Method for preparing and applying a supplement based on a polysaccharide-iron complex derived from Auricularia auricula-judae, intended for iron supplementation, improvement of iron deficiency anemia, and strengthening immunity

Abstract

The present invention falls within the technical field of iron supplement preparation, and relates more particularly to a process for preparing and applying a supplement based on a polysaccharide-iron complex derived from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity, comprising the following steps: preparation of raw materials, selection of iron-rich Auricularia auricula-judae as raw material; culture and optimization, engineering modification of fermented and cultured Auricularia auricula-judae; screening and selection of Auricularia auricula-judae strains with a high iron content;The beneficial effects are as follows: by constructing gene expression vectors, transforming fungal cells, and screening and identifying transformants with high synthetic capacity, the level of expression of genes related to the generation and conversion of the iron element is increased, allowing the cultivation of Auricularia auricula-judae varieties containing higher levels of iron, in order to improve the yield and production efficiency of the polysaccharide-iron complex.

Description

1 Method for preparing and applying a supplement based on a polysaccharide-iron complex from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity Technical field 5 The present invention falls within the technical field of vitamin preparation, and relates more particularly to a method for preparing and applying a supplement based on a polysaccharide-iron complex from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity. Background Technology 10 The polysaccharide-iron complex is available as a powder or lamellar crystals, ranging in color from brown to dark brown, and is stable in light and air. The polysaccharide-iron complex is primarily used for the prevention and treatment of iron deficiencies; in addition, it can also be used for the treatment of chronic iron deficiency anemia, restless legs syndrome, hypothyroidism, and autoimmune diseases; furthermore,15 The polysaccharide-iron complex also presents beneficial effects on cardiovascular diseases, diseases of the digestive system and diseases of the nervous system. In the state of the art, processes for preparing iron-polysaccharide complexes tend to be more efficient and environmentally friendly. Researchers are continually exploring new extraction and purification methods to improve the purity and activity of iron-polysaccharide complexes, while reducing energy consumption and emissions during the production process. For example, the use of advanced technologies such as microwave-assisted extraction or supercritical fluid extraction allows for more efficient extraction of iron-polysaccharide complexes from Auricularia auricula-judae. However, the optimization of the source of iron-polysaccharide complex extraction—namely, Auricularia auricula-judae—is neglected; that is, a lack of optimization aimed at increasing, starting from the Auricularia auricula-judae itself,the extraction yield of polysaccharide-iron complexes. Consequently, it is necessary to propose a process for preparing and applying a supplement based on a polysaccharide-iron complex derived from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity, and to solve the problem of insufficient optimization of raw materials in the extraction of polysaccharide-iron complexes in the prior art; it is therefore possible to optimize the genetic characteristics of the raw material in order to increase the extraction yield of the polysaccharide-iron complex. 5 Content of the invention The objective of the present invention is to propose a process for preparing and applying a supplement based on a polysaccharide-iron complex derived from Auricularia auricula- Judae, intended to supplement iron, improve iron-deficiency anemia and strengthen immunity, in order to resolve the problem raised in the aforementioned prior art,of the inadequacy10 of raw material optimization in the extraction of polysaccharide-iron complexes; it is thus possible to optimize the genetic characteristics of the raw material in order to increase the extraction yield of the polysaccharide-iron complex. In order to achieve the aforementioned objective, the present invention proposes the following technical solution: a process for preparing a supplement based on a polysaccharide-iron complex derived15 from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity, said process comprising the following steps: preparation of raw materials, selection of iron-rich Auricularia auricula-judae as raw material; culture and optimization, genetic modification of the fermented and cultured Auricularia auricula-judae20; Screening and selection of Auricularia auricula-judae strains exhibiting high iron content; extraction and purification, purification of the polysaccharide-iron complex by nanometric techniques. 25 Preferably,The specific operations for the preparation of the raw materials include: the selection of Auricularia auricula-judaerichesenfer as raw material for the preparation of the polysaccharide-iron complex. Preferably, the specific operations for the culture and optimization include: 30 2025 / 5805 BE2025 / 5805 3 inoculating the fungus into a culture medium containing a carbon source, a nitrogen source, as well as minerals and trace elements, the pH of the medium being controlled at a neutral or slightly acidic value, the culture vessel being placed in a thermostatic stirrer or fermenter, and the appropriate temperature being set between 25 and 30°C; the construction of gene expression vectors, the transformation of fungal cells, 5 as well as the screening and identification of transformants exhibiting high synthetic capacity,in order to increase the level of expression of genes related to iron synthesis and conversion into the iron-polysaccharide complex. The specific operations for screening and selecting Auricularia auricula-judae strains with high iron content include: 10 during fungal culture, exposure to ultraviolet light irradiation, with a light intensity of 10 W / m² and an irradiation duration of six to fourteen hours, in order to screen and select Auricularia auricula-judae strains exhibiting a high iron content. Preferably, the specific extraction and purification operations include: destruction of the fungal cell wall by grinding in order to release the iron-polysaccharide complex into the culture medium, water being chosen as the extraction solvent; Filtration and adsorption of the extract using anofiltration membranes or nanometric adsorbents, in order to eliminate impurities and improve the purity of the polysaccharide-iron complex; crystallization treatment of the polysaccharide-iron complex solution after extraction and purification,The separation of the crystals and the mother liquor by centrifugation, followed by a drying treatment to remove the water. Preferably, the process also includes: grinding the polysaccharide-iron complex after drying, then packaging the polysaccharide-iron complex in well-sealed containers to ensure hygiene and product safety. Application of the iron-polysaccharide complex prepared by the process for preparing a supplement based on an iron-polysaccharide complex from Auricularia auricula-judae, intended to supplement iron, improve iron-deficiency anemia and strengthen immunity, said application consisting of using the iron-polysaccharide complex obtained for iron supplementation. 30 2025 / 5805 BE2025 / 5805 4 Application of the iron-polysaccharide complex prepared by the process for preparing a supplement based on an iron-polysaccharide complex from Auricularia auricula-judae, intended to supplement iron, improve iron-deficiency anemia and strengthen immunity,said application consisting of using the polysaccharide-iron complex obtained to improve iron deficiency anemia. Application of the polysaccharide-iron complex prepared by the process of preparing a supplement based on the polysaccharide-iron complex from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity, said application consisting of using the polysaccharide-iron complex obtained to strengthen immunity. Application of the polysaccharide-iron complex prepared by the process of preparing a supplement based on the polysaccharide-iron complex from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity, said application consisting of using the polysaccharide-iron complex obtained to simultaneously supplement iron, improve iron deficiency anemia and strengthen immunity. Compared to the state of the art,The beneficial effects of the present invention are as follows: 15. The process for preparing and applying a supplement based on a polysaccharide-iron complex derived from Auricularia auricula-judae, intended to supplement iron, improve iron-deficiency anemia, and strengthen immunity, proposed by the present invention, allows, through the construction of gene expression vectors, the transformation of fungal cells, as well as the screening and identification of transformers exhibiting a high synthetic capacity, 20. increasing the level of expression of genes related to iron synthesis and conversion into a polysaccharide-iron complex, cultivating varieties of Auricularia auricula-judae containing higher levels of iron, and thus improving the yield and production efficiency of the polysaccharide-iron complex; this also allows us to solve the problem of insufficient optimization of raw materials in the extraction of polysaccharide-iron complexes from state 25 of the art,and to optimize the genetic characteristics of the raw material in order to increase the extraction yield of the polysaccharide-iron complex. Specific embodiments In order to describe clearly and completely the objectives, technical solutions and advantages of the present invention, it should be understood that the specific embodiments described herein constitute only a part of the embodiments of the present invention and not all of them; they are intended solely to illustrate the embodiments of the present invention and should not be interpreted as a limitation thereof. All other embodiments obtained by persons skilled in the art, without creative effort, on the basis of the embodiments of the present invention, fall within the scope of protection of the present invention. Example of implementation 1 The present invention proposes a technical solution: a process for preparing a supplement based on a polysaccharide-iron complex derived from Auricularia auricula-judae, intended to supplement iron,to improve iron deficiency anemia and strengthen immunity, said process10 comprises the following steps: I. Preparation of raw materials, selection of iron-rich Auricularia auricula-judae as raw material; first of all, it is necessary to select Auricularia auricula-judae containing a high iron content as raw material for the preparation of the iron-polysaccharide complex. The iron element constitutes the precursor of the iron-polysaccharide complex,15 which can be formed by a specific conversion process. Among the different varieties of Auricularia auricula-judae,Sarawak's Auricularia auricula-judae is often chosen as a high-quality raw material for the preparation of the iron-polysaccharide complex due to its high iron content and ease of obtaining and processing. Selected Auricularia auricula-judae must be carefully washed to remove dirt and impurities from the surface. This step is essential to guarantee the hygiene of the subsequent process and the quality of the product. Washing is carried out using running water or specific food-grade cleaning agents to ensure the cleanliness of the Auricularia auricula-judae surface. After washing, the Auricularia auricula-judae must be dried to remove Excess water. Drying can be carried out by natural air drying or with the aid of a food dehydrator. Dried Auricularia auricula-judae are easier to preserve and transport.and also facilitate subsequent grinding and extraction operations. The dried Auricularia auricula-judae is then ground into a powder to allow for more complete subsequent extraction. Grinding can be carried out by mechanical grinding or by hand grinding, so as to obtain particles fine enough to improve extraction efficiency. The Auricularia auricula-judae powder obtained must be stored correctly to avoid moisture, contamination, and deterioration. The storage environment must be dry, cool, and well-ventilated, while avoiding direct exposure to sunlight. It is also advisable to check and turn the stored powder regularly to ensure its good condition. II. Culture and optimization, genetic modification of fermented and cultured Auricularia auricula-judae5; in order to support the growth and metabolism of the fungus, a culture medium rich in carbon sources, nitrogen sources, minerals and trace elements is prepared. The carbon sources,such as glucose or sucrose, provide energy to the fungus; nitrogen sources, such as yeast extract or apetone, support cell growth; minerals and trace elements ensure the normal functioning of the fungus's vital activities. Furthermore, the pH of the culture medium is controlled to a neutral or slightly acidic value to recreate the fungus's natural growth environment. The selected fungus is inoculated into the prepared culture medium, the inoculation operation being carried out under sterile conditions to avoid any microbial contamination. The inoculated culture vessels are placed in a thermostatic shaker or fermenter, and the appropriate temperature range is generally set between 25 and 30°C to ensure good growth and reproduction of the fungus. In addition, depending on the mushroom growth characteristicsThe rotation speed of the agitator or the aeration rate of the fermenter is adjusted to meet the oxygen requirements of the fungus. To further improve the fungus's ability to synthesize the polysaccharide-iron complex, 20 genetic engineering techniques are used to modify it. First, gene expression vectors are constructed, and the genes encoding the key enzymes involved in iron synthesis and conversion to the polysaccharide-iron complex are cloned into these vectors. Next, these vectors are introduced into the fungal cells using transformation techniques, allowing the fungus to express these genes. The transformed fungal cells must then be screened and identified; by comparing the synthetic capacities of different transformants, strains exhibiting high synthetic capacities are selected. In order to further increase the yield of the polysaccharide-iron complex, the culture conditions must also be optimized. This includes adjusting the proportion of the culture medium components, the pH, and the temperature.humidity and lighting are required to allow the fungus to grow and metabolize under optimal conditions. Fungal strains obtained after culture and optimization must be properly stored to ensure the durability and sustainability of their genetic characteristics. Methods such as freezing, drying, or storage in liquid nitrogen can be used for long-term preservation of strains. Furthermore, when necessary, strains can be subcultured and reactivated to ensure their continued use for the production of the polysaccharide-iron complex. III. Screening and selection of Auricularia auricula-judae strains exhibiting high iron content. The choice of light source is crucial, and it is necessary to ensure that the wavelength of the emitted light is suitable for the conversion reaction of the iron element into the iron-polysaccharide complex within the fungal cells. Furthermore,The lighting equipment must provide a stable output and adjustable light intensity to meet the needs of different fungal strains or different growth stages. Before photo-induced conversion, the fungus is cultured to an appropriate growth stage, generally during the exponential or stationary phase, to ensure that the cells contain a sufficient amount of the precursor substance. In addition, the concentration and homogeneity of the fungal culture must be controlled to ensure the consistency of the reaction during the photo-induced conversion process. Key parameters of photo-induced conversion include light intensity and irradiation time. In the present example, The light intensity is set at 10W / m², an optimized value that guarantees the efficiency of the conversion while avoiding excessive damage to the fungal cells. The irradiation time is adjusted according to the characteristics of the fungal strain and the experimental requirements.The irradiation time is generally between six and fourteen hours. Too long an irradiation time can lead to cell damage or product degradation, while too short a time may not allow sufficient conversion. The prepared fungal culture is placed in the lighting equipment, ensuring that it receives uniform ultraviolet irradiation; the temperature and humidity of the lighting environment should also be controlled to simulate the natural growth environment of the fungus. After the irradiation is complete, the fungal culture must undergo further processing, including turning off the lighting, collecting the culture, and extracting and purifying the complex. 2025 / 5805 BE2025 / 5805 8 Iron polysaccharide. During the extraction and purification stages, it is important to avoid any degradation of the product due to light exposure in order to guarantee the quality and purity of the final product. IV. Extraction and purification,Purification of the polysaccharide-iron complex by nanometric techniques; in order to release the polysaccharide-iron complex contained in fungal cells, it is first necessary to destroy the cell wall of the fungus. This operation is generally carried out by physical or chemical methods, such as high-pressure homogenization, ultrasonic fragmentation, or the enzymatic method. The choice of a rupture method adapted to the characteristics of the fungus is essential in order to avoid the degradation of the polysaccharide-iron complex molecules. After rupture, the fungal cells release the polysaccharide-iron complex as well as other cellular constituents into the culture medium, which facilitates subsequent extraction steps. Then, an appropriate solvent is chosen to extract the polysaccharide-iron complex from the culture medium. Pure water is widely used as an extraction solvent due to its good capacity to dissolve the polysaccharide-iron complex and its relatively low cost. During extraction,The temperature and duration are appropriately controlled to ensure complete dissolution of the polysaccharide-iron complex in pure water, while avoiding the concomitant extraction of undesirable impurities. The extract contains the polysaccharide-iron complex but may also contain certain impurities. To remove these impurities and improve the purity of the polysaccharide-iron complex, anofiltration membranes or nanometric adsorbents are used for filtration and adsorption. Anofiltration membranes offer very high filtration precision and effectively remove impurity particles larger than their pore size, while nanometric adsorbents can adsorb selectively removes certain specific impurities. Thanks to these steps, the purity of the polysaccharide-iron complex is significantly improved.providing a high-quality raw material for the subsequent crystallization process. The polysaccharide-iron complex solution obtained after extraction and purification is then subjected to a crystallization treatment. By appropriately controlling the temperature, concentration, and crystallization conditions, the polysaccharide-iron complex precipitates in crystalline form. Once crystallization is complete, the crystals are separated from the mother liquor by centrifugation. During centrifugation, the high-speed rotation allows the crystals to settle at the bottom of the centrifuge tube, thus separating them from the mother liquor. Finally, the separated polysaccharide-iron complex crystals are subjected to a drying treatment, generally by Video-assisted drying by atomization, in order to eliminate the water contained in the crystals and obtain a dry final product of the polysaccharide-iron complex. 5 Example of implementation 2 Based on example of implementation 1,The following process is proposed: preparation of the polysaccharide-iron complex. I. Preparation of raw materials. Fresh Auricularia auricula-judae are selected as raw material for the preparation of the polysaccharide-iron complex. The Auricularia auricula-judae are subjected to washing, drying, and grinding to obtain a homogeneous biomass powder. II. Fungal culture. Preparation of the culture medium: glucose, yeast powder, inorganic salts, and trace elements are mixed in a specific ratio to prepare a liquid culture medium. The pH of the culture medium is adjusted to 6.5. Inoculation and culture: Streptomycetes are inoculated into the culture medium, then placed in a thermostatic shaker, the temperature being set at 28°C and the rotation speed at 150 rpm. The culture duration is 48 hours.to ensure sufficient growth and metabolism of the fungus. 20 III. Modification by genetic engineering Vector construction: the genes encoding the key enzymes involved in iron synthesis and conversion into polysaccharide-iron complexes are cloned into an expression vector. Transformation and screening: the constructed expression vector is transformed in streptomycetes, and positive transformants are obtained by screening based on resistance. 25 Verification of expression: the positive transformants are cultured and their production of iron and polysaccharide-iron complexes is analyzed, in order to select strains exhibiting a high synthesis capacity. IV. Screening and Selection of Strains The suspension of cultured fungal cells is uniformly spread on 30 culture plates (2025 / 5805 BE2025 / 5805 10), then placed under fluorescent lamps for photo-induced conversion. The light intensity is set to 5 W / m² and the irradiation duration is 8 hours. By light stimulation,The iron element present in the fungal cells is converted into a polysaccharide-iron complex. V. Extraction and Purification Extraction: After photo-induced conversion, the fungal cells are ground and then extracted using pure water as a solvent. The extraction temperature is controlled at 40°C, the extraction time is 2 hours, and the solvent / biomass ratio is 3:1. Filtration and Concentration: The extract is filtered to remove impurities and then concentrated using a rotary evaporator to obtain a crude extract of the polysaccharide-iron complex. Purification by Nanotechnology: Nanofiltration membranes are used to filter the crude extract to further remove impurities and increase the purity of the polysaccharide-iron complex. VI. Crystallization and Drying The purified solution of the polysaccharide-iron complex is subjected to a crystallization treatment. By controlling the crystallization conditions,The polysaccharide-iron complex precipitates in crystalline form. The crystals are then dried using a vacuum dryer to remove the water and obtain the final polysaccharide-iron complex product. VII. Packaging and Storage The dried polysaccharide-iron complex product is packaged using airtight packaging materials to ensure product hygiene and safety. The packaged polysaccharide-iron complex is stored in a cool, dry, and light-free environment to prevent moisture and the influence of light. VIII. Quality Control The polysaccharide-iron complex product obtained is subject to quality controls, including purity, latent strength, and activity. By comparison with reference products, the quality of the product is guaranteed to meet the relevant standards and requirements. Example of implementation 3 Application of the polysaccharide-iron complex prepared by the process for preparing a supplement based on the polysaccharide-iron complex derived from Auricularia auricula-judae, intended to supplement iron,to improve iron deficiency anemia and to strengthen immunity, said application 30 2025 / 5805 BE2025 / 5805 11 consisting of using the polysaccharide-iron complex obtained for iron supplementation. Example of embodiment 4 Application of the polysaccharide-iron complex prepared by the process of preparing a supplement based on a polysaccharide-iron complex derived from Auricularia auricula-judae, intended to supplement iron, to improve iron deficiency anemia and to strengthen immunity, said application 5 consisting of using the polysaccharide-iron complex obtained to improve iron deficiency anemia. Example of implementation 5: Application of the polysaccharide-iron complex prepared by the process of preparing a supplement based on the polysaccharide-iron complex derived from Auricularia auricula-judae, intended to supplement iron, improve iron deficiency anemia and strengthen immunity,said application 10 consisting of using the obtained iron-polysaccharide complex to strengthen immunity. Example of embodiment 6 Application of the iron-polysaccharide complex prepared by the process for preparing a supplement based on an iron-polysaccharide complex derived from Auricularia auricula-judae, intended to supplement iron, improve iron-deficiency anemia and strengthen immunity, said application 15 consisting of using the obtained iron-polysaccharide complex to simultaneously supplement iron, improve iron-deficiency anemia and strengthen immunity. Comparative example 1 Enzymatic process for preparing an iron-polysaccharide complex, comprising the following steps: 20 I. Preparation of raw materials Iron-rich yeasts are selected as raw materials. The yeasts are cultured, harvested and then dried for later use. II. Selection and preparation of the enzyme. An enzyme capable of efficiently catalyzing the conversion of the element iron into the 25-polysaccharide-iron complex is selected.such as an enzyme for the synthesis of the polysaccharide-iron complex. This enzyme can be expressed in microorganisms by genetic engineering techniques, then purified and concentrated. III. Enzymatic Conversion The prepared yeasts are mixed with an appropriate quantity of enzyme solution, and the reaction temperature, pH, and reaction time are controlled to allow the enzyme to catalyze the conversion of the element iron into a polysaccharide-iron complex. The specific reaction conditions can be optimized according to the characteristics of the enzyme and the nature of the raw materials. IV. Extraction and Purification After enzymatic conversion,The polysaccharide-iron complex is extracted from the reaction system using a suitable extraction solvent and under appropriate extraction conditions. Purification methods such as chromatography and crystallization are then used to remove impurities and improve the purity of the polysaccharide-iron complex. 10 V. Drying and Packaging The purified polysaccharide-iron complex undergoes a drying treatment to remove water and is then packaged using suitable packaging materials to ensure hygiene and product safety. The product is stored in a cool, dry environment, protected from moisture and light. 15 VI. Quality Control The obtained polysaccharide-iron complex product undergoes quality control, including purity, latency, and activity. By comparison with products of Reference, it is guaranteed that the product quality meets the relevant standards and requirements. Comparative Example 220 A process for preparing a polysaccharide-iron complex by photo-induced biosynthesis is proposed,comprising the following steps: I. Preparation of raw materials. Auricularia auricula-judae eriches enfers are selected as raw material. The raw materials are subjected to washing, grinding and pretreatment operations to enable the subsequent photo-induced biosynthesis reaction. II. Optimization of lighting conditions. The conditions of the photo-induced biosynthesis reaction are optimized by adjusting parameters such as light intensity, wavelength of illumination and duration of irradiation. An appropriate light source and lighting equipment are selected to ensure the stability and uniformity of the illumination. III. Photo-induced biosynthesis reaction. Pre-treated raw materials are placed under specific lighting conditions, such that the iron element is converted into a polysaccharide-iron complex by a high-pressure catalytic reaction. By controlling the lighting conditions and reaction time,5. An efficient photo-induced biosynthesis is carried out. IV. Extraction and Purification After the photo-induced biosynthesis reaction, an appropriate solvent and extraction method are used to extract the iron-polysaccharide complex from the raw materials. Purification techniques such as chromatographic separation and crystallization are then implemented to remove impurities and improve the purity of the iron-polysaccharide complex. V. Drying and Packaging The purified iron-polysaccharide complex undergoes a drying treatment to remove water, then is packaged using airtight packaging materials to ensure product hygiene and safety. The product is stored in a cool, dry environment, protected from moisture and light. VI. Quality Control The polysaccharide-iron complex product obtained is subject to quality controls, including purity, latency, and activity. By comparison with reference products,The quality of the product is guaranteed to meet the relevant standards and requirements. The results of the comparison show that implementation example 2 presents the best effects. Thanks to the construction of gene expression vectors, the transformation of fungal cells, and the screening and identification of transformers exhibiting high synthetic capacity, the level of expression of genes related to iron synthesis and conversion into the iron-polysaccharide complex is increased, making it possible to cultivate varieties of Auricularia auricula-judae containing higher levels of iron, and thus improve the yield and production efficiency of the iron-polysaccharide complex; this also makes it possible to solve the problem of insufficient optimization of raw materials in the extraction of iron-polysaccharide complexes in the prior art.and to optimize the genetic characteristics of the 30 raw materials in order to increase the extraction yield of the polysaccharide-iron complex. Although embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that various variations, modifications, substitutions, and transformations can be made to these embodiments without departing from the principles and spirit of the present invention; the scope of the present invention is defined by the 5 annexed claims and their equivalents. 2025 / 5805 BE2025 / 5805,