Modified biomass extract

Abstract

The present invention relates to a method for modifying a biomass extract and to a biomass extract composition obtained by this method, wherein immobilized lipases are used during the alcohol extraction process to increase the product yield and to provide fatty acid esters (mainly ethyl esters when ethanol is used as extractant). The present invention also relates to a perfuming or flavoring composition comprising the modified biomass extract, and to a perfumed consumer product comprising the modified biomass extract or the perfuming or flavoring composition.

Description

Technical Field

[0001] This invention relates to a method for modifying a biomass extract and a biomass extract composition obtained by the method. The invention also relates to a flavoring or seasoning composition comprising the modified biomass extract, and a flavored consumer product comprising the modified biomass extract or the flavoring or seasoning composition. Background Technology

[0002] To prepare a concentrated, highly aromatic fragrance blend known as "absolute," several process steps are required. First, the plant material is typically extracted using a hydrocarbon solvent (such as hexane). After the solvent evaporates, an intermediate crude product, "concrete," is obtained. The concrete is then further extracted, usually with ethanol. The ethanol extract is cooled to solidify the wax, which is then removed by methods such as filtration. After evaporating the ethanol from the filtrate, a fragrance oil known as "absolute" is obtained, which can be used to prepare flavored consumer products.

[0003] The drawback of this method is that the fats, oils, and waxes present in the extract (plant extract) can hinder the extraction of fragrance compounds because these compounds are readily soluble in these non-polar components. Therefore, fats, oils, and waxes act as a "trap," leading to a reduced absolute yield. Even with multiple extractions using ethanol, a satisfactory yield of fragrance compounds from the absolute may not be achieved. This can result in the discarded fats, oils, and waxes still containing valuable fragrance compounds that could not be fully extracted.

[0004] Therefore, increasing the crude distillate yield is a viable way to increase current crude distillate production. At the same time, this also helps reduce waste and byproducts associated with crude distillate production, thus achieving a more sustainable production process.

[0005] In addition, it is desirable to generate new fragrance characteristics in the original composition while achieving the above objectives.

[0006] Furthermore, in other commonly used fragrance composition production processes such as distillation, fats, oils, and waxes may remain as byproducts. These byproducts may still contain fragrance compounds that cannot be effectively separated from the non-polar components. In addition, in the above situations, it would be desirable to have methods to increase the yield of fragrance compounds and then dissolve them in ethanol for the preparation of scented consumer products.

[0007] Since the flavoring industry also suffers from similar drawbacks, there is a desire to find a solution that can be transferred to the food industry.

[0008] The present invention aims to achieve the above objectives. Summary of the Invention

[0009] In a first aspect, the present application relates to a method of modifying a biomass extract containing fatty acids as well as aroma and / or flavor compounds, the method comprising the steps of:

[0010] - providing a biomass extract containing fatty acids as well as aroma and / or flavor compounds, an alcoholic solvent and an immobilized lipase to form a reaction mixture;

[0011] - subjecting the reaction mixture to specific conditions to allow the fatty acids to react with the alcoholic solvent to form fatty acid esters;

[0012] - removing the immobilized lipase from the reaction mixture to obtain a modified biomass extract.

[0013] Preferably, the present application relates to a method of modifying a biomass extract containing (i) oil, fat, wax or mixtures thereof, and (ii) aroma and / or flavor compounds, wherein the method comprises the steps of:

[0014] - providing:

[0015] o a biomass extract containing (i) oil, fat, wax or mixtures thereof, and (ii) aroma and / or flavor compounds,

[0016] o an alcoholic solvent, and

[0017] o an immobilized lipase

[0018] to form a reaction mixture;

[0019] - subjecting the reaction mixture to specific conditions to allow the oil, fat, wax or mixtures thereof to react with the alcoholic solvent to form fatty acid esters;

[0020] - removing the immobilized lipase from the reaction mixture to obtain a modified biomass extract.

[0021] The method according to the present application relates to a method of modifying a biomass extract.

[0022] “Biomass” refers to a material derived from a living organism or biological source, such as a living cell, microorganism, fungus, algae or plant. The biomass can be fresh or dried. The biomass can be a whole plant, a tissue or an organ, such as a leaf, stem, flower, root, etc. Other examples include fruits or seeds, etc. “Biomass extract” refers to a material extracted from the above-mentioned biomass.

[0023] In a particular embodiment, the method according to the present application refers to a modification of the olfactory properties of the biomass extract.

[0024] In one specific embodiment, the method according to the invention refers to modifying a biomass extract to improve its extractability for fragrance and / or flavor compounds. Therefore, the method of the invention can improve the efficiency of extracting fragrance and / or flavor compounds from biomass extracts. As mentioned above, the fats and oils present in biomass extracts can hinder the extraction of fragrance and / or flavor compounds. The method of the invention overcomes this problem by converting fatty acids into fatty acid esters. Preferably, improving the extraction rate refers to using ethanol for extraction.

[0025] In one particular embodiment, the method according to the invention refers to altering the olfactory properties of a biomass extract and / or improving the extractability of fragrance and / or flavor compounds from the biomass extract.

[0026] The method according to the present invention includes the following steps: providing a biomass extract containing fatty acids and flavor compounds, an alcohol solvent, and an immobilized lipase to form a reaction mixture.

[0027] In one particular embodiment, the biomass extract can be obtained by solvent extraction, supercritical fluid extraction, carbon dioxide extraction, steam distillation extraction, pressing or cold pressing extraction, enfleurage extraction, maceration extraction, and / or fractionation extraction. Preferably, the biomass extract can be obtained by solvent extraction, supercritical fluid extraction, and / or carbon dioxide extraction.

[0028] In a preferred embodiment, the biomass extract is obtained by extraction using a nonpolar solvent (e.g., a hydrocarbon solvent, such as alkanes, particularly hexane). Preferably, the solvent used for extraction is removed after extraction, meaning the biomass extract is substantially solvent-free or completely solvent-free.

[0029] In a preferred embodiment, the biomass extract is a biomass extract that is insoluble in ethanol at 20°C. Preferably, the biomass extract is insoluble in ethanol at a concentration of at least 10% by weight at 20°C.

[0030] In one particular implementation, the biomass extract can be liquid or solid. For example, it can be a paste or powder.

[0031] In one particular embodiment, the water content of the biomass extract is less than 2% by weight, preferably less than 1% by weight, more preferably less than 0.5% by weight. In one embodiment, the biomass extract has been dried, i.e., subjected to a drying step, to achieve the aforementioned water content.

[0032] In one particular implementation, the biomass can be derived from plants.

[0033] In one particular implementation, the biomass can be either fresh or dried.

[0034] In one particular implementation, the biomass can be a whole plant, tissue, or organ, such as a leaf, stem, flower, root, fruit, or seed.

[0035] In one particular implementation, the biomass is selected from the group consisting of: Spartium junceum, Dianthus sp., Acacia farnesiana, Magnolia champaca, Narcissus jonquilla, Elaeagnus sp., Helicrysum italicum, Plumeria sp., Gardenia sp., Gardenia tahitensis, Pelargonia gravolens, Hyacinthus, Iris sp., Jasminum grandifolium, Jasminum sambac, Lavandula hybrid, Lavandula angustifolia, Lilium, and Hedychium. Magnolia sp., Tagetes minuta, Acaciade albata, Narcissus pseudonarcissus, Citrus x aurantium, Rosa sp., Agave amica, Viola sp., Mallow, Anise, Illicium verum, Elettaria cardamomum, Capsicum frutescens, Caraway, Daucus carota sativus, Ceratonia silique, Syzygium aromaticum, Piper cubaba, Foeniculum vulgare, Trigonella foenum-graecum, Zingiber Juniper (Juniperus officinalis), nutmeg (Myristica fragrans), chili pepper (Capsicum annuum), patchouli (Pogostemon cablin), pepper (Piper nigrum), saffron (Crocus sativus), coffee (Coffea sp.).(Arabic or medium-sized coffee), rock rose (Cistus ladaniferus), elderberry (Sambucus nigra), yellow grass (Anthoxanthum odoratum), laurel (Laurus nobilis), frankincense pistachio (Pistacia lentiscus), sweet snake grass (Liatris odoratissima), linden (Tilia sp.), yerba mate (Ilex paraguariensis), tonka bean (Dipterix odorata), rosemary (Rosemarinus officinalis), tea (Camelia sinensis), thyme (Thymus vulgaris), tobacco (Nicotiana tabacum), verbena (Verbena officinalis), and vanilla (Vanilla sp. (vanilla or Tahiti)). Preferably, the biomass is selected from the group consisting of: coffee (e.g., Arabica coffee), orange blossom (Citrus aurantium), rose (Rosa damascena), mimosa (Acaciade albata), tuberose (Agave polyanthes), and any mixture thereof.

[0036] In one particular embodiment, the biomass extract has undergone chemical or biotechnological treatment. This treatment step may alter / modify the chemical composition of the biomass extract.

[0037] The biomass extract contains fatty acids. In one particular embodiment, the fatty acids are free fatty acids.

[0038] In one particular embodiment, the total free fatty acid content of the biomass extract is greater than 2% by weight, preferably greater than 3% by weight, preferably greater than 5% by weight, preferably greater than 10% by weight, preferably greater than 20% by weight, preferably greater than 30% by weight, preferably greater than 40% by weight, and preferably greater than 50% by weight.

[0039] In one particular embodiment, the total content of oil, fat and / or wax in the biomass extract is greater than 2% by weight, preferably greater than 3% by weight, preferably greater than 5% by weight, preferably greater than 10% by weight, preferably greater than 20% by weight, preferably greater than 30% by weight, preferably greater than 40% by weight, preferably greater than 50% by weight.

[0040] In a preferred embodiment, the fatty acids are combined with glycerol, meaning the biomass extract comprises monoglycerides, diglycerides, triglycerides, or any mixture thereof. Preferably, the biomass extract comprises triacylglycerols. Depending on the nature of the glycerides, the biomass extract may comprise fats (solid at room temperature 20°C) and / or oils (liquid at room temperature 20°C).

[0041] In one particular embodiment, the total monoglyceride, diglyceride, and triglyceride content of the fatty acids in the biomass extract is greater than 2% by weight, preferably greater than 3% by weight, preferably greater than 5% by weight, preferably greater than 10% by weight, preferably greater than 20% by weight, preferably greater than 30% by weight, preferably greater than 40% by weight, and preferably greater than 50% by weight.

[0042] Therefore, in one embodiment, this disclosure relates to a method for modifying a biomass extract containing monoglycerides, diglycerides, triglycerides, or any mixture thereof, and fragrance and / or flavor compounds, wherein the method includes the following steps:

[0043] - Provide a biomass extract containing monoglycerides, diglycerides, triglycerides or any mixture thereof and fragrance and / or flavor compounds, an alcohol solvent and an immobilized lipase to form a reaction mixture;

[0044] - Apply specific conditions to the reaction mixture to react monoglycerides, diglycerides, triglycerides, or any mixture thereof with an alcohol solvent to produce fatty acid esters;

[0045] - The immobilized lipase was removed from the reaction mixture to obtain a modified biomass extract.

[0046] In one embodiment, this disclosure relates to a method for modifying a biomass extract containing triglycerides and fragrance and / or flavor compounds, wherein the method includes the following steps:

[0047] - Provides a biomass extract containing triglycerides and fragrance and / or flavor compounds, an alcohol solvent, and an immobilized lipase to form a reaction mixture;

[0048] - Apply specific conditions to the reaction mixture to cause triglycerides and alcohol solvents to form fatty acid esters;

[0049] - The immobilized lipase was removed from the reaction mixture to obtain a modified biomass extract.

[0050] In a preferred embodiment, the fatty acid is combined with a long-chain alcohol. The long-chain alcohol may contain 12 to 32 carbon atoms. Therefore, the biomass extract may contain wax (wax is defined as a fatty acid combined with a long-chain alcohol).

[0051] In one particular embodiment, the total long-chain fatty acid content of the biomass extract is greater than 2% by weight, preferably greater than 3% by weight, preferably greater than 5% by weight, preferably greater than 10% by weight, preferably greater than 20% by weight, preferably greater than 30% by weight, preferably greater than 40% by weight, and preferably greater than 50% by weight.

[0052] Therefore, this disclosure relates, in one embodiment, to a method for modifying a biomass extract containing waxes and fragrance and / or flavor compounds, wherein the method includes the following steps:

[0053] - Provides a biomass extract containing waxes and fragrance and / or flavor compounds, an alcohol solvent, and an immobilized lipase to form a reaction mixture;

[0054] - Apply specific conditions to the reaction mixture to cause the wax to react with an alcohol solvent to produce a fatty acid ester;

[0055] - The immobilized lipase was removed from the reaction mixture to obtain a modified biomass extract.

[0056] In a preferred embodiment, the biomass extract comprises fatty acids bound to glycerol, fatty acids bound to long-chain alcohols, or any mixture thereof. The definitions of fatty acids bound to glycerol and fatty acids bound to long-chain alcohols are as described above. Therefore, the biomass extract may comprise oils, fats, waxes, or any mixture thereof.

[0057] Therefore, this disclosure relates in one embodiment to a method for modifying a biomass extract containing fatty acids bound to glycerol, fatty acids bound to long-chain alcohols, or any mixture thereof, and fragrance and / or flavor compounds, wherein the method includes the following steps:

[0058] - Provides a biomass extract containing fatty acids bound to glycerol, fatty acids bound to long-chain alcohols, or any mixture thereof, and fragrance and / or flavor compounds, alcohol solvents, and immobilized lipases to form a reaction mixture;

[0059] - Apply specific conditions to the reaction mixture to react fatty acids with glycerol, fatty acids with long-chain alcohols or any mixture thereof, and alcohol solvents to produce fatty acid esters;

[0060] - The immobilized lipase was removed from the reaction mixture to obtain a modified biomass extract.

[0061] In one particular embodiment, the biomass extract comprises free fatty acids, fatty acids bound to glycerol, fatty acids bound to long-chain alcohols, or any mixture thereof.

[0062] If fatty acids are bound to glycerol or long-chain alcohols, the lipase treatment step performed in the presence of an alcohol solvent represents a transesterification step, in which at least a portion of the fatty acids originally bound to glycerol / long-chain alcohols are linked to the alcohol solvent, thereby forming new fatty acid ester bonds. The advantage is that the formation of new fatty acid ester bonds can be completed in a single step, unlike conventional methods which require two steps (first, the ester bond breaks to generate free fatty acids; second, the new fatty acid ester bond is formed). This also significantly reduces the operation time required by the method according to the invention. Therefore, the method according to the invention can be completed within 24 hours, preferably within 14 hours.

[0063] Therefore, in one particular embodiment, the step of applying conditions to the reaction mixture to cause the fatty acid to react with an alcohol solvent to form a fatty acid ester represents an transesterification step.

[0064] In one particular embodiment, the biomass extract comprises fatty acids (in the form of free fatty acids and / or fatty acids bound to glycerol and / or fatty acids bound to long-chain alcohols, preferably a mixture of free fatty acids, fatty acids bound to glycerols and fatty acids bound to long-chain alcohols) in an amount of at least 3% by weight, particularly at least 5% by weight, particularly at least 10% by weight, and particularly at least 20% by weight of the total dry weight of the biomass extract.

[0065] In one particular embodiment, the biomass extract comprises fatty acids (in the form of free fatty acids and / or fatty acids bound to glycerol and / or fatty acids bound to long-chain alcohols, preferably a mixture of free fatty acids, fatty acids bound to glycerol and fatty acids bound to long-chain alcohols) in a content not exceeding 97% by weight, particularly not exceeding 80% by weight, and particularly not exceeding 60% by weight.

[0066] In one particular embodiment, the biomass extract contains fatty acids in an amount of 3% to 97% by weight of the total dry weight of the biomass extract.

[0067] In one particular embodiment, the biomass extract contains fatty acids in an amount of 5% to 97% by weight of the total dry weight of the biomass extract.

[0068] In one particular embodiment, the biomass extract contains fatty acids in an amount of 10% to 80% by weight of the total dry weight of the biomass extract.

[0069] In one particular embodiment, the biomass extract contains fatty acids in an amount of 20% to 60% by weight of the total dry weight of the biomass extract.

[0070] In one particular embodiment, the biomass extract comprises unsaturated fatty acids, preferably having 12 to 54 carbon atoms, more preferably having 12 to 18 carbon atoms. In one embodiment, the ratio of saturated fatty acids to unsaturated fatty acids is 1:100 to 100:1. In another embodiment, the ratio of saturated fatty acids to unsaturated fatty acids is 1:10 to 10:1.

[0071] In one particular embodiment, the biomass extract is a fragrance concrete or the wax portion of a fragrance absolute (a byproduct of absolute production). If the biomass extract is a fragrance concrete, the method according to the invention can be used as part of a method for preparing a fragrance absolute. This method for preparing a fragrance absolute may include the further step of removing alcohol solvents from the modified biomass extract to obtain the absolute.

[0072] Therefore, this disclosure also relates to a method for preparing flavoring active ingredients, the method comprising the following steps:

[0073] - Extract biomass, preferably using hydrocarbon solvents, to obtain a fragrance extract containing fatty acids and aroma and / or flavor compounds;

[0074] - The extract is mixed with an alcohol solvent and immobilized lipase to form a reaction mixture;

[0075] - Remove immobilized lipase from the reaction mixture;

[0076] - The alcohol solvent is removed from the reaction mixture, preferably by evaporation, to obtain the flavoring essence.

[0077] All of the above implementation schemes are also applicable to the method for preparing flavoring raw materials.

[0078] In one particular embodiment, the biomass extract is a coffee extract obtained using supercritical carbon dioxide extraction. This coffee extract may contain fatty oils.

[0079] In one particular embodiment, the biomass extract is an orange blossom extract. The orange blossom extract may contain wax.

[0080] In one particular embodiment, the biomass extract is rose extract. The rose extract may contain wax.

[0081] In one particular embodiment, the biomass extract is a mimosa extract. The mimosa extract may contain fatty oils.

[0082] In one particular embodiment, the biomass extract is a tuberose extract. The tuberose extract may contain waxes or fatty oils.

[0083] In one particular embodiment, the biomass extract is an Arabian jasmine extract. The Arabian jasmine extract may contain waxes or fatty oils.

[0084] In one particular embodiment, the biomass extract is gardenia (i.e., Tahitian gardenia) extract. Gardenia extract may contain waxes or fatty oils.

[0085] In one particular embodiment, the biomass extract is a violet leaf extract. The violet leaf extract may contain waxes or fatty oils.

[0086] In one particular embodiment, the biomass extract is selected from the group consisting of: orange blossom extract, rose extract, mimosa extract, tuberose extract, jasmine extract, gardenia extract, violet leaf extract, and any mixture thereof.

[0087] Biomass extracts also contain fragrance compounds.

[0088] "Fragrance compounds" refer to compounds used as active ingredients in fragrance formulations or compositions to impart a pleasant effect; that is, their primary purpose is to give or modulate a pleasant odor. In other words, for a compound to be considered a fragrance compound, it must be recognized by professionals in the fragrance industry as capable of actively or pleasantly imparting or modifying the odor of a composition, and not merely possessing an odor. Generally, fragrance compounds belong to various chemical categories, such as alcohols, lactones, aldehydes, ketones, esters, ethers, acetates, nitriles, thiols, terpenes, and nitrogen- or sulfur-containing heterocyclic compounds.

[0089] Fragrance compounds are liquid or solid under ambient conditions and have a measurable vapor pressure at 25°C. The vapor pressure of these substances is typically greater than about 0.0000001 mmHg, or from about 0.02 mmHg to about 20 mmHg; typically, the average boiling point is below about 250°C, or below about 235°C or below about 220°C. Typically, the molecular weight of fragrance compounds is in the range of 40 to 500 Daltons, preferably from 50 to 350 Daltons.

[0090] The properties of fragrance compounds in biomass extracts depend on the biomass from which they are extracted. Many fragrance compounds are listed in references such as S. Arctander's book *Perfume and Flavor Chemicals*, 1969, Montclair, NJ, USA, or its latest edition, or other works of similar nature, such as Fenaroli's *Handbook of Flavor Ingredients*, 1975, CRC Press, or MB Jacobs' *Synthetic Food Adjuncts*, 1947, van Nostrand Co., Inc. Fragrance compounds are also described in YR Naves, *Technologie etchimie des parfums naturels*, 1974, Masson Ed, Paaris.

[0091] In one particular embodiment, the biomass extract comprises one or more fragrance compounds selected from the group consisting of: 2-furfural, 3-methylbutyric acid, furfuryl alcohol, γ-butyrolactone, 2,5-dimethylpyrazine, 5-methylfurfural, furfuryl acetate, 3-methyl-2-cyclopenten-1-one (nussol), 2-acetylpyrrole, maltol, 4-vinylguaiacol, limonene, (E)-β-ocimene, linalool, 2-phenylethanol, benzyl cyanide, α-terpenes Pinyl alcohol, geraniol, linalyl acetate, indole, methyl anthranilate, neryl acetate, geraniol, (E)-nerolidol, (E,E)-farnesol, citronellol, nerolidol, 2-phenylethyl acetate, eugenol, geraniol, methyl eugenol, tyrosol (4-hydroxyphenylethanol), eucalyptol, methyl benzoate, ethyl benzoate, methyl salicylate, (E)-isoeugenol, (Z)-jasminolide, δ-decanoolide, (E)-methylisoeugenol and (E,E)-farnesol.

[0092] In one particular embodiment, the biomass extract comprises one or more fragrance compounds selected from Table 1 below.

[0093] In one particular embodiment, the biomass extract comprises one or more fragrance compounds selected from Table 2 below.

[0094] In one particular embodiment, the biomass extract comprises one or more fragrance compounds selected from any of Tables 5 to 10 below.

[0095] In one particular embodiment, the biomass extract contains at least 0.0001% by weight of a terpene derivative, particularly at least 5% by weight, particularly at least 10% by weight, particularly at least 20% by weight.

[0096] In one particular embodiment, the biomass extract contains no more than 85% by weight of a terpene derivative, particularly no more than 60% by weight, particularly no more than 50% by weight, and particularly no more than 20% by weight.

[0097] In one particular embodiment, the biomass extract comprises 0.0001 to 85% by weight, or 10 to 60% by weight, or 20 to 50% by weight, or 5 to 20% by weight of terpene derivatives.

[0098] By the term "flavor compound," this refers to compounds capable of imparting, improving, or modifying the sensory properties (particularly their flavor and / or taste) of an edible composition. Flavoring ingredients are well known to those skilled in the art, and their properties need not be described in detail here, even if they were, they would not be exhaustive, as a skilled flavorist can identify these ingredients based on their general knowledge.

[0099] The reaction mixture contains alcohol solvents, biomass extracts, and immobilized lipase.

[0100] In one particular embodiment, the alcohol solvent is selected from methanol, ethanol, propanol, butanol, pentanol, hexanol, and any mixture thereof. Preferably, the alcohol solvent is ethanol.

[0101] In one particular embodiment, the alcohol solvent is an anhydrous alcohol solvent.

[0102] In one particular embodiment, the alcohol solvent is a high-purity alcohol solvent, such as anhydrous ethanol or ethanol 100.

[0103] In one particular embodiment, the weight ratio of biomass extract to alcohol solvent in the reaction mixture is 1:0.1 to 1:20, preferably 1:1 to 1:5.

[0104] In a particular embodiment, the reaction mixture contains another solvent in addition to the alcohol solvent. This other solvent can be a nonpolar solvent, such as cyclohexane. Therefore, in one embodiment, the reaction mixture is formed by providing a biomass extract containing fatty acids and fragrance compounds, an alcohol solvent, a nonpolar solvent (preferably cyclohexane), and an immobilized lipase. When the biomass extract contains waxes, a nonpolar solvent (such as cyclohexane) is particularly preferred.

[0105] In one particular embodiment, the alcohol solvent is the only solvent in the reaction mixture.

[0106] The reaction mixture contains immobilized lipase. The advantage of immobilized lipase is that it can be removed (e.g., by filtration) after the reaction steps are complete and reused. Furthermore, this method simplifies the operation because the adjustment steps typically required when using non-immobilized lipases can be omitted.

[0107] In one particular embodiment, the immobilized lipase is an immobilized esterase, preferably an immobilized interesterase. For example, the lipase could be NOVOZYME 435 or Lipozyme. ® 1LTM, Lipozyme ® RM IM, or any other suitable immobilized lipase. NOVOZYME 435 refers to a lipase from *Candida antarctica* immobilized on resin (e.g., Lewatit VP OC 1600). The lipase can also be an immobilized lipase from *Aspergillus niger*, such as Addzyme CalB 191G; or an immobilized lipase from *Thermomyces lanuginosus*, such as Immobead 150; or an immobilized lipase from *Rhizomucor miehei*, such as Immobead 150.

[0108] In one particular embodiment, the lipase is selected from the group consisting of: lipases from Candida antarctica, lipases from Aspergillus niger, lipases from Thermophilus spp., lipases from Rhizopus spp., and any mixture thereof.

[0109] In one particular embodiment, the lipase is selected from Novozym 435 lipase, Addzyme CalB 191G lipase, SIGMA 76546, SIGMA 52001, and any mixture thereof.

[0110] In one particular embodiment, the immobilized lipase is NOVOZYM 435, Addzyme CalB 191G, or a mixture thereof. Preferably, the immobilized lipase is NOVOZYM 435.

[0111] Immobilization of immobilized lipases can be achieved through adsorption, covalent bonding, embedding, encapsulation, or cross-linking, fixing them onto or within a solid carrier material. Suitable carrier materials include, but are not limited to, homopolymers or copolymers of natural or synthetic polymers (e.g., agarose, alginate, acrylamide, styrene, (meth)acrylate, divinylbenzene, etc.), inorganic carriers (e.g., silica, glass beads, zeolite), or mixed materials.

[0112] In one particular embodiment, based on the total weight of the reaction mixture, the content of immobilized lipase in the reaction mixture is less than 50.0% by weight, preferably less than 10% by weight, more preferably less than 5% by weight, and even more preferably less than 2% by weight.

[0113] In one particular embodiment, the immobilized lipase content in the reaction mixture is from 0.1% to 50% by weight, preferably from 1% to 10% by weight, and more preferably from 1% to 5% by weight, based on the total weight of the reaction mixture.

[0114] If the lipase is NOVOZYM 435, its dosage is based on a total weight of the reaction mixture equal to or less than 1% by weight. If the lipase is Lipozyme... ® If the amount of 1 LTM is equal to or less than 10% by weight of the total weight of the reaction mixture.

[0115] The method according to the invention includes the following steps: applying specific conditions to the reaction mixture to react fatty acids with an alcohol solvent to generate fatty acid esters. Those skilled in the art know suitable reaction conditions for reacting fatty acids with an alcohol solvent to generate (new) fatty acid esters in the presence of lipases. The fatty acid esters generated in the method steps are soluble in ethanol and can therefore be used as part of the raw material fraction in the raw material production process. Furthermore, as described above, the aroma and / or flavor compounds present in the biomass extract are no longer retained by fats, oils, or waxes, thereby allowing for more efficient extraction of aroma and / or flavor compounds.

[0116] In one particular embodiment, the reaction step is carried out at a temperature of 35°C to 70°C.

[0117] In one particular embodiment, the reaction step is carried out at a temperature of 20°C to 80°C, preferably at a temperature of 40°C to 60°C, and more preferably at a temperature of 50°C.

[0118] In one particular embodiment, the reaction step is carried out under stirring, preferably at a stirring speed of 1 to 1000 rpm, more preferably 100 to 300 rpm, and most preferably 200 rpm.

[0119] In one particular embodiment, the method according to the invention includes a further purification step. Preferably, the purification step includes cold filtration and / or decantation. If cold filtration is used, it is preferably carried out at a temperature of -15°C to 0°C. Filtration can also be performed multiple times; for example, cold filtration can be performed twice consecutively. The main purpose of the purification step is to remove residual wax from the treated biomass extract.

[0120] In one particular embodiment, the method according to the invention includes a further step of removing the alcohol solvent by evaporation. This step can be performed after the lipase reaction is complete and, optionally, after a purification step. The evaporation conditions employed depend on the nature of the alcohol solvent, which is common knowledge to those skilled in the art. Evaporation can also be carried out under reduced pressure, thereby allowing the use of lower evaporation temperatures. For example, for ethanol, evaporation can be carried out at a temperature of 55 to 60°C and a pressure of 250 mbar.

[0121] In one particular embodiment, the method according to the invention includes one or more post-processing steps.

[0122] In one particular embodiment, the post-processing step can alter, improve, and / or enhance the odor characteristics, appearance, and / or stability of the resulting biomass extract. The post-processing step may involve physical, chemical, and / or thermal treatments designed to remove unwanted components such as odor substances, acidic or polar residues, and pigments.

[0123] In one particular embodiment, the post-treatment step may include contacting the mixture with an adsorbent material (e.g., activated carbon, activated alumina, silica gel, bentonite, diatomaceous earth, ion exchange resin, zeolite, or synthetic porous polymer (e.g., styrene-divinylbenzene copolymer, preferably activated carbon) under moderate temperature conditions (e.g., 25°C to 40°C, preferably 27°C to 35°C), followed by solid-liquid separation (e.g., filtration, decantation, centrifugation, or membrane separation (e.g., microfiltration, preferably filtration)). This subsequent treatment step helps to eliminate burnt odors, ammonia odors, and / or other undesirable smells, and / or optionally, to partially or completely decolorize the mixture.

[0124] In one particular embodiment, the post-treatment step may include contacting the mixture with a pH adjuster (e.g., an alkali). In one particular embodiment, the alkali may be a weakly acidic alkali metal or alkaline earth metal salt, such as sodium carbonate, potassium bicarbonate, calcium carbonate, magnesium carbonate, or magnesium hydroxide, preferably sodium bicarbonate. This treatment may be carried out at a temperature of 25°C to 40°C, preferably 27°C to 35°C, followed by separation of insoluble substances, such as by filtration, decantation, centrifugation, or membrane separation (e.g., microfiltration), preferably filtration. This subsequent treatment step helps neutralize acidic components that contribute to a pungent or slightly acidic odor.

[0125] In one particular embodiment, the post-processing step may include water washing or phase separation techniques. In one particular embodiment, the mixture may be mixed with water or an aqueous solution. The resulting mixture may be heated (e.g., between 40°C and 60°C). The resulting mixture (heated or unheated) may be allowed to settle, and the desired phase may then be collected and subjected to solid-liquid separation treatment as appropriate (e.g., filtration, decantation, centrifugation, or membrane separation (e.g., microfiltration), preferably filtration). This post-processing helps remove hydrophilic impurities (e.g., glycerol) and / or improve the visual clarity of the extract.

[0126] In one particular embodiment, the post-treatment step may include molecular distillation of the mixture under high vacuum (e.g., greater than 0.001 mbar, preferably 0.001 to 10 mbar) and high temperature conditions (e.g., 60°C to 200°C, preferably 100°C to 130°C). This post-treatment step helps to obtain a fragrance composition with good color stability.

[0127] Post-processing steps can be used alone or in combination, with yields ranging from approximately 50% to 100%.

[0128] Another aspect of the invention relates to a biomass extract that can be obtained by the method of the invention.

[0129] In one particular embodiment, the water content of the biomass extract is less than 2% by weight, preferably less than 1% by weight, and more preferably less than 0.5% by weight.

[0130] In one particular implementation, the biomass extract is essentially solvent-free.

[0131] In one particular implementation, the biomass extract is solvent-free.

[0132] In a preferred embodiment, the biomass extract is a fragrance composition.

[0133] In one particular implementation, the biomass extract is a raw essence.

[0134] In one particular embodiment, the biomass extract comprises fatty acid esters, preferably ethyl fatty acid esters, in an amount of at least 3% by weight, preferably at least 5% by weight, based on the total biomass extract. Alternatively, the biomass extract comprises fatty acid esters, preferably ethyl fatty acid esters, in an amount of at least 30% by weight, preferably at least 40% by weight, based on the total biomass extract.

[0135] In one particular embodiment, the biomass extract comprises fatty acid esters in an amount of 3% to 10% by weight, or in an amount of 40% to 60% by weight.

[0136] In one particular embodiment, the biomass extract comprises fatty acid esters, preferably ethyl fatty acid esters, in an amount of 3% to 60% by weight.

[0137] In one particular embodiment, the biomass extract comprises fatty acid esters, preferably ethyl fatty acid esters, in an amount of 10% to 60% by weight.

[0138] In one particular embodiment, the biomass extract comprises one or more compounds selected from the group consisting of:

[0139]

[0140] In one particular embodiment, the biomass extract comprises all of the above-described compounds. The biomass extract may also comprise only one of the above-described compounds.

[0141] In one embodiment, the biomass extract comprises ethyl myristate. In a particular embodiment, the biomass extract comprises ethyl palmitate. In a particular embodiment, the biomass extract comprises ethyl linoleate. In a particular embodiment, the biomass extract comprises ethyl linolenate. In a particular embodiment, the biomass extract comprises ethyl oleate. In a particular embodiment, the biomass extract comprises ethyl stearate.

[0142] In one particular embodiment, the biomass extract comprises one or more compounds selected from the group consisting of:

[0143]

[0144] In one particular embodiment, the biomass extract comprises all of the above-described compounds. The biomass extract may also comprise only one of the above-described compounds.

[0145] In one particular embodiment, the biomass extract comprises one or more compounds selected from the group consisting of:

[0146]

[0147] In one particular embodiment, the biomass extract comprises all of the above-described compounds. The biomass extract may also comprise only one of the above-described compounds.

[0148] In one particular embodiment, the biomass extract comprises one or more compounds selected from the group consisting of:

[0149]

[0150] In one particular embodiment, the biomass extract comprises all of the above-described compounds. The biomass extract may also comprise only one of the above-described compounds.

[0151] In one particular embodiment, the biomass extract comprises one or more compounds selected from the group consisting of:

[0152]

[0153] In one particular embodiment, the biomass extract comprises all of the above-described compounds. The biomass extract may also comprise only one of the above-described compounds.

[0154] In one particular embodiment, the biomass extract is soluble in ethanol at 20°C. Preferably, the biomass extract is soluble in ethanol at a concentration of at least 10% by weight at 20°C.

[0155] Another aspect of the invention relates to a flavoring composition comprising:

[0156] - Biomass extracts as defined above;

[0157] - At least one ingredient selected from the group consisting of a spice carrier and a spice base; and

[0158] -Optionally, at least one spice adjuvant.

[0159] In one particular embodiment, the flavoring composition comprises a flavor carrier.

[0160] The term "fragrance carrier" refers to a material that is neutral from the perspective of fragrance, meaning it does not significantly alter the sensory properties of the flavoring ingredients. The carrier can be liquid or solid.

[0161] As liquid carriers, emulsion systems, i.e., solvent and surfactant systems, or solvents commonly used in the fragrance industry, can be listed as non-limiting examples. A detailed description of the properties and types of solvents commonly used in the fragrance industry is impossible to exhaustively cover. However, solvents such as butanediol or propylene glycol, glycerol, dipropylene glycol and its monoethers, 1,2,3-propanetriyltriacetate, dimethyl glutarate, dimethyl adipate, 1,3-diacetoxypropyl-2-yl acetate, diethyl phthalate, isopropyl myristate, benzyl benzoate, benzyl alcohol, 2-(2-ethoxyethoxy)-1-ethanol, triethyl citrate, or mixtures thereof, are the most commonly used. In addition to those detailed above, other suitable fragrance carriers may also be ethanol, water / ethanol mixtures, limonene or other terpenes, isoparaffins, such as those under the trademark Isopar ®Those known to the public (source: Exxon Chemical), or glycol ethers and glycol ether esters, such as those under the trademark Dowanol ® Those commonly known (source: Dow Chemical Company), or hydrogenated castor oil, such as those marketed under the brand name Cremophor ® RH 40, those who are well-known (Source: BASF).

[0162] A solid carrier is a material to which a flavored composition or certain components of a flavored composition can be chemically or physically bonded. Generally, such solid carriers are used to stabilize compositions or to control the evaporation rate of compositions or certain components. Solid carriers are currently used in the art, and those skilled in the art know how to achieve the desired effects. However, as non-limiting examples of solid carriers, absorbent adhesives or polymers or inorganic materials, such as porous polymers, cyclodextrins, dextrins, maltodextrins, wood-based materials, organic or inorganic gels, clays, gypsum, talc, or zeolites can be listed.

[0163] Other non-limiting examples of solid carriers include encapsulating materials. Examples of such materials may include wall-forming and plasticizing materials, such as monosaccharides, disaccharides or trisaccharides, natural or modified starches, hydrocolloids, cellulose derivatives, polyvinyl acetate, polyvinyl alcohol, proteins or pectins, or materials listed in references such as H. Scherz, Hydrokolloide: Stabilisatoren, Dickungs- und Geliermittel in Lebensmitteln, Band 2 der Schriftenreihe Lebensmittelchemie, Lebensmittelqualität, Behr's Verlag GmbH & Co., Hamburg, 1996. Encapsulation is a method well known to those skilled in the art and can be carried out, for example, using techniques such as spray drying, agglomeration or extrusion; or consisting of coating encapsulation including agglomeration and composite agglomeration techniques.

[0164] As a non-limiting example of a solid carrier, a core-shell capsule may be specifically cited, which uses resins of the type of amino plastics, polyamides, polyesters, polyureas, or polyurethanes, or mixtures thereof (all of which are well known to those skilled in the art), and is carried out by a phase separation method initiated by using techniques such as polymerization, interfacial polymerization, coagulation, or these techniques together (all of which have been described in the prior art), and optionally in the presence of a polymer stabilizer or a cationic copolymer.

[0165] Resins can be produced by polycondensation of aldehydes (such as formaldehyde, 2,2-dimethoxyacetaldehyde, glyoxal, glyoxylic acid, or hydroxyacetaldehyde and mixtures thereof) with amines such as urea, benzoguanidine, glycyrrhizin, melamine, hydroxymethyl melamine, methylated hydroxymethyl melamine, guanidine, etc., and mixtures thereof. Alternatively, pre-formed resins can be used to alkylate polyamines, for example, under the trademark Urac. ® (Source: Cytec Technology Corp.), Cymel ® (Source: Cytec Technology Corp., Urecoll) ® or Luracoll ® (Source: BASF) Those available for commercial purchase.

[0166] Other resins are produced by the formation of trimers from polyols such as glycerol and polyisocyanates such as hexamethylene diisocyanate, isophorone diisocyanate, or phenyl diisocyanate, or biuret of hexamethylene diisocyanate, or trimers of phenyl diisocyanate and trimethylolpropane (under the trade name Takenate). ® Those produced by the polycondensation of known (source: Mitsui Chemicals), wherein preferably, the trimer of phenyl diisocyanate with trimethylolpropane and the biuret of hexamethylene diisocyanate.

[0167] In one particular embodiment, the flavoring composition comprises a flavoring adjuvant.

[0168] The term "fragrance adjuvant" should be understood as an ingredient that can impart additional benefits, such as color, specific lightfastness, chemical stability, etc. A detailed description of the nature and type of adjuvants commonly used in fragrance and / or odor-counteracting compositions cannot be exhaustive, but it must be mentioned that the ingredients are well known to those skilled in the art. Specific, non-limiting examples include: viscosity agents (e.g., surfactants, thickeners, gelling agents, and / or rheology modifiers), stabilizers (e.g., preservatives, antioxidants, heat / light and / or buffers or chelating agents, such as BHT), colorants (e.g., dyes and / or pigments), preservatives (e.g., antibacterial or antimicrobial or antifungal or antiirritant agents), abrasives, skin coolants, fixatives, insect repellents, ointments, vitamins, and mixtures thereof. "Fixative," also known as "modifier," is understood herein as an agent that has the ability to influence, compared to the same perception without the modifier, the way an observer or user perceives the odor of a composition incorporating the modifier over time, particularly the rate and intensity of evaporation. In particular, this modifier allows for a prolonged period of time when its fragrance is perceived. Non-limiting examples of suitable modifiers may include: methyl glucoside polyol; ethyl glucoside polyol; propyl glucoside polyol; isocetyl alcohol; PPG-3 myristole; neopentyl glycol di(ethylhexanoate); sucrose laurate; sucrose dilaurate, sucrose myristate, sucrose palmitate, sucrose stearate, sucrose distearate, sucrose tristearate, sodium hyaluronate disaccharide, sodium hyaluronate, propylene glycol propyl ether; diceryl ether; polyglycerol-4 ether; isocetyl alcohol polyether-5; isocetyl alcohol polyether-7; isocetyl alcohol polyether-7; isocetyl alcohol polyether-8; isocetyl alcohol polyether-9; isocetyl alcohol polyether-10 ... Cetyl alcohol polyether-10; isocetyl alcohol polyether-12; isocetyl alcohol polyether-15; isocetyl alcohol polyether-20; isocetyl alcohol polyether-25; isocetyl alcohol polyether-30; disodium lauryl amphoteric dipropionate; hexaethylene glycol monododecyl ether; and mixtures thereof; neopentyl glycol diisonononate; cetearyl alcohol ethylhexanoate; panthenol ethyl ether; DL-panthenol; n-hexadecyl n-nonanoate; n-octadecyl n-nonanoate; fragrance precursors; cyclodextrins; encapsulating agents; and combinations thereof.

[0169] Another aspect of the present invention relates to a flavoring composition comprising:

[0170] - Biomass extracts as defined above;

[0171] - At least one ingredient selected from the group consisting of flavoring carriers and flavoring bases; and

[0172] -Optionally, at least one flavoring agent.

[0173] Flavoring carriers and flavoring adjuvants can be described separately according to the descriptions of spice carriers and spice adjuvants above.

[0174] Another aspect of the invention relates to a flavored consumer product comprising a biomass extract as defined above, or a flavoring composition as defined above.

[0175] Another aspect of the invention relates to a flavored consumer product comprising a biomass extract as defined above, or a flavoring composition as defined above.

[0176] In one particular implementation, the scented consumer product is a fine perfume, a spray perfume, or an eau de perfume; cologne, shaving lotion, or aftershave; optionally, a liquid or solid detergent in capsule or tablet form; a fabric softener; a fabric rinser; a liquid or solid fragrance enhancer; a dryer sheet; a fabric freshener; ironing lotion; paper; bleach; a carpet cleaner; curtain care products; shampoos; leave-on or rinse-off hair conditioners; colorants; color care products; or hair styling products. Products including: dental care products, disinfectants, feminine hygiene products, hair sprays (hair gels), skin creams or lotions, cold creams, deodorants or antiperspirants, hair removal products, tanning or after-sun care products, nail care products, skin cleansing products, cosmetics, soaps, shower or bath mousse, bath oils or shower gels, foot / hand care products, hygiene products, air fresheners, "ready-to-use" powder air fresheners, mold removers, furniture conditioners, wipes, dishwashing liquids or hard surface cleaners, leather care products, and car care products.

[0177] In one particular implementation, flavored consumer products are food products or beverages.

[0178] In one particular implementation, a flavored consumer product may be one of the following examples:

[0179] • Baked goods (such as bread, dry biscuits, cakes, and other baked goods).

[0180] • Cereal products (such as breakfast cereals, pre-cooked ready-made rice products, rice noodle products, millet and sorghum products, raw or pre-cooked noodles and pasta products).

[0181] • Dairy products (such as fresh cheese, soft cheese, hard cheese, milk beverages, whey, butter, partially or completely hydrolyzed milk protein products, fermented dairy products, condensed milk and the like).

[0182] • Dairy products (such as fruit or flavored yogurt, ice cream, fruit ice, frozen desserts).

[0183] • Dairy analogues (imitative dairy products) containing non-dairy ingredients (plant proteins, plant fats).

[0184] • Confectionery products (such as chewing gum, hard candy, and soft candy).

[0185] • Chocolate and compound icing,

[0186] • Products based on fats and oils or their emulsions (e.g., mayonnaise, spreads, margarine, shortening, remoulade, dressings, flavored preparations).

[0187] • Spiced, marinated, or processed fish products (such as fish intestines or fish paste).

[0188] • Eggs or egg products (dried eggs, egg whites, egg yolks, custards)

[0189] • Desserts (such as gelatin and pudding)

[0190] • Products made from soy protein or other soy components (such as soy milk and products made therefrom, preparations containing soy lecithin, fermented products such as tofu or tempeh or products made therefrom, soy sauce).

[0191] • Vegetable products (e.g., tomato sauce, dressings, processed and reconstituted vegetables, dried vegetables, deep-frozen vegetables, pre-cooked vegetables, pickled vegetables, vegetable concentrates or pastes, cooked vegetables, potato products).

[0192] • Spices or spice products (such as mustard products, horseradish products), spice blends, especially seasonings used in areas such as snacks.

[0193] • Snacks (such as baked or fried potato chips or potato dough products, bread dough products, extrusions based on corn, rice or chopped nuts),

[0194] • Ready-to-eat dishes (such as instant noodles, rice, pasta, pizza, tortillas, wraps) as well as broths and stock (such as stock, savory cube, dry stock, instant stock, pre-cooked stock, distilled stock) and sauces (instant sauce, dry sauce, prepared sauce, gravy, sweet sauce).

[0195] • Expanded range of meat products (e.g., meat patties, sausages, chili, Salisbury steak, pizza toppings, meatballs, ground meat, bolognese, chicken nuggets, Frankfurt sausages, beef). Detailed Implementation

[0196] Example

[0197] Example 1

[0198] Coffee powder was extracted using supercritical carbon dioxide extraction to obtain a coffee extract. Because this extract contains fats, it is insoluble in ethanol.

[0199] Process according to the invention

[0200] In the first step, 50.3 g of coffee extract was mixed with 0.25 L of ethanol (99.9% purity) by stirring at 40 °C. Then, 2 g of NOVOZYME 435 (4% ​​by weight) was added. The mixture was stirred at 55 °C (200 rpm) for 6 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with the ethanol (99.9%) mixture, allowing for reuse.

[0201] The ethanol filtrate was transferred to a separatory funnel to remove some of the insoluble matter (heavy phase). The clarified ethanol phase was then filtered and concentrated under vacuum (250 mbar) with moderate heating (55°C to 60°C) to give 57.3 g of the final extract. Based on the amount of original coffee extract used, the yield was 113.9% by weight. The alcohol content (ethanol) in the treated extract was less than 2% by weight. The treated extract was soluble in ethanol at 20°C at 10% by weight (96%).

[0202] Comparative process

[0203] A comparison was made with a conventional method for defatting raw coffee extract. For ease of comparison, an equal volume of the raw coffee extract provided above was defatted using ethanol washing and decantation (extract / ethanol ratio of 1:10), i.e., the raw coffee extract was extracted with ethanol to separate insoluble coffee fats. After ethanol extraction, the extraction solvent (ethanol) was evaporated, yielding a 22% by weight yield, while the raw coffee extract contained approximately 88% by weight fat.

[0204] Therefore, according to the method of the present invention, the yield of the ethanol-soluble extract is increased by five times.

[0205] Table 1 lists the compounds present in the extracts prepared according to the method of the present invention and the extracts prepared by comparative methods, and their contents. These contents were determined by gas chromatography.

[0206] Table 1. Compounds and their contents in the processed coffee extract

[0207]

[0208] As can be seen from Table 1, the method according to the present invention significantly increases the amount of ethanol-soluble fatty acid ethyl esters, particularly ethyl palmitate and ethyl linoleate. Therefore, by converting bound fatty acids into fatty acid ethyl esters, the method according to the present invention can significantly increase the yield of ethanol-soluble compounds.

[0209] Example 2

[0210] Process according to the invention (processing of neroli wax)

[0211] Orange blossom wax is collected as a byproduct of orange blossom absolute production.

[0212] In the first step, 1 kg of ethanol (99.9% purity) was slowly added to 200 g of neroli wax while stirring at 40 °C. Then, 2 g of NOVOZYME 435 (1% by weight) was added. The mixture was stirred at 45 °C (200 rpm) for 14 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with ethanol (99.9%), allowing for reuse.

[0213] The ethanol filtrate was cooled to 0°C and then filtered to remove insoluble wax. This was followed by a second cooling to -5°C and subsequent filtration to remove any remaining insoluble wax.

[0214] The clarified ethanol phase was concentrated under moderate heating (55°C to 60°C) under vacuum (250 mbar) to give 25 g of the final product. The yield was 12.5% ​​by weight based on the amount of neroli wax used. The final product was soluble in ethanol (96%) at 20°C at 10% by weight. It is an amber-brown viscous liquid with a pleasant neroli aroma.

[0215] Comparative process (production of nerol absolute)

[0216] Fresh orange blossoms were extracted with hexane, and the solvent was evaporated to obtain an orange blossom extract. Then, 100 g of the orange blossom extract was melted at 50°C and mixed with 1 L of warm ethanol (96%) in a beaker. The mixture was stirred at 50°C for 1 hour. It was then cooled to 20°C and filtered (to remove solid cold wax). The filtrate was cooled to 0°C to precipitate the residual wax. The solid fraction (wax) was extracted again with 1 L of ethanol (96%) at 50°C to obtain a second ethanol extract. The second ethanol extract was cooled twice more (to 0°C and -5°C, respectively) and then filtered. The ethanol filtrates were combined and concentrated under vacuum to obtain a liquid orange blossom absolute soluble in ethanol. Based on the initial extract used, the absolute yield was 58% by weight.

[0217] Process according to the invention (production of nerol absolute)

[0218] In the first step, 100 g of orange blossom extract (prepared as described above) was mixed with 500 g of ethanol (99.9% purity) and stirred at 40°C. Then, 1 g of NOVOZYME 435 (1% by weight) was added. The mixture was stirred at 45°C (200 rpm) for 14 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with the ethanol (99.9%) mixture, thus allowing for reuse.

[0219] The ethanol filtrate was then cooled to 0°C and filtered to remove insoluble wax. The resulting ethanol filtrate was then cooled to -5°C and filtered to remove residual insoluble wax.

[0220] The clarified ethanol phase was concentrated under moderate heating (55°C to 60°C) under vacuum (250 mbar) to obtain 67 g of liquid orange blossom extract. Based on the extract used initially, the absolute yield was 67% by weight. The resulting absolute was an amber-brown viscous liquid with a pleasant orange blossom aroma.

[0221] Therefore, compared with the comparative method, the method for preparing neroli protopor according to the present invention significantly improves the yield of ethanol-soluble compounds (58 wt% vs. 67 wt%).

[0222] Table 2 lists the compounds present in the raw semen prepared according to the method of the present invention and the raw semen prepared by a comparative method, and their contents. These contents were determined by gas chromatography. In addition, Table 2 also lists the compounds obtained after treating neroli wax as described above, and their contents.

[0223] Table 2. Compounds and their contents (%, w / w) in the treated neroli wax, the absolute prepared according to the comparative method, and the absolute prepared according to the method of the present invention.

[0224]

[0225] As shown in Table 2, the method according to the present invention can convert neroli wax into ethanol-soluble fatty acid ethyl esters. Furthermore, in the production of neroli absolute, compared with the comparative method, the method according to the present invention can obtain a higher content of fatty acid ethyl esters, thereby achieving a higher neroli absolute yield.

[0226] Example 3

[0227] Process according to the invention (processing of rose wax)

[0228] Rose wax is collected as a byproduct of rose absolute production.

[0229] In the first step, 0.2 kg of ethanol (99.9% purity) was slowly added to 100 g of rose wax while stirring at 40 °C. Then, 1–5 g of immobilized lipase (1–5% by weight) was added. Different lipases were added in different experiments to study the effect of the immobilized lipase. The lipases added in different experiments and their amounts are summarized in Table 3.

[0230] Table 3. Different lipases used and their dosage

[0231]

[0232] The mixtures were stirred at 60°C (200 rpm) for 14 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with ethanol (99.9%), allowing for reuse.

[0233] The ethanol filtrate was cooled to 0°C and filtered to remove insoluble wax. It was then cooled again to -5°C and filtered to remove any remaining insoluble wax.

[0234] The clarified ethanol phase was concentrated under moderate heating (55°C to 60°C) under vacuum (250 mbar) to obtain 37–52 g of final product. The yield was 37–52 wt% based on the amount of rose wax used. The yields of different lipases are shown in Table 4 below. The final product was dissolved in ethanol (96%) at 10 wt% at 20°C. These products were reddish-brown viscous liquids with a pleasant rose aroma.

[0235] Table 4. Yields of rose wax products depending on the lipase used

[0236]

[0237] As shown in Tables 3 and 4, although the dosage of Novozym 435 lipase was significantly lower than that of Addzyme CalB191G lipase, its yield was comparable. Furthermore, although the dosage of Addzyme CalB191G lipase was the same as that of SIGMA76546 and SIGMA 52001 lipases, its yield was higher.

[0238] Table 5 shows the compounds obtained after treating rose wax as described above and their contents.

[0239] Table 5. Compounds and their contents (%, w / w) in treated rose wax, depending on the type and amount of lipase used.

[0240]

[0241]

[0242]

[0243] As can be seen from Table 5, the method according to the present invention can convert rose wax into ethanol-soluble fatty acid ethyl esters. Furthermore, Addzyme CalB 191G lipase yields the highest amount of fatty acid ethyl esters.

[0244] Example 4

[0245] Process according to the invention (processing of mimosa oil)

[0246] Mimosa oil is collected as a byproduct of mimosa extract production.

[0247] In the first step, 400 mL of ethanol (99.9% purity) was slowly added to 400 g of mimosa oil. Then, 2 g of NOVOZYME 435 (1% by weight) was added. The mixture was stirred at 60°C (200 rpm) for 24 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with a mixture of ethanol (99.9%) and cyclohexane, allowing for reuse.

[0248] The filtrate was concentrated under vacuum (250 mbar) at a temperature of 50°C to 60°C to obtain 220 grams of crude converted mimosa oil.

[0249] The crude converted oil (220 g) was mixed with 1.1 L of ethanol (96%) and poured into a decantation flask. After decantation at 15 °C for 10 hours, the insoluble heavy phase was removed. The upper phase was filtered and concentrated under vacuum (250 mbar) at 50 °C to 60 °C to give 109 g of final product, a yield of 27% by weight based on the initial amount of Mimosa oil added.

[0250] The final product was dissolved in ethanol (96%) at 10% by weight at 20°C. It is an amber-colored viscous liquid with a pleasant mimosa scent.

[0251] Table 6 shows the compounds obtained after treating mimosa oil using the above method and their contents.

[0252] Table 6. Compounds and their contents in the treated Mimosa oil

[0253]

[0254] As can be seen from Table 6, the method according to the present invention can convert mimosa oil into ethanol-soluble fatty acid ethyl esters, wherein the contents of ethyl palmitate and ethyl linoleate are particularly high.

[0255] Example 5

[0256] Process according to the invention (processing of tuberose wax)

[0257] Tuberose wax is collected as a byproduct of tuberose absolute production.

[0258] In the first step, 26g of molten tuberose wax was added to 120g of ethanol (99.9% purity). Then, 0.26g of NOVOZYME 435 (1% by weight) was added. The mixture was stirred at 40°C (200 rpm) under normal pressure for 24 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with ethanol (99.9%), thus allowing for reuse.

[0259] The ethanol filtrate was cooled to 0°C and then filtered to remove insoluble wax. It was then cooled again to -15°C and filtered to remove any remaining insoluble wax.

[0260] The clarified ethanol phase was concentrated under moderate heating (55°C to 60°C) under vacuum (250 mbar) to give 2.5 g of the final product. Based on the amount of tuberose wax used, the yield was 9.6% by weight. The final product was soluble in ethanol (96%) at 20°C at 10% by weight. It was an orange-brown semi-liquid with a pleasant tuberose aroma.

[0261] Comparative process (production of tuberose absolute)

[0262] Fresh tuberose flowers were extracted with hexane, and the solvent was evaporated to obtain tuberose extract. Then, 100 g of tuberose extract was melted at 50°C and mixed with 1 L of warm ethanol (96%) in a beaker. The mixture was stirred at 50°C for 1 hour. It was then cooled to 20°C and filtered (to remove solid cold wax). The filtrate was cooled to 0°C to precipitate the residual wax. The solid fraction (wax) was extracted again with 1 L of ethanol (96%) at 50°C to obtain a second ethanol extract. The second ethanol extract was cooled twice more (to 0°C and -10°C, respectively) and filtered. The ethanol filtrates were combined and concentrated under vacuum to obtain liquid tuberose absolute soluble in ethanol. The absolute yield was 27% by weight based on the initial extract used.

[0263] Process according to the invention (production of tuberose absolute)

[0264] In the first step, 10 g of tuberose extract was mixed with 100 mL of ethanol (99.9% purity). Then, 0.1 g of NOVOZYME 435 (1% by weight) was added. The mixture was stirred at 40°C (200 rpm) for 24 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with ethanol (99.9%), thus allowing for reuse.

[0265] The ethanol filtrate was cooled to 0°C and filtered to remove insoluble wax. The resulting ethanol filtrate was then cooled to -15°C and filtered to remove any remaining insoluble wax.

[0266] The clarified ethanol phase was concentrated under moderate heating (55°C to 60°C) under vacuum (250 mbar) to obtain 3.6 g of liquid neroli extract. Based on the initial extract used, the absolute yield was 36% by weight. The obtained absolute was an orange-brown paste with a pleasant tuberose aroma.

[0267] Therefore, compared with the comparative method, the method for preparing tuberose absolute according to the present invention has a significantly higher absolute yield (27% by weight vs. 36% by weight).

[0268] Table 7 lists the compounds present in the raw semen prepared according to the method of the present invention and the raw semen prepared by comparative methods, and their contents. All contents were determined by gas chromatography.

[0269] Table 7. Compounds and their contents in the raw semen prepared by the comparative method and the raw semen prepared according to the method of the present invention.

[0270]

[0271] As can be seen from Table 7, in the production process of tuberose absolute, compared with the comparative method, the method according to the present invention can obtain a higher content of fatty acid ethyl esters, thereby obtaining a higher yield of tuberose absolute.

[0272] Example 6 – Reuse of Immobilized Lipase

[0273] Process according to the invention (processing of fenugreek oil)

[0274] First test

[0275] 100 g of fenugreek oil was mixed with 0.2 L of ethanol (99.9% purity) by stirring. Then 10 g of NOVOZYME435 (10% by weight) was added. The mixture was stirred at 45 °C (200 rpm) for 6 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with ethanol (99.9%), thus allowing for reuse.

[0276] The ethanol filtrate was transferred to a separatory funnel to remove some of the insoluble matter (heavy phase). The clarified ethanol phase was then filtered and concentrated under vacuum (250 mbar) with moderate heating (55°C to 60°C) to give 91.4 g of the final extract. The treated extract was soluble in ethanol at 20°C at 10% by weight (96%). Based on the initial amount of fenugreek oil used, the yield of the fenugreek-soluble extract was 91.4% by weight. The alcohol content (ethanol) in the treated extract was less than 2% by weight.

[0277] Second test

[0278] 101 g of fenugreek oil was mixed with 0.2 L of ethanol (99.9% purity) by stirring. Then, 10 g of NOVOZYME435 (10% by weight) was added. The enzyme added was the same one used in the first experiment. The mixture was stirred at 45°C (200 rpm) for 6 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with a mixture of ethanol (99.9%), thus allowing for reuse.

[0279] The ethanol filtrate was transferred to a separatory funnel to remove some of the insoluble matter (heavy phase). The clarified ethanol phase was then filtered and concentrated under vacuum (250 mbar) with moderate heating (55°C to 60°C) to give 74.4 g of the final extract. The treated extract was soluble in ethanol (96%) at 20°C at 10% by weight. Based on the initial amount of fenugreek oil used, the yield of fenugreek soluble extract was 74.4% by weight. The alcohol content (ethanol) in the treated extract was less than 2% by weight.

[0280] Third test

[0281] 101.6 g of fenugreek oil was treated according to the above method (first and second experiments) to obtain 70.0 g of soluble extract. The enzyme used was the recovered enzyme used in the second experiment. Based on the initial amount of fenugreek oil, the yield of fenugreek soluble extract was 70% by weight. The alcohol content (ethanol) in the treated extract was less than 2% by weight.

[0282] Fourth test

[0283] 100.9 g of fenugreek oil was treated according to the above method (first and second experiments) to obtain 76.0 g of soluble extract. The enzyme used was the recovered enzyme used in the third experiment. Based on the initial amount of fenugreek oil, the yield of fenugreek soluble extract was 76% by weight. The alcohol content (ethanol) in the treated extract was less than 2% by weight.

[0284] Fifth test

[0285] 100 g of fenugreek oil was mixed with 0.2 L of ethanol (99.9% purity) by stirring. Then 1 g of NOVOZYME435 (1% by weight) was added. This enzyme was the recovered enzyme used in the fourth experiment. The mixture was stirred at 45 °C (200 rpm) for 30 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with a mixture of ethanol (99.9%).

[0286] The ethanol filtrate was transferred to a separatory funnel to remove some of the insoluble matter (heavy phase). The clarified ethanol phase was then filtered and concentrated under vacuum (250 mbar) with moderate heating (55°C to 60°C) to obtain 104 g of the final extract. The treated extract was soluble in ethanol (96%) at 20°C at 10 wt%. Based on the initial amount of fenugreek oil used, the yield of the fenugreek soluble extract was 104 wt%. The alcohol content (ethanol) in the treated extract was less than 2 wt%.

[0287] Table 8 shows the compounds produced after fenugreek oil treatment as described above and their contents.

[0288] Table 8. Compounds produced after processing fenugreek oil as described above and their contents.

[0289]

[0290] All extracts were homogeneous, brownish liquids with a typical nutty / maple pyrazine fenugreek aroma. The molecules producing this aroma were below the gas chromatography threshold but potent, similar to sotolon (CAS 28664-35-9).

[0291] Example 7

[0292] Process according to the invention (processing of arabian jasmine extract)

[0293] In the first step, 92g of molten jasmine wax was concentrated under vacuum in a rotary evaporator to remove residual solvent. The water bath temperature was approximately 60°C, and the vacuum degree was 300 mBar. While stirring at 40°C, 92g of 99.9% ethanol and 185g of cyclohexane were slowly added. Then, 1g of NOVOZYME 435 was added. The mixture was stirred at 45°C (20 rpm) and atmospheric pressure for 24 hours. The solution was then filtered to recover the enzyme. The immobilized enzyme was washed with a mixture of 99.9% ethanol, allowing for reuse.

[0294] In the second step, the ethanol filtrate is cooled to -15°C and then filtered to remove residual insoluble wax.

[0295] The clarified solution obtained by filtration was concentrated under moderate heating (55°C to 60°C) under vacuum (250 mbar) to obtain 13.3 g, with a wax yield of 3.3%.

[0296] The product is soluble in anhydrous ethanol at 20°C at 10% by weight. It is a yellow-orange semi-liquid with a pleasant Arabian jasmine aroma.

[0297] Table 9. Compounds obtained after treatment with Arabian jasmine as described above and their contents.

[0298]

[0299] Example 8

[0300] Process according to the invention (processing of gardenia (Gardenia tahitensis) extract)

[0301] In the first step, 16g of molten gardenia wax was concentrated under vacuum in a rotary evaporator to remove residual solvent. The water bath temperature was approximately 60°C, and the vacuum degree was 300 mBar. While stirring at 40°C, 16g of 99.9% ethanol and 31g of cyclohexane were slowly added. Then, 0.16g of NOVOZYME 435 was added. The mixture was stirred at 45°C (200 rpm) and atmospheric pressure for 24 hours. The solution was then filtered to recover the immobilized enzyme. The immobilized enzyme was washed with 99.9% ethanol solution, allowing for reuse.

[0302] In the second step, the ethanol filtrate is cooled to -15°C and then filtered to remove residual insoluble wax.

[0303] The clarified solution obtained by filtration was concentrated under vacuum (100 mbar) with moderate heating (55°C to 60°C) to obtain 0.68 g, with a wax yield of 0.37%.

[0304] The product is soluble in anhydrous ethanol at 20°C at a concentration of 10% by weight. It is a beige semi-liquid with a pleasant gardenia scent.

[0305] Example 9

[0306] Process according to the invention (processing of viola leaf extract)

[0307] In the first step, 100g of molten violet leaf wax was concentrated under vacuum in a rotary evaporator to remove residual solvent. The water bath temperature was approximately 60°C, and the vacuum degree was 300 mbar. While stirring at 40°C, 100g of 99.9% ethanol and 200g of cyclohexane were slowly added. Then, 1g of NOVOZYME 435 was added. The mixture was stirred at 45°C (200 rpm) and atmospheric pressure for 24 hours. The solution was then filtered to recover the immobilized enzyme. The immobilized enzyme was washed with 99.9% ethanol solution, allowing for reuse.

[0308] In the second step, the ethanol filtrate is cooled to -15°C and then filtered to remove residual insoluble wax.

[0309] The clarified solution obtained by filtration was concentrated under vacuum (100 mbar) with moderate heating (55°C to 60°C) to obtain 91.5 g, with a wax yield of 91%.

[0310] The product is soluble in anhydrous ethanol at 20°C at 10% by weight. It is a greenish-brown semi-liquid with a pleasant violet leaf aroma.

[0311] Table 10. Compounds obtained after the above-mentioned treatment of violet leaves and their contents.

[0312]

[0313] In the third step, various techniques are employed to improve the properties of the obtained violet leaf extract.

[0314] 0.1g of activated charcoal powder was mixed with 10g of violet leaf extract at 30°C, then filtered to remove the burnt and amino flavors and to slightly discolor the product. This yielded 9.5g of extract, with a final extract yield of 95%.

[0315] 0.1 g of sodium bicarbonate was mixed with 10 g of violet leaf extract at 30°C, and then filtered to remove the acidic flavor. This yielded 9.6 g of extract, with a final extract yield of 96%.

[0316] 20g of water was mixed with 20g of violet leaf extract to remove glycerol and improve the appearance of the product. The mixture was heated to 50°C and decanted after 12 hours. The lower layer was retained and filtered. 18.4g of extract was obtained, with a final extract yield of 92%.

[0317] 50g of violet leaf extract was decolorized by molecular distillation. The colorless product is easier to use in fragrance manufacturing. The rotary evaporator temperature was 120℃ and the vacuum degree was 0.001mbar. 29g of extract was obtained, with a yield of 58%.

Claims

1. A method for modifying a biomass extract containing fatty acids and fragrance and / or flavor compounds, the method comprising the following steps: - Provides a biomass extract containing fatty acids and aroma and / or flavor compounds, an alcohol solvent, and an immobilized lipase to form a reaction mixture; - Apply specific conditions to the reaction mixture to cause fatty acids to react with alcohol solvents to form fatty acid esters; - The immobilized lipase was removed from the reaction mixture to obtain a modified biomass extract.

2. The method according to claim 1, wherein the biomass extract is obtained by solvent extraction, supercritical fluid extraction, carbon dioxide extraction, steam distillation extraction, pressing or cold pressing extraction, fat extraction, maceration extraction and / or fractionation extraction, preferably solvent extraction, supercritical fluid extraction and / or carbon dioxide extraction.

3. The method according to any one of claims 1 and 2, wherein the biomass extract is an extract selected from the group consisting of: Spartium junceum, Dianthus sp., Acacia farnesiana, Magnolia champaca, Narcissus jonquilla, Elaeagnus sp., Helicrysum italicum, Plumeria sp., Gardenia sp., Gardenia tahitensis, Pelargonia gravolens, Hyacinthus, Iris sp., Jasminum grandifolium, Jasminum sambac, Lavandula Hybrid, narrow-leaved lavender (Lavandula angustifolia), lily (Lilium), ginger flower (Hedychium sp.), magnolia (Magnolia sp.), small marigold (Tagetes minuta), mimosa (Acacia dealbata), yellow daffodil (Narcissus pseudonarcissus), orange blossom (Citrus x aurantium), rose (Rosa sp.), tuberose (Agave amica), violet (Viola sp.).), mallow, fennel, star anise (Illicium verum), cardamom (Elettaria cardamomum), shrub-like pepper (Capsicum frutescens), caraway (Carum carvi), carrot (Daucus carota sativus), carob (Ceratonia silique), clove (Syzygium aromaticum), long pepper (Piper cubaba), fennel (Foeniculum vulgare), fenugreek (Trigonella foenum-graecum), ginger (Zingiber officinalis), juniper (Juniperus officinalis), nutmeg (Myristica fragrans), pepper (Capsicum annuum), patchouli (Pogostemon cablin), black pepper (Piper nigrum), saffron (Crocus sativus), coffee (Coffea sp. (Arabic or medium-fruit coffee)), rockrose (Cistus ladaniferus), elderberry (Sambucus The following herbs are included: *Imperata nigra*, *Anthoxanthum odoratum* (an annual spring grass), *Laurus nobilis*, *Pistacia lentiscus*, *Liatris odoratissima*, *Tilia sp.*, *Ilex paraguariensis*, *Dipterix odorata*, *Rosemarinus officinalis*, *Camelia sinensis*, *Thymus vulgaris*, *Nicotiana tabacum*, *Verbena officinalis*, and vanilla sp. (vanilla or Tahiti).

4. The method according to any one of claims 1 to 3, wherein the biomass extract is a fragrance extract or the wax portion of a fragrance essential oil.

5. The method according to any one of claims 1 to 4, wherein the alcohol solvent is selected from methanol, ethanol, propanol, butanol, pentanol, hexanol, and any mixture thereof, preferably, the alcohol solvent is ethanol.

6. The method according to any one of claims 1 to 5, wherein the immobilized lipase is an immobilized esterase, preferably an immobilized transesterase.

7. The method according to any one of claims 1 to 6, wherein, based on the total reaction mixture, the immobilized lipase in the reaction mixture is less than 50.0% by weight, preferably less than 10% by weight, more preferably less than 5% by weight.

8. The method according to any one of claims 1 to 7, wherein the weight ratio of biomass extract to alcohol solvent in the reaction mixture is 1:0.1 to 1:20, preferably 1:1 to 1:

5.

9. The method according to any one of claims 1 to 8, wherein the method further comprises a purification step, preferably cold filtration and / or decantation.

10. A biomass extract, preferably a fragrance and / or flavor composition, which can be obtained by the method of any one of claims 1 to 9.

11. The biomass extract according to claim 10, wherein the biomass extract is a primary essence.

12. The biomass extract according to claim 10 or 11, wherein the extract comprises fatty acid esters, preferably ethyl fatty acid esters, in an amount of at least 3% by weight, preferably at least 5% by weight, based on the total biomass extract.

13. A flavoring or seasoning composition comprising: -A biomass extract as defined in any one of claims 10 to 12; - At least one ingredient selected from the group consisting of a spice carrier and a spice base; and -Optionally, at least one spice adjuvant.

14. A flavored consumer product comprising a biomass extract as defined in any one of claims 10 to 12, or a flavoring or seasoning composition as defined in claim 13.

15. The scented consumer product according to claim 14, wherein the scented consumer product is a perfume, fabric care product, body care product, cosmetic preparation, skin care product, air care product, or home care product.

16. The scented consumer product of claim 15, wherein the scented consumer product is a fine perfume, a topical perfume or eau de toilette, cologne, shaving lotion or aftershave, optionally in capsule or tablet form, a liquid or solid detergent, a fabric softener, a liquid or solid fragrance enhancer, dry cleaning sheets, a fabric freshener, ironing lotion, paper, bleach, carpet cleaner, curtain care products, shampoo, leave-on or rinse-out hair conditioner, coloring agents, color care products, hair styling products, or dental products. Personal care products, disinfectants, feminine hygiene products, hair sprays, skin creams or lotions, cold creams, deodorants or antiperspirants, hair removal products, tanning agents, sunscreens or after-sun products, nail products, skin cleansers, cosmetics, soaps, shower or bath mousse, bath oils or shower gels, foot / hand care products, hygiene products, air fresheners, "ready-to-use" powder air fresheners, mold removers, furniture care products, wipes, dishwashing liquids or hard surface cleaners, leather care products, and car care products.

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