Fungal enzymatic composition for degrading plastic products

Abstract

The invention relates to a fungal enzymatic composition for degrading plastic products and plastic polymers obtained from the simultaneous cultivation of at least three different species of Trichoderma fungi. Moreover, the present invention also relates to a method for the depolymerization and degradation of plastic-containing waste by the fungal enzymatic composition of the invention.

Description

[0001] DESCRIPTION

[0002] FUNGAL ENZYMATIC COMPOSITION FOR DEGRADING PLASTIC PRODUCTS

[0003] TECHNICAL FIELD

[0004] The invention disclosed herein relates generally to the field of biodegradation and bioremediation. More specifically, the present invention relates to plastic depolymerization, fragmentation and biodegradation. The invention relates to a method for the depolymerization and degradation of plastic-containing waste by a composition comprising encapsulated lytic enzymes obtained from the simultaneous cultivation of at least three different species of Trichoderma fungi.

[0005] STATE OF THE ART

[0006] Plastic is found in several areas of industry, with diverse applications, such as food packaging, pharmaceuticals, toys, general household products, construction, decoration and countless other industries. Each plastic material has a different chemical composition, some of which are reusable, and others have a complex recycling process.

[0007] Plastic polymers are classified into two large distinct groups according to processing and thermal behavior: thermoplastics and thermosets. The former are moldable, as they soften when heated; and the latter are not easily moldable by heating. Among the plastics that are produced and used for various commercial purposes, there are polyethylene (PE)-based products, polypropylene (PP) products, polyvinyl chloride (PVC) products, polyurethane (PU) products, polystyrene (PS, styrofoam) products, polyvinyl chloride (PVC) products, polytetrafluoroethylene (Teflon), polyethylene terephthalate (PET or PETE or polyester) products, among others.

[0008] In 2018, the United Nations (UN) started a global awareness movement about the disposal of plastic objects in the environment and its effects. One of the main aggravating factors of pollution caused by plastic materials is due to the high half-life of this material, which sometimes takes more than 200 years to decompose in nature, which causes a strong environmental impact. Around 91% of the plastic used worldwide is not recycled. According to the ASTM Standard D5033, plastic recycling can be categorized as primary, secondary, tertiary and quaternary (Standard Guide to Development of ASTM Standards Relating to Recycling and Use of Recycled Plastics. American Society for Testing and Materials (ASTM) International; West Conshohocken, PA, USA: 2000).

[0009] Each category involves distinct processes and objectives. Primary recycling, also known as closed-loop recycling, entails mechanically reprocessing plastic scrap to produce a product with properties equal to those of the original material. Secondary recycling, referred to as downgrading recycling, involves mechanically reprocessing plastic scrap to produce a product with altered properties. Tertiary recycling focuses on the recovery of chemical constituents from plastic scrap, while quaternary recycling harnesses the energy content of scrap plastic to generate steam and electricity. Mechanical recycling is used in both primary and secondary recycling. While most thermoplastics, including PET, PE, and PP, have high potential for mechanical recycling, thermosets such as unsaturated polyester and epoxy resin cannot be mechanically recycled due to their molecular structure. The varying processing requirements and molecular incompatibility of different plastic-types present challenges in the production of recycled plastic from plastic waste. The mechanical recycling process comprises several key stages, including collection, sorting, cleaning, size reduction, and compatibilization or separation.

[0010] Generally, the main recycling methods require high-temperature and high-pressure conditions (chemical methods) or consume a large amount of energy and / or generate a large amount of toxic and harmful substances to the environment, which also results in secondary pollution (biological).

[0011] To minimize these effects, based on an environmentally sustainable approach to the recycling of plastic products, biodegradation methods have been intensively applied and studied.

[0012] Recent research on biodegradation has addressed the ability of microorganisms to degrade plastic. With the current knowledge on plastic degradation through microbial means, it is known that enzymes secreted or obtained from microorganisms act mainly on PET and PU polymers (Wei R; ZimmermannW., Microb Biotechnol. 2017 Nov; 10(6): 1308-1322; Danso D., et al. Appl Environ Microbiol. 2019 Sep 17;85(19):e01095-19). Generally, microbial degradation of polymers is a slow process.

[0013] However, the best enzymes and microorganisms with activity in PU and PET still show moderate catalysis rates, therefore microbial degradation of polymers is still a slow process. This high resistance derives mainly from the fiber's high molecular weight, strong C-C interactions and the extremely hydrophobic surface, which is very difficult for enzymes to attack. Furthermore, polymers have different molecular orderings (e.g. amorphous and crystalline forms), resulting in different levels of degradability.

[0014] The first evidence of biodegradation of aromatic polyesters was reported in a study published in 2005, in which the microorganism Thermobifida fusca was used to degrade PET (MullerJR., et al. Enzymatic degradation of Poly(ethylene terephthalate): rapid hydrolyze using a hydrolase form T. fusca. Macromolecular Rapid Communication 2005, 26, 1400-1405). From then on, other microorganisms were tested in the degradation of this substrate.

[0015] Other approaches to biodegradation consist of the use of enzymes isolated from microorganisms. Currently, there are many known functional and verified enzymes, such as PETases, proteases, lipases, carboxylesterases, cutinases and esterases, most of which are biochemically broken down according to the synthetic polymer substrates or oligomers / monomers to which they bind. Cutinase enzymes represent the largest dominant fraction within the studies performed. For example, enzymes have been reported Thermobifida alba Cut1 (ADV92525), Thermobifida fusca Cut2 (CBY05530), Thermobifida fusca Cut1 (AET05798), Thermobifida halotolerans Serine hydrolase (AFA45122), Saccharomonospora viridis Cutinase (BAO42836), Thermomonospora curvata triacilglicerol lipase (WP 012851645), LCC (AEV21261), Ideonella sakaiensis PETase (GAP38373), Polyangium brachysporum triacilglicerol lipase (WP047194864), Vibrio gazogenes lipase (WP021018894), LiplAF5-2 (ACC95208), Oleispira antarctica LipA (CCk74972), Ideonella sakaiensis MHETase (WP 082368715), Humicola insolens cutinase (4OYY) and Fusarium oxysporum cutinase (5AJH) degrade polyethylene substrates. For degradation of the polyamide (PA) substrate, enzymes have been applied Arthrobacter sp NylE (WP079941038), Arthrobacter sp NylA (BAA05090), Arthrobacter sp NylB (CAA24927) and Arthrobacter sp NylC (YP001965068). The enzymes Pseudomonas oleovorans AHs (CAB54050), Pseudomonas fluorescens StyB (CAB06823), Pseudomonas fluorescens StyA (CAB06823) and Pseudomonas fluorescens StyC (CAB06825) degraded polystyrene (PS), while the degradation of polyurethane (Pll) can occur with Pseudomonas fluorescens PueA (AAC23718), Pseudomonas fluorescens PueB (AAY92474) and Pseudomonas fluorescens PulA (AAF66684) (Danso D. et al ppi Environ Microbiol. 2019 Sep 17;85(19):e01095-19). Nonetheless, the use of fungal enzymes in plastic biodegradation is not well known, in contrast to the use of fungal enzymes obtained from fungi of the genus Trichoderma for the degradation of biomass or biological materials, such as cellulosic and lignocellulosic materials. In this sense, PCT application published as WO 2021205160 describes KatG enzymes, enzyme compositions, and their uses in the enzymatic degradation of plastics, enzymes that can be produced in host microorganisms that can include Trichoderma spp. However, KatG enzymes are different from enzymes produced naturally by Trichoderma.

[0016] Therefore, there is a demand for further improved fungal enzymes capable of degrading plastic polymers for use in bioremediation for the degradation of plastic waste.

[0017] DESCRIPTION OF THE INVENTION

[0018] In a first aspect, the present disclosure provides a composition for degrading plastic products, hereinafter the composition of the invention, comprising: a) an enzymatic extract obtained from a culture of a fungal mixture comprising at least three live Trichoderma species selected from the list consisting of T. harzianum spp., T. viride spp., T. longibrachiatum spp., T. koningii spp., and any variant thereof, cultivated all together in laboratory and / or at industrial scale, on micronized oat as solid means of cultivation, wherein T. longibrachiatum spp. was transformed by means of the exposition to radiation before being cultured with the other Trichoderma species, and b) at least a phosphatidine as a surfactant, and preferably, wherein the enzymatic extract and the surfactant are encapsulated within the composition.

[0019] The fungal mixture used in the present invention was publicly available prior to the filing date and can be obtained without restrictions from the commercial suppliers AgroBT (Chile) commercial product: ‘3TAC’ (https: / / agrobt.cl / producto / 3tac / ) and IABT (Investigation y Avance Biotecnologico S.L., Spain). This mixture contains exclusively the species T. harzianum spp., T. viride spp., and T. longibrachiatum spp., as indicated in the commercial product specifications. The biological material is accessible to any person skilled in the art and enables the invention to be reproduced without the need for additional information. The inventors of the present invention found that the composition of the invention, in which the enzymatic extract is encapsulated within a phosphatidine-based amphipathic assembly, has a surprising and effective plastic-degrading activity, thereby demonstrating its suitability for use in the degradation of plastics. Equally surprising, the inventors of the present invention have demonstrated that the composition of the present invention exhibit this plastic-degrading activity without requiring the concomitant use of canonical enzymes such as PETases, proteases, lipases, carboxylesterases, cutinases and / or esterases.

[0020] In a second aspect, the present disclosure relates to the use of the composition of the invention for degrading plastic products.

[0021] In a third aspect, the present disclosure relates to a method of degrading plastic products, hereinafter the method of degrading plastic product of the present invention, wherein the method comprising a depolymerization step, performed at room temperature, wherein the plastic product is contacted with the composition of the invention; and wherein the depolymerization step is conducted in an aqueous liquid medium. The method of the present invention is carried out at room temperature and ambient pressure, which provides advantages over other enzymatic plastic degradation methods known in the prior art, which typically require elevated temperatures and high pressures. These milder conditions reduce energy consumption and simplify process implementation, while still enabling effective degradation of plastics.

[0022] BRIEF DESCRIPTION OF THE DRAWINGS

[0023] To complete the description and in order to provide for a better understanding of the invention, a set of drawings is provided. Said drawings form an integral part of the description and illustrate aspects and embodiments disclosed herein, which should not be interpreted as restricting the scope of the invention, but just as an example of how the invention can be carried out. The drawings comprise the following figures:

[0024] Figure 1 shows a photograph of an electrophoretic SDS-PAGE gel of the enzyme extract of the invention showing the presence of seven different proteins (numbers 1 to 7 shown on the right side of the figure) based on the molecular weight (kDa) of the control sample.

[0025] Figure 2A shows a scanning electron microscopy (SEM) image (500x) obtained from individual samples of the PET fragments used as a control (not treated with the composition of the invention).

[0026] Figures 2B and 2C show SEM images (500x and 1500x, respectively) obtained from individual samples of the PET fragments treated with the composition of the invention for 40 min at room temperature.

[0027] Figures 2D and 2E show SEM images (500x and 1500x, respectively) obtained from individual samples of the PET fragments treated with the composition of the invention for 60 min at room temperature.

[0028] Figures 3A and 3B show SEM images (100x and 1000x, respectively obtained from individual samples of the PET fragments used as a control (not treated with the composition of the invention).

[0029] Figures 3C and 3D show SEM images (100x and 1000x, respectively) obtained from individual samples of the PET fragments treated with the composition of the invention for 24 hours at room temperature

[0030] Figures 4A and 4B show SEM images (100x and 1000x, respectively obtained from individual samples of the PET fragments used as a control (not treated with the composition of the invention).

[0031] Figures 4C and 4D show SEM images (100x and 1000x, respectively) obtained from individual samples of the PET fragments treated with the composition of the invention for 24 hours at room temperature

[0032] DETAILED DESCRIPTION OF THE INVENTION.

[0033] The following definitions and methods are provided to better define the present invention and to guide those of ordinary skill in the art in the practice of the present invention. Unless otherwise noted, terms are to be understood according to conventional usage by those of ordinary skill in the relevant art. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.

[0034] As previously stated, the present invention relates to a composition for degrading plastic products comprising: a) an enzymatic extract obtained from a culture of a fungal mixture comprising at least three live Trichoderma species selected from the list consisting of T. harzianum spp., T. viride spp., T. longibrachiatum spp., and any variants thereof, wherein the T. longibrachiatum spp. has been transformed by exposure to radiation before being cultured together with the other Trichoderma species, and b) at least a phosphatidine, as a surfactant.

[0035] In the context of the invention, a "plastic product” refers to any item made from at least one polymer, monomer(s), hydrocarbons; such as plastic sheet, film, tube, rod, profile, shape, massive block, fiber, etc. Preferably, the plastic article is a manufactured product, such as a rigid or flexible packaging, agricultural films, bags and sacks, disposable items or the like. Preferably, the plastic article comprises a mix of semicrystalline and / or amorphous polymers, or semi- crystalline polymers and additives. The plastic articles may contain additional substances or additives, such as plasticizers, mineral or organic fillers. According to the invention, the plastic article may be selected from a plastic film or a rigid plastic article.

[0036] According to the invention, the term "plastic film" refers to a flexible sheet of plastic (i.e. , capable of being flexed without breaking). Examples of plastic films include agricultural films, plastic bags or sacks, films for flexible packaging, food films, mailing films, liner films, multipack films, industrial films, personal care films, nets, etc.

[0037] According to the invention, the term "rigid plastic article" refers to a plastic article which is not a film. These articles are preferably produced by calendaring, injection-molding, thermoforming, blow molding, or even by rotomolding and 3D printing. Examples of rigid plastic articles are thin wall packaging such as food and beverage packaging, boxes, trays, containers, food service ware, electronics casings, cosmetic cases, outdoor gardening items such as pots, rigid packaging, containers, cards, cotton swabs, irrigation pipes, etc.

[0038] A "polymer" refers to a chemical compound or mixture of compounds whose structure is constituted of multiple repeating units linked by covalent chemical bonds. Within the context of the invention, the term "polymer" includes natural or synthetic polymers, comprising a single type of repeating unit (i.e., homopolymers) or different types of repeating units (i.e., block copolymers and random copolymers). As an example, synthetic polymers include polymers derived from petroleum oil or biobased polymers, such as polyolefins, aliphatic or aromatic polyesters, polyamides, polyurethanes and polyvinyl chloride. Natural polymers include lignin and polysaccharides, such as cellulose, hemi-cellulose, starch and derivatives thereof that may or may not be plasticized. According to the invention, "oligomers" refer to molecules containing from 2 to about 20 monomer units.

[0039] Within the context of the invention, the term "polyester" can be a homopolymer or copolymer. As an example, polylactic acid is an aliphatic homopolymer composed of one monomer, lactic acid; and polyethylene terephthalate is an aliphatic-aromatic copolymer composed of two monomers, terephthalic acid and ethylene glycol. Such polyesters may be native or chemically modified. In the present description, "polyesters" encompass polyethylene terephthalate (PET), polytrimethylene terephthalate (PTT), polybutylene terephthalate (PBT), polyethylene isosorbide terephthalate (PEIT), polylactic acid (PLA), poly(L-lactic acid) (PLLA), poly(D-lactic acid) (PDLA), poly(D,L-lactic acid) (PDLLA), PLA stereocomplex (scPLA), polyhydroxy alkanoate (PHA), Poly(3-hydroxybutyrate) (P(3HB) / PHB), Poly(3-hydroxyvalerate) (P(3HV) / PHV), Poly(3- hydroxyhexanoate) (P(3HHx)), Poly(3-hydroxyoctanoate) (P(3HO)), Poly(3-hydroxydecanoate) (P(3HD)), Poly(3-hydroxybutyrate-co-3- hydroxyvalerate) (P(3HB-co-3HV) / PHBV), Poly(3- hydroxybutyrate-co-3- hydroxyhexanoate) (P(3HB-co-3HHx) / (PHBHHx)), Poly(3- hydroxybutyrate-co-5- hydroxyvalerate) (PHB5HV), Poly(3-hydroxybutyrate-co-3- hydroxypropionate) (PHB3HP), Polyhydroxybutyrate-co-hydroxyoctonoate (PHBO), polyhydroxybutyrate- co-hydroxyoctadecanoate (PHBOd), Poly(3-hydroxybutyrate-co-3- hydroxyvalerate- co-4-hydroxybutyrate) (P(3HB-co-3HV-co-4HB)), polybutylene succinate (PBS), polybutylene succinate adipate (PBSA), polyhydroxyalkanoate (PHA), polybutylene adipate terephthalate (PBAT), polyethylene furanoate (PEF), Polycaprolactone (PCL), poly(ethylene adipate) (PEA), polyethylene naphthalate (PEN) and blends / mixtures thereof.

[0040] In a preferred embodiment, the plastic is selected from the list consisting of PE, PET or polyester, PVC, PS, PP, Pll, Teflon, polybutyrate adipate terephthalate (PBAT), Polylactic acid (PLA), chlorinated polyethylene (CPE), acrylonitrile butadiene styrene (ABS), polycarbonate (PC), nylon, HDEP, LDEP, PE, and blends or mixtures thereof. In another preferred embodiment, the composition of the invention also acts on plastics that might contain a polylactic acid bond to any of its forms and / or derivatives thereof, forming bonds with hydrocarbon polymers.

[0041] A "recycling process" or "complete recycling process" in relation to a plastic article refers to a process by which at least one polymer, monomer, etc., of said plastic article is degraded to yield re-polymerizable monomers and / or oligomers, which are advantageously retrieved in order to be reused.

[0042] In another preferred embodiment, the enzymatic extract included in the composition for degrading plastic products of the invention can be obtained in laboratory or on an industrial scale. The laboratory method for obtaining the extract is disclosed in EP1384405 B1 , which is incorporated herein by reference, particularly with respect to the extract and the method for its preparation.

[0043] EP1384405 B1 discloses that the Trichoderma species are filamentous fungi that exhibits antagonistic activity towards other fungi, including conspecifics strains. The species disclosed include T. harzanium spp., T. viride spp., T. polysporum spp., T. longibrachiatum spp., and T. koningii spp., as well as laboratory-derived variants / dentified by strain designations such as T 22, Tr 115, Tr 116, KRL-Ag 2 (Rifai). The disclosure also refers to haloforms of these species, including Hypocrea and Podostroma.

[0044] In the present invention, the T. longibrachiatum strain is transformed by exposure to cobalt radiation through multiple micro-exposures over approximately 50 generations, resulting in a strain with unique characteristics, resistant to the secretions released by T. harzianum spp., and T. viride spp. These secretions are inactivated by metabolites released by the transformed T. longibrachiatum spp., which alter the pH of the medium and promote absorption of the secretions by the oat substrate. The enzymes involved are thermolabile and remain stable in the oat without degradation.

[0045] The mixture of the three above-mentioned fungal species comprising T. harzianum spp., T. viride spp., and T. longibrachiatum spp., used to produce the enzymatic extract, are co-cultivated as disclosed in EP1384405 B1 or, preferably, may be obtained from commercial suppliers. The mixture was publicly available before the filing or priority date and can be obtained without restrictions from the commercial suppliers such as AgroBT (Chile, commercial product: ‘3TAC’; https: / / aqrobt.cl / producto / 3tac / ) and IABT (Investigation y Avance Biotecnologico S.L., Spain). According to the product specifications, this mixture contains the species mentioned above, T. harzianum spp., T. viride spp., and T longibrachiatum spp. Therefore, the biological material is accessible to any person skilled in the art and enables the invention to be reproduced without the need for additional information.

[0046] In accordance with the present invention, the co-culture of the three Trichoderma spp. strains or the use of the commercial mixture is carried out in a conventional bioreactor, on a solid substrate comprising a micronized, slightly hydrated layer of oat, sterilized by controlled microwave exposure to avoid degradation of the fructose contained in the germ. The culture is fed with glucose and maltose to obtain an aqueous solution rich in secreted enzymes but low in Trichoderma spp. biomass. When the culture reaches maximum growth, the trays are harvested and immediately diluted in fresh substrate, allowing slower fungal growth and dilution of enzymes that could otherwise affect the fungal structure during product aging.

[0047] According to the present invention, the three live Trichoderma species, namely T. viride spp., T. longibrachiatum spp. and T. harzianum spp., are cultured in the proportions: 10:20:70 or 99:05:05, respectively.

[0048] In a more preferred embodiment, the enzymatic extract and the composition of the invention is obtained from a method comprising the following steps: a) Initial inoculation on trays containing the micronutrient oat, micro ionized sterile, and hydrated with at least three Trichoderma species, as mentioned above, in the form of original spores, or with the commercial mixture thereof, in the following proportions: T. harzianum spp. 50%, T. viride spp. 30% and T. longibrachiatum spp. (already irradiated) 20%. b) Critical levels: the culture is handled in the following levels, pH 5, temperature 17°C to 22°C, with expositions of interrupted photo periods; c) Ability of the nutrient to absorb, contain, and deliver (in aqueous solution) the volatile secretions, product of the mutual repulsion of the species T. harzianum and T. viride and the non-volatile secretions of T. longibrachiatum; d) Harvesting of the generated extract by mechanical means, between days 4 and 7. Subsequent re-sowing using live material in all of its vegetative growth phases representing at least 1% of the hydrated substrate in culture trays to continue vegetative growth.

[0049] According to the invention, the term “enzymatic extract” designates active enzymes or enzyme-producing microorganisms, such as sporulating microorganisms, as well as combinations thereof, obtained from the culture of at least three live Trichoderma species selected from the list consisting of T. harzianum spp., T. viride spp., T. longibrachiatum spp., as previously mentioned. According to the invention, “enzymatic extract” preferably refers to enzymes obtained as previously mentioned.

[0050] In a preferred embodiment, the enzymatic extract of the composition of the invention might be concentrated, preferably in ultrapure water. In another particular embodiment of the present invention, the concentrated enzymatic extract to ultrapure water ratio is from about 1 :1 to about 1 :100, preferably, 1 :10, 1 :20; 1 :30; 1 :40; 1 :50; 1 :60; 1 :70; 1 :80; 1 :90; 1 :100.

[0051] In another preferred embodiment, the enzymatic extract comprises enzymes having between about 15 and about 100 kDa molecular weight; more particularly between about 15 and about 75 kDa molecular weight.

[0052] In another preferred embodiment, the enzymatic extract of the composition of the present invention comprises lytic enzymes from alpha / beta hydrolase superfamily selected from the list consisting of: carboxidase-transferase-hydrolases; transferase- hydrolase-carboxidases; hydrolase-carboxidase-transferases, or any combinations thereof. In a more preferred embodiment, the enzymatic extract comprises lytic enzymes carboximethyl cellulase, chitinase, b 1 ,3 gluconase, b xylosidase, xylanase; and a high concentration of 6 pentyl alphapyrone.

[0053] In another preferred embodiment, the enzymatic extract of the composition of the present invention was analyzed by chromatographic techniques, such as Liquid Chromatography with tandem Mass Spectrometry (LC-MS-MS) which combines the separation of proteins / peptides by liquid chromatography with their subsequent analysis by mass spectrometry followed by a bioinformatics study using specialized software and protein databases such as UniProt, which allows the identification of proteins. Thus, once the peptide sequences included in the extract of the invention are obtained, these sequences are matched against all the species belonging to the genus Trichoderma known in the UniProt sequence database, but further against the entire UniProt database comprising plants, fungi and bacteria peptides. Although UniProt database has been used in the present invention for the identification of the enzymes included in the enzymatic extract of the composition of the invention, any sequence database known by the skilled person is useful in the present invention. Table 1 shows the results obtained when the sequences of the enzymes included in the enzymatic extract of the invention are matched against bacteria peptides known in the UniProt sequence database:

[0054] Table 2 shows the results obtained when the sequences of the enzymes included in the enzymatic extract of the invention are matched against fungi peptides known in the UniProt sequence database.

[0055] As can be seen from the results shown in Tables 1 and 2, no proteins with homology to any species of Trichoderma spp. have been detected, however, proteins with homology to proteins of wheat (Triticum aestivum), barley (Hordeum vulgare), oats 5 (Avena sativa) and different bacteria species of the genus Lactobacillus (L. brevis and

[0056] L. plantarum), have been found. Among the proteins detected and found in the enzymatic extract of the present invention, the proteins shown in Table 3 are the most important:

[0057] Table 3. Enzymes included in the enzymatic extract of the present invention

[0058] As can be seen from the information included in Table 3, among the enzymes included in the enzymatic extract of the present invention, enzymes such as xylanases, xylases and endochitinases are of particular interest. In addition, it can be find different 5 enzymes that are widely known for their action in PET degradation such as aspartyl / glutamyl- tRNA amidotransferase, enzymes involved in intermediary metabolisms such as phosphoglycerate kinase, glucose 6-phosphate isomerase or beta-amylase, and enzymes involved in energy storage / obtaining processes such as ATP synthase or Glyceraldehyde-3-phosphate dehydrogenase have been also 10 detected as present in the enzymatic extract of the invention.

[0059] Thus, in another preferred embodiment, the enzymatic extract of the invention is encapsulated in nanocapsules to stabilize them and to allow the reuse of the enzymatic extract, for example in continuous batches. Advantageously, the nanoencapsulation of the enzymatic extract of the invention allows that the depolymerization and degradation 15 of the plastic product by the composition of the invention to be performed at room temperature. Encapsulation techniques are well known to those skilled in the art and include, for instance, nano-emulsions. In a preferred embodiment, the encapsulation of the enzymatic extract of the invention is performed in an amphoteric enzymatic matrix.

[0060] In the context of the present invention, the term “amphoteric enzymatic matrix” refers 20 to a mixture of enzymes, preferably hydrolases exhibiting acid-base duality due to the presence of ionizable amino acid residues capable of functioning either as proton donors or acceptors depending on the local microenvironment. Owing to this amphoteric behavior, these hydrolases initiate the depolymerization process by altering the hydrophobic character of the polymer and / or plastic surface, enabling improved 25 accessibility and catalytic action of the fungal enzymatic extract described in the present invention. The amphoteric enzymatic matrix is preferably an amphoteric matrix of hydrolases, preferably hydrolases including but not limited to amylases, Betaamylases, Gamma-amylases (Glucan 1 ,4-alpha-glucosidase), among others. This amphoteric enzymatic matrix triggers the hydrolysis in the first reaction of the plastic depolymerization / degradation since disrupts or reduces the hydrophobicity of the surface of the plastic product to be depolymerized or degraded, thus allowing the subsequent attack and catalytic activity of the of the enzymes included in the main enzymatic extract included in the composition of the present invention. Table 3 shows the type of enzymes that can be used as amphoteric enzymatic matrix according to the present invention.

[0061] Table 4. Enzymes which can be used as amphoteric enzymatic matrix.

[0062] Thus, the nano-encapsulation of the enzymatic extract of the invention in an amphoteric matrix of amylase-type hydrolases of the Beta-amylase-type hydrolases, Gamma- amylase-type hydrolases (Glucan 1 ,4-alpha-glucosidase), promotes the initiation of hydrolysis in the first reactions of plastic degradation, mainly in the first phase of such degradation. The secondary post-hydrolysis reactions of the plastic by means of glycerol ester hydrolases catalyze the hydrolysis and also the synthesis of carboxylic ester groups. The generated mix allows, using the water solubility of the enzyme glycerol-ester hydrolases, acting on non-soluble substrates (specific ester) including water-oil interfaces. Post-esterase chain reactions, dominated by isomerases, convert the isomer of the compound into another chemical compound, basically, they are in charge of rearranging intramolecular conversions (see Table 4).

[0063] Due to their multiple reactions resulting from the different rearrangements of atoms in the molecule, the enzymes included in the enzymatic extract of the composition of the invention are classified in EC5 category which corresponds to isomerases (https: / / www.enzyme-database.org / - "ExplorEnz - The Enzyme Database"). The most important subcategory within the EC5 Isomerases category, for the enzymes included in the enzymatic extract of the present invention are the following: Hydroxy acid and derivatives thereof; Cis-trans-isomerases; intramolecular oxidoreductases; Keto-enol tautomerases; TT C=C bond modifiers; and S-S disulfide bond modifiers. Depending on the type of metabolic and / or catalytic activity, they present a synonymy that is different in name, but equivalent in their chemical structures, as defined in the above types of enzymes. For clarity of the point, we include the biological / chemical nomenclature:

[0064] In another preferred embodiment, the concentration of the enzymatic extract in the composition is at least 50% w / v; 51% w / v, 52% w / v; 53% w / v; 54% w / v; 55% w / v; 56% w / v; 57% w / v; 58% w / v; 59% w / v; 60% w / v; 61% w / v; 62% w / v; 63% w / v; 64% w / v; 65% w / v; 66% w / v; 67% w / v; 68% w / v; 69% w / v; 70% w / v; 71% w / v; 72% w / v; 73% w / v; 74% w / v; 75% w / v; 76% w / v; 77% w / v; 78% w / v; 79% w / v; 80% w / v; 81% w / v; 82% w / v; 83% w / v; 84% w / v; 85% w / v; 86% w / v; 87% w / v; 88% w / v; 89% w / v; and 90% w / v; preferably between at least 40%p / v to 90 %p / v; more preferably between at least 50% w / v to 90% w / v. In another preferred embodiment, the concentration of the enzymatic extract in the composition is preferably between 50% w / v to 90% w / v, more preferably, between 60% w / v to 80% w / v.

[0065] In another preferred embodiment, the at least a phosphatidine is in the composition of the invention as a micro-ionized aqueous solution comprising a salt selected from the list consisting of NaCL, KCI or any combinations thereof, and wherein the concentration of phosphatidine in the composition is at least 10% w / v; 11% w / v, 12% w / v; 13% w / v; 14% w / v; 15% w / v; 16% w / v; 17% w / v; 18% w / v; 19% w / v; 20% w / v; 21 % w / v; 22% w / v; 23% w / v; 24% w / v; 25% w / v; 26% w / v; 27% w / v; 28% w / v; 29% w / v; 30% w / v; 31% w / v; 32% w / v; 33% w / v; 34% w / v; 35% w / v; 36% w / v; 37% w / v; 38% w / v; 39% w / v; 40% w / v; 41% w / v; 42% w / v; 43% w / v; 44% w / v; 45% w / v; 46% w / v; 47% w / v; 48% w / v; 49% w / v; 50% w / v; 51 % w / v; 52% w / v; 53% w / v; 54% w / v; 55% w / v; 56% w / v; 57% w / v; 58% w / v; 59% w / v; 60% w / v; 61 % w / v; 62% w / v; 63% w / v; 64% w / v%; 65% w / v; 66% w / v; 67% w / v; 68% w / v; 69% w / v; 70% w / v; preferably between at least 10%p / v to 60 %p / v; more preferably between at least 10% w / v to 50% w / v. In another preferred embodiment, the concentration of phosphatidine in the composition is preferably between 10% w / v to 50% w / v, more preferably, between 20% w / v to 40% w / v.

[0066] In the context of the present invention, the term ‘phosphatidine’ is intended to refer to a phospholipid-type compound comprising a glycerol backbone esterified with one or more fatty acids and further comprising a phosphate group capable of conferring amphipathic and surfactant properties. As used herein, “phosphatidine” encompasses phosphatidic acids and related phosphatidyl derivatives that, due to the presence of both hydrophilic (phosphate-based) and hydrophobic (fatty acyl chain) regions, are capable of forming supramolecular assemblies such as micelles, nanoemulsions, or vesicle-like structures.

[0067] The incorporation of at least one phosphatidine in the composition of the present invention provides several technical advantages directly related to its amphipathic and surfactant properties. These supramolecular assemblies stabilize and encapsulate the enzymatic extract obtained from the Trichoderma co-culture, protecting the enzymes from premature denaturation, enhancing their dispersion in aqueous media, and allowing controlled release during the plastic degradation process. Additionally, the amphipathic nature of phosphatidines facilitates the disruption of the hydrophobic plastic surface, improving accessibility of the enzymatic extract to polymer chains and promoting the initiation of depolymerization reactions. Consequently, the presence of phosphatidine significantly increases the efficiency of the composition in degrading plastic products, particularly under room -temperature conditions.

[0068] Equally surprising, the inventors have demonstrated that the composition of the present invention exhibits plastic-degrading activity without requiring canonical enzymes such as PETases, proteases, lipases, carboxylesterases, cutinases and / or esterases, highlighting the unexpected efficacy of the composition.

[0069] Preferably, the phosphatidine is co-encapsulated with the enzymatic extract within the composition, optionally forming part of an amphoteric enzymatic matrix comprising hydrolases capable of initiating depolymerization of the plastic substrate. The amphipathic nature of the phosphatidine assemblies further facilitates disruption of the hydrophobic plastic surface, improving accessibility of the enzymatic extract to polymer chains and promoting depolymerization and degradation reactions. Advantageously, the composition exhibits efficient plastic-degrading activity at room temperature and without the need for canonical enzymes such as PETases, proteases, lipases, carboxylesterases, cutinases, or esterases.

[0070] As plastic waste is insoluble in water, fungal cultures producing biosurfactant plays a key role in the solubilization and / or emulsification of hydrocarbons. These mechanisms facilitate desorption and increase the availability of chemical group withing the plastic product by the composition of the invention, ultimately enhance the biodegradation rate. Moreover, biosurfactants are an alternative to chemical surfactants being eco-friendly, minimally toxic, biodegradable and exhibiting high specificity towards the targeted substrate.

[0071] In a preferred embodiment, the composition of the invention is a liquid composition. Advantageously, the liquid composition of the invention is stable, i.e chemically and biologically stable. In the context of the invention, "chemically stable" refers to a composition wherein the biological entities, i.e. enzymatic extract, do not show any significant loss of activity during a defined period at room temperature, in the dark. More particularly, "chemically stable" refers to a composition wherein the loss of degrading activity of the biological entities is less than 50%, preferably less than 25%, more preferably less than 10% as compared to the degrading activity of said biological entities before introduction in the composition, during a period of time of at least 30 days, preferably at least 90 days, more preferably at least 1 year. According to the invention, the composition of the invention is chemically stable during at least 90 days at 4°C. Particularly, the loss of degrading activity of the enzymatic extract in the composition of the invention is less than 10% as compared to the degrading activity of said enzymatic extract before introduction in the composition, during a period of time of at least 90 days. In the context of the invention, the term "biologically stable" refers to a composition that does not show any subsequent bacterial, yeast of fungal proliferation during a defined period of at least 30 days, preferably at least 90 days, more preferably at least 1 year, at room temperature, in the dark. In a second embodiment, the present invention provides the use of the composition of the present invention in the degradation of plastic products into monomers, oligomers, acids, additives, and / or other components that may be present in the plastic formulation.

[0072] According to the present invention, the plastic product comprises a polymer selected from the list consisting of PET, PTT, PBT, PEIT, PLA, PLLA, PDLA, PDLLA, scPLA, PHA, P(3HB) / PHB, P(3HV) / PHV, P(3HHx), P(3HO), P(3HD), P(3HB-co-3HV) / PHBV, P(3HB-co-3HHx) / (PHBHHx), PHB5HV, PHB3HP, PHBO, PHBOd, P(3HB-co-3HV-co- 4HB), PBS, PBSA, PHA, PBAT, PEF, PCL, PEA, PEN, PVC, PS, PP, PE, PU, ABS, MABS, PC, and blends / mixtures thereof.

[0073] In a preferred embodiment, the composition of the invention also acts on plastics products that might contain a polylactic acid bond to any of its forms and / or derivatives thereof, forming bonds with hydrocarbon polymers.

[0074] In another preferred embodiment, the composition of the present invention is applied to the plastic polymers in the form of a liquid composition, preferably in the form of an aqueous solution.

[0075] In a third embodiment, the present invention provides a method of degrading plastic products by the use of the composition of the present invention.

[0076] In a preferred embodiment, the method of degrading plastic of the present invention comprises a depolymerization step, performed at room temperature, wherein the plastic product is contacted with the composition of the present invention, and wherein the depolymerization step is conducted in an aqueous liquid medium.

[0077] As used herein the term "ambient temperature" or "room temperature" means a temperature between 15°C to 30°C. Such temperatures will include, 15°C, 16°C, 17°C, 8°C, 19°C, 20°C, 21 C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C and 30°C, particularly between 20°C to 28°C, more particularly between 23°C to 25°C.

[0078] According to the present invention, in the depolymerization step, the plastics are degraded into their respective monomers, oligomers, acids, additives, and / or other components that may be present in the plastic formulation. In another preferred embodiment, the pH of the aqueous liquid medium wherein the depolymerization step is conducted has a pH between pH 5.0 and pH 8.0, preferably between pH 5.5-7.5.

[0079] In another preferred embodiment of the method of the present invention, the plastic product is pretreated prior to the depolymerization step, and the pretreatment step preferably includes mechanical / physical modification of the plastic product to increase the surface of contact between the plastic polymers and the composition of the present invention.

[0080] In another preferred embodiment, the pretreatment step is preferably performed by a process selected from the list consisting of cutting, crushing, grinding, milling, fractionation, desiccating, dehydration, agglomeration, granulation, or any combinations thereof.

[0081] In another preferred embodiment, the plastic polymer polymers are degraded into monomers, oligomers, acids, additives, and / or other components that may be present in the plastic formulation.

[0082] According to the present invention, the plastic polymers are selected from the list consisting of PET, PTT, PBT, PEIT, PLA, PLLA, PDLA, PDLLA, scPLA, PHA, P(3HB) / PHB, P(3HV) / PHV, P(3HHx), P(3HO), P(3HD), P(3HB-co-3HV) / PHBV, P(3HB- co-3HHx) / (PHBHHx), PHB5HV, PHB3HP, PHBO, PHBOd, P(3HB-co-3HV-co-4HB), PBS, PBSA, PHA, PBAT, PEF, PCL, PEA, PEN, PVC, PS, PP, PE, PU, ABS, MABS, PC, and blends / mixtures thereof.

[0083] In another preferred embodiment, the composition of the invention also acts on plastics that might contain a polylactic acid bond to any of its forms and / or derivatives thereof, forming bonds with hydrocarbon polymers.

[0084] In another preferred embodiment, the rate of degradation of plastic products ranges from 1g / 1 hour wherein the plastic is a single plastic or a blend or mixture of plastics.

[0085] EXAMPLES

[0086] Following are examples of the invention by means of assays carried out by the inventors, which evidence the effectiveness of the product of the invention. The following examples serve to illustrate the invention and must not be considered to limit the scope thereof.

[0087] EXAMPLE 1. Preparation procedure for the composition of the invention comprising an enzymatic extract obtained from a culture comprising at least three live Trichoderma species selected from T. harzianum spp., T. viride spp., and T. longibrachiatum spp.

[0088] The mixture of the three fungal species comprising T. harzianum spp., T. viride spp., and T. longibrachiatum spp., used to produce the enzymatic extract are co-cultivated as disclosed in EP1384405 B1 , which is incorporated herein by reference or, preferably which is the present case, are obtained from commercial suppliers. The mixture was publicly available before the filing or priority date and can be obtained without restrictions from the commercial suppliers such as AgroBT (Chile, commercial product: ‘3TAC’; https: / / agrobt.cl / producto / 3tac / ) and IABT (Investigation y Avance Biotecnologico S.L., Spain). According to the product specifications, this mixture contains the species mentioned above, T. harzianum spp., T. viride spp., and T longibrachiatum spp,. Therefore, the biological material is accessible to any person skilled in the art and enables the invention to be reproduced without the need for additional information.

[0089] The mixture of T. viride spp., T. longibrachiatum spp., and T. harzianum spp. obtained from AgroBT in the form of the commercial product named “3TAC” were cultivated all together in a traditional bioreactor on micronized and a slightly hydrated layer of oat as solid means of cultivation and fed with glucose andmaltose to obtain an aqueous solution rich in secretions but poor in Trichoderma biomass. The micronized oat is subjected to sterilization by controlled exposition to microwaves to avoid the degradation of the fructose contained in its germ.

[0090] These fungus spores were cultivated over an oat layer previously sterilized by means of controlled exposition to microwaves and slightly hydrated until reaching to isolate each species in a separate jar.

[0091] To avoid the annihilation of the species while living together in the culture, a transformation was realized through the exposition of the T. longibrachiatum strains to radiation, transforming it by induced selection in a strain resistant to the fungicidal secretions that release the two Trichoderma species that are contained in the final product. More specifically, the original T. longibrachiatum species were subjected to micro expositions to Cobalt for 50 generations, obtaining a strain with unique characteristics.

[0092] When the culture has reached maximum growth, the fungus-containing trays are harvested and immediately diluted in the substrate, which allows the fungus to grow more slowly and the dilution of the enzymes that could affect the fungus structure during the aging of the product.

[0093] The three live Trichoderma species are cultured in the proportions: T. harzianum 50%, T. viride 30% and T. longibrachiatum (already irradiated) 20%. The culture is handled in the following levels: pH 5, temperature 17°C to 22°C, with expositions of interrupted photo periods.

[0094] The generated extract harvest implies the total collection of this by mechanical means, between days 4 and 7, its re-sowing is made from this point, using live material in all of its vegetative growth phases at 1% or more with respect to the already hydrated nutrient (micronized sterile and hydrated oat) over culture trays.

[0095] Total protein concentration was determined according to the Bradford (1976) method. The principle of this assay is that the binding of protein molecules to Coomassie dye under acidic conditions results in a color change from brown to blue. The sample analyzed has a content of 1.1 mg / ml of protein.

[0096] Once the extract is obtained, it is lyophilized until a powder is obtained. Then, a solution is prepared with deionized water and amphoteric surfactants in a proportion of 1-5% with respect to the deionized water. As amphoteric surfactant any amphoteric surfactants, chemical and / or biological, known in the art can be used, but preferably the surfactant used is a phosphatidine. A pH between 6.5 and 8 is required, preferably a pH 7. Once said solution is obtained, the powder enzymatic extract and the phosphatidine are slowly added, forming small micelles that during a process of agitation and sonification at RT temperature between 18°C to 25°C for 30 min, nanomicelles are obtained. Depending on the pH, an additional esterification step with calcium carbonate may be required.

[0097] Thus, the composition of the invention comprising the encapsulated extract into nanomicelles obtained as previously disclosed, is used for further analysis.

[0098] EXAMPLE 2. Identification of the type of enzymes included in the enzymatic extract of the composition of the invention by electrophoresis, chromatography and proteomic analysis.

[0099] A sample of the enzymatic extract of the composition of the invention was submitted to electrophoresis in an SDS-Page gel (Western blot) for the identification of the type of enzymes included in the enzymatic extract of the composition of the invention. The results obtained by Western Blot show that the concentration of proteins was low and to identify in a better way the enzymes a silver staining in the SDS-Page gel was carried out. The results obtained show the presence of seven different proteins based on the molecular weight in view of the control (Figure 1). The molecular weight of the enzymes detected is in the range of 15 kDa to 75 KDa (Figure 1).

[0100] Due to the limitation of the electrophoresis technique and in order to obtain a better characterization of the composition of the enzymes included in the enzymatic extract of the invention, a proteomic analysis was performed by Liquid Chromatography with tandem Mass Spectrometry (LC-MS-MS). This technique combines the separation of proteins / peptides by liquid chromatography with their subsequent analysis by mass spectrometry followed by a bioinformatics study using specialized software and protein databases such as UniProt, which allows the identification of proteins. The analysis of the samples of the enzymatic extract of the present invention was performed by the Proteomic Laboratory of the Centro Nacional de Biotecnologia (CNB-CSIC).

[0101] The results obtained by LC-MS-MS shows a similar protein pattern to that obtained by electrophoresis. Table 5 shows the molecular weights of the peaks detected by LC- MS-MS.

[0102] Table 5. Molecular Weight (Da) of the peaks detected by LC-MS-MS in the enzymatic extract of the invention.

[0103] After obtaining the peptide sequences of the analyzed / detected proteins, these sequences were matched with all the species belonging to the genus Trichoderma known in the sequences databases (UniProt).

[0104] 5 Once the results were obtained, and due to the specific absence of proteins of Trichoderma strains in the sequences databases, the protein sequences were matched with the entire UniProt database comprising plants, fungi and bacteria peptides. The results obtained are shown in Tables 1 and 2. No proteins with homology to any species of Trichoderma have been detected, however, proteins with homology to 10 proteins of wheat (Triticum aestivum), barley (Hordeum vulgare), oats (Avena sativa) and different bacteria species of the genus Lactobacillus (L. brevis and L. plantarum), have been found. Among the proteins detected and found in the enzymatic extract of the present invention, the proteins shown in Table 3 reproduced again below, are the most important:

[0105] 15 Table 3. Enzymes included in the enzymatic extract of the present invention. As can be seen from the information included in Table 3, among the enzymes included in the enzymatic extract of the present invention, enzymes such as xylanases, xylases and endochitinases are of particular interest. In addition, it can be find different enzymes that are widely known for their action in PET degradation such as aspartyl / glutamyl- tRNA amidotransferase, enzymes involved in intermediary metabolisms such as phosphoglycerate kinase, glucose 6-phosphate isomerase or beta-amylase, and enzymes involved in energy storage / obtaining processes such as ATP synthase or Glyceraldehyde-3-phosphate dehydrogenase have been also detected as present in the enzymatic extract of the invention.

[0106] EXAMPLE 3. Depolymerization and biodegradation of plastic material by the composition of the invention.

[0107] Blue PET plastic fragments from a post-consumer water bottle, crushed with a particle size between 5x5 mm2 and 10x10 mm2 were put in contact with the liquid composition of the invention, in a concentration of at least 25%w / w.

[0108] Firstly, dilution of the composition of the invention comprising the enzymatic extract concentrates was performed in ultrapure water at a 1 :50 ratio.

[0109] Subsequently, 2 gr of the crushed PET is mixed with 20 ml of the diluted composition of the invention. It is kept for 7 days under constant agitation and temperature, 170 rpm and 25°C, respectively, in an incubator with orbital shaking.

[0110] To monitor the depolymerizing effect by scanning electron microscopy (SEM), individual samples of the PET fragments were taken every 20 min up to a total of 80 min, and a final sample after 7 days of processing. In this last sample, a volume of 1 ml of the solution was collected for analysis by High-Performance Liquid Chromatography (HPLC) and Gas Chromatography / Mass Spectrometry (GC / MS).

[0111] Each time samples of PET fragments are extracted, the plastics are washed with ultrapure water and reserved for further analysis.

[0112] The results show that the surface of PET plastic fragments treated with the composition of the invention for 40 min (Figures 2B and 2C) and for 60 min (Figures 2D and 2E), have been completely transformed with respect to the surface of control PET plastic fragments, not treated with the composition of the invention (Figure 2A). Therefore, after 40 min of being in contact with the composition of the invention, the PET plastic fragment surface has been completely transformed.

[0113] Moreover, Figures 3A and 3B show SEM images (100x and 1000x, respectively) from the PET plastic fragment of a control sample, not treated with the composition of the present invention. As can be seen from these images, the PET plastic frag me nt- control sample shows a uniform and homogeneous surface. However, after 24 hours of being in contact with the composition of the invention, the PET plastic fragment surface has been completely transformed (Figures 3C and 3D).

[0114] Figures 3B and 3D show in more detail how the composition of the invention operates on the PET polymer surface. While the surface of the control sample, Figure 1B, remains virtually unchanged and unmarked, the surface of the PET-treated sample, Figure 3D, shows a complete surface change. The composition of the invention acts on the polymer chains at the molecular level to degrade the plastic product.

[0115] Furthermore, an additional assay with different PET plastic fragments was also performed. The samples were mechanically crushed to reduce the size of the PET plastic fragments as above-mentioned.

[0116] Figure 4A and 4B show SEM images (100x and 1000x, respectively) from the PET plastic fragment of a control sample, not treated with the composition of the present invention. As can be seen from these images, the PET plastic fragment-control sample shows a uniform and homogeneous surface. However, after 24 hours of being in contact with the composition of the invention, the PET plastic fragment surface has been completely transformed (Figures 4C and 4D).

[0117] Figures 4B and 4D show in more detail how the composition of the invention operates on the PET polymer surface. While the surface of the control sample, Figure 4B, remains virtually unchanged and unmarked, the surface of the PET-treated sample, Figure 4D, shows a complete surface change. The composition of the invention acts on the polymer chains at the molecular level to degrade the plastic product.

[0118] Finally, an Energy-dispersive X-ray spectroscopy (EDS) analysis of the PET fragment samples after being 24h in contact with the composition of the invention revealed the presence of free bromine as a constituent of the complexes present in the original PET fragment (See Table 6). Table 6. EDS analysis of PET fragments after being 24h in contact with the composition of the invention. The technical result herein confirms that the composition of the invention can depolymerize and degrade plastic articles at room temperature.

Claims

1. CLAIMS1. Composition for degrading plastic products comprising an enzymatic extract obtained from a co-culture of a fungal mixture comprising at least three different live Trichoderma species selected from the list consisting of T. harzianum spp., T. viride spp., T.polysporum spp., T. longibrachiatum spp., T. koningii spp., and variants thereof, cultivated all together on micronised oat as solid substrate, wherein T. longibrachiatum spp. was transformed by means of the exposition to radiation before being cultured with the other Trichoderma species, and at least a phosphatidine as a surfactant, wherein the enzymatic extract and the at least a phosphatidine are encapsulated within the composition.

2. The composition of the invention according to claim 1 wherein the encapsulation is a nano-encapsulation in an amphotheric matrix.

3. The composition of the invention according to claim 2, wherein the amphotheric matrix is a biological amphoteric matrix, preferably an amylase-type hydrolase enzyme.

4. The composition according to any one of claims 1 to 3, wherein the at least a phosphatidine is in the composition as a micro-ionized aqueous solution comprising a salt selected from the list consisting of NaCL, KCI or any combinations thereof, and wherein the concentration of phosphatidine in the composition is preferably between 10% p / v to 50%p / v.

5. The composition according to any one of claims 1 to 4, wherein the enzymatic extract is obtained from a culture comprising the three Trichoderma species T. viride spp., T. longibrachiatum spp., and T. harzianum spp., present in the culture in the proportions of 10:20:70 or 99:05:05, respectively.

6. The composition according to any one of claims 1 to 5, wherein the enzymatic extract comprises enzymes from alpha / beta hydrolase superfamily selected from the list consisting of: carboxidase-transferase-hydrolases; transferase-hydrolase- carboxidases; hydrolase-carboxidase-transferases, or any combinations thereof.

7. The composition according to any one of claims 1 to 6, wherein the plastic is selected from the list consisting of polyethylene terephthalate (PET), polyvinyl chloride (PVC), polystyrene (PS), polypropylene (PP), polyethylene (PE), polylactic acid (PLA), acrylonitrile butadiene styrene (ABS), methylmethacrylate acrylonitrilebutadiene styrene (MABS), polycarbonate (PC), polyurethane (Pll), nylon, and blends or mixtures thereof.

8. Use of the composition according to any one of claims 1 to 7, for degrading plastic products into monomers, oligomers, acids, additives, and / or other components that may be present in the plastic formulation.

9. Use according to claim 8, wherein the plastic is selected from the list consisting of PET, PVC, PS, PP, PE, PLA, PU, ABS, MABS, PC and blends or mixtures thereof.

10. Use according to claim 8 or 9 wherein the plastic comprises at least a polylactic acid bond to any of its forms and / or derivatives thereof, forming bonds with hydrocarbon polymers.

11. Use according to any one of claims 8 to 10, wherein the composition is applied to the plastic product in the form of a liquid composition, preferably in the form of an aqueous solution.

12. Method of degrading plastic products, wherein the method comprising a depolymerization step, performed at room temperature, preferably between 15°C and 30°C, wherein the plastic product is contacted with the composition according to any one of claims 1 to 7; and wherein the depolymerization step is conducted in an aqueous liquid medium, wherein the pH of aqueous liquid medium is preferably between pH 5.0 to 8.0.

13. The method according to claim 12, wherein the plastic product is pretreated prior to the depolymerization step, the pretreatment preferably including mechanical / physical modification of the plastic product to increase the surface of contact between the polymers and the composition, and wherein the pretreatment preferably is performed by a process selected from the list consisting of cutting, crushing, grinding, milling, fractionation, desiccating, dehydration, agglomeration, granulation, or any combinations thereof.

14. The method according to any one of claims 12 to 13, wherein the plastic is selected from the list consisting of PET, PVC, PS, PP, PE, PLA, PU, ABS, MABS, PC, and blends or mixtures thereof.

15. The method according to any one of claims 12 to 14, wherein the rate of degradation of plastic from 1g / 1 hour wherein the plastic is a single plastic or a blend or mixture of plastics.

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