NL2031945B1 - Fully soluble edible fungus protein and preparation method thereof - Google Patents

Fully soluble edible fungus protein and preparation method thereof Download PDF

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NL2031945B1
NL2031945B1 NL2031945A NL2031945A NL2031945B1 NL 2031945 B1 NL2031945 B1 NL 2031945B1 NL 2031945 A NL2031945 A NL 2031945A NL 2031945 A NL2031945 A NL 2031945A NL 2031945 B1 NL2031945 B1 NL 2031945B1
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protein
edible fungus
consumable
fungal protein
preparation
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Wang Tao
Zong Yucheng
Chen Zhengxing
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Univ Jiangnan
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    • 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
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/008—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/20—Proteins from microorganisms or unicellular algae
    • 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
    • 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
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/195—Proteins from microorganisms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
    • C07K14/375—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from Basidiomycetes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus

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Abstract

The present invention discloses a fully soluble edible fungus protein and a preparation method thereof. The fully soluble edible fungus protein is prepared by unfolding an unmodified edible fungus protein under a basic condition and adjusting pH to neutral by adding acid. The preparation method of the present invention does not alter the protein primary structure and can retain the nutritional value of protein to the greatest extent. In addition, the obtained edible fungus protein has no exogenous ingredients added, which improves the acceptance of consumers to the greatest extent, achieving the rational utilization of by-products of edible fungus, and effectively avoiding resource waste and environmental pollution.

Description

FULLY SOLUBLE EDIBLE FUNGUS PROTEIN AND PREPARATION METHOD THEREOF
TECHNICAL FIELD The present invention relates to the field of edible mycoprotein in-depth processing technologies, and in particular, to a fully soluble edible fungus protein and a preparation method thereof.
BACKGROUND As world population explodes, the demand for protein is growing. On one hand, there is still a serious shortage of protein resources in developing countries. On the other hand, the upgraded nutritional needs in developed countries require refined and in-depth processing with high value-added for existing proteins. At present, there still are many high-quality protein resources waiting to be developed across the world, especially plant-based protein resources, which have not been widely utilized due to their low solubility. Fungi are a kind of fast-growing and high-yielding biological resources. In China, well-developed technologies for the artificial cultivation of a variety of large edible fungi already exist. However, most of the stems and stipes of the large edible fungi remain after the fungi's harvest, and are used only as fungus residue in animal feeding. The fungus residue however has the same physicochemical composition as the bulk of the edible fungus, while containing an equivalent amount of protein (about 30%), and it has a protein extraction rate no different from other parts (about 80%) as measured by an alkali- solution and acid-isolation method. Due to its inferior quality and low protein solubility, the fungus residue cannot be sold as a commodity, resulting in a great waste of resources and environmental pollution. Therefore, by increasing the solubility of edible fungus protein, the edible fungus can be comprehensively utilized, the application scope of the edible fungus protein can be expanded, the industrial chain of the edible fungus can be deepened, and the economic value- added of the edible fungus can be increased. At present, the solubilization technologies for proteins mainly focus on biological enzymic modification, chemical modification, and physical modification. The enzymic modification of protein is characterized by the enzymolysis of large protein molecule, which breaks up its peptide chain to form small molecular peptide sections, exposing more polar groups to disperse in water, thereby achieving the solubilization of hydrophobic proteins. However, enzymolysis will bring about changes in the nutritional values of protein, changes in the functional properties, and production of bitter peptides. In contrast, the chemical modification is characterized by chemically enabled reaction of protein groups, which enhances the polarity of their chemical groups, thereby achieving the solubilization. However, strong chemical reactions may damage the protein's structure, reducing its nutritional values. In addition, residual chemical reagents may also lead to potential food safety issues. The physical modification method is often used in combination with the enzymic modification and chemical modification, and it is similar in nature to the two methods described above. Moreover, the high investment and high energy consumption of the physical modification greatly increase the difficulty of its popularization.
Overall, although the existing protein solubilization technologies can affect protein solubilization, it is still difficult to overcome the disadvantage of protein structure damage in the preparation process. Therefore, there remains a need to find a new alternative the conventional solubilization technologies.
SUMMARY In view of the disadvantages of the related art, the present invention provides a fully soluble edible fungus protein and a preparation method thereof. The present invention achieves the solubilization of edible fungus protein by changing the hydrophobicity of the protein secondary structure, to prepare a fully soluble edible fungus protein. The method of the present invention does not alter the protein primary structure and can retain the nutritional value of protein to the greatest extent. In addition, the obtained edible fungus protein has no exogenous ingredients added, which improves the acceptance of consumers to the greatest extent, achieving the rational utilization of by-products of edible fungus, and effectively avoiding resource waste and environmental pollution.
The technical solution of the present invention is as follows: Provided is a fully soluble edible fungus protein. The fully soluble edible fungus protein is prepared by unfolding an unmadified edible fungus protein under a basic condition and adjusting pH to neutral by adding acid, achieving the embedding of hydrophobic groups and the exposure of hydrophilic groups, thereby forming a hydrocolloid with a hydrophobic core and achieving the solubilization of edible fungus protein. The unmodified edible fungus protein is an edible fungus protein with a hydrophilic-hydrophobic staggered flexible chain structure.
The edible fungus is one of Pleurotus eryngii, Lentinula edodes, and Hericium erinaceus (Bull.) Pers. The edible fungus protein may be extracted from the edible part of the edible fungus, or may be extracted from the by-product of the edible fungus, that is, the stems and stipes of the edible fungus used in feeding.
Provided is a method for preparing a fully soluble edible fungus protein, including the following steps: (1) extracting an edible fungus protein; (2) adding water into the edible fungus protein extracted in step (1) and stirring, and adding an alkaline solution to adjust pH; (3) stirring the solution obtained after the pH adjustment in step (2) to expose the secondary structure of the protein; (4) adding a hydrochloric acid solution dropwise to adjust pH; (5) heating then cooling the solution obtained after the pH adjustment in step (4); (6) placing the cooled solution into a dialysis bag for dialysis; and (7) centrifuging the dialyzed solution to obtain a supernatant, and freeze- drying the supernatant to obtain the fully soluble edible fungus protein.
Further, in step (1), the edible fungus protein is extracted by an alkali-solution and acid-isolation method.
Further, in step (2}, the mass ratio of edible fungus protein to water is {(0.1-10):100; the stirring is carried out at 500-900 r/min for 10-30 min; the alkaline solution is NaOH solution with a molar concentration of 0.1-1 mol/L; and the pH = 11. Further, in step (8), the stirring is carried out at 500-900 r/min for 80-180 min.
Further, in step (4), the molar concentration of hydrochloric acid in the hydrochloric acid solution is 0.1-4 mol/L; and the pH value is 7.0. Further, in step (5), the heating is carried out at 95-121°C for 5-45 min.
Further, in step (6), the molecular weight cut-off of the dialysis bag is 3500 Da, and the dialysis is carried out for 18-36 h. Further, in step (7), the centrifugation is carried out at 4000-8000 x g for 15-25 min; and the freeze-drying is carried out at -60--40°C for 2-4 days. The beneficial technical effects of the present invention are as follows: (1) The present invention can achieve hydrophilic modification only through the hydrophobized self-assembly of the edible fungus protein structure without the help of foreign protein, ensuring the purity of the product from the protein source.
(2) In the present invention, the surface hydrophobic groups are passivated by heating, leading to a lower turbidity, increasing the stability of the fully soluble protein.
(3) Compared with conventional protein modification methods, the method for preparing the fully soluble edible fungus protein of the present invention has the advantages of simple operation, low energy consumption, low requirements on process and equipment, and high feasibility of industrial application; the conditions are mild, and the reagents used are all food aids with no risk of chemical reagent residues.
(4) In the present invention, the preparation principle of structurally modifying the hydrophobic protein secondary structure is novel. First, the structure of the insoluble protein is sufficiently unfolded in a basic environment through hydration and dispersion; secondly, the hydrophobic secondary structure of the edible fungus protein is altered under hydrophobic interaction during acid-base neutralization; and finally, during neutralization, the hydrophobic protein is reconstructed to form new secondary and tertiary structures, making its hydrophilic regions more exposed and hydrophobic regions embedded, achieving strong stability of the water dispersion system.
(5) The fully soluble edible fungus protein prepared by the method in the present invention can completely retain its main subunit structure, so that its primary structure is not damaged, indicating that the amino acid composition of the edible fungus protein is complete, so that the nutritional properties and functional properties are well preserved, which has broad application prospects. (6) In the present invention, the fully soluble edible fungus protein is 5 prepared by sufficiently unfolding an unmodified edible fungus protein under a basic condition and adjusting pH back to neutrality. Through this process, the edible fungus protein achieves the embedding of hydrophobic groups and the exposure of hydrophilic groups under the drive of hydrophobic interaction, thereby forming a hydrocolloid with a hydrophobic core and increasing the solubility of the protein.
BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing comparison of turbidity between the fully soluble Pleurotus eryngii protein prepared in Example 1 of the present invention and the unmodified Pleurotus eryngii protein. FIG. 2 is a graph showing comparison of turbidity between the fully soluble Lentinula edodes protein prepared in Example 2 of the present invention and the unmodified Lentinula edodes protein. FIG. 3 is a graph showing comparison of turbidity between the fully soluble Hericium erinaceus (Bull.) Pers protein prepared in Example 3 of the present invention and the unmodified Hericium erinaceus (Bull.) Pers protein. FIG. 4 shows images for comparison between the unmodified and modified edible fungus proteins prepared in Examples 1-3 of the present invention, where: a shows the modified fully soluble Pleurotus eryngii protein; b shows the modified fully soluble Lentinula edodes protein; c shows the modified fully soluble Hericium erinaceus {Bull.) Pers protein; d shows the unmodified Pleurotus eryngii protein; e shows the unmodified Lentinula edodes protein; and f shows the unmodified Hericium erinaceus (Bull) Pers protein.
DETAILED DESCRIPTION The present invention is specifically described below with reference to the accompanying drawings and examples, but the present invention is not limited to these examples. Unless otherwise specified, the experimental methods used in the following examples are all conventional methods.
Unless otherwise specified, the materials and reagents used in the following examples are all commercially available. Example 1 A method for preparing a fully soluble edible fungus protein includes the following steps: (1) Unmodified protein extraction: The remaining stems and stipes of Pleurotus eryngii picked were ground by a blade grinder and then filtered by an 80- mesh filter to obtain a powder, and the powder and deionized water were mixed in aratio of 1:10 (w/v), adjusted with 1 mol/L NaOH to pH 12, and stirred at room temperature for 4 h, and then centrifuged at 10000 x g at 4°C for 30 min to obtain a supernatant. Then, the supernatant was adjusted with 1 mol/L HCI while stirring to pH 3.6, and centrifuged at 10000 x g at 4°C for 20 min, to obtain a protein precipitate. The collected protein was washed with deionized water three times, then dispersed in a proper amount of deionized water, and adjusted with 1 mol/L NaOH to pH 7, and then freeze-dried to obtain an unmodified Pleurotus eryngii protein.
(2) The unmodified Pleurotus eryngii protein prepared in step (1) and distilled water were mixed in a mass ratio of 2:100 and stirred at 500 r/min for 10 min for hydration. 0.1 mol/L NaOH solution was added to adjust pH to 11.
(3) The solution obtained in step (2) was stirred at 900 r/min for 120 min, so that the high-level spatial structure of the Pleurotus eryngii protein is sufficiently unfolded to expose the secondary structure of the protein.
(4) 0.1 mol/L HCI solution was added dropwise to adjust pH to 7.0, so that a new spatial structure is formed through hydrophobic interaction after the molecular spatial structure of the Pleurotus eryngii protein is unfolded, so as to achieve the embedding of hydrophobic groups and the exposure of hydrophilic groups.
(5) The solution obtained in step (4) was heated at 121°C for 5 min and then rapidly cooled in an ice bath.
(6) The solution obtained in step (5) was dialyzed in a dialysis bag with a molecular weight cut-off of 3500 Da for 24 h.
(7) The solution obtained in step (6) was centrifuged at 8000 x g for 20 min to obtain a supernatant.
(8) The supernatant obtained in step (7) was freeze-dried at —40°C for 2 days to obtain a fully soluble Pleurotus eryngii protein.
Example 2 A method for preparing a fully soluble edible fungus protein includes the following steps: (1) Unmodified protein extraction: The remaining stems and stipes of Lentinula edodes picked were ground by a blade grinder and then filtered by an 80- mesh filter to obtain a powder, and the powder and deionized water were mixed in a ratio of 1:10 (w/v), adjusted with 1 mol/L NaOH to pH 12, and stirred at room temperature for 4 h, and then centrifuged at 10000 x g at 4°C for 30 min to obtain a supernatant. Then, the supernatant was adjusted with 1 mol/L HCI while stirring to pH 4.4, and centrifuged at 10000 x g at 4°C for 20 min, to obtain a protein precipitate. The collected protein was washed with deionized water three times, then dispersed in a proper amount of deionized water, and adjusted with 1 mol/L NaOH to pH 7, and then freeze-dried to obtain an unmodified Lentinula edodes protein.
(2) The unmodified Lentinula edodes protein prepared in step (1) and distilled water were mixed in a mass ratio of 1:100 and stirred at 700 r/min for 20 min for hydration. 0.5 mol/L NaOH solution was added to adjust pH to 11.
(3) The solution obtained in step (2) was stirred at 900 r/min for 120 min, so that the high-level spatial structure of the Lentinula edodes protein is sufficiently unfolded to expose the secondary structure of the protein.
(4) 0.1 mol/L HC! solution was added dropwise to adjust pH to 7.0, so that a new spatial structure is formed through hydrophobic interaction after the molecular spatial structure of the Lentinula edodes protein is unfolded, so as to achieve the embedding of hydrophobic groups and the exposure of hydrophilic groups.
(5) The solution obtained in step (4) was heated at 100°C for 30 min and then rapidly cooled in an ice bath.
(8) The solution obtained in step (5) was dialyzed in a dialysis bag with a molecular weight cut-off of 3500 Da for 24 h.
(7) The solution obtained in step (6) was centrifuged at 8000 x g for 20 min to obtain a supernatant.
(8) The supernatant obtained in step (7) was freeze-dried at -50°C for 3 days to obtain a fully soluble Lentinula edodes protein.
Example 3
A method for preparing a fully soluble edible fungus protein includes the following steps: (1) Unmodified protein extraction: The remaining stems and stipes of Hericium erinaceus (Bull.) Pers picked were ground by a blade grinder and then filtered by an 80-mesh filter to obtain a powder, and the powder and deionized water were mixed in a mass ratio of 1:10 (w/v), adjusted with 1 mol/L NaOH to pH 12, and stirred at room temperature for 4 h, and then centrifuged at 10000 x g at 4°C for 30 min to obtain a supernatant. Then, the supernatant was adjusted with 1 mol/L HCI while stirring to pH 3.0, and centrifuged at 10000 x g at 4°C for 20 min, to obtain a protein precipitate. The collected protein was washed with deionized water three times, then dispersed in a proper amount of deionized water, and adjusted with 1 mol/L NaOH to pH 7, and then freeze-dried to obtain an unmodified Hericium erinaceus (Bull.) Pers protein.
(2) The unmodified Hericium erinaceus (Bull.) Pers protein prepared in step (1) and distilled water were mixed in a mass ratio of 1:100 and stirred at 900 r/min for 30 min for hydration. 1 mol/L NaOH solution was added to adjust pH to 11.
(8) The solution obtained in step (2) was stirred at 900 r/min for 120 min, so that the high-level spatial structure of the Hericium erinaceus (Bull) Pers protein is sufficiently unfolded to expose the secondary structure of the protein.
(4) 0.1 mol/L HCI solution was added dropwise to adjust pH to 7.0, so that a new spatial structure is formed through hydrophobic interaction after the molecular spatial structure of the Hericium erinaceus (Bull.} Pers protein is unfolded, so as to achieve the embedding of hydrophobic groups and the exposure of hydrophilic groups.
(5) The solution obtained in step (4) was heated at 95°C for 45 min and then rapidly cooled in an ice bath.
(6) The solution obtained in step (5) was dialyzed in a dialysis bag with a molecular weight cut-off of 3500 Da for 24 h.
(7) The solution obtained in step (6) was centrifuged at 8000 x g for 20 min to obtain a supernatant.
(8) The supernatant obtained in step (7) was freeze-dried at —60°C for 4 days to obtain a fully soluble Hericium erinaceus (Bull) Pers protein.
Example 4
A method for preparing a fully soluble edible fungus protein includes the following steps: (1) Unmodified protein extraction: The remaining stems and stipes of Pleurotus eryngii picked were ground by a blade grinder and then filtered by an 80- mesh filter to obtain a powder, and the powder and deionized water were mixed in a ratio of 1:10 (w/v), adjusted with 1 mol/L NaOH to pH 12, and stirred at room temperature for 4 h, and then centrifuged at 10000 x g at 4°C for 30 min to obtain a supernatant. Then, the supernatant was adjusted with 1 mol/L HCI while stirring to pH 3.6, and centrifuged at 10000 x g at 4°C for 20 min, to obtain a protein precipitate. The collected protein was washed with deionized water three times, then dispersed in a proper amount of deionized water, and adjusted with 1 mol/L NaOH to pH 7, and then freeze-dried to obtain an unmodified Pleurotus eryngii protein.
(2) The unmodified Pleurotus eryngii protein prepared in step (1) and distilled water were mixed in a mass ratio of 10:100 and stirred at 600 r/min for 25 min for hydration. 0.6 mol/L NaOH solution was added to adjust pH to 12.
(3) The solution obtained in step (2) was stirred at 500 r/min for 180 min, so that the high-level spatial structure of the Pleurotus eryngii protein is sufficiently unfolded to expose the secondary structure of the protein.
(4) 0.4 mol/L HCI solution was added dropwise to adjust pH to 7.0, so that a new spatial structure is formed through hydrophobic interaction after the molecular spatial structure of the Pleurotus eryngii protein is unfolded, so as to achieve the embedding of hydrophobic groups and the exposure of hydrophilic groups.
(5) The solution obtained in step (4) was heated at 95°C for 25 min and then rapidly cooled in an ice bath.
(6) The solution obtained in step (5) was dialyzed in a dialysis bag with a molecular weight cut-off of 3500 Da for 18 h.
(7) The solution obtained in step (6) was centrifuged at 4000 x g for 25 min to obtain a supernatant.
(8) The supernatant obtained in step (7) was freeze-dried at —45°C for 3 days to obtain a fully soluble Pleurotus eryngii protein.
Example 5 A method for preparing a fully soluble edible fungus protein includes the following steps:
(1) Unmodified protein extraction: The remaining stems and stipes of Pleurotus eryngii picked were ground by a blade grinder and then filtered by an 80- mesh filter to obtain a powder, and the powder and deionized water were mixed in a ratio of 1:10 (w/v), adjusted with 1 mol/L NaOH to pH 12, and stirred at room temperature for 4 h, and then centrifuged at 10000 x g at 4°C for 30 min to obtain a supernatant. Then, the supernatant was adjusted with 1 mol/L HCI while stirring to pH 3.8, and centrifuged at 10000 x g at 4°C for 20 min, to obtain a protein precipitate. The collected protein was washed with deionized water three times, then dispersed in a proper amount of deionized water, and adjusted with 1 mol/L NaOH to pH 7, and then freeze-dried to obtain an unmodified Pleurotus eryngii protein.
(2) The unmodified Pleurotus eryngii protein prepared in step (1) and distilled water were mixed in a mass ratio of 0.1:100 and stirred at 650 r/min for 25 min for hydration. 0.8 mol/L NaOH solution was added to adjust pH to 11.
(3) The solution obtained in step (2) was stirred at 600 r/min for 80 min, so that the high-level spatial structure of the Pleurotus eryngii protein is sufficiently unfolded to expose the secondary structure of the protein.
(4) 0.2 mol/L HCI solution was added dropwise to adjust pH to 7.0, so that a new spatial structure is formed through hydrophobic interaction after the molecular spatial structure of the Pleurotus eryngii protein is unfolded, so as to achieve the embedding of hydrophobic groups and the exposure of hydrophilic groups.
(5) The solution obtained in step (4) was heated at 110°C for 10 min and then rapidly cooled in an ice bath. (6) The solution obtained in step (5) was dialyzed in a dialysis bag with a molecular weight cut-off of 3500 Da for 36 h.
(7) The solution obtained in step (6) was centrifuged at 6000 x g for 15 min to obtain a supernatant.
(8) The supernatant obtained in step (7) was freeze-dried at -55°C for 3 days to obtain a fully soluble Pleurotus eryngii protein.
Test Examples (1) Test of solubility of edible fungus protein: The solubility of edible fungus protein involved in the present invention is represented by the content of soluble edible fungus protein, that is, the proportion of the mass m1 of the soluble edible fungus protein in the soluble edible fungus protein solution to the total mass m0 of the edible fungus protein in the raw material in percentage, as shown in formula (1). The total mass moO of the edible fungus protein in the raw material and the mass m1 of the soluble edible fungus protein are determined by Kjeldahl method.
In the present invention, the solubility of the edible fungus protein is calculated based on the following formula (1): Solubility of edible fungus protein (0%) = mi, 100% "lo (1) In this formula: mo (g) is the total mass of the edible fungus protein in the raw material; and m: (g) is the mass of the soluble edible fungus protein.
The solubility calculated based on formula (1) of the fully soluble edible fungus protein prepared in Examples 1-3 is shown in Table 1. The unmodified protein refers to an edible fungus protein extracted from an edible fungus by an alkali-solution and acid-isolation method.
Table 1 Unmodified protein xam ple El Tear Pleurotus | Lentinula | erinaceus | Pleurotus | Lentinula | erinaceus eryngii edodes (Bull) eryngii edodes (Bull) protein protein Pers protein protein Pers protein protein || non of protein 50.2 40.6 38.2 100 100 100 (%) it can be learned from Table 1 that, after modification for solubilization, the protein with low solubility becomes fully soluble. 1 g of each of the fully soluble edible fungus protein and the unmodified edible fungus protein prepared in Examples 1-3 of the present invention was dissolved in 100 of water and left to stand for 24 h, to obtain images of the edible fungus protein before and after modification, as shown in FIG. 4. It can be learned from FIG. 4 that, after dissolved in water and left to stand, the modified protein in Examples 1-3 of the present invention is clear and transparent, and evenly dispersed, and the unmodified protein precipitates, indicating that the present invention achieves the complete dissolution of the edible fungus protein through modification. (2) Test of turbidity of edible fungus protein: The turbidity of the edible fungus protein involved in the present invention refers to the concentration of the protein sample diluted to 0.2% (w/v) with distilled water.
The pH was adjusted to 7, 8, 9, 10, 11, and 12 with 0.8 mol/L NaOH solution, and the transmittance corresponding to each pH value was measured at a wavelength of 500 nm.
The comparison of turbidity between the fully soluble edible fungus protein prepared in Examples 1-3 of the present invention and the unmodified edible fungus protein is shown in FIG. 1 to FIG. 3. It can be learned from FIG. 1 to FIG. 3 that, under the same pH condition, the transmittance of the edible fungus protein modified by the method of the present invention is significantly higher than the transmittance of the unmodified edible fungus protein, indicating that the modified protein is increased in solubility and transmittance, and is reduced in turbidity.
Although the present invention has been disclosed above with the preferred examples, the examples are not intended to limit the present invention.
A person skilled in the art may make variations and modifications without departing from the spirit and scope of the present invention.
Therefore, the protection scope of the present invention shall be defined by the appended claims.

Claims (10)

ConclusiesConclusions 1. Een volledig oplosbaar, consumeerbaar schimmeleiwit, bereid door een ongemodificeerd consumeerbaar schimmeleiwit te ontvouwen onder een basische conditie en door het aanpassen van de pH naar neutraal door een zuur toe te voegen, waarbij het ongemodificeerde, consumeerbare schimmeleiwit een consumeerbaar schimmeleiwit is met een hydrofiel-hydrofoob gespreid, flexibele ketenstructuur.1. A fully soluble consumable fungal protein prepared by unfolding an unmodified consumable fungal protein under a basic condition and adjusting the pH to neutral by adding an acid, wherein the unmodified consumable fungal protein is a consumable fungal protein having a hydrophilic hydrophobically spread, flexible chain structure. 2. Een volledig oplosbaar, consumeerbaar schimmeleiwit volgens conclusie 1, waarbij het consumeerbare schimmeleiwit één van Pleurotus eryngii, Lentinula edodes, en Hericium erinaceus (Bull.) Pers is.A fully soluble, consumable fungal protein according to claim 1, wherein the consumable fungal protein is one of Pleurotus eryngii, Lentinula edodes, and Hericium erinaceus (Bull.) Pers. 3. Een werkwijze voor het bereiden van het volledig oplosbare, consumeerbare schimmeleiwit volgens conclusie 1, omvattende de volgende stappen: (a) het extraheren van een consumeerbaar schimmeleiwit; (b) het toevoegen van water aan het consumeerbare schimmeleiwit dat is geëxtraheerd in stap {a} en het roeren van, en het toevoegen van een alkalische oplossing voor het aanpassen van de pH; (€) het roeren van de oplossing verkregen na de aanpassing van de pH in stap (b} voor het openbaren van de secundaire structuur van het eiwit; (d) het druppelsgewijs toevoegen van een zoutzuuroplossing voor het aanpassen van de pH; (e) het verwarmen en vervolgens verkoelen van de oplossing verkregen na de aanpassing van de pH in stap (d); (f) het plaatsen van de verkoelde oplossing in een dialysezak voor het dialyseren; en (9) het centrifugeren van de gedialyseerde oplossing voor het verkrijgen van een supernatant, en het vriesdrogen van de supernatant voor het verkrijgen van het volledig oplosbare, consumeerbare schimmeleiwit.A method for preparing the fully soluble, consumable fungal protein according to claim 1, comprising the steps of: (a) extracting a consumable fungal protein; (b) adding water to the consumable fungal protein extracted in step {a} and stirring and adding an alkaline solution for pH adjustment; (€) stirring the solution obtained after the pH adjustment in step (b} to reveal the secondary structure of the protein; (d) adding hydrochloric acid solution dropwise to adjust the pH; (e) heating and then cooling the solution obtained after the pH adjustment in step (d), (f) placing the cooled solution in a dialysis bag for dialysis, and (9) centrifuging the dialyzed solution to obtain of a supernatant, and freeze-drying the supernatant to obtain the fully soluble, consumable fungal protein. 4. De bereidingswerkwijze volgens conclusie 3, waarbij in stap (a), het consumeerbare schimmeleiwit wordt geëxtraheerd door een alkalische oplossing en een zuurisolatiewerkwijze.The preparation method according to claim 3, wherein in step (a), the consumable fungal protein is extracted by an alkaline solution and an acid isolation method. 5. De bereidingswerkwijze volgens conclusie 3, waarbij in stap (b), de massaratio van het consumeerbare schimmeleiwit tot water (0,1-10):100 is; het roeren wordt uitgevoerd op 500-900 r/min voor 10-30 min; de alkalische oplossing een NaOH oplossing is met een molaire concentratie van 0,1-1 mol/L; en de pH 2The preparation method according to claim 3, wherein in step (b), the mass ratio of the consumable fungal protein to water is (0.1-10):100; stirring is carried out at 500-900 r/min for 10-30 min; the alkaline solution is a NaOH solution with a molar concentration of 0.1-1 mol/L; and the pH 2 11.11. 6. De bereidingswerkwijze volgens conclusie 3, waarbij in stap (c), het roeren wordt uitgevoerd op 500-900 r/min voor 80-180 min.The preparation method according to claim 3, wherein in step (c), stirring is performed at 500-900 r/min for 80-180 min. 7. De bereidingswerkwijze volgens conclusie 3, waarbij in stap (d), de molaire concentratie van zoutzuur in de zoutzuuroplossing 0,1-4 mol/L is; en de pH-waardeThe preparation method according to claim 3, wherein in step (d), the molar concentration of hydrochloric acid in the hydrochloric acid solution is 0.1-4 mol/L; and the pH value 7.0 is.7.0. 8. De bereidingswerkwijze volgens conclusie 3, waarbij in stap (e), het verwarmen wordt uitgevoerd op 95-121°C voor 5-45 min. a. The preparation method according to claim 3, wherein in step (e), the heating is performed at 95-121°C for 5-45 min. a. De bereidingswerkwijze volgens conclusie 3, waarbij in stap (f), de moleculaire gewichtsafname van de dialysezak 3500 Da is, en de dialyse wordt uitgevoerd voor 18-36 uur.The manufacturing method according to claim 3, wherein in step (f), the molecular weight decrease of the dialysis bag is 3500 Da, and the dialysis is performed for 18-36 hours. 10. De bereidingswerkwijze volgens conclusie 3, waarbij in stap (g), het centrifugeren wordt uitgevoerd op 4000-8000 x g voor 15-25 min; en het vriesdrogen wordt uitgevoerd op een temperatuur gelegen tussen de -60 graden Celsius en -40 graden Celsius voor een 2-4 dagen.The preparation method according to claim 3, wherein in step (g), the centrifugation is performed at 4000-8000 x g for 15-25 min; and the freeze-drying is carried out at a temperature between -60 degrees Celsius and -40 degrees Celsius for 2-4 days.
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