WO2023010227A1 - Procédé d'obtention d'un extrait de substances aromatisantes naturelles, saveur umami - Google Patents

Procédé d'obtention d'un extrait de substances aromatisantes naturelles, saveur umami Download PDF

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Publication number
WO2023010227A1
WO2023010227A1 PCT/CL2022/050070 CL2022050070W WO2023010227A1 WO 2023010227 A1 WO2023010227 A1 WO 2023010227A1 CL 2022050070 W CL2022050070 W CL 2022050070W WO 2023010227 A1 WO2023010227 A1 WO 2023010227A1
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WIPO (PCT)
Prior art keywords
umami
mushroom
mushrooms
liquid phase
flour
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PCT/CL2022/050070
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English (en)
Spanish (es)
Inventor
Freddy BOEHMWALD THIELEMANN
Denisse BENAVIDES JIMÉNEZ
Paulina FERNÁNDEZ VILLALOBOS
Miguel MARTÍNEZ COVARRUBIAS
María Ignacia FIERRO RUBILAR
Eduardo ZAVALA PULGAR
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Done Properly Company Spa
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Publication of WO2023010227A1 publication Critical patent/WO2023010227A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
    • A23L31/10—Yeasts or derivatives thereof
    • A23L31/15—Extracts
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives

Definitions

  • the present invention is part of the development of an extract of natural flavors, specifically UMAMI flavor based on mushrooms.
  • the UMAMI flavor has recently been identified as such, and it has been defined as the fifth flavor, that is, people could recognize between 5 different types of flavors: sweet, salty, bitter, acid and UMAMI.
  • the word UMAMI is a Japanese term meaning savory, and is described by its pleasant salty or meaty flavors naturally present in many plant and animal foods. Physiologically we know that UMAMI has effects on the tongue: it stimulates salivation and generates a physiological sensation associated with pleasant stimuli. Chemically we know that the Umami flavor is due to the specific receptors of the molecules responsible for the UMAMI flavor on the tongue, among which glutamate, aspartate and 5' nucleotides have been identified.
  • the UMAMI flavor has great applications in the food industry since. It is a flavor enhancer, as we indicated, it makes food “tasty”, but advantageously it does not contain sodium, so it constitutes a healthy replacement for salt, which has such a negative impact on people's health. Similar to salt, UMAMI enhances the flavors of food, so the variety of uses is very wide.
  • UMAMI flavor has been obtained from different sources, such as various vegetables including tomato, onion, mushrooms and seaweed.
  • the invention points to a new method for obtaining UMAMI from mushrooms, which stands out for its speed and product quality.
  • the process of the invention allows obtaining a concentrated product, without contaminating flavors of the source material, which gives greater versatility. to the composition obtained.
  • Zhao Zao, Yonggan, et al. "Progresses on processing methods of umami substances: A review.” Trends in Food
  • the subject of the invention is found within the enzymatic hydrolysis processes to obtain UMAMI, which differs from what is known in the prior art, in improving both the effectiveness and efficiency of the process.
  • the inventors have developed a rapid method to obtain UMAMI of excellent quality.
  • Said quality of the UMAMI flavoring extract obtained according to the process of the invention can be evaluated, for example, in the degree of hydrolysis of the peptides present in the product, which directly determines the organoleptic characteristics of the composition when used in the food processing. Additionally, the method of the invention is highly efficient in separating the flavoring extract from solid or discarded plant material. These improved characteristics are achieved by applying the steps and conditions of the method protected in the present invention.
  • FIG. 1 Process Diagram, stages of 1. Processing of Raw Materials are shown,
  • F1 Feeding of raw materials
  • F2 Discharge of hydrolyzate towards solid-liquid separation (Filtration)
  • F3 Filter discharge to Inactivation
  • F4 Inactivated filtering discharge towards Focus
  • F5 Discharge of concentrate to container
  • F7 discharge of concentrate to container
  • Figure 3 Graph of concentration of msg-like molecules according to type of hydrolysis.
  • the present invention aims at a two-step “flash” (fast) hydrolytic process that allows the production of natural food flavorings with a high content of umami molecules.
  • This process uses as raw materials different types of mushrooms and also mycelium of edible mushrooms. All these fungal materials have in common a high content of umami molecules and a similar cell wall structure, so they can be processed in a similar way.
  • the process consists in its first stage in providing dry fungal material crushed irregularly to a particle size that must vary between 10 and 500 pm, especially between 50 and
  • the fungal material processed in this way is mixed with water to a concentration between 20 and 200 g/L, especially between 50 and 150 g/L, and preferably between 50 and 100 g/L.
  • Figure 1 illustrates the natural umami flavor and biomaterial production process.
  • the processing of the raw material is carried out from fungal material, either mushrooms and/or mycelium. Where the main requirement is that they are edible fungi, by way of non-exhaustive example, the fungal material can be chosen from fungi of any of the species named below, either as homogeneous material, or as mixtures of them. Scientific name and common name included.
  • Flammulina velutipes Flammulina velutipes; enoki
  • Hericium erinaceus Hericium
  • Lactarius deliciosus N ⁇ scalo, Revolton
  • Pleurotus citripinoleatus yellow oyster
  • Pleurotus ostreatus oyster mushroom
  • the cleaned mushrooms are sliced and/or crushed, and are subjected to a drying or dehydration stage, for example, in a tray oven with air circulation, at 55 QC for 8 hours and then at 45oC for an additional 8 hours; or in a roller dryer or any other food drying-dehydrating system available in the art.
  • a drying or dehydration stage for example, in a tray oven with air circulation, at 55 QC for 8 hours and then at 45oC for an additional 8 hours; or in a roller dryer or any other food drying-dehydrating system available in the art.
  • mycelium this corresponds to fungi that grow in the form of hyphae that together form the mycelium, without developing a fruiting body or mushroom.
  • This mycelium is obtained through liquid fermentation.
  • the resulting biomass is separated from the fermentation medium, crushed to break it up and then dried in a similar way to that indicated for mushrooms.
  • the invention process begins. Where the first stage is to grind the fungal material irregularly, for example, in a mill that combines impact and shear principles.
  • the final particle size of the flour and the regrind must be heterogeneous and range between 50 and 500 ⁇ m, especially between 50 and 400 pm, and preferably between 100-300 pm. This condition must be fulfilled in more than 80% of the flour particles, preferably in more than 90%.
  • the irregular flour obtained in the first stage is subjected to 2 successive enzymatic digestions, for which the flour is mixed with water to a concentration of between 50 and 200 g/L of substrate, preferably between 50 and 100 g/L, in a reactor with constant stirring and anaerobic environment given, for example, by saturation with gaseous N2. Maintaining anaerobic conditions is crucial for the organoleptic properties of the composition of
  • the first has the objective of degrading the wall of the fungus, with an enzymatic cocktail of polysaccharide degrading enzymes, a high enzyme dose of between 0.5 to 2% w/w is used. , which is equivalent to a concentration ranging from 5,000 to 100,000 units per gram.
  • the operating parameters of this phase are: temperature between 30"C to 60'C, pH between 4.5 to 6.0, stirring speed between 200 RPM to 1500 RPM.
  • the reaction time of this digestion stage develops in a range of 2 to 4 hours.
  • the enzyme cocktail consists of p-glucanase, in one or more varieties and optionally cellulases, hemicellulases, xylanases, arabanase. Any combination of these enzymes that exists in the art can be used.
  • the second phase consists of the degradation of the protein components of the fungus by means of an enzymatic cocktail of proteases. For this, a 5% dose and a concentration ranging from 50,000 to 100,000 units per gram or liter are used.
  • the operation parameters of this phase are: temperature between 40"C to 60°C, pH between 5.0 to 6.5, reaction rate between
  • reaction time of this digestion stage takes place in a range of
  • the enzyme cocktail consists of endo and/or exo peptidases. Any combination of proteases that exists in the art can be used.
  • the liquid phase comprising the flavoring composition
  • the liquid phase must be separated from the discarded solid phase.
  • this process could be carried out by any method that allows an adequate separation of phases, on an industrial scale, the most convenient is filtration, where the irregular particle size becomes relevant since it allows this process to be accelerated, avoiding filter clogging by too compact sediment, and at the same time reduce operating costs.
  • Filtration can be carried out by any system known in the state of the art, such as a horizontal hydraulic filter press or a vertical filter press, for example. Filtering has an estimated operating time of 30 to 60 minutes, and is conveniently carried out at a temperature between 30-50oC, until the liquid phase is completely separated and a discard dry cake is obtained.
  • the liquid phase is then subjected to enzyme inactivation, which at the same time makes it possible to eliminate possible contamination with microorganisms or cells of the processed fungus. Stopping the enzymatic activity is essential so that the product does not interact unduly with the food with which it will be used, preventing it from modifying its polysaccharide or protein content, and concomitantly its organoleptic properties.
  • sonication of the liquid phase is performed, in conjunction with a moderate increase in temperature. This produces sonoporation, where cell walls and membranes are perforated and cavitation generates shock wave differences and shear stress.
  • This sonication at moderately high temperature is carried out for a time of between 10 and
  • composition of the invention is subjected to concentration by evaporation.
  • a negative pressure between 60 and 80 kPa is preferably used at temperatures between 40-
  • the evaporated water is recirculated, as an input to be mixed with the mushroom meal for the mixture that is subjected to enzymatic digestion.
  • the product obtained is the composition or extract of the invention, which has an umami flavor, with salty notes, being mushroom-based, it has carbohydrates that provide a slightly sweet nuance, giving a slight hint of caramel. It gives a feeling of filling in the mouth and also produces an increase in salivation.
  • the product obtains a variable color in a range of earth colors, from honey to dark brown, depending on the type of fungus or mycelium used as starting material.
  • Example 1 Obtaining UMAMI from mycelium.
  • a liquid fermentation of Aspergillus oryzae fungus mycelium was obtained, with a growth medium based on potato extract supplemented with glucose and minerals, at a temperature of 32oC.
  • the resulting biomass was separated from the fermentation medium, obtaining 1 kg of wet biomass, which was laminated using an industrial vegetable and fruit processor.
  • the laminated material was dehydrated in a tray oven with air circulation, at 55oC for 8 hours and then at 45oC for an additional 8 hours.
  • the dry biomass was processed with a mill that combines impact and shear principles.
  • the final particle size of the flour and the grinding ranged between 100-300 ⁇ m, 150 g of flour was obtained.
  • the flour was brought to a suspension of 80 g/L with distilled water, in a reactor with a nitrogen atmosphere and constant stirring at 900 rpm, temperature of 55*C and pH 5 controlled by means of a pH sensor coupled to peristaltic pumps for the supply of alkaline (NaOH, KOH) or acid (HCI) solutions as required.
  • An enzyme cocktail was added to obtain a concentration of 1000 units per gram of reaction and incubated for 2.5 hours.
  • the solid-liquid separation was carried out with a vertical filter press equipped with a cloth
  • the liquid phase was transferred to a jacketed tank that kept the temperature at 60*C and was sonicated at 18-26 kHz for 12 minutes. (Ultrasonic power: 200-400
  • the concentration of the product was carried out at a reduced pressure of 70 kPa, and 60*C, until reaching a product of 30o Brix.
  • Example 2 Obtaining UMAMI from mushrooms.
  • One kilo of champignon mushrooms (Agaricus campestr ⁇ s) was obtained, which is laminated using an industrial vegetable and fruit processor slicer.
  • the laminated material was dehydrated in a tray oven with air circulation, at 55oC for 8 hours and then at 45oC for an additional 8 hours.
  • the crushed and dried biomass was processed with a mill that combines impact and shear principles.
  • the final particle size of the flour and the grinding ranged between 100-300 ⁇ m, 100 g of flour was obtained.
  • the flour was brought to a suspension of 80 g/L with distilled water, in a reactor with a nitrogen atmosphere and constant stirring at 900 rpm, a temperature of 55*C and pH 5, controlled by means of a pH sensor coupled to peristaltic pumps to supply of alkaline (NaOH, KOH) or acid (HCI) solutions as required.
  • An enzyme cocktail was added to obtain a concentration of 1000 units per gram of reaction and incubated for 3 hours.
  • An enzyme cocktail containing a mixture of p-glucanases was used. (1.3-1.4-B- glucanase (EC 3.2.1.6), 1.3-B-glucanase (EC 3.2.1.6), 1.2-1.4-B-glucanase, 1.4-B-glucanase (EC
  • the solid-liquid separation was carried out with a vertical filter press equipped with a cloth
  • the liquid phase was transferred to a jacketed tank that kept the temperature at 90'C with constant agitation for 30 minutes.
  • the concentration of the product was carried out at a reduced pressure of 80 kPa, and 60°C, until reaching an amount of 30" Brix.
  • a first comparative example was carried out between the method of the invention that uses dry and irregularly ground fungal material, mushrooms or mycelia versus the same method, but using dry and regularly ground fungal material, mushrooms or mycelia. And we proceeded to compare the effects of different particle sizes of mushroom flour and/or mycelium flour on the umami manufacturing process, the availability of free amino acids after filtration and the organoleptics when applying the UMAMI extract obtained in masses. fermented white bread type, with a panel of experts. l-a) Materials and methods.
  • the size distribution for homogeneous and small flour is:
  • the size distribution for the irregular flour of the invention is:
  • Enzymatic digestion A Paris mushroom meal (Agaricus bisporus) was suspended in water at a concentration of 80 g/L and pH 5.5.
  • the first enzyme set ⁇ -glucanase, arabanase, cellulase, hemicellulase and xylanase was added at a concentration of 1% w/w and incubated at 50°C and 500 rpm for 2 hours.
  • the pH was adjusted to 6.5 and the second enzyme group (endo and exo proteases) was added to a final concentration of 5% w/w and incubated at 50°C and 500 rpm for 2 hours.
  • the second enzyme group endo and exo proteases
  • h corresponds to the total mEq of leucine in the hydrolyzed sample and ht corresponds to the amount of mEq of leucine in the fully hydrolyzed sample.
  • Concentration The liquid resulting from the inactivation was concentrated in a rotary evaporator coupled to a vacuum pump at 60°C and 600 mmHg until reaching 30 *Brix. The concentrated product was stored at 4°C until its application.
  • Table 1 Unloading time by gravity of filtrate and weight of wet cake obtained after filtration, for each particle size distribution analyzed for mushroom meal.
  • the characteristic of the process of the invention of using flour of irregular size makes it more efficient in separating the extract or supernatant from the solid waste material, even applying pressure.
  • the degree of hydrolysis was determined in triplicate in each case.
  • the average GH in both cases was 70.5% for the regular-small particle size flour (size distribution No. 1) and 81.9% for the umami obtained from the irregular size flour ( size distribution No. 2) of the invention. This means that the inventive feature of an irregular particle size in the flour used surprisingly allows better hydrolysis of plant material.
  • Table 2 shows the standard deviation of the tasting test, where sample 313 (100% salt) is the reference with which the two samples to be evaluated are compared.
  • Sample 398 regular flour
  • 366 irregular flour, invention
  • the particle size of the mushroom flour directly influenced the power of the umami flavor obtained at the end of the process.
  • An irregular particle size favored the production of UMAMI by decreasing the filtering time and increasing the amount of free amino acids. This is complemented by the results obtained in the organoleptic tests where a greater power of flavor is obtained, detection of aromas and flavors with notes of mushroom with the flour of irregular particle size.
  • the bottle was incubated with orbital shaking at 50°C and 150 rpm for 4 hours.
  • Enzymatic digestion Two similar solutions of paris mushroom meal in water were prepared at a concentration of 80 g/L and pH 5.5. An enzyme group (arabanase, cellulase, b-glucanase, hemicellulase and xylanase) was added to one of the solutions at a concentration of 1% w/py was incubated at 50°C and 500 rpm for 2 hours. The second enzymatic group (endo and exo proteases) was then added at a final concentration of 5% w/p and incubated at
  • the other mushroom meal solution was treated with both sets of enzymes simultaneously, the first containing arabanase, cellulase, b-glucanase, hemicellulase and xylanase, the second containing endo and exo proteases, at a concentration of 1% w/w and
  • Enzymatic digestion was carried out at 50°C and 500 rpm for 4 hours.
  • msg-like molecules were quantified by analyzing the samples of chemical digestion and enzymatic digestion, using liquid chromatography coupled to a mass spectrometer (LC/MS) equipped with a quadrupole. This technique makes it possible to quantify the free amino acids present in the sample (mg free aas/g sample).
  • LC/MS mass spectrometer
  • the enzymatic and chemical digestion processes were carried out with the aim of degrading the cell content and releasing free amino acids, specifically msg-like molecules such as aspartic acid, glutamate and 5'-ribonucleotides. Once these amino acids were released, their content was analyzed using two methods: liquid chromatography coupled with Mass Spectrometer (LS/MS) and spectrophotometry using the OPA method. Chemical hydrolysis was used as a control to compare the effects of 1-step and 2-step enzymatic hydrolysis.
  • LS/MS Mass Spectrometer
  • the degree of hydrolysis (GH) was determined in each case in triplicate in order to complement the previous result.
  • the %GH average was 55.9% for the one-step hydrolysis and 81.6% for the two-step hydrolysis according to the method of the invention. That is, the efficiency of the process improves practically by 50% when carrying out the hydrolysis as determined by the method of the invention.
  • 1 step has a lower organoleptic evaluation in terms of flavor power and less detection of aromas and flavors with mushroom notes compared to the 2-step process of the invention (Table 4). It can be seen that the score of sample 4532 (umami with 2-step digestion of the invention) is higher than the score of sample 4886 (umami with 1-step digestion) in both salty and umami flavors. In addition, sample 4886 presents a higher degree of acidity as indicated by the judges, which is an important point to consider in any matrix.
  • the application of enzymes in the hydrolysis stage directly influences the amount of umami molecules that are obtained at the end of the process.
  • it favors the obtaining of umami molecules and improves the organoleptic quality of the applied product in terms of flavor and aroma. This is evidenced in the comparison of msg-like molecules between one process or another, compared to a hydrolysis chemistry.
  • enzymatic digestion in two steps allows obtaining a higher degree of hydrolysis (GH) and increasing the amino acid content.

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  • Health & Medical Sciences (AREA)
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Abstract

La présente invention concerne une méthode pour élaborer un extrait de substance aromatisante saveur umami, à partir de champignons, qui comprend les étapes suivantes consistant à: (a) fournir de la matière fongique, de champignons ou de mycéliums, sèche et broyée de manière irrégulière ; (b) soumettre la matière à 2 digestions enzymatiques successives dans un environnement anaérobie, chacune durant de 2 à 4 heures; (c) filtrer, de manière à séparer la phase liquide; (d) inactiver les enzymes de la phase liquide; et (e) concentrer par évaporation la phase liquide pour obtenir la composition aromatisante saveur umami à partir de champignons. On protège également l'extrait aromatisant saveur umami provenant de champignons qui est obtenu pr cette méthode.
PCT/CL2022/050070 2021-08-06 2022-07-18 Procédé d'obtention d'un extrait de substances aromatisantes naturelles, saveur umami WO2023010227A1 (fr)

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CL2086-2021 2021-08-06
CL2021002086A CL2021002086A1 (es) 2021-08-06 2021-08-06 Proceso para la producción de un extracto de saborizantes naturales, umami.

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104223045A (zh) * 2014-09-16 2014-12-24 广东东阳光药业有限公司 一种食用菌鲜味物质的提取方法
WO2019002550A1 (fr) * 2017-06-30 2019-01-03 Oterap Holding B.V. Exhausteur de goût culinaire
CN110447877A (zh) * 2019-09-12 2019-11-15 浙江工业大学 一种功能性食用菌调味粉及其制备方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104223045A (zh) * 2014-09-16 2014-12-24 广东东阳光药业有限公司 一种食用菌鲜味物质的提取方法
WO2019002550A1 (fr) * 2017-06-30 2019-01-03 Oterap Holding B.V. Exhausteur de goût culinaire
CN110447877A (zh) * 2019-09-12 2019-11-15 浙江工业大学 一种功能性食用菌调味粉及其制备方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
POOJARY MAHESHA M., ORLIEN VIBEKE, PASSAMONTI PAOLO, OLSEN KARSTEN: "Enzyme-assisted extraction enhancing the umami taste amino acids recovery from several cultivated mushrooms", FOOD CHEMISTRY, ELSEVIER LTD., NL, vol. 234, 1 November 2017 (2017-11-01), NL , pages 236 - 244, XP093033331, ISSN: 0308-8146, DOI: 10.1016/j.foodchem.2017.04.157 *

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