Waste management using biological and non-biological compositions

Abstract

The present invention relates to an inoculant composition comprising a first fungal species or strain and a second fungal species or strain different therefrom. [Solution] At least one of the first fungal species or strain is a saprophytic fungus. The first and second fungal species or strains are each contained in the composition at a relative concentration that allows them to be inoculated into one or more polymer materials at an inoculation ratio such that the mass ratio of fungal biomass to polymer material is 1:1 or less. Furthermore, the inoculant may include a first modified fungal species or strain that recognizes plastic products as a nutrient source and is designed to digest persistent long-chain carbon more efficiently than its naturally derived counterpart of the same species or strain. The first modified fungal species or strain may be immobilized to be in a metabolically dormant state. The invention also relates to a method for converting long-chain carbon material in plastic waste, the method comprising the step of adding an inoculant to a plastic substrate.

Description

[Technical Field]

[0001] The present invention relates to the field of decomposition in general, and more specifically to compositions and methods for promoting the digestion of persistent long-chain carbon materials such as plastics, as well as other materials present with plastic waste products. [Background technology]

[0002] In recent years, the negative environmental impact of plastic waste has become an increasingly serious problem. Conventional plastic materials can take hundreds of years to decompose, causing environmental pollution and damage to ecosystems. This invention addresses these problems by accelerating the digestion of plastics and other synthetic or organic materials. [Prior art documents] [Patent Documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 2006-158237 [Overview of the project]

[0004] The present invention provides methods for accelerating the digestion of the aforementioned materials. These methods involve the use of pre-selected fungal species or strains trained to digest refractory polymers. The selected fungal species are immobilized in a manner that maximizes efficiency, effectiveness, and ease of use, and can also be combined as a mixture of multiple fungal species to achieve the desired digestion result.

[0005] When fungi are inoculated as a mixture in the presence of plastic material, they first grow and spread on the material surface using mycelium. Mycelium is a network of filamentous structures that make up the body of the fungus. This growth and spread is called "hyphal growth." During hyphal growth, the fungi secrete several enzymes that can break down various compounds present in the plastic material, such as cellulose and lignin.

[0006] This invention relates to the field of fluid soft plastics such as films and nonwoven fabrics, as well as product structures such as absorbent articles, particularly diapers and other personal waste disposal products. The invention relates to the development of novel absorbent materials and structures usable in a variety of absorbent products. The performance objectives of absorbent products are to improve absorbency, liquid retention, and overall performance, while ensuring comfort, fit, and ease of use. The methods provided by this invention eliminate the trade-off between sustainability and these performance indicators, resulting in more sustainable products without the need for new materials.

[0007] Absorbent items such as diapers typically consist of multiple layers of materials designed to manage and retain human or animal waste. These layers include a top sheet, a collection layer, an absorbent core, a back sheet, and various materials such as adhesives, elastic components, and fasteners.

[0008] The top sheet layer is typically made of a nonwoven fabric material consisting of PE, PP, BYCO (composite fiber), or other polymers, and is designed to come into contact with the skin. The acquisition layer is designed to rapidly transfer the liquid from the skin to the absorbent core. The absorbent core is typically composed of cellulose fibers and superabsorbent polymers (SAP) to capture and retain the liquid.

[0009] The backsheet layer is typically made of a film of PE, PP, or other polymers and is designed to prevent liquid leakage. Other materials included in diapers include hot melt adhesives, surfactants and dyes for moisture indicators, and various fasteners such as Velcro and elastic materials.

[0010] Various polymers used in diapers play specific roles in diaper design, such as absorbency, liquid management, and containment. SAP is one of the main polymers used in diapers and is extremely effective at capturing and retaining liquids because it can absorb hundreds of times its own weight in liquid. [Means for solving the problem]

[0011] Plastic pollution has become a global problem affecting almost all ecosystems. Given the usefulness, widespread use, and ultimate environmental impact of plastic polymers, research into plastic degradation is an extremely promising field. The plastic polymers used are diverse in their chemical composition, and a diverse range of enzymes are needed to completely decompose them into microbial biomass, water, and carbon dioxide.

[0012] The rate of plastic degradation depends on the properties of each individual plastic, such as hydrophobicity, chemical structure, crystallinity, and molecular weight. Plastics with high molecular weights or a high proportion of crystalline regions are more difficult to decompose and digest. On the other hand, the relationship between chemical structure and degradability is not always linear. Broadly speaking, plastic polymers can be classified into two categories: polymers composed of repeating chains of identical chemical elements (referred to herein as "homochains") and polymers composed of a mixture of more heterogeneous elements (referred to herein as "heterochains"). Homochain polymers such as polyethylene (PE) and polypropylene (PP) have higher resistance to microbial degradation compared to heterochain polymers such as polyurethane (PU), polyethylene terephthalate (PET), and polystyrene (PS). PET and PU are two of the most studied plastics, but PE (approximately 30%) and PP (approximately 19%) are the plastics with the highest annual production volumes, indicating that further research is needed on their degradation pathways. The examples of heterochains, homochains, and crosslinked polymers presented herein are illustrative and intended to illustrate the digestion mechanisms specific to each polymer group. These non-limiting examples are categorized based on their respective digestion mechanisms, and the methods described herein are applicable to a broader range of materials belonging to these polymer groups, as will be readily apparent to those skilled in the art.

[0013] For reference, Figure 1A shows the chemical structures of some of the most common plastics. These are the main plastics found in diapers. The cross-linked heterochain, composed of polyethylene glycol (PEG) and sodium polyacrylate (NaPA), is a component of the superabsorbent polymer (SAP) found in all absorbent products. This large variation in chemical composition is one reason why mixing and recycling different types of plastics is difficult. Generally, homochains are polymers with a main chain consisting only of carbon and hydrogen atoms. This simple structure suggests a uniform bonding pattern centered on carbon-carbon bonds. In enzymatic digestion, homochains require an initial oxidation process before hydrolysis occurs. Due to the lack of heteroatoms in the main chain, direct hydrolysis is difficult, and an oxidation step is necessary to introduce functional groups for enzymatic attack. Oxidation can be abiotic (e.g., heating or chemical treatment) or biological. Given this composition and digestion requirements, homochains are inherently more resistant to digestion, and the time required for complete degradation may be longer.

[0014] Heterochains are characterized by the inclusion of elements other than carbon and hydrogen in the polymer backbone, such as oxygen, sodium (Na), and nitrogen (N). This diversity introduces a variety of functional groups and bonding modes within the polymer structure. The presence of oxygen and other heteroatoms allows for hydrolysis at the inoculation site without the need for a prior oxidation step. This structural complexity can facilitate enzyme access and attack, potentially streamlining the digestion process. Generally, heterochains are considered to be more easily degraded than homochains due to their diverse and heterogeneous structures.

[0015] It is known that specific fungal communities can colonize plastics (i.e., plastic acts as a community filter), and these microbial communities are called "plastispheres." The elements that function as community filters within the plastisphere, and the characteristics that allow fungi to overcome this barrier, are not yet fully understood. However, certain fungal characteristics are known to promote colonization and digestion of plastics. For example, hydrophobin production assists colonization of hydrophobic plastic surfaces through the formation of aerial structures. Hydrophobins have the function of attaching hyphae to hydrophobic plastic surfaces, and some have been suggested to significantly promote hydrolysis. Furthermore, the production of extracellular polysaccharides further promotes adhesion to the substrate surface and forms a fungal-substrate interface for enzyme secretion. Much of the research to date on the degradation and / or digestion of plastics has been conducted in highly controlled laboratory environments using "uncontaminated" or pre-treated plastics (i.e., those without additives and with no differences in surface properties), and few studies have focused on consumer plastic products under natural conditions. Therefore, there are limitations to inferences made in real-world environments. In general, fungi possess diverse enzymatic capabilities to digest and / or decompose conventional plastics, but there is still a need for effective methods to improve the digestion rate of commercially available plastic waste.

[0016] Embodiments of this disclosure provide a method for degrading various plastic waste products, including absorbent articles, using pre-selected fungi capable of degrading persistent long-chain carbons derived from both organic and synthetic materials. These embodiments enable the widespread commercialization of the method in intended use areas such as diapers and absorbent products. The fungi can be stored in a form with storage stability and can be added to plastic-containing products during manufacturing or after use. This reduces plastic waste and promotes improved environmental sustainability.

[0017] Furthermore, this specification provides a composition comprising an immobilized and storage-stable fungal inoculum trained to digest polymers present in polymeric materials. The composition comprises one or more fungal species or strains, which may be encapsulated within an immobilized material, thereby ensuring stability and protection from contamination, and providing storage stability. Upon activation by exposure to moisture, the encapsulated fungi can resume metabolism and accelerate the digestion process, providing an environmentally friendly solution for the digestion of various polymeric materials.

[0018] This specification also provides a method for industrial-scale digestion of polymeric materials, including organic waste as needed, using a pre-selected fungal inoculum. Waste materials are collected and accumulated from multiple flows, and the fungal inoculum is introduced. Environmental conditions within an industrial-scale waste treatment system, such as a bioreactor, are controlled to optimize fungal activity. Optional process aids may be introduced to promote the growth and establishment of the rehydrated fungal inoculum. The digestion process results in the production of biomass, water, carbon dioxide, and other by-products. The resulting biomass can be used for further applications such as composting, soil remediation, or as a raw material for renewable energy generation. This method provides an efficient, environmentally friendly, and scalable approach to industrial-scale management of polymeric and organic waste.

[0019] In another embodiment, a method is provided for incorporating an immobilized fungal inoculum into a product before or after use. The fungal inoculum is configured to enable significantly improved digestion of the product. The inoculation process is flexible, allowing consumers to apply the inoculum in various ways, such as by applying it directly to a used product, by placing it in a waste container, or by placing it in a dedicated digestion bag. The inoculum can also be incorporated during the product manufacturing process.

[0020] In another embodiment, a composition comprising a mixture of adapted fungal species and strains is provided. The mixture is configured as a plurality of encapsulated fungal inocula or other immobilized materials, wherein each bead contains a single fungal species or strain. The mixing of these fungal species forms a novel "consortium" that does not exist in nature, promotes fungal growth and plastic digestion, and imparts resistance to abiotic stresses including feces, urine and other conditions. Said fungal species are trained to recognize plastic products as a nutrient source and digest the persistent long-chain carbons present in synthetic polymers more efficiently compared to naturally occurring forms.

[0021] Furthermore, the present specification provides a composition comprising an immobilized, storage-stable fungal inoculum trained to target and digest polymers present in polymer-based materials. The composition comprises one or more fungal species or strains and is encapsulated within an immobilization material, whereby stability and protection from contamination are ensured, and storage stability is promoted. When activated by exposure to moisture, the encapsulated fungi resume metabolism and are able to accelerate the digestion process.

[0022] The fungal species used in the embodiments of the present disclosure are selected from white rot fungi, brown rot fungi and soft rot fungi (species W, species B and species S, respectively). The classification of these species and strains is based on properties observed in nature, morphological characteristics and genomic potential. To date, only a small fraction of the enzymatic potential for plastic degradation possessed by these fungal species has been characterized. Accordingly, the process of screening and adapting these fungi can enhance the gene expression of known enzymes and activate the expression of enzymes that have not yet been characterized. This complex mixture of enzymes derived from single and multiple species promotes the digestion of complex polymers. The process involves an iterative approach including experimental optimization and systematic manipulation. The process of selection, adaptation and combination of fungal species, measured by experimental characterization of digestion markers on polymer substrates, forms the basis of the inventive embodiments according to the present disclosure.

[0023] Further, the present specification provides a method for controlling rehydration and activation of dormant immobilized fungal inoculum in an absorbent product. The process optimizes fungal performance to improve biodegradability of the product while maintaining absorbency. The emergence of the fungal inoculum is carefully controlled, enabling effective digestion of the absorbent product after use, and contributing to a more sustainable waste management solution.

[0024] The present specification also provides a method for encapsulating and immobilizing a preselected fungus as discrete beads or on a continuous substrate. The method enables the use of the fungus for digestion of persistent long-chain carbon materials. The process comprises the steps of homogenizing fungal mycelium, optionally filtering the homogenized mycelium, mixing the homogenized mycelium with an alginate polymer solution, shaping the mixture into beads or hydrogel, and performing a dehydration treatment. The encapsulated fungus may be coated for additional protection and extended shelf life, and is applicable to digestion applications in various environments.

[0025] In another embodiment, a method of using an immobilized fungal inoculum is provided, wherein the fungal inoculum is incorporated into the product before or after use of the product, and is configured to enable efficient digestion of the product within an improved (non-limiting) period, such as, preferably, 6 to 12 months. The inoculation process is flexible: consumers can apply the inoculum in various manners, including applying it directly to a used product, putting it into a waste container, or putting it into a dedicated digestion bag. It is also possible to incorporate the inoculum during the manufacturing process of the product.

[0026] Furthermore, this specification provides a highly absorbent hydrogel embedded with a fungal inoculum, designed for use in absorbent products. This hydrogel exhibits high swelling capacity, rapid hydration rate, and excellent liquid retention capacity, and possesses the unique characteristic of containing a bioactive fungal inoculum. This combination results in absorbent products with improved fluid handling performance and enhanced biodegradability, contributing to more sustainable and environmentally friendly absorbent products.

[0027] Furthermore, embodiments of methods for incorporating fungal inoculants into the manufacturing process of absorbent products are also provided to improve the biodegradability of absorbent products. In one embodiment, the fungal inoculant is prepared as particles having a particle size similar to that of a superabsorbent polymer (SAP). These particles are mixed with a pulp fiber mixture in a hopper and then molded into an absorbent core for the product. This process is designed so that the majority of the fungal inoculant particles are not exposed to temperatures that kill dormant fungi.

[0028] In an alternative embodiment, a fungal inoculum is embedded in a nonwoven fabric substrate, cut into patches, and then applied to an absorbent product partially assembled using a vacuum rotating drum. These patches are secured by tackifiers, adhesives, or mechanical bonding and further covered with an additional protective layer.

[0029] Furthermore, this specification provides a method for industrial-scale digestion of polymeric materials, including organic waste as needed, using a pre-selected fungal inoculum. In this embodiment, waste materials are collected and accumulated from multiple flows, and the fungal inoculum is introduced. Environmental conditions within the industrial-scale waste treatment system, such as anaerobic or aerobic digesters or bioreactor-type landfills, are controlled to optimize fungal activity. Process aids may optionally be introduced to promote the growth and establishment of the rehydrated fungal inoculum. As a result of the digestion process, biomass, water, carbon dioxide, and other by-products are generated. The resulting biomass can be used for applications such as composting, soil remediation, or as a raw material for renewable energy production. This novel method provides an efficient, environmentally friendly, and scalable approach to industrial-scale management of polymeric and organic waste. [Brief explanation of the drawing]

[0030] [Figure 1A] Figure 1A shows some of the most common plastic chemical structures as background to the embodiments of this disclosure. [Figure 1B] Figure 1B is a graph showing that the inoculum according to the embodiment of this disclosure can effectively inoculate plastic materials with an inoculum:plastic weight ratio of less than 1:1. [Figure 1C] Figure 1C shows the flowchart, a photograph of the petri dish, and the growth rate data of seed W for Example 2. [Figure 2] Figure 2 shows a colorimetric analysis of lignin peroxidase production from species S and laccase and manganese peroxidase [C] production from species W. [Figure 3] Figure 3 shows an SEM image illustrating the effectiveness of the embodiment of the present disclosure in Example 3. [Figure 4] Figure 4 is an SEM image showing that BSNW was effectively inoculated with seed W in Example 3. [Figure 5]Figure 5 is a diagram including a TGA plot showing the effectiveness of the embodiment of the present disclosure in Example 3. [Figure 6] Figure 6 is a diagram including a DSC plot showing the effectiveness of the embodiment of the present disclosure in Example 3. [Figure 7] Figure 7 is a diagram including a DSC plot showing the effectiveness of the embodiment of the present disclosure in Example 3. [Figure 8] Figure 8 is a diagram including a DSC plot showing the effectiveness of the embodiment of the present disclosure in Example 3. [Figure 9] Figure 9 is a figure including an FTIR plot showing the effectiveness of the embodiment of the present disclosure in Example 3. [Figure 10] Figure 10 shows a typical adaptation process related to this disclosure. [Figure 11] Figure 11 is a figure including a carbonyl index plot showing the digestion efficiency according to the embodiment of this disclosure. [Figure 12] Figure 12 is a diagram including an FTIR plot showing significant differences in performance between the adapted species and a non-adapted control of the same species. [Figure 13] Figure 13 shows a typical adaptation process related to this disclosure. [Figure 14] Figure 14 is a graph illustrating that embodiments of the inoculum of this disclosure, including modified fungal species, can effectively inoculate plastic waste with other waste, including natural polymer waste, under disposal conditions, at an inoculation rate of less than 1:1 fungal inoculum:waste. [Figure 15] Figure 15 is a diagram including FTIR data showing the growth area on a diaper under different conditions for the species relating to this disclosure. [Figure 16] Figure 16 is a diagram including FTIR data showing the growth area on a diaper under different conditions for the species relating to this disclosure. [Figure 17] Figure 17 is a photograph showing fungal growth on a diaper when an embodiment of the inoculant of this disclosure is used. [Figure 18]Figure 18 is a diagram including FTIR data comparing an adapted species embodiment with a non-adapted corresponding species according to this disclosure. [Figure 19] Figure 19 is a schematic diagram illustrating an embodiment of the method according to this disclosure. [Figure 20] Figure 20 is a schematic diagram illustrating an embodiment of the fungal consortium relating to this disclosure for decomposing waste. [Figure 21] Figure 21 is a diagram containing photographs showing the growth of each inoculant blend in Example 8. [Figure 22] Figure 22 is a diagram including a photograph showing the seed blend in a petri dish according to Example 8. [Figure 23] Figure 23 shows the carbonyl index data for samples when multiple modified fungal species were used individually and when they were used in combination. [Figure 24] Figure 24 shows the emergence data from beads according to this disclosure and Example 8. [Figure 25] Figure 25 is a schematic diagram illustrating various forms (form factors) applicable to the embodiments of this disclosure. [Figure 26] Figure 26 is a graph illustrating the combination of a polymer product contaminated at a low inoculation rate and a dry inoculum in the presence of wet contaminants. [Figure 27] Figure 27 is a photograph showing the emergence of fungi from immobilized and encapsulated mycelial beads in a typical waste product environment according to Example 11. [Figure 28] Figure 28 is a photograph of a petri dish showing the emergence of fungi from an immobilized fungal inoculum attached to a nonwoven fabric, according to Example 13. [Figure 29] Figure 29 is a diagram including photographs showing the rehydration process and appearance of the inoculum relating to this disclosure. [Figure 30] Figure 30 is a diagram including a photograph showing the appearance of fungal hyphae related to this disclosure on diaper material and vigorous colony formation. [Figure 31]Figure 31 is a schematic diagram illustrating the process of growing filamentous fungi on a polymer scaffold to form the shelf-stable product according to this disclosure. [Figure 32] Figure 32 is a diagram including a photograph showing the emergence of the modified fungal species relating to this disclosure from a fiber scaffold inoculum. [Figure 33] Figure 33 is a diagram including a photograph showing the emergence of the modified fungal species relating to this disclosure from a coated fiber inoculum. [Figure 34] Figure 34 is a schematic diagram illustrating the method for manufacturing the absorbable product according to this disclosure. [Figure 35] Figure 35 is a diagram including a photograph showing the appearance of modified fungal inoculum from the absorbable product relating to this disclosure. [Figure 36] Figure 36 is a diagram including a photograph showing the emergence of the fungal inoculum from the film container relating to this disclosure. [Figure 37] Figure 37 is a photograph showing the results of Example 16. [Figure 38] Figure 38 is a photograph showing the absorption results in Example 17. [Figure 39] Figure 39 schematically illustrates the unexpected synergistic absorption effect that occurs when natural hydrogels and synthetic hydrogels are combined. [Figure 40] Figure 40 is a schematic diagram showing a fungal inoculum treatment regime for plastic waste according to an embodiment of the present disclosure. [Modes for carrying out the invention]

[0031] Polymer digestion can be described using a first-order reaction kinetics model. This model is represented by an exponential decay equation that shows the decomposition rate is proportional to the amount of remaining material. C = C0 × e^(-kt)

[0032] Here, C represents the concentration of the residual material, C0 represents the initial concentration, k represents the decomposition rate constant, and t represents time.

[0033] Based on experimental studies, the estimated degradation times for some common polymer materials are as follows: Superabsorbent polymer (SAP): These polymers are designed to be highly stable and may take hundreds of years to decompose. For example, one study showed that the half-life of highly absorbent polymers in soil is approximately 300 years. Thin film PE film: The estimated degradation time of thin PE film in the environment is several hundred years. In a study on the degradation of PE film in seawater, the weight loss of the film was less than 3% even after 28 months. PE non-woven fabric: Similar to thin-film PE films, PE nonwoven fabrics may take several hundred years to decompose in the environment. In studies on the decomposition of nonwoven fabrics, the weight loss of PE nonwoven fabric after 90 days was less than 2%. PP non-woven fabric: Even PP nonwoven fabrics may take several hundred years to decompose. In a study on the decomposition of PP nonwoven fabrics in soil, the weight loss of the material after 180 days was less than 5%. Cellulose fiber: Cellulose fibers are biodegradable and decompose much faster than synthetic polymers. The time required for decomposition varies depending on the conditions, but it is usually several months to several years. In a study on the decomposition of cellulose fibers in soil, the weight reduction of the fibers after 90 days was approximately 30%.

[0034] In this specification, “immobilize” means fixing fungi or bacteria in a predetermined location within a material in order to facilitate targeted and controlled digestion. In this specification, "shelf stable" means an inoculum that is prepared to maintain viability over time without requiring refrigeration or other specific storage conditions. In this specification, "web conversion stable" means an inoculant that maintains viability throughout the entire manufacturing process of an absorbable product, even under stress conditions such as high temperatures. In this specification, "rehydrate" refers to the process by which a dehydrated fungal inoculum absorbs water and reactivates the metabolic processes necessary for digestion. In this specification, “emerge” means the process by which, after rehydration or under suitable environmental conditions, the fungal inoculum breaks out of its encapsulation state, begins to reactivate, grows, and recognizes the polymer matrix. In this specification, "fungi" refers to microorganisms used in digestion processes to break down polymer materials. In this specification, “preselected” means fungi that have been selected and screened based on their ability to digest specific types of polymer materials in a waste environment, as well as their ability to effectively compete in the presence of biological and chemical contaminants. In this specification, “adapted” or “engineered” means a fungal strain or microbial community that has been genetically or environmentally conditioned to improve its digestive capacity for a particular substrate. In this specification, "colonize" means the ability of fungi or microorganisms to establish themselves and grow on a polymer-based material, thereby initiating a digestive process. In this specification, “degrade” means a biochemical process in which fungi or microorganisms break down complex polymer chains into simpler, less harmful substances. In this specification, "waste stabilization" refers to technologies for managing waste by minimizing the leaching of toxins and microplastics into the environment. In this specification, “enzymatic digestion” or “enzymatic degradation” refers to the process by which fungi produce a series of enzymes (and other metabolites) to chemically break down polymers into simpler compounds. In this specification, "metabolize" refers to the process by which fungi absorb carbon derived from polymers during digestion and convert it into energy and cellular components. In this specification, "scalable" means a process or technology that can be scaled from laboratory scale to industrial scale without compromising its functionality or effectiveness. In this specification, "consortia" means a group of different microbial species that cooperate to digest polymers more efficiently than a single species. In this specification, "consortia composition" refers to the specific composition of a microbial consortium, as measured by the proportions of each species. In this specification, “contaminate” means that the contaminated material or waste product, including biological waste, is subject to digestion. In this specification, “disposal environment” means the environment or conditions under which polymer products are disposed of and subjected to a digestion process. In this specification, “inoculation” means active fungal or microbial cells introduced into a polymer material to initiate digestion. In this specification, "inoculation rate" means the density of microorganisms or fungal cells introduced per unit weight or unit volume of a substrate, and is an important indicator for ensuring effective and scalable digestion. In this specification, "inoculate" means the act of introducing a microbial or fungal inoculum into a polymer material in order to initiate a digestive process. In this specification, "absorbency" refers to a material's ability to absorb and retain liquids, and is relevant to the design of polymer-based materials such as diapers that are intended for digestion. In this specification, "retention" refers to a product's ability to retain absorbed liquid under pressure, and is important for the performance of products such as sanitary napkins and diapers. In this specification, "permeable" refers to the property of a material to allow the passage of liquids, which may affect the rate and efficiency of plastic consumption by fungal inoculants. In this specification, "integration" means the process of incorporating a fungal inoculant into a polymer matrix during or after the manufacturing process in order to facilitate digestion. In this specification, "embed" means a process that ensures close contact and effective digestion by directly incorporating a fungal inoculum into the structure of a polymer-based material. In this specification, "controlled time release" refers to a formulation characteristic that enables the time-controlled release of fungal inoculum from the product in order to synchronize waste management schedules and digestion. In this specification, "scaffold" refers to a structural framework into which a fungal inoculant is incorporated, for the purpose of promoting uniform distribution and effective colonization for digestion. In this specification, “pretreatment” means a treatment applied to a polymer material before the introduction of an inoculant in order to enhance its susceptibility to digestion or decomposition. In this specification, “biological additive” means nutrients or other compounds added to a substrate to support the growth and metabolic activity of degrading organisms. In this specification, "bead" means granules whose shape or size is not specified. In this specification, "filler" means a material other than fungal biomass, including alginate, trehalose, and nutrient mixtures, which is included in the embodiments of the inoculant of this disclosure. In this specification, "hydrogel" means a naturally derived or synthetic material having a three-dimensional network of hydrophilic polymers formed by chemical or physical crosslinking, and capable of absorbing water. In this specification, “natural hydrogel” means a material that includes, but is not limited to, cellulose, chitosan, collagen, alginate, agarose, hyaluronic acid, gelatin, and fibrin. In this specification, "synthetic hydrogel" means a material including, but not limited to, poly(hydroxyethyl methacrylate) (PHEMA), polyethylene glycol (PEG) hydrogel, polyacrylic acid (PAA), and superabsorbent polymer (SAP). In this specification, “micro-colony” or “micro-colonies” means an isolated cluster of fungal cells. In embodiments of the present invention, micro-colonies contain individually encapsulated modified fungal cells, each exhibiting different properties and behaviors, thereby providing multiple inoculation sites and improving adaptability and functionality compared to natural fungal colonies.

[0035] Fungal modification to improve the digestion of polymer-based materials using pre-selected fungi and the resulting species aggregates.

[0036] This disclosure discloses a method for developing fungal species adapted to digest plastic materials and to be resistant to abiotic stress. The development process includes screening candidate fungi, modifying them to improve digestion efficiency, and mixing the fungal inoculum with plastic waste products.

[0037] Fungal species are first screened for their inherent ability to digest plastic materials. Selected fungi are then modified to utilize plastic as a carbon source in their metabolic processes and to withstand abiotic stress conditions such as feces and urine, which are commonly present in used absorbent articles and other polymer-based commercial products.

[0038] These modified fungi can grow even under harsh conditions and have the ability to utilize plastic as a carbon source. Further optimization, including additional modifications to improve the rate of plastic digestion, enables efficient and rapid digestion of plastic waste in various environments.

[0039] The aforementioned modification method and the modified fungal products obtained thereby possess extremely high versatility and scalability, and are applicable to a wide range of polymer types commonly used in absorbent articles and other polymer-based materials.

[0040] Embodiments of this disclosure also include methods for improving the digestion of non-plastic materials, cellulose, and human waste, in addition to the decomposition of plastics. Immobilized fungi can be used in combination with other technologies, thereby enabling the removal of environmental toxins, reduction of methane production, and carbon sequestration.

[0041] Embodiments of the present disclosure for degrading plastics using fungi offer advantages over known methods of incorporating enzymes into products. Specifically, embodiments of the present disclosure utilize fungi as living organisms that can survive at relatively low inoculation rates, and these fungi are modified to grow continuously using polymers as a carbon source, even in the presence of abiotic stressors or other fungal species. Known methods and techniques, on the other hand, generally rely on enzyme production in laboratories or controlled environments, which is time-consuming and costly, and difficult to use in typical commercial settings.

[0042] In contrast, embodiments of this disclosure utilize the enzyme-producing ability and plastic material digestion ability inherent in fungi. Fungi can grow and spread rapidly even at low inoculation rates, enabling their application to large-scale applications. Unlike laboratory-produced enzymes, fungi can adapt to changes in environmental conditions and continue to grow and produce enzymes over long periods.

[0043] The following examples demonstrate that fungi modified in accordance with this disclosure digest polymers in diapers more rapidly than unmodified fungi. The natural decomposition time of typical diaper materials (without using the embodiments of this disclosure), based on experimental studies, is as described above.

[0044] Embodiments of this disclosure focus on the digestion of absorbent products such as diapers and the various materials used in their composition. Specifically, they demonstrate the ability of fungal species / strains or combinations thereof to target and digest various polymers commonly found in absorbent products, including films, elastic materials, nonwovens, hot melts, and superabsorbent materials. As disclosed herein, highly efficient and effective products and technologies are provided for improving the degradation of absorbent products by selecting and modifying fungal strains to specifically target and digest these polymers, even in the presence of abiotic stressors.

[0045] The ability of fungal strains to target and digest various polymers in the presence of human excrement and other abiotic stressors present in absorbent products is applicable to other types of absorbent products, including adult diapers, feminine hygiene products, and other absorbent products intended for use by humans or animals. Fungal strains can be modified to specifically target and digest polymers used in various absorbent products, thereby providing a highly effective digestion and disposal method.

[0046] The ability of fungal species / strains or combinations thereof to digest absorbent products extends to other product categories, including plastic bags, packaging materials, textile products, and other common polymer-based materials. As shown herein, the methods and compositions of this disclosure are adaptable to target and digest polymers contained in these materials, thereby providing highly effective and versatile digestion and disposal methods.

[0047] The methods and products of this disclosure represent a significant advance over existing technologies in the field of biodegradable absorbent products. Unlike some published studies, the embodiments of this disclosure are operable outside of laboratory environments, enabling commercial applications at the consumer scale. Furthermore, the embodiments of this disclosure achieve the digestion of materials such as superabsorbent polymers (SAPs). In addition, the embodiments of this disclosure do not require material pretreatment, thus providing a more practical and efficient solution in plastic waste management. Further advantages of the embodiments of this disclosure include: By using modified fungal inoculants, a scalable solution is provided for degrading polymer components in absorbent products without any modification of the plastic material. Compositions of fungal species modified to digest long-chain carbons in plastics enable effective digestion of a wide variety of polymer types. The embodiments of this disclosure are effective for a wide range of plastic waste types in their commercially available form (i.e., without any plastic pretreatment steps). Embodiments of this disclosure are modified to target and digest plastic components within absorbent products, and unlike existing studies that show fungi growing on natural polymer components in diapers such as cellulose, they actually digest plastics present within them.

[0048] Furthermore, this specification discloses a method for scaling up pre-selected fungi by bioprocessing, i.e., mycelial expansion including biofermentation, to obtain large-scale, supplyable fungi that improve the digestion of recalcitrant long-chain carbon materials.

[0049] A seed inoculum, which is a clone or direct descendant of a modified saprophytic fungal species, is provided. This seed inoculum is combined with a nutrient mixture tailored to the specific metabolic requirements of the selected fungal species. A bioreactor promotes maximum hyphal expansion. After maximum expansion is achieved, the resulting fungal biomass is homogenized, followed by a filtration process as needed. The homogenized hyphae are then combined with a polymer solution, such as alginate, and subjected to further processing. Subsequently, mounting and immobilization processes are applied to ensure the stability of the fungal inoculum. These processing techniques used for modified saprophytic fungi enable methodologies for improving digestion and / or degradation to be developed from laboratory-level concepts into commercially viable and scalable applications.

[0050] Embodiments of this disclosure relate to the digestion of absorbent products such as diapers and various materials used in their composition. Specifically, they demonstrate the ability of modified fungal species / strains or combinations thereof to target and digest various polymers contained in absorbent products, including films, elastic materials, nonwoven fabrics, hot melts, and superabsorbent materials.

[0051] Films are commonly used in absorbent products as a layer to prevent liquid leakage and are typically composed of polyethylene (PE), polypropylene (PP), or other suitable polymers. Elastic components are used to provide stretchability and fit and are typically composed of synthetic rubber or other suitable materials. Nonwoven fabrics are used as the top sheet or acquisition layer of absorbent products and can be composed of various polymers such as PE, PP, or bicomponent (Baico) fibers. Hot melts are used as adhesives in absorbent products and are typically composed of polymers such as polyethylene or polypropylene. Superabsorbent materials are the main components of absorbent products and are typically composed of cross-linked polymers such as sodium polyacrylate.

[0052] By selecting and modifying fungal strains to specifically target and digest these polymers, highly efficient and effective digestion of absorbable products can be achieved.

[0053] Importantly, the ability of fungal strains to target and digest various polymers in the presence of human waste and other abiotic stressors present in absorbent products makes them applicable to a wide range of absorbent products, including adult diapers, feminine hygiene products, and other absorbent products intended for human or animal use. Fungal strains can be modified to specifically target and digest polymers used in various absorbent products, thereby providing highly effective digestion and disposal methods.

[0054] The ability of fungal species / strains or combinations thereof to digest absorbent products extends to related fields such as plastic bags, packaging materials, textile products, and other polymer-based materials. The methods and compositions of the present invention are adaptable to target and digest polymers contained in these materials, thereby realizing highly effective and versatile digestion and disposal methods.

[0055] Furthermore, the examples demonstrate that the fungal inoculum maintains its effectiveness even after rehydration following immobilization. Scaled-up fungal inoculum were prepared using 2% medium-viscosity sodium alginate and homogenized hyphae, with alginate content ranging from 1% to 4% (low and medium viscosity). This mixture was used in Examples 1 to 3 below.

[0056] Example 1: Identification and screening of fungal species that digest diaper materials

[0057] This embodiment involves the use of fungal species that have been identified and evaluated for their effectiveness. These fungi exhibit saprotrophic capabilities suitable for the development of sustainable biological processes. Fungi play an important role in natural ecosystems by decomposing organic matter, including complex polymers found in wood, such as lignin, cellulose, and hemicellulose. Embodiments of this disclosure leverage the unique saprotrophic capabilities of specific fungal species, which are classified as white rot, brown rot, and soft rot fungi (referred to herein as Species W, Species B, and Species S, respectively). The use of these uniquely capable fungal species in unique ways forms the basis of the inventive embodiments of this disclosure, and fungal species are frequently described herein based on these classifications.

[0058] White rot fungi (Species W) White rot fungi are characterized by the production of high levels of oxidizing enzymes, which enable them to break down lignin, a complex aromatic polymer in wood. These fungi have robust hyphae and secrete hydrophobic metabolites that aid in colonization and stability. These species exhibit a moderate delay in emergence, but their growth and colonization are rapid. Taxonomically, white rot fungi belong to the phylum Basidiomycota and are mainly classified under the class Agaricomycetes, which includes orders such as Agaricales, Hymenochaetales, and Polyporales. The specific species used in this embodiment include Pleurotus ostreatus, Pleurotus djamor, Dichomitus squalens, Trametes versicolor, Phanerochaete chrysosporium, Lentinula edodes, and Phellinus pini.

[0059] Brown rot fungi (Species B) Brown rot fungi are notable for producing high levels of hydrolytic enzymes, which efficiently digest cellulose and hemicellulose in wood, leaving behind a brown, brittle lignin residue. These fungi have excellent competitive capabilities against microbial contaminants and generally have a long emergence delay, but once established, they exhibit moderate growth and establishment rates. Brown rot fungi also belong to the phylum Basidiomycota and class Agaricomycetes, and include species belonging to orders such as Gloeophyllales and Polyporales. Species used in this example include Inonotus obliquus and Fomitopsis spraguei.

[0060] Soft rot fungi (Species S) Soft rot fungi can grow in harsh environments where other fungi struggle to survive, such as in wood with high moisture content, extreme temperature conditions, or pH conditions. These fungi exhibit high compatibility with other fungal species and are useful in diverse microbial communities. Soft rot fungi exhibit moderate to rapid emergence delays, while their growth and establishment are relatively slow. Taxonomically, soft rot fungi belong to the phylum Ascomycota, mainly classified into the classes Dothideomycetes and Sordariomycetes, and include orders such as Hypocreales, Xylariales, and Eurotiales. The specific species used in this example include Aureobasidium pullulans, Pestalotiopsis microspora, Aspergillus sp., Fusarium sp., Aspergillus versicolor, and Aspergillus fumigatus.

[0061] Table 1 Taxonomic and functional classification of species [Table 1]

[0062] Table 2 Examples of pre-selected fungal species [Table 2] Fungal digestion of polymers refers to the extracellular process by which fungi break down complex polymer materials. This process involves multiple stages. First, the fungi sense and recognize the polymer as a nutrient source, then they colonize the polymer and surround it with a biofilm. Within this biofilm, the fungi secrete extracellular enzymes such as oxidases and hydrolases, which initiate the breakdown of the polymer, producing intermediate compounds. These intermediates are then taken up and assimilated by the fungi. Therefore, as used herein, the term "digest" encompasses the entire process of recognizing, colonizing, enzymatically breaking down, and assimilating polymer materials.

[0063] Table 3. Fungal digestion of polymers [Table 3]

[0064] The stages of the digestion process, as well as the related modification processes and procedures, used to design the embodiments of the modified fungi relating to this disclosure are described in detail below.

[0065] Recognition Fungi recognize polymers as a potential nutrient source through chemical signals or structural elements specific to the polymer. The modification process disclosed herein involves using a pre-digested polymer, which encourages the fungi to extend their hyphae towards the polymer rather than ignoring it. To determine whether the recognition stage is occurring, the growth of the fungi toward the polymer can be observed by inoculating fungi into a nutrient medium petri dish and placing a small amount of polymer at a distance from the inoculation point. In field tests using complete polymer products, fungal colonization on the polymer surface can be monitored over time by visual and microscopic observation to confirm directional growth toward the polymer.

[0066] Colonization After recognition, the fungi begin to colonize the polymer surface, attaching themselves to it and growing. At this stage, hyphae develop to establish a stable environment, preparing for complete digestion. Whether or not colonization has occurred can be evaluated by visually and microscopically observing the attachment and growth of mycelia on the polymer surface. The fungal growth area can be calculated by measuring the colonization range, and the growth rate can be determined by tracking the expansion of the fungal network over time.

[0067] Biofilm secretion Within biofilms, fungi secrete various extracellular substances, including enzymes and hydrophobins. These adaptations allow fungi to rapidly secrete mixtures suitable for specific polymers they sense. In addition to visual and microscopic observation, colorimetric analysis kits can be used to detect the presence of enzyme production when fungi colonize plastics. These techniques confirm the active involvement of fungi in biofilm formation and digestion processes.

[0068] Biodeterioration Fungi weaken polymers through mechanical action and oxidases, forming carbonyl groups. This process targets crosslinking sites and amorphous regions of the polymer to promote further degradation, particularly in homochains that are resistant to immediate hydrolysis. Tensile testing can be used to identify changes in polymer strength and elasticity, serving as an indicator of structural weakening. Furthermore, oxidation can be identified by analyzing changes in functional groups within the polymer structure using FTIR and detecting an increase in the strength of the OH band compared to a control.

[0069] Biofragmentation Hydrolytic enzymes cleave the carbon chains of polymers, producing intermediate products such as oligomers. Fungi are adapted to produce the enzymes necessary to break down these polymers into intermediates suitable for assimilation. Biological fragmentation can be identified by various analytical techniques for polymers. For homochains, cleavage of the polymer backbone can be indicated by detecting an increase in the carbonyl index using FTIR. For heterochains, properties such as thermal stability and crystallinity can be evaluated using DSC. For crosslinked polymers such as superabsorbent polymers (SAPs), the most appropriate method is to identify the mass loss of the sample during heating using TGA, which demonstrates degradation and fragmentation.

[0070] Bioassimilation Intermediate products such as oligomers generated by biofragmentation are taken up into fungal cells. These intermediates are metabolized, supplying energy and building blocks for fungal growth. Fungi possess a unique ability to initiate and catalyze biodegradation and biofragmentation. Once these steps are confirmed, the intermediate byproducts are taken up by the fungi. This uptake is demonstrated by the persistence of metabolism even when the polymer is inoculated at low inoculation rates, demonstrating that the fungi are dependent on this carbon source for survival.

[0071] Mineralization Hydrolysis products are transported into the cell wall and converted into microbial biomass. The sustained life cycle and continuous growth on the polymer substrate indicate that the fungi effectively utilize the plastic as a nutrient source and continue all the aforementioned processes. Mineralization is determined by the persistence of growth on the polymer product even at low inoculation rates. This is an indicator that the fungi effectively utilize the degraded polymer components as a carbon source, leading to continued metabolic activity and further colonization of the substrate.

[0072] Polymer Testing Embodiments of this disclosure primarily, but not limited to, those relating to the field of fluid-absorbent articles, particularly diapers and other personal waste disposal products. Furthermore, this disclosure also provides embodiments of novel absorbent materials and structures for use in various absorbent products that can improve absorbency, liquid retention, and overall performance while ensuring comfort, fit, and ease of use. The methods provided herein do not require novel materials to achieve more sustainable products and eliminate the trade-off between performance metrics and sustainability.

[0073] The polymers used in the methods and products described herein are classified into homochain, heterochain, crosslinked polymers, and copolymers. Homochain polymers such as polyethylene (PE) and polypropylene (PP) are less susceptible to hydrolysis and require oxidation for digestion and decomposition. These materials are tested in various components of diapers, such as backsheet nonwovens, backsheet films, and leg cuffs. Heterochain polymers, including polyethylene terephthalate (PET) and polyurethane (PU), are relatively more susceptible to hydrolysis but have complex structures. These are tested in acquisition and distribution layers (ADLs) and elastic strands, respectively.

[0074] Crosslinked polymers and copolymers also play important roles in the methods and products of this disclosure. The superabsorbent polymer (SAP) used is a hydrogel consisting of sodium polyacrylate (NaPA) crosslinked with approximately 5% polyethylene glycol (PEG), and is designed to improve liquid retention capacity. Furthermore, copolymers such as acrylonitrile butadiene styrene (ABS) are used in adhesive applications, improving adhesive performance by combining the properties of different monomers.

[0075] The materials tested in this specification include a diverse group of polymers used in various components of diapers. For example, backsheet nonwoven fabrics made of polypropylene (PP) and backsheet films made of polyethylene (PE) are representative of homochain polymers. On the other hand, acquisition and distribution layers made of polyethylene terephthalate (PET) and elastic strands made of polyurethane (PU) are representative of heterochain polymers. Furthermore, adhesives made of cross-linked hydrogels such as SAP and copolymers such as ABS demonstrate the breadth of materials used in this invention.

[0076] Absorbent products such as diapers contain multiple layers designed to perform specific functions. The top sheet is typically a nonwoven fabric made of PE or PP and is intended to come into contact with the skin. The acquisition layer rapidly transfers liquid to the absorbent core. The absorbent core is made of cellulose fibers and SAP and captures and retains the liquid. The back sheet, made of PE or PP, prevents liquid leakage. In addition, additional components such as adhesives, elastic members, and fasteners are included to ensure functionality and fit.

[0077] This versatile approach allows these materials to be scaled up and adapted to other combinations within the layer structure of absorbent products. Furthermore, the methodology and materials developed in this invention are applicable to a wide range of plastic products, not limited to absorbent products, providing a scalable solution for waste management.

[0078] Table 4 Diaper materials [Table 4]

[0079] Example 2: Digestion of diaper material In Example 2, 30 fungi belonging to the classifications of white rot, brown rot, and soft rot were used and mixed with superabsorbent polymer (SAP) at progressively decreasing inoculation rates. Specifically, the weight ratio of fungal inoculum to plastic was 99:1, then 10:1, then 4:1, and finally 1:1. As a result of this adaptation process, 12 fungi were selected that showed the ability to effectively recognize and colonize SAP as a nutrient source. These adapted cultures were then applied at a 1:1 inoculation rate to the following diaper materials: top sheet nonwoven fabric (PP), back sheet nonwoven fabric (PP), core wrap (PP), back sheet film (PE), and SAP. The average growth rate was observed over 30 days. Figure 1C shows the flowchart of Example 2, petri dish photographs showing significant growth in soft rot fungi (species S) and white rot fungi (species W), and growth rate data of white rot fungi (species W) on different plastic materials.

[0080] To confirm biofilm secretion and enzyme production, fungal species and strains were compared with known polymer-degrading proteins, and their ability and gene expression were evaluated when cultured in the presence of TS. Specific enzyme activity was qualitatively evaluated by the following colorimetric analysis. Laccase: The formation of a dark green halo on a culture medium to which ABTS has been added indicates positive laccase secretion. Lignin peroxidase: The formation of a clear halo on a culture medium supplemented with azur-B indicates positive lignin peroxidase secretion. Manganese peroxidase: The formation of a reddish-brown halo on a culture medium supplemented with guaiacol indicates positive manganese peroxidase secretion.

[0081] Qualitative analysis revealed that the adapted fungal species exhibited diverse and abundant enzyme production in the presence of plastic. Figure 2 shows colorimetric analysis results illustrating lignin peroxidase production in soft rot fungi (species S) and laccase and manganese peroxidase [C] production in white rot fungi (species W).

[0082] Example 3: Optimization of the inoculant for diaper material digestion

[0083] In Example 3, the weight ratio of inoculant to plastic was set to 1:1, meaning that the weight of the plastic waste to be treated was equal to the weight of the fungal biomass and nutrient compounds. This ratio is important to ensure that the fungi have sufficient resources to efficiently colonize and digest the plastic material. In the preferred inoculant embodiments described in detail below, as typically shown in Figure 1B, it has been shown that inoculation into plastic materials is effective even at inoculant:plastic weight ratios of less than 1:1.

[0084] The wet inoculum used in Example 3 consisted of 4% fungal biomass and 96% cross-linked sodium polyacrylate. The fungal biomass was cultured from three species (two of species S and one of species W) of soft rot fungi (species S) and white rot fungi (species W) that had been screened and adapted. This inoculum was applied to each individual layer of the diaper and tested for 30 days on the following materials: backsheet nonwoven fabric, backsheet film, front ear nonwoven fabric, landing zone, leg cuff nonwoven fabric, topsheet nonwoven fabric, adhesive, acquisition / distribution layer, elastic strand, and SAP. Environmental control samples were tested simultaneously with the fungal-treated samples, and the results were compared to undegraded / undigested raw material (virgin) material. The digestion results for each material layer were analyzed, including both biological activity and polymer characterization. The data obtained indicate that the enzymes produced by the adapted focal fungal strains digest polymers belonging to the homochain, heterochain, and cross-linked polymer groups. Furthermore, compared to environmental controls and untreated raw materials, fungal-treated samples showed changes in physical and chemical structure (oxidation), as well as cleavage of polymer bonds (hydrolysis), indicating successful recognition, colonization, secretion, biodegradation, and biofragmentation by the adapted fungal samples.

[0085] Furthermore, the presence of mycelial growth and enzymatic activity on the surface of various diaper materials provides evidence that digestion is occurring. All diaper materials tested were recognized and colonized by one or more adapted fungal species. Figure 3 is an SEM image showing that the top sheet (TS) was effectively inoculated with soft rot fungus (species S), and the fine filamentous structures observed throughout the matrix are mycelia. Figure 4 is an SEM image showing that the back sheet nonwoven fabric (BSNW) was effectively inoculated with white rot fungus (species W), and digestion of the polypropylene nonwoven material can be confirmed.

[0086] Thermogravimetric analysis (TGA) results showed that fungal treatment samples of SAP (PEG-crosslinked NaPA) with white rot fungus (species W) exhibited lower thermal stability at all stages of the test compared to untreated SAP control and environmental control samples. As shown in Figure 5, the fungal treatment samples showed a significant initial weight loss of approximately 95-96% at around 100°C, which is thought to be due to increased porosity or surface area resulting from fungal digestion of the crosslinked structure, leading to high hydrophilicity. Between 100°C and 500°C, further weight loss was observed, indicating the decomposition of polymer bonds, and a stepwise weight loss was observed, indicating that shorter polymer chains were decomposed at lower temperatures. In the high-temperature range (500°C to 1000°C), the fungal treatment samples left only about 2.08% residue, the lowest amount among the samples, indicating thorough decomposition of the organic structure. These results clearly demonstrate the significant effect of fungal digestion on SAP.

[0087] Furthermore, differential scanning calorimetry (DSC) results showed significant digestion in hot-melt adhesives subjected to fungal treatment with white rot fungus (species W) and environmental exposure. As shown in Figure 6, both the environmental control and fungal-treated samples exhibited lower glass transition temperatures (Tg) compared to the untreated adhesive, suggesting digestion due to a decrease in molecular weight or structural changes. In the environmental control sample, crystallization behavior was observed during the first heating cycle with a start temperature of 37.74°C and a peak temperature of 71.76°C, whereas this behavior changed significantly in the fungal-treated sample, indicating structural changes. Moreover, in the environmental control sample, a decrease in crystallinity was observed during the second heating cycle, with a start temperature of 92.53°C, a peak temperature of 100.09°C, and a specific energy of 1.712 J / g, while no melting peak was observed in the fungal-treated sample, indicating widespread digestion and loss of crystalline regions. These results indicate that both environmental exposure and fungal treatment digest the adhesive, but fungal treatment causes more pronounced structural changes and complete loss of crystallinity.

[0088] Similarly, as shown in Figure 7, the DSC results for the elastic material showed significant digestion in the fungal-treated (species W) sample compared to the untreated sample and the environmental control sample. The untreated elastic material showed almost no digestion between heating cycles and exhibited a stable structure. On the other hand, the environmental control sample showed high crystallinity with a specific energy of 8.129 J / g in the first heating cycle, suggesting that the environmental conditions strengthened the polymer structure while causing slight digestion between cycles. In contrast, the fungal-treated sample showed moderate crystallinity with a specific energy of 3.921 J / g in the first heating cycle, indicating significant digestion compared to the untreated sample and the environmental control sample. The stable structure between heating cycles in the fungal-treated sample indicates that a large structural change occurred during the initial fungal treatment, resulting in significant digestion and moderate crystallinity compared to the other samples.

[0089] Figure 8 shows the DSC results for the PET nonwoven fabric layer, indicating significant digestion in the fungal-treated (species S) sample compared to the untreated and environmental control samples. In the environmental control sample, a slight decrease in specific energy at the endothermic peak (melting) and limited digestion with a minor decrease in crystallinity were observed. On the other hand, in the fungal-treated sample, a high specific energy was maintained at the endothermic peak, and although the effect on crystallinity was small, a secondary melting peak was observed, suggesting complex thermal behavior and the formation of different crystalline phases or structures due to the fungal treatment. These findings indicate that while the untreated sample exhibits stable crystallinity and thermal properties, the fungal-treated sample undergoes significant structural changes and more complex thermal behavior.

[0090] Fourier transform infrared spectroscopy (FTIR) was used to analyze diaper material before and after inoculation with white rot fungus (species W), and the results are shown in Figure 9. The FTIR results indicate a reduction in the carbon skeleton and oxidation of the material after fungal exposure, providing additional evidence that digestion occurs that cannot be explained by environmental factors alone. In Figure 9, plot A shows undigested samples, environmentally digested samples, and fungally digested samples for polyethylene and plot B shows samples for polypropylene, respectively. Environmental digestion is caused by ultraviolet light and heat. For both polypropylene and polyethylene, at 2900 cm² -1 The sharp peaks in the vicinity correspond to the CH stretching vibration of the methyl group, representing a long carbon skeleton. Furthermore, at 1460 cm² -1 and 1375cm -1 The peaks represent the CH2 and CH3 side chain groups in polypropylene. The decrease in these peaks, as well as the peaks at 1740 cm², indicates that... -1 (Carbonyl group), 1100 cm -1 (Ether and alcohol groups), and 3350 cm -1 The formation of a new peak at the (alcohol group) was confirmed as evidence of oxidation.

[0091] Example 4: Fungal adaptation to polymers A typical adaptation process is schematically shown in Figure 10, illustrating the steps in which an untreated fungal species 100 first undergoes adaptation to plastic 112, followed by further adaptation to abiotic stress 114. Subsequently, the biomass of the adapted fungal species is recovered from the agar plate and used to form the inoculum. In Example 4, a soft rot fungus (species S) was adapted to digest polyethylene (PE), and a white rot fungus (species W) was adapted to digest polypropylene (PP). In this adaptation modification, the fungi were gradually adjusted to utilize these complex polymers as their primary carbon source by systematically reducing their access to simple carbon sources such as glucose and sucrose.

[0092] The adaptation process began with the preparation of a mixture containing a selected polymer (PE in species S, PP in species W) and a carbon nutrient source. In the initial stages, the fungi grew and metabolized using both the polymer and the additional nutrient source. Over successive generations, the concentration of the additional carbon nutrient source was gradually reduced. This forced the fungi to become more dependent on the polymer as the primary carbon source, and the expression of enzymes necessary for digesting the polymer was enhanced. This systematic process yields a modified fungal species that grows significantly faster using plastic carbon compared to the naturally occurring counterpart strain. The fungal species can be further modified using a similar sequential process to better accept specific types of polymer materials as a carbon source for metabolism.

[0093] Each generation was cultured for two weeks before being transferred to the next culture (a medium with reduced carbon concentration). The culture composition of each generation in Example 4 was as follows: First generation (100% carbon): 5g glucose, 5g malt extract, 2.5g yeast extract, 0.5g MgSO4·7H2O, and 1g KH2PO4 were dissolved in 500mL of ultrapure water. Second generation (75% carbon): 3.75g glucose, 3.75g malt extract, 1.875g yeast extract, 0.5g MgSO4·7H2O, and 1g KH2PO4 were dissolved in 500mL of ultrapure water. Third generation (50% carbon): 2.5g glucose, 2.5g malt extract, 1.25g yeast extract, 0.5g MgSO4·7H2O, and 1g KH2PO4 are dissolved in 500mL of ultrapure water. Fourth generation (25% carbon): 1.25g glucose, 1.25g malt extract, 0.625g yeast extract, 0.5g MgSO4·7H2O, and 1g KH2PO4 were dissolved in 500mL of ultrapure water. 4.1st generation (15% carbon): 0.75g glucose, 0.75g malt extract, 0.375g yeast extract, 0.5g MgSO4·7H2O, and 1g KH2PO4 were dissolved in 500mL of ultrapure water. 4.2nd generation (10% carbon): 0.5g glucose, 0.5g malt extract, 0.25g yeast extract, 0.5g MgSO4·7H2O, and 1g KH2PO4 were dissolved in 500mL of ultrapure water. 4.3rd generation: Dissolve 0.5g of glucose and 0.5g of yeast extract in 500mL of ultrapure water. Fifth generation (0% carbon): 0.0g glucose, 0.0g malt extract, 0.0g yeast extract, 0.5g MgSO4·7H2O, and 1g KH2PO4 were dissolved in 500mL of ultrapure water.

[0094] In each generation, the fungal strains were cultured with their corresponding plastic polymer pieces for two weeks before being transferred to the next sequential culture generation. During the transfer, the plastic polymer was first removed, all hyphae were scraped off, and the cultures were dried in a 95°C oven for 12 hours. FTIR data obtained during this process showed improvement in the carbonyl index, an indicator of polymer digestion, in both PE (species S) and PP (species W), as shown in Figure 11. The FTIR results indicate a significant increase in digestive activity in each adapted fungal species as the simple carbon source was reduced, demonstrating an enhanced ability of these fungi to decompose complex polymers.

[0095] Example 5: Field Test

[0096] The effectiveness of the modifications / adaptations was verified by field testing (Example 5). In these tests, 0.3 g of inoculum of fungi identified as soft rot fungi (species S1) and white rot fungi (species W1) was mixed with 30 g of polymer product. Conditions for both adapted and non-adapted inoculums were tested. These mixtures were left in an open outdoor environment and collected after 6 weeks. The collected materials were backsheet film (BSF) made of polyethylene (PE) and backsheet nonwoven fabric material made of polypropylene (PP), which were then tested.

[0097] The FTIR plots in Figure 12 show significant differences between the adapted species S1 and W1 and the unadapted strains. The adapted inoculum showed increased oxidation peaks and more pronounced changes in polymer structure. The adapted soft rot fungi (species S1) and white rot fungi (species W1) successfully digested polyethylene and polypropylene, respectively, clearly demonstrating that these can be scaled to real-world waste environments by using fungi that maintain enzyme expression even outside of laboratory conditions.

[0098] Example 6: Application to contaminants

[0099] Example 6 focuses on the adaptation of fungi to abiotic stressors such as urine, aiming to improve performance on substrates such as plastics, urea, and feces. This example demonstrates the adaptation of fungi to saline environments by focusing on growth, development, and enzyme activity under different salt concentration conditions. An example of the abiotic stress adaptation process used in Example 6 is schematically shown in Figure 13. White rot fungi (species W) adapted according to Example 4 were cultured on potato dextrose agar (PDA) and liquid medium. In liquid culture, the fungi were progressively exposed to salt concentrations of 0.45%, 0.9%, and 1.8%. Positive controls (no salt added) and negative controls (no inoculum) were also prepared. Cultures were carried out at 30°C and monitored every 3 days for 43 days, with biomass and enzyme activity measured. Every 3 days, 15 mL of liquid was taken from each container and subjected to analysis. Parameters such as pH, electrical conductivity, salinity, oxidation-reduction potential (ORP-mV), and total dissolved solids (TDS) were measured from the liquid culture to monitor environmental conditions and fungal metabolic activity. These parameters indicated that the fungi were growing sustainably without death. The fungi adapted well to increasing salinity and showed significant growth and enzymatic activity even under the highest salinity conditions. These modified fungal species adapted to plastic and abiotic stress formed the basis for the embodiments of this disclosure and were used in the following examples.

[0100] Example 7: Field test of contaminated product and inoculant combinations

[0101] As schematically shown in Figure 14, embodiments of the inoculum containing modified fungal species according to this disclosure are effective for inoculating plastic waste under “field” conditions, coexisting with other waste, including natural polymer waste, even when the weight ratio of fungal inoculum to waste is less than 1:1. Example 7 demonstrates the effectiveness of the inoculum even under conditions of fully contaminated products and fungal inoculum left in an open environment. 650 contaminated diapers of the same composition were collected from a childcare facility. The diapers were inoculated with fungal inoculum in different proportions. The type of contamination (urine or feces) was recorded. Nine modified fungi were tested for each of white rot fungi (species W), soft rot fungi (species S), and brown rot fungi (species B), with three replicates placed in one bucket for each condition. The wet inoculum used in Example 7 consisted of 4% fungal biomass and 96% cross-linked sodium polyacrylate. Fungal biomass was cultured from fungal species screened and adapted according to the above examples and scaled up by liquid fermentation. The mycelial biomass was recovered, homogeneously mixed with 2% sodium polyacrylate, and then extruded into a calcium chloride solution to crosslink and form beads approximately 3 mm in diameter. In particular, the recorded inoculation rates are based only on the weight of the dry diaper, excluding the weight of the contaminated material.

[0102] Samples for FTIR analysis were collected one month later, and each sample was observed and retrieved again three months later for further analysis. Figure 15 shows the total growth area on the diaper for all fungal species, along with box plots indicating the type of contamination (urine, urine, and feces), demonstrating that the modified fungi in the inoculum can compete and colonize against both biological and abiotic contaminants.

[0103] Figure 16 shows the total growth area at inoculation rates of 40%, 20%, and 10% (as defined in Figure 14), demonstrating that the fungal inoculum can effectively establish itself even at low inoculation rates.

[0104] Significant fungal growth was observed after one month, and after two months, it was confirmed that the white rot fungus (species W) had grown completely throughout the bucket filled with diapers, as shown in the photograph in Figure 17.

[0105] The carbonyl index (intensity or area change of carbonyl groups in the FTIR spectrum) was used as quantitative data to measure the degree of plastic digestion (a general surrogate indicator of digestion). The presence of carbonyl groups indicates that the polymer is digested, and further digestion is promoted due to the instability of the carbonyl groups. The carbonyl index is calculated from the FTIR spectrum by analyzing the peaks indicating the presence of carbonyl and methylene groups (see Equation 1 below). As oxidation progresses during digestion, the methylene groups decrease and are eventually converted into carbonyl groups. The degree of digestion can be evaluated by comparing the carbonyl index (CI) of the digested sample with that of the undigested control.

number

[0106] FTIR data confirmed that all fungal species showed signs of polymer digestion after one month, and the calculated carbonyl index (CI > 1) indicated better digestion compared to the control sample. After two months, the carbonyl index significantly increased to an average of 6. On the other hand, polypropylene (PP) continued to maintain a CI > 1, but the increase was relatively small. As shown in Figure 18, all modified fungal species (species S, species W) showed higher digestion efficiency compared to their respective unmodified controls.

[0107] Two months later, differential scanning calorimetry (DSC) analysis was also performed. This sample was a composite sample consisting of backsheet film (PE) and backsheet nonwoven fabric (PP), and crystallinity was calculated using one heating cycle in this method. A significant decrease in crystallinity was observed in two samples of white rot fungus (species W), indicating that substantial digestion had occurred. Specifically, for PE, species W3 showed 79.96% crystallinity, and species W4 showed 69.08% crystallinity. For PP, species W3 showed 31.59% crystallinity, and species W4 showed 92.25% crystallinity.

[0108] Table 5: Decrease in crystallinity in white rot fungus (species W) samples [Table 5]

[0109] Furthermore, this specification discloses embodiments of complex fungal consortia configured to enhance the digestion of polymer waste products. The pre-selected fungi are not found in nature in any particular combination, thereby enabling adaptation to both biological and abiotic factors. The consortia is configured to recognize and utilize the symbiotic relationships among its members, providing an effective and robust inoculum under various environmental conditions. The importance of such a consortium lies in its ability to synergistically promote the digestion of refractory polymers and efficiently decompose complex polymer mixtures often found in waste products.

[0110] Figure 19 shows the composition of a fungal consortium consisting of two or more pre-selected fungi. These fungi are selected based on their ability to form a viable consortium in a waste environment and synergistically enhance the digestion of recalcitrant polymer materials. This consortium can handle complex polymer products containing multiple types of polymers. The selection characteristics of the consortium members are shown in a way that highlights the strengths of each member in a specific product mixing environment. These fungi are selected from a pre-screened catalog adapted based on characteristics observed in nature, and the enzymatic and metabolic capabilities of each member are optimized for polymer digestion.

[0111] Figure 20 shows a unique blend of fungal consortia, indicating that it is tailored to target specific polymer products. For example, products with a high proportion of homochains require a higher proportion of oxidase-producing fungi compared to products consisting primarily of heterochains or natural polymers. The consortia members are blended in appropriate ratios to ensure efficient colonization and digestion. This encapsulated blend is configured to allow all members to emerge and colonize even under reduced inoculation conditions. The ratio is a function of factors such as doubling acceleration and emergence delay time, highlighting the complexity of constructing effective species blends at inoculation rates of less than 1:1, and in some cases even less than 0.1:1. Embodiments of this disclosure ensure a consortia that can effectively digest polymers even under reduced inoculum concentrations.

[0112] Example 8: Formulation of a seed consortium inoculum blend

[0113] Example 8 demonstrates the formation of a blend of fungal species that effectively colonize and synergistically interact without competing with each other. For each fungal species, the lag time from emergence, growth rate, and suitability were considered and analyzed, and these factors were reflected in the various mixing ratios within the inoculum blend to ensure overall effectiveness. As shown in Table 6 below, four species of soft rot fungi (species S) and two species of white rot fungi (species W) were blended in various ratios. Each petri dish was inoculated with a total weight of 10 g of beads. Table 6: Seed ratio blend [Table 6]

[0114] Observations revealed that in blends 1 and 2, effective growth was observed from soft rot fungi (species S1) and white rot fungi (species W1), while little to no growth was observed from soft rot fungi (species S2) and white rot fungi (species W2). In blend 3, effective growth was observed from all species except soft rot fungi (species S2). The results for blend 4 were inconsistent, but significant growth was observed from all four species in one replicate test. Figure 21 includes photographs showing the growth after one week for each blend shown in Table 6.

[0115] Subsequently, the blend was optimized so that three species—white rot fungi (species W) and soft rot fungi (species S)—grow proportionally. Figure 22 includes photographs showing fungal growth from these optimized blends, demonstrating that all selected species exhibit balanced and effective growth.

[0116] Example 9A: Testing of seed consortia inoculum blends

[0117] In Example 9A, two species of white rot fungi (species W) and two species of soft rot fungi (species S) were used and tested on contaminated diapers at an inoculation rate of 10%. Each fungal species was tested individually and in mixed blends, with 12 replicates set up for each treatment condition. The diapers were placed in buckets for each replicate sample and buried in outdoor soil. Samples of polyethylene (PE) film were collected after 1 month and 2 months and subjected to FTIR analysis. Figure 23 shows the carbonyl index for each fungal species individually and for all species combined. The box plot shows that although some individual fungal species reached carbonyl index values ​​above 1, the species blends showed the most consistent and effective results, demonstrating the synergistic effect achieved by the fungal consortium. This result highlights the importance of species diversity in real-world environments where fungi must compete with biological and abiotic stressors.

[0118] Example 9B: Comparative test of seed consortia inoculum blends In Example 9B, the individual fungal species that showed the highest performance in Example 9A were tested against the highest-performing three-species blend and the highest-performing four-species blend. After three months, the samples were collected, cut for DSC analysis, and obtained PP nonwoven, PE film, and PET nonwoven layers. As shown in Table 7, the white rot fungi (species W3 and W4) showed effective digestion of PE and PP, but failed to colonize and digest the PET layer. The three-species blend showed moderate effectiveness in degrading PE and was effective in degrading the PET layer, but did not colonize the PP nonwoven fabric. The four-species blend was effective against all three layers, showing the highest effectiveness particularly against PE and PET, while its effectiveness against PP was lower than that of species W3 alone. Example 9B demonstrates that embodiments of inoculum configured as a consortium containing multiple modified fungal species can achieve a synergistic effect of degrading multiple polymers without inhibiting the digestion of other polymers. White rot fungus (species W) showed higher efficacy against homochains but relatively lower efficacy against heterochains. On the other hand, soft rot fungus (species S) was effective against heterochains. A blend of all four species was necessary to digest all three polymers.

[0119] Table 7: Decrease in crystallinity after 3 months [Table 7]

[0120] In fact, inoculations containing a fungal consortium consisting of modified fungal species that have undergone the treatment described herein, resulting in a more readily accepting plastic material as a carbon source for metabolism and improved resistance to abiotic stress, have been shown to effectively inoculate contaminated diapers even when the initial fungal biomass to plastic ratio is less than 1%. Preferably, the modified fungal species and strains, and / or the consortium consisting of modified fungal species and strains, are modified to enable inoculation at an initial mass ratio of fungal biomass to plastic of 1:1 or less. More preferably, the modified fungal species and strains, and / or the consortium consisting of modified fungal species and strains, are enable inoculation at an initial mass ratio of fungal biomass to plastic of 1:10 or less. Furthermore, embodiments of the present disclosure have shown that effective inoculation is possible even in the initial mass ratio range of 1:10-1:200 for fungal biomass to plastic. Unlike known treatment methods that rely on the direct application of enzymes to plastic materials, embodiments of the present disclosure utilizing a consortium of modified fungal species and strains are self-sustaining. In other words, the fungal species can continue to grow effectively, and as a result, it can further express enzymes that are effective in utilizing plastic materials as a carbon source, enabling further proliferation and growth.

[0121] Immobilized fungal inoculum: Preparation, control, and integration for the digestion of polymer materials

[0122] As described above, in another embodiment, the immobilized composition includes a modified fungal inoculum capable of targeting and degrading polymers present in the polymer-based material. These compositions have been shown to be shelf-stable and, importantly, reactivatable upon use. In certain embodiments, the composition comprises multiple fungal species, strains, or combinations thereof, which are maintained in a stasis state by dehydration or freeze-drying. This configuration enables long-term storage without impairing the effectiveness of promoting the digestion of polymer-based materials even after reactivation from the immobilized state.

[0123] This composition can be used as a standalone product for additives, or it can be incorporated into various polymer-based materials such as disposable diapers, sanitary napkins, incontinence pads, wound dressings, wipes, and other liquid or solid waste management products. The composition can be added in the form of encapsulated alginate beads or other substrates, which allows for precise dosage units and protects fungi from contamination.

[0124] This specification describes a method for forming a storage-stable fungal inoculum specifically designed for digestive applications. This formulation involves embedding filamentous fungi in a stable form using an encapsulating material, which allows for gradual rehydration and activation in the presence of moisture. This approach allows the fungi to remain metabolically dormant until environmental conditions are suitable for fungal activity, facilitating their use in various application scenarios. The fungal species used are primarily filamentous fungi, selected based on their high robustness and broad applicability to environments with sufficient moisture.

[0125] The encapsulation techniques used herein are crucial for the stability and efficacy of the inoculum. Fungi can be encapsulated with or without a polymer matrix, and may also be immobilized solely by preservatives such as trehalose. In this case, the fungi are maintained in an anhydrous state (anahydrobiosis), i.e., a "waterless state of life." To prevent premature emergence, the encapsulated particles are formed into beads using alginate (1 to 5%). Factors such as biomass ratio, fragment size, and potential nutrients are optimized to ensure the viability and functionality of the fungi. Furthermore, the crosslinking density is adjusted to affect the strength and porosity of the beads, enabling proper growth and respiration, while ensuring genetic stability at room temperature. Delivery forms include individual beads 122, compressed tablets 124, sheets 126 (dryer sheet-like form due to fiber deposition), Velcro strips, capsules 128 (detergent capsules or pharmaceutical capsule-like forms), nonwoven sachets, foam deposits 130, film deposits 132, and absorbent nonwoven assemblies, which are schematically shown in Figure 25.

[0126] After rehydration and activation, the inoculum gradually absorbs moisture through a controlled microenvironment formed by the complex. Adding hydrophilic abiotic components to the composition promotes active moisture draw-in, and a mixture of living hyphae fragments and hygroscopic components further draws moisture from the surrounding environment, including water vapor. Spores may be added to the composition to delay activation and optimize microenvironment formation. The rehydration process safely rehydrates the fungal cell walls, restarting metabolic processes and allowing hyphae to grow and emerge from the encapsulating matrix. This enables the fungi to begin searching for food sources in the surrounding environment, ultimately leading to the effective digestion of waste material. Table 8 below details each step of preparation, storage, waste mixing, rehydration, and metabolic activation, highlighting the interaction of biotic and abiotic factors in the inventive approach.

[0127] [Table 8]

[0128] Example 10: Preparation and testing of fungal inoculum In Example 10, a fungal inoculum of modified species W was prepared according to the method described above. Specifically, biomass obtained from liquid fermentation was recovered and homogenized with a 2% sodium alginate solution. This mixture was crosslinked by extruding it into a calcium chloride solution and formed into beads. The resulting beads were then coated with trehalose and freeze-dried until moisture was removed. The fungal biomass concentration in the mixture was tested in various ranges from 2% to 20%.

[0129] Figure 24 shows the percentage of beads that appeared after 7 and 10 days for each mixture, indicating that the appearance rate increased with increasing biomass concentration. This data demonstrates that fungal appearance can be controlled by the mixture composition. Quality assurance and quality control (QA / QC) were performed on 2% and 4% fungal mixtures, with the lower concentration selected as a representative example showing effectiveness at the minimum biomass dose. When these mixtures were allowed to appear for 14 days, it was confirmed that more than 90% of the beads had appeared normally after 14 days. This example demonstrates the effectiveness of the inoculum preparation method and the ability to control fungal appearance based on the composition of the inoculum mixture.

[0130] Wet formulations contain essential biological elements such as fungal hyphae or spores, which are mixed with preservatives such as trehalose to stabilize and protect the fungi. Furthermore, depending on the application, additional components such as conditioning media, polymer encapsulation matrices, process aids, and secondary nutrients are incorporated to ensure optimal fungal growth and activity during the digestion process. These components are described in more detail in Table 9 below.

[0131] Table 9: Components of the wet inoculum mixture [Table 9]

[0132] Immobilization techniques are used to dry wet mixtures, maintaining the viability and functionality of fungal components. Methods such as dehydration, freeze-drying, sublimation drying (lyophilization), and spray drying are employed to remove moisture from the mixture, resulting in stable dried products suitable for storage and various applications. These immobilization techniques are described in more detail in Table 10 below. Table 10: Immobilization techniques [Table 10]

[0133] The dried mixture obtained as a result of the immobilization process can be used as is, or its functionality can be further enhanced by mixing it with additional compounds. This includes incorporating other immobilized biological elements, hydrophilic and hygroscopic compounding elements, fillers, disintegrants, and desiccants, making it possible to form comprehensive compositions suitable for specific digestion and / or decomposition applications. The components of the dried inoculum are described in more detail in Table 11 below. Table 11: Components of dried inoculum [Table 11]

[0134] The dried inoculum according to the embodiments of this disclosure is storage-stable and, unlike any known technique, can be mixed with waste products at a lower weight ratio. This approach enables efficient digestion and / or decomposition not only under clean laboratory conditions, but also under real-world environmental conditions where synthetic and natural polymers are mixed and contaminated with waste. The method ensures practical effectiveness in non-laboratory environments by taking into account the complexity of such products. For example, significant digestion capacity has been demonstrated even when the dried inoculum is combined with a contaminated polymer product at an inoculation rate of less than 10% relative to the weight of the synthetic polymer, under conditions where wet contaminants are present, as shown in Figure 26.

[0135] Example 11: Encapsulated polymer matrix In Example 11, the preparation of encapsulated filamentous fungi for storage stability was established, with emphasis on the use of a protective polymer matrix and a carefully formulated fungal hyphal mixture. In this example, 1% moist fungal biomass obtained from a modified basidiomycete belonging to species W was used, although in embodiments of this disclosure, the concentration can be reasonably set within the range of 0.1% to 50%. The fungal biomass was mixed with a 2% medium viscosity sodium alginate solution to form a homogeneous mixture. To form the encapsulation matrix, this alginate-fungal mixture was crosslinked with calcium chloride to form uniform beads with a moist diameter of approximately 5 mm. These moist beads were then freeze-dried and stored with a desiccant to improve storage stability and prevent moisture absorption during storage. Tests confirmed that the freeze-dried encapsulated fungi emerged normally within 14 days, both on a petri dish and in a contaminated diaper environment. The specific proportions of the protective polymer matrix and fungal hyphal mixture were important for maintaining the fungi in a dormant state. This composition enables slow and controlled rehydration, ensuring a gradual resumption of fungal metabolic activity. This process prevents cell damage caused by rapid rehydration and protects the fungi from contamination by biological or abiotic moisture present in the waste environment.

[0136] The above procedure was repeated using the same components and techniques, and the dried encapsulated beads were placed inside the contaminated diaper core. Figure 27 is a photograph showing the emergence of fungi from immobilized encapsulated mycelial beads in a typical waste product environment.

[0137] Notably, this method successfully produced storage-stable encapsulated fungi even when using non-adaptive basidiomycete species, further confirming the effectiveness of this procedure for basidiomycetes in general.

[0138] Example 12: Encapsulation using cryopreservatives Example 12 demonstrates a method for encapsulating filamentous fungi without crosslinking, using trehalose as a cryopreservative (freeze-protective agent) to improve the stability and viability of the fungal biomass. In this example, 1% moist fungal biomass obtained from a modified basidiomycete belonging to species W was used, but as with Example 11, in embodiments of this disclosure, the concentration can be reasonably set within the range of 0.1% to 50%. Instead of crosslinking, the fungal biomass was mixed with trehalose to form a cryoprotective environment. This mixture was then processed to form a stable composition. In this example, instead of forming beads, the fungal biomass bound to trehalose was integrated with a hydrophilic filler component to ensure a uniform and protective matrix. This mixture was then freeze-dried and stored with a desiccant to maintain the fungi in a stable dormant state and to keep them dry. Tests confirmed that this embodiment rehydrates well when exposed to moisture and combined with a hydrophilic filler component. The presence of trehalose as a cryopreservative allowed fungal cells to be rehydrated slowly and appropriately, avoiding cell damage caused by rapid water absorption. Furthermore, the hydrophilic filler played a crucial role in ensuring a controlled rehydration process, which is essential for the gradual resumption of fungal metabolic activity.

[0139] Example 13: Encapsulated mycelium in a dry state for appearance and adhesion In Example 13, the effectiveness of trehalose and alginic acid as methods for encapsulating fungal inoculants was evaluated, with particular focus on the appearance and adhesion of encapsulated inoculants on various nonwoven fabric materials. This example demonstrates that dried, dormant encapsulated / immobilized hyphae can be effectively reactivated and made usable, and demonstrates applicability to both polymer matrix encapsulation and trehalose encapsulation forms. Fungal inoculants consisting of 96% alginic acid and 4% fungal biomass (species S and W) were mixed with trehalose powder and alginic acid powder, respectively, to prepare mixtures with a 20% inoculum concentration. These mixtures were thoroughly mixed to ensure uniform dispersion of the inoculum within the powder. Subsequently, the encapsulated inoculum powder was seeded onto multiple agar plates and labeled as appropriate. Different nonwoven fabric materials (loop material, backsheet, and baby wipe) were selected as substrates for adhesion tests. Each material was partitioned, labeled, and then coated with adhesive spray before the encapsulated inoculum powder was applied. A control plate without adhesive was also prepared. After incubation, the appearance of fungal colonies was observed and recorded. As a result, both trehalose-encapsulated and alginate-encapsulated inoculum powder adhered well to the nonwoven fabric material and showed significant fungal growth. While the trehalose and alginate encapsulation methods showed equivalent effectiveness in promoting fungal growth, slight differences in growth patterns and adhesion efficiency were observed depending on the type of substrate. From the control test, it was confirmed that adhesive spray promotes the adhesion and subsequent growth of encapsulated inoculum.

[0140] Therefore, Example 13 demonstrates that the emergence and adhesion of fungi on a nonwoven fabric substrate can be promoted using encapsulated mycelium that is dry and has storage stability. These results indicate that both trehalose and alginic acid are effective encapsulation methods and can be applied to applications that promote digestion by incorporating fungal inoculants into polymer-based products (see Figure 28: emergence of immobilized fungal inoculant attached to nonwoven fabric).

[0141] As described above, the dried inoculum described herein can be molded into various final product forms (form factors) depending on the application, examples of which are shown in Figure 25. These include individual beads or powder forms 122, compressed tablets 124, dryer sheets, Velcro strips, detergent capsules 134, nonwoven sachets, etc. Furthermore, the inoculum can also be integrated into foam / sponge assemblies 130, absorbent nonwoven assemblies 126, and Listerine strip-like nonwoven ultrasonic bonding 132. These diverse delivery forms enhance the versatility and scalability of the inoculum, enabling its effective use in multiple waste digestion applications. In all of these forms, it has been shown that the fungal inoculum can be reactivated by slow rehydration with time-controlled release. Furthermore, the product may contain nutrients that provide a controlled microenvironment to assist the rehydration process. In addition, the inclusion of hygroscopic trehalose can achieve higher activity concentrations. The presence of live hyphae fragments and hygroscopic components has the effect of attracting moisture from the surrounding environment. Furthermore, spores may be included for the purpose of delayed activation. The composition product described herein has been confirmed to exhibit genetic stability at room temperature when combined with a desiccant.

[0142] The fungal microcolonies used within the beads can be as small as 0.5 micrometers, representing the smallest viable hyphae. While hyphae of this size are unviable in nature, novel engineering designs of the inoculum enable their survival and proliferation in this embodiment. By isolating and modifying such microcolonies, the inoculum can be distributed over a wide area with minimal material, resulting in cost reduction and improved efficiency. This approach enables a controlled and consistent inoculation process, making it highly suitable for industrial and commercial applications in polymer degradation.

[0143] In nature, fungal growth typically arises as an interconnected hyphae network originating from spores or as large, diffuse colonies. These natural colonies exhibit high uniformity in their development and interaction with the environment. In contrast, the microcolonies in this inoculum composition are not derived from spores, but are engineered and recovered from biomass harvested from specific modified fungi. Each encapsulated microcolony is isolated individually, allowing each to exhibit different properties and behaviors. This engineered process creates multiple microcolonies with diverse properties, each adapting to its own microenvironment. This diversity among microcolonies forms a more robust and adaptable fungal consortium, improving the overall effectiveness and functionality of the inoculum composition in the digestion of polymer materials.

[0144] Example 14A: Appearance of fungal inoculum composition In Example 14A, nonwoven fabric materials embedded with encapsulated mycelium and compressed tablets containing encapsulated mycelium were prepared, both encapsulated with the modified species S. The absorbent pads were hydrated, but no additional liquid was added to the mixture. These combinations were then placed in an aquarium. Figure 29 includes photographs showing the rehydration process and the appearance of both inoculum after 5 and 14 days, demonstrating the water absorption for rehydration and the subsequent appearance of the fungus.

[0145] Example 14B: Appearance of fungal inoculum composition In Example 14B, compressed tablets with the same composition as those in Example 14A were tested on contaminated diapers placed in an outdoor bucket environment. After 18 days, the bucket was opened, and as shown in the photograph in Figure 30, the appearance of fungal hyphae in the diaper material and vigorous colony formation were confirmed.

[0146] Scaffold encapsulation for fungal immobilization This embodiment relates to a process for growing filamentous fungi on a polymer scaffold and then encapsulating them to produce a storage-stable product, a schematic of which is shown in Figure 31. This method and the resulting product allow the fungi to adapt to various polymer types, improving emergence efficiency and growth activity. The encapsulated fungi are applicable to films, nonwovens, foams, and other polymer-based materials.

[0147] The scaffold types include nonwoven materials that come into direct contact with fibers or films, and their absorbency is enhanced when combined with encapsulated live fungi. The growth and encapsulation process involves growing filamentous fungi within the nonwoven scaffold, utilizing novel technologies that provide a larger surface area to enhance adaptability. This process is characterized by the combination of multiple polymers to promote efficient fungal metabolism.

[0148] The scaffold can take various forms, such as foam, particles, or actual plastic resin, and fungal spores can be embedded within the scaffold. Both solid-phase and liquid-phase encapsulation methods are used to ensure the viability and effectiveness of the fungi. This approach enables a wide range of applications, including the processing of films, nonwovens, foams, and other polymer materials. The process of growing filamentous fungi within nonwovens and encapsulating them, with or without embedding, provides a versatile and efficient method for digestion and / or degradation under diverse environmental conditions.

[0149] Example 15: Solid-phase fermentation with nonwoven fabric integration In Example 15, modified filamentous fungi were colonized by solid-phase fermentation. The fungi were induced to form aerial hyphae, which then bonded to the fibers of a nonwoven fabric placed on top of the fungal culture. This process allowed the fungal hyphae to integrate well with the nonwoven fabric fibers, forming a robust physical scaffold. The entire nonwoven fabric sheet, colonized by the fungal hyphae, was then combined with a sodium alginate mixture. This mixture provided additional structural support and protection. Subsequently, this composite sheet was crosslinked in a calcium chloride solution to form a robust immobilized structure. This approach provided the fungal hyphae with a physical scaffold for attachment, structural support for immobilization, and a microenvironment with directionality for controlled rehydration. Test results confirmed that this modification rehydrated well, and the fungi emerged and effectively colonized on the material. Furthermore, the reactivation of the fungal metabolic processes that had been dormant during immobilization accelerated enzyme activity, promoting rapid digestion of the polymer scaffold. Figure 32 includes a photograph showing the appearance of the fibrous scaffold inoculum of modified strain W after 7 days.

[0150] Example 16: Enrobing of fibers with alginate-mycelial mixture In Example 16, instead of simply growing mycelium within the nonwoven fabric, a sodium alginate-mycelial mixture with specific viscosity and mycelial size was prepared. This preparation ensured that the alginate-mycelial mixture could adequately enrode the fibers of the nonwoven fabric. This alginate-mycelial mixture was then applied to the nonwoven fabric, allowing the fungal mycelium to integrate with the fibers. This enroving process formed a protective layer around the fibers. Subsequently, the entire composition was crosslinked using calcium chloride to obtain a solid, immobilized structure. This method and the resulting product provided the fungi with a physical scaffold for support and a directional microenvironment for controlled rehydration. Test results confirmed that this modification, similar to Example 15, rehydrated well, and the fungi emerged and effectively colonized on the material. Furthermore, the fungal metabolic processes were preserved during immobilization and reactivated upon rehydration, accelerating the digestion of the polymer scaffold via enzymatic activity. Figure 33 includes a photograph showing the appearance of the enrobed fiber inoculum of modified strain W after 7 days.

[0151] Example 17: Absorbable product incorporating a storage-stable fungal inoculant In Example 17, a liquid absorbent product similar to a diaper was prepared with a multi-layer structure. The product included a liquid-impermeable backsheet, an absorbent core made of fluff pulp and superabsorbent polymer (SAP) (covered with a core-wrap nonwoven fabric), an acquisition and distribution layer (ADL), and a polypropylene topsheet. Its schematic structure is shown in Figure 34. A control product had the same structure but was prepared without immobilized live microorganisms, and incorporated 1 g of superabsorbent polymer and 2 g of film, nonwoven fabric, and adhesive. Furthermore, several test samples were prepared to evaluate the various form factors described above. Sample obtained by adding 1 g of dried alginate bead-shaped inoculum containing fungus variant S to a core mixture. Sample containing 3g of dried modified mycelium of modified strain S embedded in a core wrap. Samples in which 1g of the same alginate beads were placed as the core and 3g of the same mycelium were placed as the core wrap simultaneously. A sample consisting of a polymer film layer, 1g of encapsulated mycelium, and a nonwoven fabric layer, attached to the backsheet, served as a "landing zone" on the backsheet.

[0152] To confirm that these absorbent products possessed the performance required for practical use, liquid retention tests and re-wetting tests were performed on all samples. In the liquid retention test, the product was immersed in water for 1 minute, then suspended to dry for 2 minutes, and the amount of liquid retained was calculated by subtracting the dry weight from the wet weight. In the re-wetting test, a dry filter paper was placed on the product, a 1 kg load was applied for 10 seconds, and the moisture retention was evaluated by comparing the wet and dry weights of the filter paper.

[0153] Furthermore, each sample was placed in a tank box, and the appearance of fungi (1 cm 2 (defined as visible growth exceeding a certain threshold) was observed. As expected, no fungal growth was observed in the control sample. On the other hand, in samples containing encapsulated modified fungi, differences in the time to emergence were observed depending on the form factor; emergence was confirmed in 11 days with alginate beads and in 5 days with embedded hyphae, but no growth was observed in the layer attached to the outer layer. These results are summarized in Table 12 below.

[0154] Table 12: Absorption amount and appearance time of embedded fungal inoculum [Table 12]

[0155] The fluid retention test showed that the control sample absorbed 43.79 g of liquid. Samples with fungi embedded in the polymer layer showed similar liquid absorption within ±2% of the control. On the other hand, the sample with encapsulated mycelial beads absorbed approximately 6 g more liquid, indicating that the encapsulated fungi placed in the nonwoven fabric inside and outside the absorbent product did not adversely affect the absorbency itself, and that the addition of alginate beads improved fluid retention by approximately 6:1. In the re-wetting test, all samples showed similar moisture retention in the range of 0.50 to 0.56 g, confirming that the presence of fungal inoculation did not affect the product's ability to maintain dryness. Regarding emergence, no fungal growth was observed in the control sample, while emergence was observed in the sample with embedded mycelium after approximately 5 days. In contrast, emergence was observed in the alginate bead sample after approximately 11 days, and no fungal growth was observed in the outer layer (chassis part). These results demonstrate that even when immobilized viable microorganisms (fungi) are incorporated into absorbable products, effective fungal growth and emergence are possible without compromising absorption performance. Figure 35 shows a photograph illustrating the emergence of modified fungal (Species S) inoculum from an absorbable product.

[0156] Example 18: Liquid-impermeable container containing biological factors for waste treatment In Example 18, 1 g of hyphae of a non-encapsulated and immobilized modified fungus (Species S) was attached to a plastic film bag. A size 4 disposable diaper without hyphae was moistened with 200 mL of a saline and urea mixture, and the moistened diaper was placed inside the bag with the hyphae attached. After 7 days, fungal emergence and growth were observed in both the bag and the outer layer of the diaper. Figure 36 shows a photograph illustrating the emergence from the film container using Species S inoculant. The results of Example 18 demonstrate that fungi can be incorporated into waste bags and packaging materials commonly used for feminine hygiene products, for example, leading to effective colonization and digestion. By incorporating fungal inoculants into the waste stream after use, the fungi utilize the contaminated product as a water source for rehydration and emergence, promoting digestion within the waste environment.

[0157] In one embodiment, a method for controlling the rehydration and appearance of an immobilized fungal inoculum in an absorbent product includes identifying the rehydration conditions for the immobilized fungal inoculum. This method makes it possible to strategically control the rehydration of the fungal inoculum. The immobilized fungal inoculum is typically maintained in a metabolic stasis state and is configured to be rehydrated when the absorbent product becomes contaminated. Examples include an environment in which the hydrogel swells within the absorbent core of a disposable diaper, or a configuration in which it is embedded within a nonwoven fabric substrate that attracts liquid to the encapsulating material.

[0158] Due to their unique physical properties, hydrogels are used as immobilization media for fungal inoculants. Hydrogels have a network structure in which polymer chains are cross-linked in three dimensions, and possess the ability to absorb and retain large amounts of water in the voids between polymer chains. In addition to these properties, hydrogels are highly suitable materials for biological applications, including the embodiments of this disclosure, due to their flexibility, hydrophilicity, superabsorbency, viscoelasticity, biodegradability, and biocompatibility.

[0159] Of particular note is the application of smart hydrogels (stimulus-responsive hydrogels), which constitute an important variation. Smart hydrogels are sensitive to specific environmental changes and respond by changing their shape or volume when exposed to those conditions. This sensitivity to stimuli can be classified as internal or external stimuli, depending on the source of the stimulus when the hydrogel is used in vivo. External stimuli that can be used to control the rehydration and emergence of immobilized fungal inoculum include physical conditions such as temperature, pressure, light, electric fields, magnetic fields, and ultrasonic irradiation. Alternatively, hydrogels can also be configured to respond to chemical stimuli such as pH, ionic strength, and CO2.

[0160] pH-sensitive hydrogels can be designed to allow rehydration and reappearance of immobilized fungal inoculants only at specific pH values. For example, a hydrogel can be configured to remain unresponsive in the presence of water, and for the fungal inoculant to be rehydrated and activated only upon exposure to urine. This approach enables the safe and practical use of immobilized fungal inoculants in consumer absorbent products such as diapers, sanitary napkins, and incontinence pads.

[0161] The hydrogel coating used herein is intended to ensure that immobilized fungal inoculants are rehydrated only under specific conditions, thereby enabling the safe and effective use of fungi in consumer absorbable products. By precisely controlling the rehydration and reappearance of fungal inoculants, this product brings greater sophistication and specificity to fungal applications in the absorbable product field.

[0162] Furthermore, methods for embedding immobilized fungal inoculants within nonwoven or foam substrates are also provided. These substrates possess fluid handling capabilities due to capillary action and permeability, ensuring that liquids effectively move to the immobilized fungal inoculants. In this specification, capillary action refers to the phenomenon in which liquids rise within a substrate against gravity due to intermolecular forces. Similarly, permeability refers to the ability of a substrate to allow liquids to pass through. Nonwoven or foam substrates with high capillary properties and high permeability efficiently absorb liquids and ensure that they reach the encapsulated fungal inoculants, thereby inducing their rehydration and appearance. Such strategic placement within the substrate, and hydration that occurs only when exposed to specific fluid conditions such as urine, provides a highly targeted and efficient mechanism for the activation of fungal inoculants, further enhancing the safety and usefulness of the present invention in consumer absorbent products.

[0163] Water Absorbent Capacity (WAC) is governed by the following relationship: (Osmotic pressure) 2 + Affinity) / Rubber elasticity

[0164] Osmotic pressure In this formulation, osmotic pressure is influenced by the ion concentrations inside the hydrogel and in the surrounding fluid. Sodium alginate is a polymer that forms a gel in the presence of calcium ions (derived from CaCl2). Sodium ions from sodium alginate and calcium ions from CaCl2 contribute to the ion concentration inside the hydrogel. The difference in ion concentration between the inside of the hydrogel and the surrounding fluid creates osmotic pressure, which drives the absorption of water.

[0165] Affinity The affinity in this formulation is determined by the hydrophilic groups present in the hydrogel. Sodium alginate is a hydrophilic polymer and has a high affinity for water. This high affinity contributes to the water absorption capacity of the hydrogel. Depending on the fungal species used, the fungal hyphae may also contribute to the hydrophilicity of the hydrogel.

[0166] Rubber elasticity The rubber elasticity in this formulation is a function of the crosslink density. In this case, crosslinking is provided by calcium ions derived from CaCl2, and these calcium ions crosslink sodium alginate to form a gel. The density of these crosslinks determines the rubber elasticity of the hydrogel. By adjusting the concentration of CaCl2, it is possible to control the crosslink density and, consequently, the rubber elasticity of the hydrogel.

[0167] Osmotic pressure, caused by the ion concentration difference, is the primary driving force of swelling, and its magnitude increases proportionally to the square of the ion concentration difference. Affinity is a factor related to hydrophilic groups and plays a smaller role compared to osmotic pressure. Rubber elasticity is determined by the degree of crosslinking and acts as a factor that resists swelling. Granules swell to the point where osmotic pressure, affinity, and rubber elasticity are in equilibrium.

[0168] Synthetic Absorbent Hydrogel It is a synthetic polymer produced by the polymerization of monomers. Synthetic superabsorbent polymers (SAPs) are polymerized from monomers such as NaPA (sodium polyacrylate) through multiple polymerization steps, and then crosslinked with polymers.

[0169] Natural Absorbent Hydrogel It is a naturally derived polymer that undergoes ionic crosslinking or chemical crosslinking. Naturally derived polymers such as alginic acid (or cellulose, chitosan, agar, pectin, starch, hyaluronic acid, carrageenan, guar gum, xanthan gum, etc.) have improved water absorption capacity when they are grafted or crosslinked (ionically crosslinked or chemically crosslinked).

[0170] When sodium polyacrylate (NaPA) crosslinked with 5% polyethylene glycol (PEG) is mixed with sodium alginate crosslinked with CaCl₂, ion exchange occurs. That is, Na in NaPA + ions displace Ca in the alginate network 2+ ions. This ion exchange reduces the crosslinking density in alginic acid (because Ca 2+ ions form stronger crosslinks than Na + ions), consequently, rubber elasticity decreases, allowing greater swelling. On the other hand, on the SAP side, the inflow of Ca 2+ ions increases the crosslinking degree, which increases rubber elasticity and decreases water absorption capacity.

[0171] Changes in osmotic pressure, which increase proportionally to the square of the ion concentration difference, have a great influence on the water absorption of the entire system. SAP becomes more elastic and its water absorption decreases, while alginate swells more efficiently due to reduced crosslinking. This ion exchange mechanism optimally utilizes the respective advantages of natural hydrogel (alginate) and synthetic hydrogel (SAP). Alginate can absorb more fluid, improving the efficiency of the entire system, while SAP provides structural support through increased elasticity. This balanced design makes it possible to utilize the high swelling capacity of natural polymer hydrogels while reducing the required amount of SAP.

[0172] Example 19: Liquid retention property depending on the difference in amounts of dry SAP and alginate-encapsulated hydrogel In Example 19, the fluid retention of dry superabsorbent polymer (SAP), dry alginate encapsulated hydrogel, and combinations thereof was evaluated. Three samples were prepared: Sample A contained 1 g of alginate hydrogel only, Sample B contained 1 g of SAP only, and Sample C contained a combination of 1 g of SAP and 1 g of alginate beads. Each mixture was placed in a tray, 100 g of 1% NaCl solution was added, and the tray was then placed in a resealable bag. The samples were exposed to the fluid for three different contact times: 2 minutes, 1 hour, and 4 hours. Each mixture was tested in three replicates. After the predetermined contact time, the mixture was filtered for 2 minutes, and the fluid retention was calculated by subtracting the initial dry weight from the wet weight after filtration.

[0173] Sample A: After 2 minutes, it held 5.8g of fluid; after 1 hour, it held 4.6g; and after 4 hours, it held 4.4g of fluid. Sample B: Regardless of the contact time with water, it consistently absorbed and retained 40-45g of fluid. Sample C: The fluid held 49.1g after 2 minutes, 78.7g after 1 hour, and 98.7g (almost the entire amount) after 4 hours.

[0174] These results indicate that the mixture of SAP and alginate beads (Sample C) exhibited an unexpected synergistic effect, significantly improving fluid retention over time. While SAP alone stably retained a certain amount of fluid, the addition of alginate beads significantly increased the total fluid retention capacity, especially over long periods. This result suggests that combining synthetic hydrogels like SAP with natural hydrogels like alginate-encapsulated hydrogels greatly improves absorption and fluid retention, leading to higher performance in absorbent products during prolonged use. Figure 37 includes a photograph showing the results of Example 19.

[0175] Example 20: Field test of alginate-encapsulated hydrogel in diapers In Example 20, a size 4 diaper was used to evaluate the effectiveness of alginate-encapsulated hydrogel in a real-world environment. A diaper contaminated with 225 g of urine was cut open to access the absorbent core, and 1 g of dry sodium alginate beads, cross-linked with calcium chloride and having a diameter of 0.9-1.1 mm in a dry state, was added to the core.

[0176] After one month, these beads had swelled to a diameter of over 3 mm. In contrast, the same beads, when mixed only with water or urine without contact with SAP, only swelled to a diameter of 1.5–1.6 mm. This significant difference in swelling indicates that the SAP present in the contaminated diaper core promoted the rehydration and swelling of the alginate beads.

[0177] The water that rehydrated the beads was supplied through interaction with the swollen SAP, allowing the natural polymer hydrogel to access the water within the SAP. This interaction resulted in ion exchange from the SAP, enabling the alginate beads to swell beyond normal osmotic equilibrium. This field test demonstrates that incorporating alginate beads into the absorbent core of a used diaper effectively utilizes the water retained in the SAP, maximizing the swelling and functionality of the hydrogel. Figure 38 is a photograph showing the water absorption of a mixture of natural and synthetic hydrogels in Example 17 after one month. Figure 39 also shows this unexpected synergistic effect resulting from the combination of natural hydrogel 140 and synthetic hydrogel 142.

[0178] Example 21 In Example 21, an encapsulated modified fungus / alginate mixture was prepared. This mixture was designed to encapsulate the fungus within a protective alginate matrix, thereby immobilizing the fungus while maintaining its viability and the possibility of re-emergence under specific conditions.

[0179] Next, this encapsulated fungus / alginate mixture was embedded on a cellulose-based nonwoven fabric substrate. This substrate was selected for its excellent fluid handling capabilities, such as permeability and capillary action, which allows for the effective absorption and transport of fluid to the encapsulated fungus.

[0180] After embedding, the mixture-substrate complex was dehydrated, which caused the fungi to enter a stasis state and further enhanced immobilization. This dehydration process effectively preserved the embedded fungus / alginate mixture, preparing it for future rehydration and emergence. Subsequently, the dehydrated mixture-substrate complex was rehydrated under controlled conditions. These conditions were specifically designed to simulate the fluid exposure the complex would encounter in absorbent product applications. After successful rehydration, the previously immobilized fungi were observed to emerge from their encapsulated state and begin colonizing on the supplied agar medium. This colonization demonstrated not only that the fungi successfully re-emerged from their encapsulated and immobilized state, but also that they maintained their viability and functionality after rehydration.

[0181] This example demonstrates the controlled rehydration and emergence of immobilized fungi under specific conditions. This capability can be utilized in the development of highly functional consumer absorbent products that leverage the potential of fungi in digesting or decomposing materials.

[0182] In yet another embodiment, a process is provided for encapsulating and immobilizing pre-selected fungi for commercial use, for the purpose of digesting recalcitrant long-chain carbon materials, including plastics and other polymeric waste. This process is a natural development step that takes place after scaling up the pre-selected fungal species in a bioreactor.

[0183] After the mycelial growth reaches its maximum expansion, the fungal hyphae are subjected to a bioprocessing step. This step typically involves homogenization, and filtration as needed, to form a uniform biomass. This homogenized biomass is then mixed with a polymer solution, such as alginate, to form a stable and handleable material suitable for subsequent processes.

[0184] Next, the fungal hyphae-alginate mixture is mixed with a curing solution to form a hydrogel, which is then subjected to spray drying. Spray drying can be performed towards a designated recovery area for future use, or it can be sprayed directly onto the substrate on which the fungus acts. Alternatively, the fungal hyphae-alginate mixture can be extruded into a calcium carbonate solution to form beads, which are then recovered.

[0185] These beads or spray-dried materials, while containing immobilized fungi, can undergo additional treatments to improve storage stability and resistance. This treatment may include coating with trehalose or other compounds that provide protection during drying. This additional treatment step is optional and may be omitted depending on the product's end use and storage conditions.

[0186] Processed or unprocessed fungal products are then dehydrated by freeze-drying or oven heating. During this process, the temperature is maintained below the heat tolerance threshold for each fungal species to prevent denaturation. This dehydration process significantly reduces the product weight to 1 / 10-1 / 75th of its original weight, improving handling and lowering transportation costs.

[0187] Dehydrated fungal products are packaged for storage and distribution. Encapsulated and immobilized fungi are storage-stable and retain their ability to digest persistent long-chain carbon materials, making them suitable for commercial deployment in waste management applications.

[0188] The process of adapting and optimizing methods developed for small-scale waste digestion to large-scale industrial applications begins with the collection and aggregation of waste containing a mixture of polymer materials and organic waste (food waste, garden waste, and other organic materials). These waste flows can be collected from various sources, including municipal waste treatment facilities, manufacturing plants, and other industries that generate polymer waste.

[0189] The collected waste material is then subjected to a process similar to that described in the aforementioned invention, and a pre-selected fungal inoculum is introduced. A fungal species with superior ability to digest specific polymeric materials is selected and introduced into the waste mixture. For industrial-scale applications, the amount of waste to be processed is larger, requiring a larger amount of fungal inoculum.

[0190] The waste material is placed in a designated waste container. In this case, the container is an industrial-scale bioreactor or a similar large-scale waste treatment system. Within these systems, an optimal environment for fungal growth and digestion is created and maintained by controlling factors such as temperature, humidity, pH, and aeration.

[0191] Furthermore, the digestive performance of rehydrated fungal inoculants can be improved by introducing process aids such as nutrients, bio-derived enzymes, chemical acids, and other additives.

[0192] The resulting biomass can be further utilized for composting, soil improvement, or as a raw material for renewable energy production such as biofuels and biogas.

[0193] Ultimately, this process leverages the digestive capacity of pre-selected fungi for resilient long-chain carbon materials, thereby reducing the environmental impact of polymeric waste and providing an efficient, sustainable, and scalable industrial waste management solution.

[0194] 1. Pre-Use Incorporation In this embodiment, the fungal inoculant is incorporated into the polymer product itself during the manufacturing process. This includes one or more of the following methods:

[0195] Incorporation of fungal inoculants into the core mixture of absorbable products: This method involves directly incorporating a fungal inoculant into an absorbent core material. Inclusion of fungal inoculants as components in a mixture of product material ingredients: The fungal inoculant is included as part of the mixture of material components that make up the product. Offline processing for the incorporation of fungal inoculants into substrates: In a separate processing step, the fungal inoculant is incorporated into the substrate material by mixing it with the substrate material. Content of fungal inoculum in a non-dehydrated state: The fungal inoculum is incorporated while it is still in an undehydrated state. Incorporation of fungal inoculants into the external environment of products requiring activation: Here, the fungal inoculum is placed on the outside of the product and provided as crushable beads, powder, or a similar structure or form. This inoculum can be activated by the user as needed, such as through contact with a fluid.

[0196] 2. Post-Use Incorporation In this alternative embodiment, when a used polymer product is disposed of, the consumer or waste container comes into contact with the product with a fungal inoculum. This can be done by several methods or actions that are easily performed in everyday use.

[0197] Consumers can directly apply fungal inoculants to used polymer products using one or more form factors, including (but not limited to) crushable beads, capsules, wipes, and powders. This active involvement in the degradation process enables a personalized and controllable approach to waste management. In another aspect, a consumer may introduce a fungal inoculum into a disposal container or disposal bag designed to hold one or more used products after use. This step comprises introducing the fungal inoculum into the container, and allowing the fungal inoculum to contact the used polymer product to initiate the degradation process. In another embodiment, the waste container or bag itself has the function of releasing or retaining the fungal inoculum. These containers are pre-filled with the fungal inoculum, and are configured such that the degradation process can be initiated when used products are loaded therein. This integrated approach minimizes user operation and maximizes convenience. In yet another embodiment, the fungal inoculum can be incorporated at any stage of a waste management workflow. In such scenarios, a waste management facility or system can provide the necessary inoculum for used products, which further reduces the burden on consumers.

[0198] As mentioned above, the present invention also provides a superabsorbent hydrogel formulated to incorporate a fungal inoculum, and this hydrogel has the unique feature of mimicking the properties of a superabsorbent polymer (SAP). This hydrogel product exhibits high swelling capacity, rapid hydration rate, and excellent liquid retention performance, while encapsulating the fungal inoculum for bioactive applications. The inventive embodiments described herein inherently also encompass novel biodegradable superabsorbent hydrogel formulations.

[0199] The superabsorbency of this hydrogel derives from a special formulation including optimized crosslinking density, carefully selected polymers, introduction of ionizable groups, and advanced processing techniques. Incorporation of the fungal inoculum is a central element of its effectiveness. Fungal strains are selected based on the desired biological activity. These fungi are embedded within the hydrogel matrix and remain metabolically dormant until rehydration. The superabsorbency of the hydrogel ensures controlled, sustained rehydration of the fungal inoculum and promotes biological activity under specific conditions.

[0200] Furthermore, the size and dispersibility of individual fungal hydrogels provide an optimal effective surface area for improving absorption rate and total absorption capacity. This achieves an effective means of rehydrating fungal inoculants, and provides an alternative absorption capacity source that is essential to the performance of absorbent products such as diapers. In one embodiment, by completely replacing SAP with the novel fungal hydrogel of the present invention, it is possible to achieve the performance characteristics required for an absorbent product.

[0201] The product of the present invention combines the superabsorbent properties of SAP with the biological activity of fungal inoculants, providing a versatile and environmentally friendly material. This superabsorbent hydrogel containing fungal inoculants can be adapted for a wide range of applications including but not limited to agriculture, bioremediation, wastewater treatment, and personal care products.

[0202] As yet another embodiment, there is provided a method of incorporating a fungal inoculum into the manufacturing process of an absorbent product. This innovative process is designed to be seamlessly integrated into existing production lines, particularly lines that use core forming assemblies.

[0203] The process starts with the preparation of an immobilized fungal inoculum. The fungal inoculum is prepared in a form compatible with existing equipment, specifically as particles having a particle size range similar to that of superabsorbent polymer (SAP), usually 0.5 mm to 1.5 mm, with an average particle size of less than 1 mm. This particle size compatibility allows the fungal inoculum to be handled and processed in the same manner as SAP, facilitating integration into the manufacturing process.

[0204] The immobilized fungal inoculum is then fed into a hopper arranged upstream of the core forming assembly. This hopper is provided exclusively for the fungal inoculum, enabling accurate and controlled addition to the absorbent product.

[0205] The fungal inoculum is then mixed with a pulp fiber mixture. The ratio of fungal inoculum to pulp fiber is carefully adjusted to maintain the absorbency of the final product while ensuring sufficient fungal activity for the digestion of polymer products.

[0206] After thorough mixing with pulp fibers, the mixture is transferred to a core molding assembly. Here, the mixture is placed in a rotating drum and molded into an absorbent core for the absorbent product.

[0207] This process is designed so that the majority (over 90%) of the fungal inoculum particles do not reach temperatures that would kill dormant fungi. This is important for maintaining the biological activity of the fungal inoculum in the final product.

[0208] This embodiment uniquely integrates a bioactive fungal inoculum into the manufacturing process of absorbable products. The fungal inoculum is specially designed to be compatible with existing core molding equipment, either alone or in combination with SAP. This compatibility enables the production of biodegradable absorbable products without significant changes to existing manufacturing processes.

[0209] The process of the present invention provides a practical and efficient method for incorporating fungal inoculants into absorbable products, contributing to the development of more sustainable and environmentally friendly absorbable products.

[0210] In another embodiment, the fungal inoculant is embedded and immobilized within a nonwoven fabric substrate that serves as a carrier. This nonwoven fabric substrate can be placed between any layers of an absorbent product, providing a versatile method for incorporating a fungal inoculant into a product.

[0211] This process begins with the preparation of the inoculation patch. The fungal inoculation is embedded in a nonwoven fabric substrate to form a roll-shaped material that can be cut to the desired size. This roll is manufactured in advance and can be stored until use, serving as a source of readily available inoculation patches during the manufacturing process.

[0212] When it comes time to incorporate the inoculation into the absorbable product, a roll is dispensed and the patch is cut to the specified size. This cutting process can be automated to ensure consistency in patch size and to improve the efficiency of the manufacturing process.

[0213] Once the patches are cut, they are grasped by a vacuum rotating drum. This drum suctions and rotates the patches, transporting them to the partially assembled absorbent product. The use of a vacuum rotating drum allows for precise placement of the patches, minimizing the risk of misalignment or detachment during the application process.

[0214] Next, the patch is applied to the absorbent product. This application is facilitated by the use of a tackifier or adhesive to securely fix the patch in place. The choice of tackifier or adhesive can be adjusted according to the requirements of the absorbent product, taking into account the materials used in the product, the required product performance, and the environmental conditions in which the product will be used.

[0215] After the patch is applied, an additional layer (film or nonwoven fabric) is added to the absorbent product. This layer holds the patch in place and protects the fungal inoculation from damage or migration during the remaining stages of the manufacturing process and subsequent product use. This alternative embodiment of the present invention provides a practical and efficient method for incorporating fungal inoculants into absorbent products. By embedding the inoculant within a nonwoven fabric substrate and applying it as a patch, precise placement of the fungal inoculant within the product becomes possible, contributing to the development of more sustainable and environmentally friendly absorbent products.

[0216] Storage stability of living organisms, discrete capsules, and related morphologies.

[0217] In another embodiment, a particle delivery system for delivering stable living organisms (live microorganisms) is configured to be integrated into existing absorbent product manufacturing processes. This system utilizes hydrogel particles that can be easily mixed with multiple biological species after drying / curing, enabling purpose-specific applications in absorbent products. These embodiments offer flexible delivery formats, such as capsule-like final packaging similar to dry detergent pods, or gel suspensions for post-use applications. This approach allows for the customization of digestive agents (degradants) according to consumer needs and environmental conditions.

[0218] This embodiment utilizes a controlled release system using hydrogels to deliver a stable living organism. These hydrogels are preferably incorporated into polymer absorbent products such as diapers and sanitary napkins in a dehydrated state, and are positioned in a location that comes into contact with liquid after use. Upon contact with liquid, the hydrogels rehydrate, inducing the emergence of the encapsulated live fungal organisms. This controlled release mechanism ensures that the activation of the digestive agent occurs only in the presence of water, thus aligning the system's operation with the product's functional lifecycle.

[0219] While the encapsulation of these organisms can be achieved using multiple technologies, alginate microparticles are particularly preferred. These microparticles provide a stable environment that protects the fungi until rehydration. Furthermore, this system allows for the incorporation of additional nutrients and enzymes into the formulation, improving the efficacy of the fungal inoculum after activation. This facilitates a robust digestion process after hydrogel rehydration. In addition, the formulations of these particles can be modified to include a delayed-release coating, depending on the embodiment. These coatings are designed to prevent premature activation of the fungi, ensuring that activation occurs only after a sufficient amount of time has elapsed since product use. This allows for the maximum possible digestive capacity of the fungi.

[0220] An important feature of these embodiments is their high compatibility with a variety of existing absorbent product designs. Discrete capsules formulated as alginate microparticles or similarly encapsulated hydrogels can be seamlessly integrated into the structure of these products. Strategically placed, these capsules remain inert and inconspicuous before and during use of the product. However, they activate upon exposure to moisture and contribute to the digestion (degradation) of the product. This integration represents a significant advance in that it can add digestibility to a wide range of products without changing the basic design of the product or the user experience.

[0221] In another embodiment, a storage-stable fungal inoculum is embedded in a web substrate to build an advanced system for efficient encapsulation and rehydration. In certain embodiments, a fiber material is impregnated with alginate to ensure adhesion and curing for effective encapsulation. In alternative embodiments, encapsulated beads are embedded within the textile, providing options for microcapsule functionalization or adhesion. The method utilizes a fibrous scaffold to deliver organisms for digestion, improving efficiency and effectiveness in absorbent products and other applications.

[0222] Furthermore, live fungal inoculum with storage stability can be embedded into a nonwoven web substrate, which acts as an efficient scaffold for encapsulation and rehydration. This embodiment utilizes advanced techniques such as screen printing, slot coating, and microgravure roll-to-roll printing. These methods allow alginate, a biocompatible and biodegradable hydrogel, to precisely impregnate fibrous nonwoven materials, achieving effective encapsulation of fungal organisms within the fiber matrix. This process not only provides a stable environment for the fungi, but also ensures immediate contact with the nonwoven scaffold upon rehydration, which is important for accelerating growth and digestive activity.

[0223] In yet another embodiment, alginate is impregnated into a fibrous nonwoven fabric material. This process is similar to screen printing, and the alginate mixed with the fungal inoculant is uniformly distributed throughout the nonwoven fabric. This distribution allows for controlled and efficient encapsulation of the fungi, ensuring uniform dispersion throughout the material. After impregnation, the alginate undergoes a curing process to fix the encapsulation, leaving the fungi in a dormant but viable state. This curing can be carried out by various known techniques, such as UV curing or heat treatment, providing flexibility in the manufacturing process.

[0224] In yet another embodiment, slot coating or microgravure printing processes are used. These techniques enable high-precision coating of the alginate-fungal mixture, ensuring consistent and effective encapsulation across various types of nonwoven materials. Upon rehydration, the fungi come into immediate contact with the nonwoven scaffold. This proximity provides an ideal environment for the fungi to rapidly rehydrate and initiate growth and digestion processes. The scaffold not only physically supports the fungi but also facilitates the efficient transport of water and nutrients, playing a crucial role in activation and growth.

[0225] The design of this system emphasizes accelerating fungal growth after rehydration. The nonwoven fabric scaffold impregnated with encapsulated fungi becomes an active site for digestion the moment it comes into contact with moisture. This immediate activation is particularly advantageous in absorbent products where the presence of fluid triggers fungal emergence, allowing the digestion process to begin without delay. The activated fungi utilize the nonwoven fabric material as a growth medium, rapidly expanding and beginning to digest the product.

[0226] The above description is merely illustrative of various embodiments. Those skilled in the art can devise various alternatives, modifications, and variations without departing from the spirit of this disclosure. Therefore, this disclosure is intended to encompass all alternatives, modifications, and variations included within the scope of this disclosure.

Claims

1. The first fungal species or strain, An inoculant composition comprising a second fungal species or strain different from the first fungal species or strain, At least one of the first fungal species or strain is a saprophytic fungus, An inoculant composition in which each of the first fungal species or strain and the second fungal species or strain is contained such that they can be inoculated into one or more polymer materials in terms of their relative concentrations and concentrations in the composition, such that the mass ratio of fungal biomass to polymer material is 1:1 or less.

2. The inoculant composition according to claim 1, wherein at least one of the first fungal species or strain is classified as a white rot fungus.

3. The inoculant composition according to claim 1, wherein each of the first fungal species or strain and the second fungal species or strain is contained in such a way that they can be inoculated into one or more polymer materials, in terms of their relative concentrations and concentrations in the composition, at an inoculation ratio such that the mass ratio of fungal biomass to polymer material is 0.5:1 or less.

4. The inoculant composition according to claim 1, wherein each of the first fungal species or strain and the second fungal species or strain is contained in such a way that they can be inoculated into one or more polymer materials in terms of their relative concentrations and concentrations in the composition, such that the mass ratio of fungal biomass to polymer material is 0.1:1 or less.

5. The inoculant composition according to claim 2, wherein the second fungal species or strain is classified as a brown rot fungus or a soft rot fungus.

6. The inoculant composition according to claim 2, wherein the second fungal species or strain is classified as a white rot fungus.

7. The first fungal species or strain is a modified fungal species or strain that recognizes plastic products as a nutrient source and is designed to digest refractory polymers more efficiently through the initiation of enzymatic digestion compared to its naturally occurring counterpart of the same fungal species or strain. The inoculant composition according to claim 1, wherein the second fungal species or strain is a modified fungal species or strain designed to recognize plastic products as a nutrient source and to digest persistent long-chain carbons more efficiently through the initiation of enzymatic digestion compared to a naturally occurring counterpart of the same fungal species or strain.

8. The inoculant composition according to claim 6, wherein each of the first fungal species or strain and the second fungal species or strain is designed by a modification process such that it exhibits improved resistance to abiotic stress compared to a naturally occurring counterpart of the same fungal species or strain.

9. The inoculant composition according to claim 1, wherein each of the first fungal species or strain and the second fungal species or strain is designed by a modification process such that it exhibits improved resistance to biological and / or abiotic stress compared to a naturally occurring counterpart of the same fungal species or strain.

10. Furthermore, it includes a fixed material, The inoculant composition according to claim 1, wherein each of the first fungal species or strain and the second fungal species or strain is immobilized in the composition and thus placed in a metabolically resting state.

11. The inoculant composition according to claim 7, wherein one of the first fungal species or strain and the second fungal species or strain belongs to the phylum Basidiomycota.

12. The first fungal species or strain and the second fungal species or strain are Aureobasidium pullulans, Pestalotiopsis microspora, Aspergillus spp., Fusarium spp., Aspergillus versicolor, Aspergillus fumigatus, Pleurotus ostreatus, Pleurotus djamor, Dichomitus squalens, Trametes versicolor, Phanerochaete chrysosporium, Lentinula edodes, Phellinus pini, Inonotus obliquus and Fomitopsis spraguei An inoculant composition according to claim 11, selected from the group consisting of the following.

13. The inoculant composition according to claim 1, wherein one of the first fungal species or strain and the second fungal species or strain belongs to the phylum Basidiomycota.

14. The inoculant composition according to claim 1, wherein the first fungal species or strain belongs to a different phylum from the second fungal species or strain.

15. The first fungal species or strain and the second fungal species or strain are Aureobasidium pullulans, Pestalotiopsis microspora, Aspergillus spp., Fusarium spp., Aspergillus versicolor, Aspergillus fumigatus, Pleurotus ostreatus, Pleurotus djamor, Dichomitus squalens, Trametes versicolor, Phanerochaete chrysosporium, Lentinula edodes, Phellinus pini, Inonotus obliquus and Fomitopsis spraguei An inoculant composition according to claim 1, selected from the group consisting of the following.

16. Each of the first fungal species or strain, the second fungal species or strain, and the third fungal species or strain is, Aureobasidium pullulans, Pestalotiopsis microspora, Aspergillus spp., Fusarium spp., Aspergillus versicolor, Aspergillus fumigatus, Pleurotus ostreatus, Pleurotus djamor, Dichomitus squalens, Trametes versicolor, Phanerochaete chrysosporium, Lentinula edodes, Phellinus pini, Inonotus obliquus and Fomitopsis spraguei An inoculant composition according to claim 15, selected from the group consisting of the following.

17. The inoculant composition according to claim 1, wherein one or both of the first fungal species or strain and the second fungal species or strain are present in the composition in the form of spores of the fungal species or strain.

18. The inoculant composition according to claim 1, wherein one or both of the first fungal species or strain and the second fungal species or strain are filamentous fungi belonging to any phylum of fungi that have a hyphae stage in their life cycle.

19. When the inoculant is dehydrated, the fungus is in a dormant state in its initial state. The inoculant composition according to claim 1, wherein the inoculant is configured to be rehydrated upon exposure to water, thereby reactivating the fungus from its initial state.

20. An inoculant composition comprising a first modified fungal species or strain, The first modified fungal species or strain is designed to recognize plastic products as a nutrient source and to digest persistent long-chain carbon more efficiently than its naturally occurring counterpart of the same fungal species or strain. An inoculant composition in which the first modified fungal species or strain is immobilized within an immobilization material and thus placed in a metabolically resting state.

21. The aforementioned modified fungal species or strain A step of supplying fungal mycelium to a growth medium containing initial concentrations of simple carbon and plastic material, A step of reducing the concentration of the simple carbon from the initial concentration over time while maintaining the presence of the plastic material in the culture medium, The inoculant composition according to claim 20, which is designed by a process including the following.

22. The inoculant composition according to claim 21, wherein the first modified fungal species or strain is designed to have improved resistance to abiotic stress compared to a naturally occurring counterpart of the same fungal species or strain.

23. The inoculant composition according to claim 20, wherein the first modified fungal species or strain is designed to have improved resistance to abiotic stress compared to a naturally occurring counterpart of the same fungal species or strain.

24. The inoculant composition according to claim 20, wherein the immobilization material is configured to absorb moisture, thereby initiating the first modified fungal species or strain from its immobilized state to a mobile state.

25. The inoculant composition according to claim 22, wherein the immobilization material is configured to absorb moisture, thereby initiating the first modified fungal species or strain from its immobilized state to a mobile state.

26. The inoculant composition according to claim 21, wherein the immobilization material is configured to absorb moisture, thereby initiating the first modified fungal species or strain from its immobilized state to a mobile state.

27. Furthermore, it includes a second modified fungal species or strain, The aforementioned second modified fungal species or strain is designed to recognize plastic products as a nutrient source and to digest persistent long-chain carbon more efficiently than its naturally occurring counterpart of the same fungal species or strain. The inoculant composition according to claim 20, wherein the second modified fungal species or strain is different from the first modified fungal species or strain.

28. The inoculant composition according to claim 20, wherein the first fungal species or strain belongs to either the phylum Basidiomycota or the phylum Ascomycota.

29. The inoculant composition according to claim 20, wherein the first fungal species or strain belongs to the phylum Basidiomycota.

30. The inoculant composition according to claim 20, wherein the first fungal species or strain is a filamentous fungus belonging to any of the phyla of fungi that have a hyphae stage in their life cycle.

31. The inoculant composition according to claim 20, wherein the first fungal species or strain is present in the composition in the form of spores of the first fungal species or strain.

32. A method for reducing the mass of plastic waste, a) A step of forming an inoculum containing one or more pre-selected fungal species, pre-selected fungal strains, or combinations thereof, which can metabolize and consume the long-chain carbon molecules in the polymer material; b) A step of providing a plastic substrate containing long-chain carbon molecules; c) A step of adding the inoculant to the plastic substrate to form an inoculant / substrate composite; and, d) A method comprising the step of digesting the plastic substrate with one or more fungal species, fungal strains, or combinations thereof in the inoculum / substrate composite, thereby converting the long-chain carbon material in the composite.

33. The method according to claim 32, wherein at least one of the pre-selected fungal species or pre-selected fungal strains is present in the inoculum in the form of spores of the pre-selected fungal species or fungal strain.

34. The method according to claim 32, wherein the pre-selected fungal species or fungal strain is cloned and scaled up via bioprocessing by mycelial growth before the formation of the inoculum in step a).

35. The method according to claim 32, wherein the plastic substrate is part of an absorbent consumer product.

36. The method according to claim 35, wherein the absorbent consumer product is selected from the group consisting of diapers, hygiene products and absorbent pads.

37. The inoculation is formed by immobilizing one or more fungal species, fungal strains, or combinations thereof. The method according to claim 32, wherein the inoculum is activated by contact with water in the inoculum / substrate complex, thereby causing one or more fungal species, fungal strains, or combinations thereof to appear.

38. The method according to claim 32, wherein the inoculant / substrate complex is formed by the integration of the inoculant and the substrate through a product manufacturing process.

39. The method according to claim 32, wherein the pre-selected fungal species, pre-selected fungal strain, or combination thereof includes a filamentous fungus belonging to any of the phyla of fungi that have a hyphae stage in their life cycle.

40. The aforementioned inoculation One or more fungal species classified as white rot fungi, The method according to claim 32, comprising one or more fungal species classified as soft rot fungi or one or more fungal species classified as brown rot fungi.

41. The aforementioned inoculation One or more fungal species classified as soft rot fungi, The method according to claim 32, comprising one or more fungal species classified as white rot fungi or one or more fungal species classified as brown rot fungi.

42. The aforementioned inoculation One or more fungal species classified as brown rot fungi, The method according to claim 32, comprising one or more fungal species classified as soft rot fungi or one or more fungal species classified as white rot fungi.

43. The aforementioned inoculation The method according to claim 32, comprising one or more fungal species classified as white rot fungi, one or more fungal species classified as soft rot fungi, and one or more fungal species classified as brown rot fungi.

44. The aforementioned pre-selected one or more fungal species The method according to claim 43, selected from the group consisting of Aureobasidium pullulans, Pestalotiopsis microspora, the genus Aspergillus, the genus Fusarium, Aspergillus versicolor, Aspergillus fumigatus, Pleurotus ostreatus, Pleurotus djamor, Dichomitus squalens, Trametes versicolor, Phanerochaete chrysosporium, Lentinula edodes, Phellinus pini, Inonotus obliquus, and Fomitopsis spraguei.

45. The aforementioned pre-selected one or more fungal species The method according to claim 32, selected from the group consisting of Aureobasidium pullulans, Pestalotiopsis microspora, the genus Aspergillus, the genus Fusarium, Aspergillus versicolor, Aspergillus fumigatus, Pleurotus ostreatus, Pleurotus djamor, Dichomitus squalens, Trametes versicolor, Phanerochaete chrysosporium, Lentinula edodes, Phellinus pini, Inonotus obliquus, and Fomitopsis spraguei.

46. The method according to claim 32, wherein the one or more pre-selected fungal species include at least one species belonging to the phylum Basidiomycota.

47. The method according to claim 46, wherein the pre-selected fungal species, pre-selected fungal strain, or combination thereof includes filamentous fungi.

48. The method according to claim 32, wherein the inoculation is in a dehydrated form configured to be reactivated upon contact with water.

49. The aforementioned inoculation The fungal inoculant is embedded in a substrate selected from the group consisting of natural fiber nonwoven materials, foamed materials, and combinations thereof, thereby being added to the substrate. As the inoculated material is dehydrated, the fungus, which is in a dormant state in its initial state, The method according to claim 48, wherein the inoculum is rehydrated by exposure to water, causing the fungus to reappear prior to step (d).

50. The method according to claim 49, wherein the inoculant / substrate composite is incorporated into the absorbable product in the initial state.

51. moreover, A step of changing the environment variables of the inoculant / substrate complex, selected from the group consisting of temperature, humidity, pH, and oxygen content; A step of introducing one or more chemical or biological additives for the pretreatment of the plastic substrate; The method according to claim 32, comprising one or more steps of applying ultraviolet (UV) pretreatment to the substrate.

52. The aforementioned chemical additive is selected from the group consisting of oxygen-releasing compounds, nitrates, pH buffers, and acid treatments. The biological adjuvant is selected from the group consisting of an enzymatic treatment for forming microsites on the surface of the plastic substrate. The method according to claim 51, wherein the UV pretreatment sterilizes the plastic substrate and alters its surface properties to increase its susceptibility to digestion by fungi.

53. An inoculant composition in which multiple different pre-selected fungal microcolonies are immobilized within multiple encapsulated fungal beads, Each bead contains a single fungal species or strain thereof. An inoculant composition in which the aforementioned multiple different modified fungal species or strains form a consortium of species or strains that do not coexist in nature.

54. The inoculant composition according to claim 53, wherein at least one of the plurality of pre-selected fungal species or strains thereof recognizes plastic products as a nutrient source and is designed to digest persistent long-chain carbon more efficiently than a naturally derived counterpart of the same fungal species or strain.

55. The inoculant composition according to claim 54, wherein at least one of the plurality of pre-selected fungal species or strains thereof is designed to have improved resistance to abiotic stress compared to a naturally derived counterpart of the same fungal species or strain.

56. The inoculant composition according to claim 53, wherein at least one of the plurality of pre-selected fungal species or strains thereof is designed to have improved resistance to abiotic stress compared to a naturally derived counterpart of the same fungal species or strain.

57. The inoculant composition according to claim 32, wherein the pre-selected fungal species or strain thereof is selected from a plurality of different fungal phyla.

58. The aforementioned pre-selected fungal species or strain thereof The inoculant composition according to claim 53, selected from the group consisting of Aureobasidium pullulans, Pestalotiopsis microspora, Aspergillus, Fusarium, Aspergillus versicolor, Aspergillus fumigatus, Pleurotus ostreatus, Pleurotus djamor, Dichomitus squalens, Trametes versicolor, Phanerochaete chrysosporium, Lentinula edodes, Phellinus pini, Inonotus obliquus, and Fomitopsis spraguei.

59. The inoculant composition according to claim 53, wherein the beads contain an alginate.

60. The inoculant composition according to claim 53, wherein at least one of the plurality of first fungal species or strains thereof is present in the composition in the form of spores of the fungal species or strain thereof.

61. A fungal inoculant composition that is immobilized and has storage stability, A fungal inoculant selected from one or more fungal species or strains, which recognizes plastic products as a nutrient source and is designed to digest refractory long-chain carbon more efficiently than its naturally derived counterpart of the same fungal species or strain, A fungal inoculant composition comprising: an immobilization material that encapsulates the fungal inoculant, keeps the fungal inoculant in a dormant state, and protects it from contamination.

62. The composition according to claim 32, wherein the fungal inoculant is stored by dehydration, cryopreservation, or freeze-drying.

63. The composition according to claim 61, wherein the immobilization material is alginate beads.

64. Furthermore, the composition according to claim 61, comprising a cryoprotectant.

65. The composition according to claim 61, wherein the composition is rehydrated by exposure to water, thereby enabling the release of the fungal inoculant.

66. The composition according to claim 61, wherein the composition is formed as beads, a nonwoven fabric substrate, or a foamed structure.

67. The composition according to claim 61, wherein the immobilized material encapsulates the fungal inoculant via a crosslinking reaction with calcium ions.

68. The composition according to claim 67, wherein the composition is porous and maintains a dehydrated state until reactivation by contact with moisture, while allowing oxygen to reach the fungal inoculant.

69. The composition according to claim 61, comprising a coating layer containing trehalose for improving storage stability.

70. A method for producing an inoculant composition, A step of selecting one or more fungal species or strains thereof; A step of immobilizing one or more selected fungal species or strains thereof by dehydration or freeze-drying; and A method comprising the steps of: forming an encapsulated fungal inoculant by encapsulating one or more immobilized fungal species or strains thereof within an immobilization material; and the following steps:

71. The method according to claim 70, wherein the one or more fungal species or strains thereof include at least one species belonging to the phylum Basidiomycota.

72. The method according to claim 70, wherein the one or more fungal species or strains thereof include filamentous fungi.

73. The method according to claim 70, wherein the enclosed fungal inoculant further comprises a modified medium containing transcription factors such as cytokinins and hormones recovered from fungal biomass during fermentation to assist in the emergence of fungi.

74. The method according to claim 70, wherein the encapsulated fungal inoculant further comprises a prepared medium containing one or more recovered metabolites, hydrophobin, a competing compound, and β-glucan, thereby improving fungal activity and stability.

75. A method for controlling the biotransformation of polymer products using a fungal inoculant, a. A process of incorporating a fungal inoculant into a polymer-based product before or after use; b. A step of keeping the fungal inoculant in a dormant state until exposed to a specific environmental stimulus or condition; and c. A method comprising the step of activating the dormant fungal inoculant to initiate the bioconversion of the polymer product at a controlled rate.

76. A hydrogel complex, One or more natural hydrogels having a first fluid absorption capacity C1 and a first mass M1, One or more synthetic hydrogels having a second fluid absorption capacity C2 and a second mass M2, Includes, A hydrogel composite wherein the composite exhibits a total fluid absorption capacity CT equal to or greater than the fluid absorption capacity C2 of the one or more synthetic hydrogels individually.

77. The hydrogel composite according to claim 76, wherein each of the one or more natural hydrogels and the one or more synthetic hydrogels is a crosslinked polymer.

78. Furthermore, the hydrogel complex according to claim 76, comprising one or more fungal inoculants embedded within the complex.

79. The hydrogel complex according to claim 76, wherein the natural hydrogel is an alginate and the synthetic hydrogel is SAP.

80. The hydrogel composite according to claim 76, wherein the natural hydrogel and the synthetic hydrogel are combined to form a microenvironment that enables ionic interactions in the presence of a fluid, thereby affecting one or both of the rubber elasticity and / or osmotic pressure of the hydrogel composite.

81. An absorbent article having at least one layer comprising the hydrogel composite described in claim 76.

82. Furthermore, the hydrogel complex contains one or more fungal inoculants embedded within it. The absorbable article according to claim 81, wherein the fungi in the inoculant are in a metabolically resting state by immobilizing one or more fungal inoculants.

83. The absorbable article according to claim 82, wherein the one or more fungal inoculants include an immobilization material configured to absorb moisture, thereby initiating the movement of the fungi in the inoculant from their immobilized state.

84. A method for manufacturing an inoculant product, a) A step of providing a spawn inoculant from a pre-selected fungal species having the ability to decompose long-chain carbon compounds; b) A step of introducing the inoculant into a nutrient mixture prepared to satisfy the metabolic requirements of the pre-selected fungal species; c) A step of obtaining proliferated mycelial biomass by promoting the mycelial growth of the pre-selected fungal species under controlled conditions in a bioreactor and allowing it to reach the maximum growth level; d) A step of obtaining a mycelial polymer solution by homogenizing the proliferated mycelial biomass and then mixing it with a polymer solution; e) A step of obtaining treated fungal biomass by introducing a prepared culture medium into the mycelial polymer solution, wherein the prepared culture medium contains one or more secondary biological elements useful for supporting fungal growth and activity, selected from the group consisting of biomass, enzymes, polysaccharides, transcription factors, and antimicrobial agents; and, f) A method comprising the step of producing an inoculant product having storage stability by recovering, encapsulating, and immobilizing the treated fungal biomass.

85. The method according to claim 84, wherein the prepared medium further comprises cytokinins and transcription factors such as hormones recovered from fungal biomass during fermentation to assist in the emergence of fungi.

86. The method according to claim 84, wherein the prepared medium comprises one or more recovered metabolites, hydrophobins, competing compounds, and β-glucans, thereby improving fungal activity and stability.

87. The method according to claim 84, wherein the inoculant is a progeny or clone obtained directly from the pre-selected fungal species, thereby assisting in the transmission of the desired trait.

88. The method according to claim 84, wherein the formulation of the nutrient mixture is based on the specific metabolic requirements of the pre-selected fungal species, thereby improving growth and digestive capacity.

89. The method according to claim 84, wherein the conditions within the bioreactor are controlled with consideration to factors such as nutrients, temperature, pH, and moisture, thereby achieving maximum mycelial growth.

90. The method according to claim 84, wherein the homogenization treatment includes a filtration step prior to mixing with the polymer solution.

91. The method according to claim 84, wherein the polymer solution is an alginate solution.

92. The method according to claim 84, wherein the encapsulation and immobilization steps provide stability and shelf life to the fungal inoculant product.

93. The method according to claim 84, wherein the fungal inoculant product is intended for the digestion of persistent long-chain carbon materials in waste management applications.

94. The method according to claim 84, wherein the pre-selected fungal species belongs to the saprophytic fungi.

95. The method according to claim 84, wherein the persistently decomposable long-chain carbon material includes plastics or other polymeric waste.

96. A method for producing a fungal composition that has storage stability, a) A step of providing a fungal inoculum from a pre-selected filamentous fungus; b) A step of preparing a polymer scaffold selected from the group consisting of nonwoven fabrics, foams, particles, and plastic resins; c) A step of introducing the fungal inoculum into the polymer scaffold by solid-phase fermentation or liquid-phase fermentation to obtain a fungal scaffold complex; d) A step of encapsulating the fungal scaffold complex with an encapsulation preservative and a polymer matrix to obtain an encapsulated scaffold; and, e) A method comprising the step of immobilizing the encapsulation scaffold to form a fungal composition having storage stability.

97. The method according to claim 96, wherein the fungal inoculum comprises fungal mycelial biomass or spores.

98. The method according to claim 96, wherein the polymer scaffold is a nonwoven fabric material that comes into direct contact with fibers or a film to enhance absorbency.

99. The method according to claim 96, wherein the fungal inoculum is introduced into the polymer scaffold by solid-phase fermentation, which includes growing filamentous fungi within the nonwoven fabric scaffold.

100. The method according to claim 96, wherein the fungal inoculant is introduced into the polymer scaffold by liquid-phase fermentation, which includes mixing a moist mycelial mixture with the polymer scaffold and immobilizing it.

101. The method according to claim 96, wherein the encapsulating preservative is selected from the group consisting of trehalose, alginate, carrageenan, dextran, maltodextrin, sucrose, and sorbitol.

102. The method according to claim 96, wherein the polymer matrix is ​​sodium alginate crosslinked with calcium chloride.

103. The method according to claim 96, wherein the enclosed scaffold forms an immobilized structure by being crosslinked in a calcium chloride solution.

104. The method according to claim 96, wherein the fungal inoculant is introduced into the polymer scaffold by preparing a sodium alginate mixture that coats the fibers of the nonwoven material.

105. The method according to claim 96, which makes it possible to adapt the fungus to various polymer types, thereby improving digestive efficiency and vitality.

106. The method according to claim 96, wherein the fungal inoculum comprises a prepared medium containing one or more secondary biological elements selected from the group consisting of biomass, enzymes, polysaccharides, transcription factors, and antimicrobial agents.

107. The method according to claim 96, wherein the prepared medium further comprises transcription factors such as cytokinins and hormones recovered from fungal biomass during fermentation to enable the emergence of more rapid and active fungi.

108. The method according to claim 96, wherein the scaffold is selected from the group consisting of foam, particles, and plastic resin, and fungal spores are embedded within the scaffold.

109. The method according to claim 96, wherein the encapsulated fungus is applicable to films, nonwoven fabrics, foams and other polymer materials and is used for digestion.

110. The method according to claim 96, wherein the fungal metabolic processes are preserved during the immobilization and can be reactivated upon rehydration, thereby accelerating the digestion of the polymer scaffold via enzymatic activity.

111. The method according to claim 96, wherein the encapsulating scaffold is used in the product to cause the digestion of a persistent long-chain carbon material.

112. A composition, a) A product selected from absorbent products or waste containers, b) comprising a fungal inoculant having storage stability integrated into the product, The fungal inoculum having storage stability maintains a metabolically dormant state during use of the product. A composition comprising a fungal inoculant having storage stability, which is activated upon exposure to moisture after use.

113. The aforementioned fungal inoculum, The composition according to claim 112, which is incorporated into one or more parts of an absorbent product, selected from the group consisting of a hydrogel in an absorbent layer, a portion laminated to a film of an impermeable layer without impairing breathability, a portion incorporated into a nonwoven permeable layer without impairing strike-through or rewetting, a cuff portion, and an outer landing zone.

114. The composition according to claim 112, wherein the fungal inoculant is enclosed, thereby ensuring a dormant state until activation by moisture.

115. The composition according to claim 113, wherein the hydrogel containing the fungal inoculant is integrated into the absorbent core mixture of the product.

116. The composition according to claim 113, wherein the fungal inoculant is laminated to a film to form an impermeable layer used as a backsheet, packaging, or wrapper.

117. The composition according to claim 113, wherein the fungal inoculant is incorporated into one or more nonwoven fabric permeable layers selected from the group consisting of a top sheet, a distribution layer, a back sheet nonwoven fabric, and a cuff portion.

118. The composition according to claim 113, wherein the fungal inoculant is incorporated into a landing zone, an expandable ear portion, or other structural member of the absorbent product.

119. The composition according to claim 113, wherein the fungal inoculation is integrated into a film to form a substantially impermeable waste container for waste disposal purposes.

120. The composition according to claim 112, wherein the encapsulation structure of the fungal inoculant is configured to delay its appearance during product use.

121. The composition according to claim 112, wherein the fungal inoculant is applied in combination with other biological agents to enhance waste treatment.

122. The composition according to claim 112, wherein the product is an impermeable waste container, and the fungal inoculant maintains a metabolically dormant state until it is activated upon disposal.

123. The composition according to claim 112, wherein the fungal inoculant promotes the digestion process of the product after use.

124. A method for manufacturing an absorbent product, a) A step of preparing a fungal inoculum that has storage stability while retaining its biological elements; and b) A method comprising the step of incorporating the fungal inoculum into an absorbable product by embedding the fungal inoculum in a web substrate or by adding the fungal inoculum as discrete beads to an absorbent core.

125. The method according to claim 124, wherein the fungal inoculant is incorporated as discrete beads consisting of immobilized fungal inoculant particles having absorption properties.

126. The method according to claim 124, wherein the fungal inoculum is embedded in a web substrate, and then a roll of the web substrate is prepared and the roll is cut into patches of a desired size to be integrated into an absorbable product.

127. The method according to claim 124, wherein the fungal inoculant is incorporated as discrete beads, then introduced into a hopper located upstream of the core-forming assembly, mixed with a pulp fiber mixture, and transported to the core-forming assembly to form an absorbent core.

128. The method according to claim 124, wherein the fungal inoculum is embedded in a web substrate applied to an absorbent product using a vacuum rotating drum, fixed with a tackifier or adhesive, and further covered with an additional protective layer.

129. The method according to claim 124, wherein the discrete beads or the web substrate are encapsulated so as to remain dormant until activated by moisture after product use.

130. The method according to claim 124, wherein the fungal inoculum comprises a modified medium containing one or more secondary biological elements selected from the group consisting of enzymes, polysaccharides, and transcription factors to support the growth and activity of the fungus after activation.

131. The method according to claim 124, wherein the encapsulation structure of the fungal inoculant is configured to delay its appearance during product use and to ensure activation only upon exposure to moisture.

132. The method according to claim 124, wherein the fungal inoculant promotes the biodegradation process of the absorbable product after use.

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