] Another suitable amylase is the amylase of SEQ ID NO: 2 having WO 09 / 061380 or a variant thereof having 90% sequence identity with SEQ ID NO: 2. Preferred variants of SEQ ID NO: 2 are those having C-terminal truncation and / or substitution, deletion, or insertion at one or more of the following positions: Q87, Q98, S125, N128, T131, T165, K178, R180, S181, T182, G183, M201, F202, N225, S243, N272, N282, Y305, R309, D319, Q320, Q359, K444, and G475. More preferred variants of SEQ ID NO: 2 are those having substitutions at one or more of the following positions: Q87E,R, Q98R, S125A, N128C, T131I, T165I, K178L, T182G, M201L, F202Y, N225E,R, N272E,R, S243Q,A,E,D, Y305R, R309A, Q320R, Q359E, K444E, and G475K, and / or those having deletions at positions R180 and / or S181 or T182 and / or G183. The most preferred amylase variants of SEQ ID NO: 2 are those having the following substitutions:

[0075] N128C + K178L + T182G + Y305R + G475K;

[0076] N128C + K178L + T182G + F202Y + Y305R + D319T + G475K;

[0077] S125A + N128C + K178L + T182G + Y305R + G475K; or

[0078] S125A + N128C + T131I + T165I + K178L + T182G + Y305R + G475K, wherein the variant is C-terminated and optionally further includes a substitution at position 243 and / or a deletion at positions 180 and / or 181.

[0079] Other suitable amylases are α-amylases having SEQ ID NO: 12 in WO 01 / 66712 or variants having at least 90% sequence identity with SEQ ID NO: 12. Preferred amylase variants are those having substitutions, deletions, or insertions at one or more of the following positions in SEQ ID NO: 12 in WO 01 / 66712: R28, R118, N174; R181, G182, D183, G184, G186, W189, N195, M202, Y298, N299, K302, S303, N306, R310, N314; R320, H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471, N484. Particularly preferred amylases include variants having deletions of D183 and G184 and having substitutions for R118K, N195F, R320K, and R458K, as well as variants having substitutions at one or more positions selected from the group consisting of M9, G149, G182, G186, M202, T257, Y295, N299, M323, E345, and A339, with the most preferred being variants having substitutions at all of these positions.

[0080] Other examples are amylase variants, such as those described in WO 2011 / 098531, WO 2013 / 001078 and WO 2013 / 001087.

[0081] Commercially available amylases include Duramyl™, Termammyl™, Fungammyl™, Stainzyme™, StainzymePlus™, Natalase™, Liquozyme X, and BAN™ (from Novozymes), as well as Rapidase™, Purastar™ / Effectenz™, Powerase, and Preferenz S100 (from Genencor International Inc. / DuPont).

[0082] Lysing enzyme:The lyase can be a pectin lyase of bacterial or fungal origin. This includes chemically or genetically modified mutants. In a preferred embodiment, the pectic acid lyase is derived from Bacillus, particularly Bacillus subtilis, Bacillus licheniformis, or Bacillus agaradhaerens, or a variant derived from any of these sources. For example, commercially available pectin lyases include XPect, Pectawash, and Pectaway (Novozymes), as described in US 6,124,127, WO 1999 / 027083, WO 1999 / 027084, WO 2002 / 006442, WO 2002 / 092741, and WO 2003 / 095638.

[0083] Mannanase: Suitable mannanases include those of bacterial or fungal origin. This includes chemically or genetically modified mutants. Mannanases can be basic mannanases of families 5 or 26. They can be wild-type from the genera *Bacillus* or *Pythium*, particularly *Bacillus mucosa*, *Bacillus licheniformis*, *Bacillus alkalophilus*, *Bacillus croceae*, or specific *Pythium* species. Suitable mannanases are described in WO 1999 / 064619. Commercially available mannanases are from Mannaway (Novozymes).

[0084] Deoxyribonuclease (DNAase): Suitable deoxyribonucleases (DNases) are any enzymes that catalyze the hydrolysis and cleavage of phosphodiester bonds in the DNA backbone, thereby degrading DNA. According to the invention, DNases obtainable from bacteria are preferred; particularly, DNases obtainable from the genus *Bacillus* are preferred; especially, DNases obtainable from *Bacillus subtilis* or *Bacillus licheniformis* are preferred. Examples of such DNases are described in patent applications WO 2011 / 098579 or PCT / EP2013 / 075922.

[0085] Overhydrolytic enzymes: Suitable perhydrolases catalyze perhydrolysis reactions, which result in the production of peracids from carboxylic acid ester (acyl) substrates in the presence of a peroxide source (e.g., hydrogen peroxide). While many enzymes carry out this reaction at low levels, perhydrolases exhibit high perhydrolysis:hydrolysis ratios (typically greater than 1). Suitable perhydrolases can be of plant, bacterial, or fungal origin. This includes chemically modified mutants or protein-engineered mutants.

[0086] Examples of useful perhydrolysins include naturally occurring mycobacterial perhydrolysins or variants thereof. Exemplary enzymes are derived from *Mycobacterium smegmatis*. The enzymatic properties, structure, and variants of such enzymes are described in WO 2005 / 056782, WO 2008 / 063400, US 2008 / 145353, and US 2007167344.

[0087] Peroxidase / oxidase: Suitable peroxidases comprise enzymes classified as EC 1.11.1.7 as stated by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB), or any fragment derived therefrom that exhibits peroxidase activity.

[0088] Suitable peroxidases include those of plant, bacterial, or fungal origin. This includes chemically modified mutants or protein-engineered mutants. Examples of useful peroxidases include peroxidases from the genus *Coprinus*, such as those from *C. cinerea* (EP 179,486), and their variants, such as those described in WO 93 / 24618, WO 95 / 10602, and WO 98 / 15257.

[0089] These peroxidases also include halogen peroxidases, such as chloride peroxidase, bromoperoxidase, and compounds exhibiting chloride or bromoperoxidase activity. Halogen peroxidases are classified according to their specificity for halide ions. Chloride peroxidase (EC1.11.1.10) catalyzes the formation of hypochlorite from chloride ions.

[0090] In the embodiments, the halogen peroxidase of the present invention is chloride peroxidase. Preferably, the halogen peroxidase is vanadium halide peroxidase, i.e., a vanadate-containing halogen peroxidase. In a preferred method of the present invention, the vanadate-containing halogen peroxidase is combined with a chloride ion source.

[0091] Halogen peroxidases have been isolated from many different fungi, particularly from the dematiaceous hyphomycete fungal group, such as Caldariomyces (e.g., C. fumago), Alternaria, Curvularia (e.g., C. verruculosa and C. inaequalis), Helicobacter, Fibrosporium, and Botrytis.

[0092] Halogen peroxidases have also been isolated from bacteria such as Pseudomonas (e.g., P. pyrrocinia) and Streptomyces (e.g., Streptomyces aureofaciens).

[0093] In a preferred embodiment, the halogen peroxidase may be derived from the genus *Curvularia*, particularly *Curvularia verruculosa* or *Curvularia inaequalis*, for example, *Curvularia inaequalis* CBS 102.42 described in WO 95 / 27046; or *Curvularia verruculosa* CBS 147.63 or *Curvularia verruculosa* CBS 444.70 described in WO 97 / 04102; or derived from species of the genus *Drechslera hartlebii* as described in WO 01 / 79459, *Dendryphiella salina* as described in WO 01 / 79458, *Phaeotrichoconis crotalarie* as described in WO 01 / 79461, or *Geniculosporium* as described in WO 01 / 79460.

[0094] Suitable oxidases specifically include any laccase contained in enzyme classification EC 1.10.3.2 or any fragment thereof exhibiting laccase activity, or compounds exhibiting similar activity, such as catechol oxidase (EC 1.10.3.1), o-aminophenol oxidase (EC 1.10.3.4), or bilirubin oxidase (EC 1.3.3.5).

[0095] The preferred laccase is a microbial enzyme. This enzyme can be derived from plants, bacteria, or fungi (including filamentous fungi and yeast).

[0096] Suitable examples of fungi include laccases derived from the following strains: *Aspergillus*, *Neurospora* (e.g., *Neurospora crassa*), *Stenocystis*, *Botrytis*, *Collybia*, *Fomes*, *Lentinula*, *Pleurotus*, *Coralus* (e.g., *Coralus longifolius* and *Coralus discolor*), *Rhizoctonia* (e.g., *Rhizoctonia solani*), *Coprinus* (e.g., *Coprinus comatus*, *Coprinus friesii*, and *Coprinus plicatilis*), *Psathyrella* (e.g., *P. condelleana*), *P. papilionaceus* (e.g., *P. papilionaceus*), *Schytalidium* (e.g., *S. thermophilum*), and *Polyporus* (e.g., *P.*). Pinsitus), genus P. radiata (e.g., P. radiata) (WO 92 / 01046) or genus C. hirsutus (e.g., C. hirsutus) (JP 2238885).

[0097] Suitable examples of bacteria include laccases derived from strains of the genus Bacillus. Laccases derived from the genera *Coprinus* or *Heterophytes* are preferred; particularly those derived from *Coprinus spp.*, as disclosed in WO 97 / 08325; or from *Heterophytes thermophilus*, as disclosed in WO 95 / 33836.

[0098] microorganism

[0099] The bioactive substance may also be one or more microorganisms, such as one or more fungi, yeasts, or bacteria. In a preferred embodiment, the one or more microorganisms are one or more dehydrated bacteria or yeasts.

[0100] In certain embodiments, the bioactive substance is one or more microbial spores (as opposed to vegetative cells), such as bacterial spores; or fungal spores, conidia, hyphae. Preferably, the one or more spores are Bacillus endospores; even more preferably, the one or more spores are spores of Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, and / or Bacillus megaterium.

[0101] Particles

[0102] The layered particles of the present invention are small particles containing bioactive substances and non-volatile liquids. These particles can be (generally) spherical.

[0103] These particles typically have a (weight / volume average) diameter of 20 µm-3000 µm, specifically 50 µm-2000 µm, 100 µm-1500 µm, or 250 µm-1200 µm.

[0104] In a particularly preferred embodiment, these particles have a (weight / volume average) diameter of 200 µm to 700 µm.

[0105] These particles consist of a core and one or more coatings (outer layers) surrounding the core. At least one coating is a matrix layer containing a bioactive substance (e.g., an enzyme) and a non-volatile liquid as described below. The core and the surrounding matrix are made of different compositions.

[0106] In one embodiment, these particles do not include surfactants, detergent builders, and / or bleach.

[0107] core

[0108] Suitable cores for use in this invention are any materials suitable for stratification in a fluid bed process. The core can be insoluble, dispersible, or soluble in water. The core material is preferably dispersible in water (disintegrates upon hydration) or dissolved in water by entering a real aqueous solution. Clays (e.g., layered silicate bentonite, kaolin, montmorillonite, hydropyrite, soapstone, bedesite, attapulgite, and magnesia), silicates (such as sand (sodium silicate), nonpareils), and aggregated potato starch or flour, or other starch granule sources (e.g., wheat and corn cobs) are considered dispersible. Cores can be produced by various methods known in the art, such as by granulation.

[0109] The core can be an organic particulate compound, such as a natural compound, or a condensed carbohydrate (e.g., sugar, starch, dextrin, flour (e.g., vegetable flour)). The material may have undergone steam treatment.

[0110] Napril is a spherical particle made from seed crystals, which are created by combining a layer of powder and a layer of solute with the seed crystal in a rotating spherical container and then rolling it into a spherical shape. Napril is typically made from a combination of sugar (such as sucrose) and powder (such as corn starch).

[0111] In one embodiment of this teaching, the core is sodium chloride or sodium sulfate crystals (or aggregated crystals), also known as seed crystals, or other inorganic salt crystals. In another embodiment of this teaching, the core is sucrose crystals. Particles composed of inorganic salts and / or sugars and / or small organic molecules can be used as the core of this teaching. Suitable water-soluble components incorporated into the core include: inorganic salts, such as sodium chloride, ammonium sulfate, sodium sulfate, magnesium sulfate, zinc sulfate; or urea, citric acid, sugars (e.g., sucrose, lactose), etc.

[0112] The core of this teaching may further include one or more of the following: activators, polymers, fillers, plasticizers, fibrous materials, extenders, and other compounds known to be used in the core.

[0113] Suitable polymers include polyvinyl alcohol (PVA), which includes partially and fully hydrolyzed PVA, polyethylene glycol, polyethylene oxide, polyvinylpyrrolidone, and carbohydrate polymers (e.g., starch, amylose, amylopectin, α-glucan and β-glucan, pectin, glycogen), including mixtures and derivatives thereof.

[0114] Suitable fillers that can be used in the core include inert materials, which are used to increase volume and reduce cost, or to adjust the desired enzyme activity in the final particles. Examples of such fillers include, but are not limited to, water-soluble agents such as salts, sugars, and water-dispersible agents such as clay, talc, silicates, cellulose and starch, and cellulose and starch derivatives.

[0115] Suitable plasticizers that can be used at the heart of this teaching are low molecular weight organic compounds and are highly specific to the polymer being plasticized. Examples include, but are not limited to, sugars (e.g., glucose, fructose, and sucrose), sugar alcohols (e.g., glycerol, low molecular weight polyethylene glycol, sorbitol, xylitol, and maltitol and other diols), polar low molecular weight organic compounds (e.g., urea), or other known plasticizers (e.g., water).

[0116] Suitable fibrous materials that can be used in the core of this teaching include, but are not limited to: cellulose and cellulose derivatives, such as HPMC (hydroxypropyl methylcellulose), CMC (carboxymethyl cellulose), and HEC (hydroxyethyl cellulose).

[0117] In one embodiment of this teaching, particularly for feed applications, the core is a water-soluble or dispersible corn cob material or sugar or salt crystals. In another embodiment, particularly suitable for household cleaning applications, the core is a water-soluble or dispersible sugar or salt crystal or napril.

[0118] Those skilled in the art will recognize that, for feed and food applications, the core (and any polymers, fillers, plasticizers, fibrous materials, and extenders) is acceptable for food and / or feed applications. For household cleaning applications, no such restrictions need to be imposed.

[0119] In a preferred embodiment, the core is substantially free of bioactive substances.

[0120] The core can have an average diameter of 20 µm-3000 µm, specifically 50 µm-2000 µm, 100 µm-1500 µm, or 250 µm-1200 µm.

[0121] matrix

[0122] The matrix layer surrounding the core is a substantially homogeneous mixture containing bioactive substances and non-volatile liquids. More precisely, the bioactive substances and non-volatile liquids are separated, partitioned, or arranged in discrete layers.

[0123] The amount of non-volatile liquid is at least 1% w / w of the matrix; preferably, at least 2% w / w of the matrix; more preferably, at least 4% w / w of the matrix; even more preferably, at least 5% w / w of the matrix; and most preferably, at least 7% w / w of the matrix.

[0124] The matrix must retain its overall non-liquid physical structure. For example, the amount of non-volatile liquid can be less than 50% w / w of the matrix; preferably, less than 40% w / w; more preferably, less than 30% w / w; most preferably, less than 25% w / w; and particularly less than 20% w / w.

[0125] The matrix may include one or more other granulating materials, such as binders (e.g., synthetic polymers, waxes, fats, or carbohydrates), fillers, fibrous materials (cellulose or synthetic fibers), stabilizers, solubilizers, suspending agents, viscosity modifiers, light spheres, plasticizers, salts, lubricants, and / or fragrances. The matrix may also include crystalline materials or mixtures of crystalline materials. Examples of crystalline materials are silicates, such as mica or clays like kaolin, montmorillonite, bentonite, and talc; and inorganic salts, such as alkali metal sulfates, carbonates, nitrates, and halides; alkaline earth metal sulfates, carbonates, nitrates, and halides; transition metal sulfates, carbonates, nitrates, and halides; and ammonium sulfates, carbonates, nitrates, and halides; for example, Na₂SO₄, K₂SO₄, CaSO₄, MgSO₄, ZnSO₄, (NH₄)₂ 2SO4, Na2CO3, NaHCO3, K2CO3, KHCO3, CaCO3, MgCO3, ZnCO3, (NH4)2CO3, NaNO3, KNO3, Ca(NO3)2, Mg(NO3)2, Zn(NO3)2, NH4NO3, NaCl, KCl, CaCl2, MgCl2, ZnCl2, and NH4Cl; or crystals, like citrates, such as sodium citrate or potassium citrate. Hydrates are also included.

[0126] The matrix, typically as a homogeneous blend, may contain salts of polyvalent cations, reducing agents, antioxidants, peroxide decomposition catalysts, and / or acidic buffer components.

[0127] Coating

[0128] The particle may optionally contain at least one additional coating, for example, to improve storage stability, reduce dust formation during processing, enhance matrix adhesion to the core, or color the particle. The one or more optional coatings may include salt coatings or other suitable coating materials, such as polyethylene glycol (PEG), methyl hydroxypropyl cellulose (MHPC), and polyvinyl alcohol (PVA). Examples of enzyme particles with multiple coatings are shown in WO 93 / 07263 and WO 97 / 23606. One or more coatings may also include functional components, such as bleaching catalysts (e.g., manganese bleaching catalyst; MnTACN) and / or bleaching activators (e.g., TAED, NOBS).

[0129] The coating may be applied in an amount of at least 0.1% (e.g., at least 0.5%, 1%, or 5%) of the core weight. This amount may be at most 100%, 70%, 50%, 40%, or 30%.

[0130] The coating is preferably at least 0.1 µm thick, specifically at least 0.5 µm, at least 1 µm, or at least 5 µm. In a specific embodiment, the coating thickness is less than 100 µm. In a more specific embodiment, the coating thickness is less than 60 µm. In even more specific embodiments, the total coating thickness is less than 40 µm.

[0131] The coating should seal the core (and matrix layer) by forming a substantially continuous layer. A substantially continuous layer should be understood as a coating with very few or no pores, such that the sealed / enclosed core unit has very few or no uncoated areas. The layer or coating should be particularly uniform in thickness.

[0132] The coating may further contain other materials known in the art, such as fillers, anti-sticking agents, pigments, dyes, plasticizers and / or adhesives, such as titanium dioxide, kaolin, calcium carbonate or talc.

[0133] Salt coating may include at least 60% salt by weight w / w, for example, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% by weight w / w.

[0134] The salt can be added from a salt solution (where the salt is completely dissolved) or from a salt suspension (where the fine particles are less than 50 µm, for example less than 10 µm or less than 5 µm).

[0135] The salt coating may contain a single salt or a mixture of two or more salts. The salt may be water-soluble, specifically having a solubility of at least 0.1 g in 100 g of water at 20°C, preferably at least 0.5 g / 100 g of water, for example, at least 1 g / 100 g of water, for example, at least 5 g / 100 g of water.

[0136] The salt can be an inorganic salt, such as a sulfate, sulfite, phosphate, phosphonate, nitrate, chloride, or carbonate, or a salt of a simple organic acid (less than 10 carbon atoms, such as 6 or fewer carbon atoms), such as a citrate, malonate, or acetate. Examples of cations in these salts are alkali or alkaline earth metal ions, ammonium ions, or first transition metal ions, such as sodium, potassium, magnesium, calcium, zinc, or aluminum. Examples of anions include chloride, bromine, iodine, sulfate, sulfite, bisulfite, thiosulfate, phosphate, dihydrogen phosphate, dibasic phosphate, hypophosphite, dihydrogen pyrophosphate, tetraborate, borate, carbonate, bicarbonate, silicate, citrate, malate, maleate, malonic acid, succinate, lactate, formate, acetate, butyrate, propionate, benzoate, tartrate, ascorbate, or gluconate. Specifically, base or alkaline earth metal salts of sulfate, sulfite, phosphate, phosphonate, nitrate, chloride, or carbonate, or salts of simple organic acids such as citrate, malonate, or acetate can be used.

[0137] The salt in the coating may have a constant humidity of more than 60% at 20°C, specifically more than 70%, more than 80%, or more than 85%, or it may be another hydrated form of the salt (e.g., anhydrous). The salt coating may be as described in WO00 / 01793 or WO 2006 / 034710.

[0138] A specific example of a suitable salt is NaCl (CH4). 20℃ =76%), Na2CO3 (CH 20℃ =92%), NaNO3 (CH 20℃ =73%), Na2HPO4 (CH 20℃ =95%), Na3PO4 (CH 25℃ =92%), NH4Cl (CH 20℃ =79.5%), (NH4)2HPO4 (CH 20℃ =93.0%), NH4H2PO4 (CH 20℃ =93.1%), (NH4)2SO4 (CH 20℃ =81.1%), KCl (CH 20℃ =85%), K2HPO4 (CH 20℃ =92%), KH2PO4 (CH 20℃ =96.5%), KNO3 (CH 20℃ =93.5%), Na2SO4 (CH 20℃ =93%), K2SO4 (CH 20℃ =98%), KHSO4 (CH 20℃ =86%), MgSO4 (CH20℃ =90%), ZnSO4 (CH 20℃ =90%) and sodium citrate (CH 25℃ =86%). Other examples include NaH2PO4, (NH4)H2PO4, CuSO4, Mg(NO3)2, and magnesium acetate.

[0139] The salt can be in anhydrous form, or it can be a hydrated salt, i.e., a crystalline salt hydrate with one or more bound water crystals, as described, for example, in WO 99 / 32595. Specific examples include anhydrous sodium sulfate (Na₂SO₄), anhydrous magnesium sulfate (MgSO₄), and magnesium sulfate heptahydrate (MgSO₄). . 7H2O), zinc sulfate heptahydrate (ZnSO4) . 7H2O), disodium hydrogen phosphate heptahydrate (Na2HPO4) . 7H2O), magnesium nitrate hexahydrate (Mg(NO3)2(6H2O)), sodium citrate dihydrate and magnesium acetate tetrahydrate.

[0140] Preferably, the salt is applied as a salt solution, for example using a fluidized bed.

[0141] Detergent composition

[0142] The particles of the present invention can be added to detergent compositions and thus become a component of the detergent composition. When used in detergent compositions, the bioactive component of the particles is preferably a (detergent) enzyme or bacterial spore.

[0143] For example, the detergent compositions of the present invention can be formulated as hand-wash or machine-wash detergent compositions, including laundry detergent additive compositions suitable for pre-treating stained fabrics, and rinse-additive fabric softener compositions, or formulated as detergent compositions for general household hard surface cleaning operations, or formulated for hand-wash or machine-wash dishwashing operations.

[0144] In a specific aspect, the present invention provides a detergent additive comprising particles as described herein.

[0145] In one embodiment, the present invention relates to detergent compositions comprising particles of the invention in combination with one or more additional cleaning composition components. The selection of additional components is within the capabilities of a person skilled in the art and includes conventional ingredients, including exemplary non-limiting components described below.

[0146] For textile care, the selection of components may include considerations such as the type of textile to be cleaned, the type and / or extent of soiling, the temperature at which cleaning is performed, and the formulation of the detergent product. Although the components mentioned below are classified under a general heading according to their specific functionality, this is not to be construed as limiting, as components may contain additional functionality as would be understood by a person skilled in the art.

[0147] In one embodiment of the invention, enzyme-containing particles may be added to the detergent composition in amounts corresponding to: 0.001 mg to 200 mg of enzyme protein per liter of wash liquor, for example, 0.005 mg to 100 mg of enzyme protein, preferably 0.01 mg to 50 mg of enzyme protein, more preferably 0.05 mg to 20 mg of enzyme protein, and even more preferably 0.1 mg to 10 mg of enzyme protein.

[0148] surfactants

[0149] The detergent composition may contain one or more surfactants, which may be anionic and / or cationic and / or nonionic and / or semi-polar and / or zwitterionic, or mixtures thereof. In a specific embodiment, the detergent composition contains a mixture of one or more nonionic surfactants and one or more anionic surfactants. The one or more surfactants are typically present at levels from about 0.1% to 60% by weight (e.g., about 1% to about 40%, or about 3% to about 20%, or about 3% to about 10%). The one or more surfactants are selected based on the desired cleaning application, and the surfactants include any one or more conventional surfactants known in the art. Any surfactant known in the art for use in detergents may be used.

[0150] When included therein, the detergent will typically contain from about 1% to about 40% by weight (e.g., from about 5% to about 30%, including from about 5% to about 15%, or from about 20% to about 25%) anionic surfactant. Non-limiting examples of anionic surfactants include sulfates and sulfonates, particularly linear alkylbenzene sulfonates (LAS), isomers of LAS, branched alkylbenzene sulfonates (BABS), phenyl alkyl sulfonates, α-olefin sulfonates (AOS), olefin sulfonates, chain olefin sulfonates, alkyl-2,3-dimethylbis(sulfate), hydroxyalkyl sulfonates, and disulfonates, alkyl sulfates (AS) (e.g., sodium dodecyl sulfate (SDS)), fatty alcohol sulfates (FAS), and primary alcohol sulfates (PAS). Alcohol ether sulfates (AES or AEOS or FES, also known as alcohol ethoxy sulfates or fatty alcohol ether sulfates), secondary alkyl sulfonates (SAS), paraffinic sulfonates (PS), ester sulfonates, sulfonated fatty acid glycerides, α-sulfonic acid fatty acid methyl esters (α-SFMe or SES) (including methyl sulfonate (MES)), alkyl succinic acids or alkenyl succinic acids, dodecenyl / tetradecenyl succinic acids (DTSA), fatty acid derivatives of amino acids, diesters and monoesters of sulfonic acid succinic acids or soaps, and combinations thereof.

[0151] When included therein, the detergent will typically contain from about 0.1% to about 10% by weight of a cationic surfactant. Non-limiting examples of cationic surfactants include alkyl dimethyl ethanol quaternary ammonium (ADMEAQ), hexadecyl trimethyl ammonium bromide (CTAB), dimethyl distearate ammonium chloride (DSDMAC), and alkyl benzyl dimethyl ammonium, alkyl quaternary ammonium compounds, alkoxylated quaternary ammonium (AQA) compounds, and combinations thereof.

[0152] When included therein, the detergent will typically contain from about 0.2% to about 40% by weight (e.g. from about 0.5% to about 30%, particularly from about 1% to about 20%, from about 3% to about 10%, such as from about 3% to about 5%, or from about 8% to about 12%) of a nonionic surfactant. Non-limiting examples of nonionic surfactants include alcohol ethoxylates (AE or AEO), alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters (e.g., ethoxylated and / or propoxylated fatty acid alkyl esters), alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkyl polysaccharides (APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine (glucosamide (GA) or fatty acid glucosamide (FAGA)), together with products available under the trade names SPAN and TWEEN, and combinations thereof.

[0153] When included therein, the detergent will typically contain from about 0.1% to about 20% by weight of a semi-polar surfactant. Non-limiting examples of semi-polar surfactants include amine oxides (AOs) (e.g., alkyl dimethyl amine oxides), N-(cocoylalkyl)-N,N-dimethyl amine oxides and N-(butter-alkyl)-N,N-bis(2-hydroxyethyl) amine oxides, fatty acid alkanolamides and ethoxylated fatty acid alkanolamides, and combinations thereof.

[0154] When included therein, the detergent will typically contain from about 0.1% to about 10% by weight of a facultative zwitterionic surfactant. Non-limiting examples of facultative zwitterionic surfactants include betaine, alkyl dimethyl betaine, sulfobetaine, and combinations thereof.

[0155] Water-soluble additives

[0156] Water-soluble additives are compounds that dissolve hydrophobic compounds in aqueous solutions (or conversely, polar substances in nonpolar environments). Typically, water-soluble additives exhibit both hydrophilic and hydrophobic characteristics (as known from surfactants); however, the molecular structure of water-soluble additives generally does not favor spontaneous self-aggregation, see, for example, the review by Hodgdon and Kaler (2007), Current Opinion in Colloid & Interface Science 12: 121-128. Water-soluble additives do not exhibit a critical concentration above which self-aggregation, as observed with surfactants, and the formation of micelles, thin layers, or other well-defined intermediate phases by lipids occur. Instead, many water-soluble additives exhibit a continuous type of aggregation process in which the size of the aggregates increases with increasing concentration. However, many water-soluble additives alter the phase behavior, stability, and colloidal properties of systems containing substances with both polar and nonpolar characteristics, including mixtures of water, oils, surfactants, and polymers. Water-soluble additives are routinely used in a variety of industries, from pharmaceuticals and personal care to food and technical applications. The use of water-soluble additives in detergent compositions allows for, for example, more concentrated surfactant formulations (such as in the process of compressing liquid detergents by removing water) without causing undesirable phenomena such as phase separation or high viscosity.

[0157] The detergent may contain 0%-5% by weight, for example, about 0.5% to about 5%, or about 3% to about 5%, of a water-soluble additive. Any water-soluble additive known in the art for use in detergents may be used. Non-limiting examples of water-soluble additives include sodium benzenesulfonate, sodium p-toluenesulfonate (STS), sodium xylenesulfonate (SXS), sodium cumenesulfonate (SCS), sodium cymene sulfonate, amine oxides, alcohols and polyethylene glycol ethers, sodium hydroxynaphthylformate, sodium hydroxynaphthylsulfonate, sodium ethylhexyl sulfate, and combinations thereof.

[0158] Builders and co-builders

[0159] The detergent composition may contain about 0% to 65% (e.g., about 5% to about 50%) by weight of a detergent builder or co-builder, or a mixture thereof. In dishwashing detergents, the level of the builder is typically 40% to 65%, particularly 50% to 65%. The builder and / or co-builder may specifically be chelating agents that form water-soluble complexes with calcium and magnesium ions. Any builder and / or co-builder known in the art for use in laundry detergents may be used. Non-limiting examples of detergent builders include citrates, zeolites, diphosphates (pyrophosphates), triphosphates such as sodium triphosphate (STP or STPP), carbonates such as sodium carbonate, soluble silicates such as sodium silicate, layered silicates (e.g., SKS-6 from Hoechst), ethanolamines such as 2-aminoethanol (MEA), diethanolamine (DEA, also known as iminodiethanol), triethanolamine (TEA, also known as 2,2',2”-nitrotriethanol), and carboxymethyl inulin (CMI) and combinations thereof.

[0160] The detergent composition may also contain 0%-50%, for example, about 5% to about 30% by weight, a detergent co-adjuvant or a mixture thereof. The detergent composition may include a co-adjuvant alone, or in combination with an builder, such as a zeolite builder. Non-limiting examples of co-adjuvants include homopolymers of polyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) or copolymers of (acrylic acid / maleic acid) (PAA / PMA). Other non-limiting examples include citrates, chelating agents (e.g., aminocarboxylates, aminopolycarboxylates, and phosphates), and alkylsuccinic acids or alkenylsuccinic acids. Other specific examples include 2,2',2”-N-aminotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid (IDS), ethylenediamine-N,N'-dibutanoic acid (EDDS), methylglycine diacetic acid (MGDA), glutamic acid-N,N-diacetic acid (GLDA), 1-hydroxyethane-1,1-diphosphonic acid (HEDP), ethylenediaminetetra-(methylenephosphonic acid) (EDTMPA), diethylenetriaminepenta(methylenephosphonic acid) (DTMPA or DTPMPA), N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic-N-monoacetic acid (ASMA), aspartic-N,N-diacetic acid (ASDA), aspartic-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA), N-(2-sulfomethyl)-aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid (SEAS), N- (2-Sulfomethyl)-glutamic acid (SMGL), N-(2-sulfoethyl)-glutamic acid (SEGL), N-methyliminodiacetic acid (MIDA), α-alanine-N,N-diacetic acid (α-ALDA), serine-N,N-diacetic acid (SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine-N,N-diacetic acid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilic acid-N,N-diacetic acid (SLDA), taurine-N,N-diacetic acid (TUDA), and sulfomethyl-N,N-diacetic acid (SMDA), N-(2-hydroxyethyl)-ethylenediamine-N,N',N'-triacetic acid (HEDTA), diethanolamine glycine (DEG), diethylenetriaminepenta(methylene phosphate) (DTPMP), aminotri(methylene phosphate) (ATMP), and combinations thereof and salts thereof. Further exemplary builders and / or co-builders are described in, for example, WO 2009 / 102854, US 5,977,053.

[0161] bleaching system

[0162] This detergent may contain 0%-50% by weight of a bleaching system. Any bleaching system known in the art for use in laundry detergents can be utilized. Suitable bleaching system components include bleaching catalysts, photobleaching agents, bleaching activators, hydrogen peroxide sources such as sodium percarbonate and sodium perborate, preformed peracids, and mixtures thereof. Suitable preformed peracids include, but are not limited to: peroxycarboxylic acids and their salts, percarbonates and their salts, perimidic acids and their salts, and peroxymonosulfate and its salts (e.g., potassium peroxymonosulfate). (R) and mixtures thereof. Non-limiting examples of bleaching systems include peroxide-based bleaching systems that combine with peracids to form bleaching activators, which may contain, for example, inorganic salts, including alkali metal salts such as sodium salts of perborates (usually monohydrates or tetrahydrates), percarbonates, persulfates, superphosphates, and persilicates. The term bleaching activator, as used herein, means a compound that reacts with a peroxide bleaching agent (like hydrogen peroxide) to form a peracid. The peracid formed in this manner constitutes the activated bleaching agent. The term used herein... Suitable bleaching activators include those belonging to the ester amide, imide, or acid anhydride classes. Suitable examples are tetraacetylethylenediamine (TAED), sodium 4-[(3,5,5-trimethylhexanoyl)oxy]benzenesulfonate (ISONOBS), dipperoxylauric acid, 4-(dodecyloxy)benzenesulfonate (LOBS), 4-(decyloxy)benzenesulfonate, 4-(decyloxy)benzoate (DOBS), 4-(nonanoyloxy)benzenesulfonate (NOBS), and / or those disclosed in WO Those in 98 / 17767. Specific families of bleaching activators of interest are disclosed in EP 624154, and particularly preferred from this family is triethyl acetylacetic acid (ATC). ATC, or short-chain triglycerides (like triacetin), has the advantage of being environmentally friendly because it eventually degrades into citric acid and alcohol. Furthermore, triethyl acetylacetic acid and triacetin exhibit good hydrolytic stability in the product during storage, and are effective bleaching activators. Finally, ATC provides good washing ability for laundry detergent additives. Alternatively, the bleaching system may contain peroxy acids of the type of amides, imides, or sulfones, for example. The bleaching system may also contain peracids, such as 6-(phthalimide)percapanoic acid (PAP). The bleaching system may also include a bleaching catalyst. In some embodiments, the bleaching component may be an organic catalyst selected from the group consisting of organic catalysts having the following formula:

[0163]

[0164] and its mixtures; wherein each R 1 Independently, it is a branched alkyl group containing 9 to 24 carbons or a straight-chain alkyl group containing 11 to 24 carbons, preferably, each R 1Independently, it is a branched alkyl group containing 9 to 18 carbons or a straight-chain alkyl group containing 11 to 18 carbons; more preferably, each R 1 Independently selected from the group consisting of: 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, n-dodecyl, n-tetradecyl, n-hexadecyl, n-octadecyl, isonyl, isodecyl, iso-tridecyl, and iso-pentadecanyl. Other exemplary bleaching systems are described, for example, in WO 2007 / 087258, WO 2007 / 087244, WO 2007 / 087259, and WO 2007 / 087242. Suitable photobleaching agents may be, for example, sulfonated zinc phthalocyanine.

[0165] polymer

[0166] The detergent may contain 0%-10% (e.g., 0.5%-5%, 2%-5%, 0.5%-2%, or 0.2%-1%) of a polymer by weight. Any polymer known in the art for use in detergents may be used. The polymer may function as a co-adjuvant as mentioned above, or may provide anti-redeposition, fiber protection, dirt release, dye transfer inhibition, oil stain removal, and / or antifoaming properties. Some polymers may have more than one of the properties mentioned above and / or more than one of the motifs mentioned below. Exemplary polymers include (carboxymethyl) cellulose (CMC), poly(vinyl alcohol) (PVA), poly(vinylpyrrolidone) (PVP), poly(ethylene glycol) or poly(ethylene oxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin (CMI), and polycarboxylates such as PAA, PAA / PMA, poly-aspartic acid, and lauryl methacrylate / acrylic acid copolymers, hydrophobically modified CMC (HM-CMC) and silicone, copolymers of terephthalic acid and oligomeric polyethylene glycol, copolymers of poly(ethylene terephthalate) and poly(ethylene oxyterephthalate) (PET-POET), PVP, poly(vinylimidazolium) (PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO), and polyvinylpyrrolidone-vinylimidazolium (PVPVI). Other exemplary polymers include sulfonated polycarboxylate esters, polyethylene oxide and polypropylene oxide (PEO-PPO), and diquaternary ammonium ethoxysulfate. Other exemplary polymers are disclosed, for example, in WO 2006 / 130575 and US 5,955,415. Salts of the polymers mentioned above are also considered.

[0167] Fabric colorant

[0168] The detergent compositions of the present invention may further include fabric colorants, such as dyes or pigments, which, when formulated in the detergent composition, can deposit on the fabric upon contact with a washing liquid containing the detergent composition, and thus alter the color of the fabric through absorption / reflection of visible light. Fluorescent whitening agents emit at least some visible light. In contrast, fabric colorants alter the color of a surface when they absorb at least a portion of the visible spectrum. Suitable fabric colorants include dyes and dye-clay conjugates, and may also include pigments. Suitable dyes include small molecule dyes and polymer dyes. Suitable small molecule dyes include those selected from the group consisting of dyes falling into the Colour Index (CI) classification: Direct Blue, Direct Red, Direct Violet, Acid Blue, Acid Red, Acid Violet, Basic Blue, Basic Violet, and Basic Red, or mixtures thereof, such as those described in WO 2005 / 03274, WO 2005 / 03275, WO 2005 / 03276, and EP 1876226 (incorporated herein by reference). The detergent composition preferably comprises from about 0.00003 wt% to about 0.2 wt%, from about 0.00008 wt% to about 0.05 wt%, or even from about 0.0001 wt% to about 0.04 wt% of a fabric toner. The composition may contain from 0.0001 wt% to 0.2 wt% of a fabric toner, which may be particularly preferred when the composition is in the form of a unit-dose sachet. Suitable toners are also disclosed, for example, in WO 2007 / 087257 and WO 2007 / 087243.

[0169] One or more detergent enzymes

[0170] Detergent additives, together with detergent compositions, may include one or more (additional) enzymes, such as those mentioned above under the heading "Enzymes".

[0171] Generally, the properties of one or more selected enzymes should be compatible with the selected detergent (i.e., optimal pH, compatibility with other enzymes and non-enzyme components, etc.), and the one or more enzymes should be present in an effective amount.

[0172] One or more detergent enzymes can be incorporated into a detergent composition by adding a single additive containing one or more enzymes, or by adding a combination of additives containing all of these enzymes. The detergent additives of the present invention, i.e., single or combined additives, can be formulated into, for example, granules, liquids, slurries, etc. Preferred detergent additive formulations are granules, especially non-dusted granules; liquids, especially stabilized liquids; or slurries.

[0173] One or more detergent enzymes can be incorporated into a detergent composition by adding a single additive containing one or more enzymes, or by adding a combination of additives containing all of these enzymes. The detergent additives of the present invention, i.e., separate additives or combined additives, are formulated as granules of the present invention.

[0174] auxiliary materials

[0175] Any detergent component known in the art for use in laundry detergents may also be used. Other optional detergent components include corrosion inhibitors, shrinkage inhibitors, anti-fouling redeposition agents, anti-wrinkle agents, bactericides, adhesives, corrosion inhibitors, disintegrants / disintegration agents, dyes, enzyme stabilizers (including boric acid, borates, CMC and / or polyols, such as propylene glycol), fabric conditioners (including clays), fillers / processing aids, optical brighteners / brighteners, foaming agents, foam (foam) regulators, fragrances, soil suspending agents, softeners, defoaming agents, dulling inhibitors, and wicking agents, used alone or in combination. Any ingredient known in the art for use in laundry detergents may be used. The selection of such ingredients is entirely within the skill of a person skilled in the art.

[0176] dispersant - The detergent compositions of the present invention may also contain dispersants. Specifically, powder detergents may contain dispersants. Suitable water-soluble organic materials include homopolymerized or copolymerized acids or salts thereof, wherein the polycarboxylic acid contains at least two carboxyl groups separated from each other by no more than two carbon atoms. Suitable dispersants are described, for example, in Powdered Detergents, Surfactant Science Series, Volume 71, Marcel Dekker, Inc.

[0177] Dye transfer inhibitors - The detergent compositions of the present invention may further include one or more dye transfer inhibitors. Suitable polymeric dye transfer inhibitors include, but are not limited to, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidinone and polyvinylimidazole, or mixtures thereof. When present in the subject composition, the dye transfer inhibitor may be present at a level from about 0.0001% to about 10%, from about 0.01% to about 5%, or even from about 0.1% to about 3% by weight of the composition.

[0178] Fluorescent whitening agent- The detergent compositions of the present invention will also preferably contain additional components that can color the cleaned item, such as optical brighteners or fluorescent brighteners. Optical brighteners, also known as fluorescent brighteners, are dyes that absorb light in the ultraviolet and violet regions of the electromagnetic spectrum (typically 340 nm-370 nm) and re-emit light in the blue region (typically 420 nm-470 nm). These agents are commonly used to enhance the appearance of fabric and paper colors, causing a "whitening" effect by increasing the total amount of reflected blue light, making the material appear less yellow.

[0179] Fluorescent brighteners are well known in the art, and many such brighteners are commercially available. Typically, brighteners are provided and used in the form of their alkali metal salts, such as sodium salts.

[0180] Preferred fluorescent agents are selected from the following categories: stilbene biphenyl, triazine aminostilbene, bis(1,2,3-triazol-2-yl)stilbene, bis(benzo[b]furan-2-yl)biphenyl, 1,3-diphenyl-2-pyrazoline, thiophene dibenzoxazole, and coumarin. The fluorescent agent is preferably sulfonated.

[0181] Preferred categories of fluorescent agents are stilbene biphenyl compounds, such as Tinopal™ CBS-X; diamine stilbene disulfonic acid compounds, such as Tinopal DMS-X and Blankophor™ HRH; pyrazoline compounds, such as Blankophor SN; and thiophene dibenzoxazole compounds, such as Tinopal OB.

[0182] Fluorescent agents are also described in McElhone, HJ (2009), “Fluorescent Whitening Agents”, Kirk-Othmer Encyclopedia of Chemical Technology, 1-16, DOI: 10.1002 / 0471238961.0612211513030512.a01.pub2.

[0183] Suitable levels of fluorescent brighteners include those from about 0.01 wt%, from 0.05 wt%, from about 0.1 wt% or even from a lower level of about 0.2 wt% to a higher level of 0.5 wt% or even 0.75 wt%; for example, from 0.01 wt% to 0.5 wt%.

[0184] Fouling release polymers- The detergent compositions of the present invention may further comprise one or more soil-releasing polymers that help remove soiling from fabrics, such as cotton or polyester-based fabrics, particularly hydrophobic soiling from polyester-based fabrics. Soil-releasing polymers may be, for example, polymers based on nonionic or anionic terephthalic acid, polyvinylcaprolactam and related copolymers, vinyl graft copolymers, polyester polyamides, see, for example, Powdered Detergents, Surfactant Science Series, Volume 71, Chapter 7, Marcel Dekker, Inc. Another type of soil-releasing polymer is an amphiphilic alkoxylated oil stain cleaning polymer comprising a core structure and a plurality of alkoxylated groups attached to said core structure. The core structure may comprise a polyalkylimide structure or a polyalkanolamine structure, as described in detail in WO 2009 / 087523 (incorporated herein by reference). Furthermore, random graft copolymers are suitable soil-releasing polymers. Suitable graft copolymers are described in more detail in WO 2007 / 138054, WO 2006 / 108856, and WO 2006 / 113314 (both incorporated herein by reference). Other fouling-releasing polymers are substituted polysaccharide structures, particularly substituted cellulose structures, such as modified cellulose derivatives, as described in EP 1867808 or WO 2003 / 040279 (both incorporated herein by reference). Suitable cellulose polymers include cellulose, cellulose ethers, cellulose esters, cellulose amides, and mixtures thereof. Suitable cellulose polymers include anionic modified cellulose, nonionic modified cellulose, cationic modified cellulose, zwitterionic modified cellulose, and mixtures thereof. Suitable cellulose polymers include methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose, ester carboxymethylcellulose, and mixtures thereof.

[0185] Anti-redeposition agents – The detergent compositions of the present invention may further include one or more anti-redeposition agents, such as carboxymethyl cellulose (CMC), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyoxyethylene and / or polyethylene glycol (PEG), homopolymers of acrylic acid, copolymers of acrylic acid and maleic acid, and ethoxylated polyethyleneimine. The cellulose-based polymers described above under the category of dirt-releasing polymers may also function as anti-redeposition agents.

[0186] Other suitable excipients Including but not limited to shrink-proof agents, wrinkle-resistant agents, bactericides, adhesives, carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foam regulators, fragrances, pigments, defoamers, solvents, and structural agents and / or structural elastic agents used in liquid detergents.

[0187] Laundry soap bars

[0188] The granules of this invention can be added to laundry soap bars and used for hand washing of fabrics and / or textiles. The term laundry soap bar includes laundry bars, soap bars, combo bars, synthetic detergent bars, and detergent bars. The types of bars are generally distinguished by the type of surfactant they contain, and the term laundry soap bar includes those containing soaps derived from fatty acids and / or synthetic soaps. Laundry soap bars have a physical form that is solid at room temperature, rather than liquid, gel, or powder. The term solid is defined as a physical form that does not change significantly over time, i.e., if a solid object (e.g., a laundry soap bar) is placed inside a container, the solid object does not change to fill the container in which it is placed. The bar is typically in the form of a bar when it is solid, but it can also be other solid shapes, such as round or oval.

[0189] The laundry soap bar may contain one or more additional enzymes, protease inhibitors such as peptide aldehydes (or sulfide adducts or hemiacetal adducts), boric acid, borates, borax and / or phenylboronic acid derivatives such as 4-carboxyphenylboronic acid, one or more soaps or synthetic surfactants, polyols such as glycerol, pH-controlling compounds such as fatty acids, citric acid, acetic acid and / or formic acid, and / or salts of monovalent cations and organic anions, wherein the monovalent cation may be, for example, Na+. + K + or NH4 + Furthermore, the organic anion can be, for example, a formate, acetate, citrate, or lactate, such that the salt of the monovalent cation and the organic anion can be, for example, sodium formate.

[0190] Laundry soap bars may also contain complexing agents such as EDTA and HEDP, fragrances and / or different types of fillers, surfactants such as anionic synthetic surfactants, builders, polymerized dirt releasers, detergent chelators, stabilizers, fillers, dyes, colorants, dye transfer inhibitors, alkoxylated polycarbonates, defoamers, structural agents, binders, leachants, bleach activators, clay detergents, anti-redeposition agents, polymeric dispersants, brighteners, fabric softeners, fragrances and / or other compounds known in the art.

[0191] Laundry soap bars can be processed in conventional laundry soap bar manufacturing equipment, such as, but not limited to, mixers, pressing machines (e.g., two-stage vacuum pressing machines), extruders, cutters, marking molds, cooling tunnels, and packaging machines. The invention is not limited to preparing laundry soap bars by any single method. The premix of the invention can be added to the soap at different stages of the process. For example, a premix containing soap, the granules of the invention, optionally one or more additional enzymes, protease inhibitors, and salts of monovalent cations and organic anions can be prepared and then the mixture can be pressed into bars. Enzymes and optionally additional enzymes can be added simultaneously as protease inhibitors, for example, in liquid form. In addition to the mixing and pressing steps, the process may further include grinding, extrusion, cutting, molding, cooling, and / or packaging steps.

[0192] The present invention also relates to the following embodiments:

[0193] 1. A layered particle comprising a core and a matrix layer surrounding the core, wherein the matrix comprises a bioactive substance and 1%-50% non-volatile liquid.

[0194] 2. The particles as described in embodiment 1, wherein the bioactive substance is an enzyme or a microorganism.

[0195] 3. The particles as described in embodiment 1 or 2, wherein the bioactive substance is an enzyme selected from the group consisting of: proteases, lipases, keratinases, amylases, glycoses, cellulases, pectinases, mannanases, arabinoseases, galactanases, xylanases, DNases, hydrolases, oxidases, laccases, peroxygenases, halogen peroxidases, and peroxidases.

[0196] 4. The particles as described in any one of embodiments 1-3, wherein the bioactive substance is a bacterial spore, such as Bacillus endospore.

[0197] 5. The particles as described in any one of embodiments 1-4, wherein the bioactive substance is dehydrated yeast cells or dehydrated bacterial cells.

[0198] 6. The particles as described in any one of embodiments 1-5, wherein the vapor pressure of the non-volatile liquid is less than 1 kPa at 25°C.

[0199] 7. The particles as described in any one of embodiments 1-6, wherein the non-volatile liquid is a polyol.

[0200] 8. The particles as described in any one of embodiments 1-7, wherein the non-volatile liquid is a polyol selected from the group consisting of glycerol, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, tripropylene glycol, and polyethylene glycol.

[0201] 9. The particles as described in any one of embodiments 1-8, wherein the matrix comprises 5% w / w-30% w / w of the non-volatile liquid.

[0202] 10. The particles as described in any one of embodiments 1-9, wherein the core comprises at least 10% w / w of crystalline material relative to the non-liquid portion of the composition, the crystalline material being one or more inorganic salts or clays, such as one or more salts of sulfates, carbonates, nitrates, chlorides; and / or kaolin, montmorillonite, bentonite, talc.

[0203] 11. The particles as described in any one of embodiments 1-10, wherein the matrix layer surrounding the core is applied in a fluidized bed sprayer.

[0204] 12. The particles as described in any one of embodiments 1-11, wherein the core is substantially free of the bioactive substance.

[0205] 13. Particles as described in any one of embodiments 1-12, wherein the core is nonpareils or is composed of inorganic salts.

[0206] 14. The particles as described in any one of embodiments 1-13, wherein the particles comprise an additional coating, wherein the coating constitutes 5% w / w to 70% w / w relative to the particles and comprises at least 60% w / w of salt having a constant humidity of at least 60% at 20°C.

[0207] 15. A particulate detergent composition comprising a detergent builder, a surfactant, and particles as described in any one of embodiments 1-14.

[0208] Further embodiments of the present invention include:

[0209] Example 1. A layered particle comprising a core surrounded by a matrix layer, wherein the matrix comprises a bioactive substance and 1%-50% non-volatile liquid.

[0210] Example 2. Particles as described in Example 1, wherein the bioactive substance is an enzyme or a microorganism.

[0211] Example 3. Particles as described in Example 1 or 2, wherein the bioactive substance is an enzyme.

[0212] Example 4. Particles as described in any one of Examples 1-3, wherein the bioactive substance is an enzyme selected from the group consisting of: proteases, lipases, keratinases, amylases, glycoses, cellulases, pectinases, mannanases, arabinoseases, galactanases, xylanases, DNases, hydrolases, oxidases, laccases, peroxygenases, halogen peroxidases, and peroxidases.

[0213] Example 5. Particles as described in any one of Examples 1-4, wherein the bioactive substance is a detergent enzyme selected from the group consisting of: protease, lipase, amylase, cellulase, pectinase, mannanase, xylanase, DNase, hydrolase, and oxidase.

[0214] Example 6. Particles as described in any one of Examples 1-5, wherein the bioactive substance is a bacterial spore, such as Bacillus spores.

[0215] Example 7. Particles as described in any one of Examples 1-6, wherein the bioactive substance is dehydrated yeast cells or dehydrated bacterial cells.

[0216] Example 8. Particles as described in any one of Examples 1-7, wherein the vapor pressure of the non-volatile liquid is less than 1 kPa at 25°C.

[0217] Example 9. Particles as described in any one of Examples 1-8, wherein the vapor pressure of the non-volatile liquid is less than 0.5 kPa at 25°C.

[0218] Example 10. Particles as described in any one of Examples 1-9, wherein the vapor pressure of the non-volatile liquid is less than 0.1 kPa at 25°C.

[0219] Example 11. Particles as described in any one of Examples 1-10, wherein the non-volatile liquid is water-soluble at 25°C.

[0220] Example 12. Particles as described in any one of Examples 1-11, wherein the surface tension of the non-volatile liquid is at least 30 mN / m at 20°C.

[0221] Example 13. Particles as described in any one of Examples 1-12, wherein the surface tension of the non-volatile liquid is at least 40 mN / m at 20°C.

[0222] Example 14. Particles as described in any one of Examples 1-13, wherein the surface tension of the non-volatile liquid is at least 50 mN / m at 20°C.

[0223] Example 15. Particles as described in any one of Examples 1-14, wherein the non-volatile liquid has a dynamic viscosity of at least 0.001 Pa at 25°C.

[0224] Example 16. Particles as described in any one of Examples 1-15, wherein the non-volatile liquid has a dynamic viscosity of at least 0.01 Pa at 25°C.

[0225] Example 17. Particles as described in any one of Examples 1-16, wherein the non-volatile liquid has a dynamic viscosity of at least 0.1 Pa at 25°C.

[0226] Example 18. Particles as described in any one of Examples 1-17, wherein the non-volatile liquid is a polyol.

[0227] Example 19. Particles as described in any one of Examples 1-18, wherein the non-volatile liquid is a polyol selected from the group consisting of: glycerol, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, tripropylene glycol, and polyethylene glycol.

[0228] Example 20. Particles as described in any one of Examples 1-19, wherein the non-volatile liquid is glycerol.

[0229] Example 21. Particles as described in any one of Examples 1-20, wherein the matrix comprises 2% w / w-40% w / w of the non-volatile liquid.

[0230] Example 22. Particles as described in any one of Examples 1-21, wherein the matrix comprises 5% w / w-30% w / w of the non-volatile liquid.

[0231] Example 23. Particles as described in any one of Examples 1-22, wherein the matrix comprises 6% w / w-25% w / w of the non-volatile liquid.

[0232] Example 24. Particles as described in any one of Examples 1-23, wherein the matrix comprises 7% w / w-20% w / w of the non-volatile liquid.

[0233] Example 25. Particles as described in any one of Examples 1-24, wherein the matrix comprises a crystalline material, which is one or more inorganic salts, silicates or clays.

[0234] Example 26. Particles as described in any one of Examples 1-25, wherein the matrix comprises a crystalline material, which is one or more salts of sulfate, carbonate, nitrate, or chloride; and / or one or more silicates; and / or kaolin, montmorillonite, bentonite, or talc.

[0235] Example 27. Particles as described in any one of Examples 1-26, wherein the matrix comprises a crystalline material, which is one or more alkali metal sulfates, carbonates, nitrates, chlorides; alkaline earth metal sulfates, carbonates, nitrates, chlorides; transition metal sulfates, carbonates, nitrates, chlorides; or ammonium sulfates, carbonates, nitrates, chlorides.

[0236] Example 28. Particles as described in any one of Examples 1-27, wherein the matrix comprises at least 10% w / w of crystalline material relative to the non-liquid portion of the matrix.

[0237] Example 29. Particles as described in any one of Examples 1-28, wherein the matrix comprises at least 20% w / w of crystalline material relative to the non-liquid portion of the matrix.

[0238] Example 30. Particles as described in any one of Examples 1-29, wherein the matrix comprises at least 30% w / w of crystalline material relative to the non-liquid portion of the matrix.

[0239] Example 31. Particles as described in any one of Examples 1-30, wherein the matrix comprises at least 40% w / w of crystalline material relative to the non-liquid portion of the matrix.

[0240] Example 32. Particles as described in any one of Examples 1-31, wherein the matrix comprises at least 50% w / w of crystalline material relative to the non-liquid portion of the matrix.

[0241] Example 33. Particles as described in any one of Examples 1-32, comprising applying an additional salt coating to these particles.

[0242] Example 34. Particles as described in any one of Examples 1-33, comprising applying an additional coating to the particles, wherein the coating constitutes 5% w / w-70% w / w relative to the particles and comprises at least 60% w / w of a salt having a constant humidity of at least 60% at 20°C.

[0243] Example 35. Particles as described in any one of Examples 1-34, comprising applying an additional coating to the particles, wherein the coating constitutes 5% w / w to 70% w / w relative to the particles and comprises at least 75% w / w of a salt having a constant humidity of at least 80% at 20°C.

[0244] Example 36. Particles as described in any one of Examples 1-35, comprising applying an additional coating to the particles, wherein the coating constitutes 10% w / w-30% w / w relative to the particles and comprises at least 75% w / w of a salt having a constant humidity of at least 80% at 20°C.

[0245] Example 37. Particles as described in any one of Examples 1-36, comprising applying an additional coating to the particles, wherein the coating comprises sodium sulfate.

[0246] Example 38. Particles as described in any one of Examples 1-37, wherein the diameter is 200 µm-3000 µm.

[0247] Example 39. Particles as described in any one of Examples 1-38, wherein the diameter is 200 µm-2000 µm.

[0248] Example 40. Particles as described in any one of Examples 1-39, wherein the diameter is 200 µm-1000 µm.

[0249] Example 41. Particles as described in any one of Examples 1-40, wherein the diameter is 200 µm-700 µm.

[0250] Example 42. Particles as described in any one of Examples 1-41, wherein the core is substantially free of the bioactive substance.

[0251] Example 43. Particles as described in any one of Examples 1-42, wherein the core is Napriel.

[0252] Example 44. Particles as described in any one of Examples 1-43, wherein the core is composed of one or more inorganic salts, such as sodium chloride or sodium sulfate.

[0253] Example 45. Particles as described in any one of Examples 1-44, wherein the matrix layer surrounding the core is applied in a fluidized bed sprayer.

[0254] Example 46. A detergent composition comprising a detergent builder, a surfactant, and particles as described in any one of Examples 1-45.

[0255] Example 47. A detergent composition as described in Example 46, wherein the detergent composition is a particulate composition.

[0256] Example 48. Use of the granules as described in any one of Examples 1-45 as a component in a method for manufacturing a detergent composition.

[0257] Example 49. Use as described in Example 48, wherein the detergent composition is a particulate composition.

[0258] The invention is further described through the following examples, which should not be construed as limiting the scope of the invention.

[0259] Example

[0260] Chemicals are at least reagent-grade commodities.

[0261] Test methods

[0262] To assess whether the release of reactive dust increases after particles are subjected to mechanical forces that would cause reference particles to break or break, a grinding method was applied. The test method uses a grinding apparatus as a pre-analysis step before measuring reactive dust release, thus providing a more robust and realistic description of the particles' robustness against mechanical stress. Reactive dust release from coated particles before and after the application of mechanical force was analyzed using a grinding apparatus via well-known washing methods (as described in reactive dust analysis). In both Examples 1 and 2, the coating process is identical, but this is not intended to limit the scope of the invention.

[0263] This grinding device is a MillMaster Grain Mill manufactured by Mashmaster Private Limited (Francis Smith, PO Box 1768, Coorparoo DC, Qld 4151, Australia). The specific specifications of this instrument are as follows:

[0264] - 130 mm precision-machined roller;

[0265] - Stainless steel rollers with a diameter of 38 mm; and

[0266] - Infinitely adjustable roll gap setting from 0.1 mm to 1.9 mm for precision control and accuracy.

[0267] The milling apparatus (MillMaster Grain Mill) has two graduated dials that act as eccentric adjusters for the desired gap. These eccentricity adjusters have been modified to achieve gaps as low as 0 mm (from the initially available 0.1 mm to 1.9 mm). The gap is adjusted before grinding tests by measuring it and ensuring it is significantly below D10, which is the 10th percentile of the particle size distribution (meaning 10% of the particle volume has a size equal to or less than a given value). In reported instances, the gap was adjusted to 150 micrometers to ensure the mentioned safety margin requirement, as the product to be analyzed was sieved between 425 and 600 micrometers. In this way, most particles shrink as they pass through the mill, subjecting them to high mechanical stress, leading to particle deformation and / or breakage.

[0268] The grinding equipment was used at a roller speed of 30 rpm to 40 rpm and the sample was fed at a rate of 4 g / min to 6 g / min.

[0269] Other types of similar grinding devices can be used to grind the particles of the present invention. Importantly, the reference particles (similar to the particle composition but without non-volatile liquids) are ground in the same manner as the particles of the present invention in order to compare dust levels.

[0270] Sample preparation

[0271] The test methods and active dust analysis were applied to a mixture of 2.5% w / w active particles and 97.5% unbleached detergent powder to simulate the interaction of active particles with other particles of different properties, as this would be the case in the application of the product.

[0272] The mixture was fed into the grinding apparatus in a 40 g sample size. Based on active dust analysis, 30 g of the resulting ground product was analyzed for active dust to determine the amount of "reacting dust after grinding." Similarly, 30 g of the undisturbed mixture (unground particles containing active ingredients) was analyzed for active dust to determine the amount of "active dust before grinding."

[0273] Active dust analysis

[0274] The release of active dust was analyzed using a well-known washout dust meter: the activity of bioactive substances on the dust filter was analyzed, and the results were converted into the nanograms of bioactive substances divided by the grams of the sample. In this way, the results are independent of inactive dust that may be generated by detergent powder.

[0275] The enzyme particles are fluidized using air within a glass column. Released dust is collected on a glass fiber filter.

[0276] The amount of bioactive dust on the filter was determined by analytical methods for the dust filter containing the bioactive substances under discussion.

[0277] Analysis conditions:

[0278] Temperature: Room temperature

[0279] Sample quantity: 30.0 + / - 0.05 g

[0280] Airflow velocity: 1.06 m 3 / h (~ 0.3 m / s)

[0281] Analysis time: 40 min.

[0282] Air humidity: 0%RH-1%RH

[0283] Fiberglass filter: 15 cm Fisherbrand™, Class 261

[0284] Example 1

[0285] Reference particles were generated in a fluidized bed, as shown in Table 1. These particles do not conform to the composition described in the claims. The content of the protease (Savinase™) and the release of active enzyme dust before and after applying the "Test Method" are shown in Table 2. The results show that the lack of non-volatile liquids led to a high release of active enzyme dust.

[0286] Example 2

[0287] Particles containing 15% glycerol were generated in the enzyme layer in a fluidized bed, as shown in Table 1. The content of the protease (Savinase™) and the release of active dust before and after applying the "Test Method" are shown in Table 2. As indicated, this amount of non-volatile liquid reduced the release of active enzyme dust.

[0288] Table 1. The formulations used in Examples 1 and 2 were granulated by fluidized bed granulation. Subsequently, the particles from both Examples 1 and 2 were coated with Na2SO4, PEG4000, and kaolin, with the same coating formulation.

[0289]

[0290] *The glycerol content of the enzyme layer is approximately 15%.

[0291] Table 2. Release of active enzyme dust before / after application of the test method to the particles.

[0292]

[0293] The "fraction of active dust after grinding" is calculated as the ratio between the "active dust after grinding" and the "protease content".

Claims

1. A layered particle comprising a core and a matrix layer surrounding the core, wherein the matrix comprises a bioactive substance and 1%-50% non-volatile liquid.

2. The particles of claim 1, wherein the bioactive substance is an enzyme or a microorganism.

3. The particles of claim 1 or 2, wherein the bioactive substance is an enzyme selected from the group consisting of: Protease, lipase, keratinase, amylase, glycosylase, cellulase, pectinase, mannanase, arabinosease, galactanase, xylanase, DNase, hydrolase, oxidase, laccase, peroxygenase, halogen peroxidase, and peroxidase.

4. The particles according to any one of claims 1-3, wherein the bioactive substance is a bacterial spore, such as Bacillus endospore.

5. The particles according to any one of claims 1-4, wherein the bioactive substance is dehydrated yeast cells or dehydrated bacterial cells.

6. The particles according to any one of claims 1-5, wherein the vapor pressure of the non-volatile liquid is less than 1 kPa at 25°C.

7. The particles according to any one of claims 1-6, wherein the non-volatile liquid is a polyol.

8. The particles according to any one of claims 1-7, wherein the non-volatile liquid is a polyol selected from the group consisting of: Glycerin, ethylene glycol, diethylene glycol, triethylene glycol, propylene glycol, dipropylene glycol, tripropylene glycol, and polyethylene glycol.

9. The particles according to any one of claims 1-8, wherein the matrix comprises 5% w / w-30% w / w of the non-volatile liquid; preferably, wherein the core comprises at least 10% w / w of a crystalline material relative to the non-liquid portion of the composition, the crystalline material being one or more inorganic salts or clays, such as one or more salts of sulfates, carbonates, nitrates, chlorides; and / or kaolin, montmorillonite, bentonite, talc; preferably, wherein the matrix layer surrounding the core is applied in a fluidized bed sprayer; preferably, wherein the core is substantially free of the bioactive substance; preferably, wherein the core is nonpareils or composed of inorganic salts; preferably, the particles comprise an additional coating, wherein the coating constitutes 5% w / w-70% w / w relative to the particles and comprises at least 60% w / w of a salt having a constant humidity of at least 60% at 20°C.

10. A particulate detergent composition comprising a detergent builder, a surfactant, and particles as described in any one of claims 1-9.

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