Use of isolated Fusarium solani IIa and substrate staining

Abstract

The present invention relates to the field of fungal production of dyes and pigments and the application of said fungal production of dyes for dyeing substrates. More specifically, the present invention relates to an isolated fungus belonging to the species Fusarium solani deposited under the number DSM 34187, a dye produced by said fungus, a method for dyeing a substrate using said fungus, a method for dyeing a substrate using a dye produced by said fungus, a method for producing said dyes, and also the further application of said dyes as antimicrobial substances.

Description

Technical Field

[0001] The present invention relates to the field of fungal production of dyes and pigments and the application of said fungal production of dyes for staining substrates. More specifically, the present invention relates to an isolated fungus belonging to the species Fusarium solani, a dye produced by said fungus, a method for staining a substrate using said fungus, a method for staining a substrate using a dye produced by said fungus, a method for producing said dye, and also to a further application of said dye as an antimicrobial substance.

Background Art

[0002] The textile industry is one of the largest global industrial pollution causes and has one of the largest water footprints. The dyeing process is one of the main causes of river and lake pollution and poses occupational risks to textile workers and ultimately to end consumers. By estimation, 79 billion cubic meters of water use within the global textile and clothing industry was revealed in 2015, which corresponds to one third of the total economy of the EU in 2017.

[0003] Slama HB et al. (2021) outline synthetic dyes in the textile industry, their emission impacts, and treatment methods. Dyes are used to color various types of substrates, such as textile fibers, paper, and cosmetics, but are also used in food and pharmaceuticals. The textile industry alone accounts for approximately 75% of the global dye market and includes about 10,000 different dyes. The textile industry produces fibers to form yarn, which is then converted into fabric. Various types of dyeing processes are used to color textile materials, including uniformly coating the fabric with dye, printing dye on specific areas of the textile material, bleaching, and finishing, which includes crosslinking, softening, and waterproofing the textile material. Two main categories of dyes are known: natural dyes, which are mainly derived from plants, and synthetic dyes, which are artificially synthesized from chemical compounds. Synthetic dyes are further classified into cellulose fiber dyes, such as reactive dyes, direct dyes, indigo dyes, and sulfur dyes; protein fiber dyes, such as azo dyes, anthraquinone dyes, triarylmethane dyes, and phthalocyanine dyes; and synthetic fiber dyes, such as disperse dyes and basic dyes.

[0004] As described by Slama HB et al. (2021), synthetic dyes are derived primarily from petrochemical compounds and are commercialized in liquid, powder, paste, or granular forms. The majority of these synthetic dyes cause harmful effects when released into the environment in untreated or partially treated forms, causing multiple pollution effects on the air, soil, plants, and water resources, but they also cause serious human diseases.

[0005] Furthermore, the production of synthetic dyes from petrochemical compounds has a substantial impact on the environment due to the significant and widespread environmental impacts of the petrochemical industry. The production of pigments and dyes by microorganisms, as well as the subsequent dyeing methods, represent a promising alternative for a more environmentally friendly and sustainable dyeing industry.

[0006] Kristensen SB et al. (2021) describe how Fusarium solani strain 77-13-4 OE:fsr6 G418R, developed by Nielsen MR et al. (2019), produces pigments such as aurofusarin, bicavelin, and fusarvin under selected culture conditions. Nielsen MR et al. (2019) describe a vector system for targeted integration and overexpression of genes in Fusarium solani, in which the Zn(II)2Cys6 transcription factor fsr6, which controls mycelial pigmentation, is cloned and overexpressed. Thereafter, Nielsen et al. (2019) targeted and activated the fusarvin (PKS3:fsr) gene cluster.

[0007] Menezes Bruna S. et al. (2020) describe pigment production by Fusarium solani BRM054066. Molelekoa Tumisi Beiri J. et al. (2021) describe the production of pigments from cultured filamentous fungi of agricultural and industrial by-products using liquid-phase fermentation and solid-state fermentation methods.

[0008] Rathna Janarthanam et al. (2016) describe the production of naphthoquinones and phenols by Fusarium solani PSC-R originating from Palk Bay. Venil CK et al. (2020) describe fungal dyes as potential coloring compounds for textile dyeing. However, the authors also point out the need to explore novel pigment-producing fungi to meet the existing demand for natural dyes. Thus, there is an urgent need for new biological and sustainable production systems for dyes, and for biological and sustainable dyeing of substrates such as textiles. [Overview of the project] [Problems that the invention aims to solve]

[0009] The object of the present invention is to provide a biological system for the sustainable production of dyes and pigments, as well as for a sustainable method of coloring substrates. [Means for solving the problem]

[0010] This objective is solved by the subject matter of the present invention. Surprisingly, it was shown that a specific isolated fungus belonging to the species Fusarium solani can produce a red dye that can be used to alter the color of various substrates. This isolated fungus lacks artificially targeted activation of the gene cluster involved in dye production; for example, the isolated fungus of the present invention lacks artificial activation of the PKS3:fsr gene cluster. Even more surprisingly, it was shown that substrates can be colored either by incubating them with the fungus or by incubating the isolated dye with the substrate. Even more surprisingly, it was shown that when FeCl3 is added to the fungal dye, the fungal dye is converted into a darkened dye or pigment, which can also be used to color substrates.

[0011] According to the present invention, an isolated fungus belonging to the species Fusarium solani is provided, deposited under the number DSM34187 at the Leibnitz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH on February 24, 2022.

[0012] According to the present invention, the isolated Fusarium solani is used, in particular, for staining substrates. According to further embodiments of the present invention, i. A step of providing inoculum of isolated fungus Fusarium solani deposited under number DSM34187, ii. Step of inoculating the culture medium with the inoculum, iii. Optionally, pre-culture the inoculated culture medium, and optionally, inactivate the fungus after pre-culture. iv. The step of contacting the substrate with the culture medium of ii. or iii. until the desired color is obtained, and v. Step of heating the substrate A method for changing the color of a substrate is provided, which includes a series of steps.

[0013] According to certain embodiments, the culture medium preferably contains a carbohydrate source, particularly glucose, in the range of 1-4% (m / v). In particular, pre-culture in iii. and / or contact in iv. should be carried out at a pH in the range of 4.8–6.7, especially in the range of 5.0–6.5.

[0014] In particular, pre-culturing in iii. and / or contact in iv. are carried out at temperatures in the range of 15°C to 35°C, especially in the range of 25°C to 28°C. More specifically, pre-culturing in iii. and / or contact in iv. are carried out under aerobic conditions.

[0015] In particular, the method for altering the color of a substrate described herein further comprises the step of contacting the substrate with FeCl3, the contact resulting in a decrease in lightness (L) according to the HSL color model.

[0016] A further embodiment of the present invention provides a method for altering the color of a substrate, including a method described herein that further comprises the step of contacting the substrate with FeCl3. In particular, contact of the substrate with FeCl3 results in a decrease in lightness (L) according to the HSL color model.

[0017] According to further embodiments, i. A step of providing inoculum of isolated fungus Fusarium solani deposited under number DSM34187, ii. Step of inoculating the culture medium with the inoculum, iii. A step of culturing the inoculated culture medium, iv. Steps to collect biomass and / or culture medium, and v. Optionally, step iv. Extract the dye from the collected material. A method for producing a fungal dye, comprising a series of steps, is also provided herein.

[0018] According to certain embodiments, the culture medium contains a carbohydrate source, preferably glucose, and optionally further contains peptone. In particular, the culture in iii. is carried out at a pH in the range of pH 5.0 ± 0.2 to 6.5 ± 0.2.

[0019] In particular, the culture in iii. is carried out at a temperature in the range of 15°C to 35°C, preferably in the range of 25°C to 28°C. In certain embodiments, the culture in iii. is carried out under aerobic conditions.

[0020] More specifically, the culture in iii. is carried out at least until a red / violet color development is detected in the culture medium and / or biomass. According to an embodiment, the culture medium is a liquid or solid medium.

[0021] According to certain embodiments, the method for producing the fungal dye described herein further comprises the step of adding FeCl3 to produce a blackened pigment, and in particular, the pigment has a lightness (L) in the range of 0 to 20%.

[0022] i. drying the collected biomass and / or the collected culture medium containing the fungus Fusarium solani deposited under the number DSM 34187, ii. suspending the dried material from i. in a solvent, preferably an alcohol, more preferably ethanol, iii. evaporating the solvent of ii., iv. resuspending the residue of iii. in a solvent different from the solvent of ii., preferably an ester, more preferably ethyl acetate, v. evaporating the solvent of iv., and vi. optionally, resuspending the residue of v. A method for extracting a fungal dye is also provided herein, which comprises the consecutive steps of

[0023] According to the present invention, the isolated Fusarium solani of the present invention may be used to produce a dye, or the dye may be extracted from the fungus as described herein. The dye has a color in the range of RGB 170±50, 10±5, 39±10, ranging from cherry red, dark red, brown red, and burgundy red or any variation thereof.

[0024] The present invention also, i. The step of contacting the substrate with the dye described herein until the desired color is obtained, and ii. Step of heating the substrate The present invention provides a method for changing the color of a substrate, comprising a series of steps.

[0025] In particular, the heating of the substrate is carried out at a temperature in the range of 60°C to 121°C, and for at least 20 minutes. A method for producing a darkened pigment is also provided herein, comprising the step of adding FeCl3 to a dye described herein, or the step of adding FeCl3 to the above-described production method.

[0026] The present invention further provides a darkened pigment produced by the method described herein, wherein the pigment has a lightness (L) in the range of 0 to 20%. A further embodiment of the present invention also provides a method for changing the color of a substrate, comprising the steps of contacting the substrate with a dye described herein and heating the substrate until a desired color is obtained.

[0027] According to one embodiment of the present invention, the substrate is a textile material selected from the group consisting of natural textile materials, synthetic textile materials, and combinations of natural textile materials and synthetic textile materials. In particular, natural textile materials are not limited to cotton, silk, wool, abaca, coir, linen, flax, wood, cashmere, and mohair.

[0028] In particular, synthetic woven materials may include, but are not limited to, polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal.

[0029] To stain the substrate, it may be heated to a temperature in the range of 60°C to 121°C, and in particular, the heating is continued for at least 20 minutes. Furthermore, according to a further embodiment, the dye produced by Fusarium solani of the present invention, deposited under number DSM34187, has antimicrobial activity.

[0030] Substrates having antimicrobial activity, wherein the substrate is stained with a dye produced by Fusarium solani of the present invention deposited under number DSM34187, are also particularly included herein. [Brief explanation of the drawing]

[0031] [Figure 1A] These are photographs showing phase-contrast microscopy images of FS IIa at 40× magnification (A) and 100× magnification (B), and a phase-contrast microscopy image of FS IIa at 40× magnification (C) for comparison, showing a microconidia with three septa. [Figure 1B] These are photographs showing phase-contrast microscopy images of FS IIa at 40× magnification (A) and 100× magnification (B), and a phase-contrast microscopy image of FS IIa at 40× magnification (C) for comparison, showing a microconidia with three septa. [Figure 1C] These are photographs showing phase-contrast microscopy images of FS IIa at 40× magnification (A) and 100× magnification (B), and a phase-contrast microscopy image of FS IIa at 40× magnification (C) for comparison, showing a microconidia with three septa. [Figure 2A] These are photographs showing the incubation of Fusarium solani on Sabouraud agar (A) and the incubation of Fusarium solani in Sabouraud broth (side view: B, top view: C). [Figure 2B]These are photographs showing the incubation of Fusarium solani on Sabouraud agar (A) and the incubation of Fusarium solani in Sabouraud broth (side view: B, top view: C). [Figure 2C] These are photographs showing the incubation of Fusarium solani on Sabouraud agar (A) and the incubation of Fusarium solani in Sabouraud broth (side view: B, top view: C). [Figure 3] The first photograph shows extraction with ethyl acetate yielding a red dye / pigment (left flask). The second photograph shows the addition of FeCl3 causing the formation of a dark, insoluble complex (right flask). [Figure 4] This is a photograph showing multifiber (MF) stained with FS IIa. 1 - MF is incubated in a 72-hour culture of FS IIa at room temperature for 3 hours, followed by dye fixation at 60°C for 25 minutes. 2 - Subsequent overnight incubation in FeCl3*6H2O (0.1M, 0.03g ml-1) changes the color due to complex formation between the dye and FeCl3. 3 - MF is incubated in a dye staining bath for 1.5 hours, followed by heat treatment at 60°C for 25 minutes. After cooling, a second incubation in FeCl3 solution results in a gray / black color change for cotton, cellulose, and silk. 4 - MF is incubated in a highly concentrated dye staining bath for 1.5 hours, followed by heat treatment at 60°C for 25 minutes. [Figure 5] This figure shows color measurement using the RGB system. [Figure 6] This figure shows the color measurement using the HSL system. [Figure 7] This figure shows a comparison of the red dye of the present invention (the three images in the middle) with commercially available red dyes / colors (the image on the left showing burgundy, located outside the figure, and the image on the right showing Barbados cherry). [Figure 8] This is a photograph showing the results of a disk test regarding the antimicrobial activity of the dye of the present invention. [Figure 9]This photograph shows N. lichenicola IIa (DSM34187, left) versus N. solani (DSM62805, right). Both strains were grown on Sabouraud agar at 28°C for 4 days. Both strains produced a reddish-brown pigment, with isolate IIa showing higher color intensity or a higher amount of pigment. Both reached a diameter of approximately 4 cm. [Figure 10] This photograph shows N. lichenicola IIa (DSM34187, left) versus N. solani DSM62805 (right). The organisms were grown on Sabouraud agar at +4°C for 45 days. While the aerial mycelium of the isolated strain IIa showed increased pigment deposition, both strains released pigment into the agar layer. [Figure 11] This photograph shows N. lichenicola IIa (DSM34187, left) versus N. solani DSM62805 (right). Both strains were grown in Saboubou broth at 28°C for 48 hours. Both strains released red pigment into the surrounding medium, but again, isolated strain IIa exhibited a deeper reddish color. [Figure 12] This photograph shows N. lichenicola IIa (DSM34187, left) versus N. solani DSM62805 (right). Both strains were grown in Saboubou broth at 28°C for 72 hours. Both strains released red pigment into the surrounding medium, but again, isolated strain IIa exhibited a deeper reddish color. [Figure 13] This is a photograph showing a phase-contrast microscopy examination of N. lichenicola IIa (DSM34187). [Figure 14] This is a photograph showing a phase-contrast microscopy examination of N. Solani DSM62805. [Figure 15] This is a graph showing a chromatogram for molecular identification. [Modes for carrying out the invention]

[0032] Unless otherwise indicated or defined, all terms used herein have their ordinary meanings in the art, as will be obvious to those skilled in the art. See, for example, Sambrook et al., "Molecular Cloning: A Laboratory Manual" (4th edition), Volumes 1-3, Cold Spring Harbor Laboratory Press (2012); Krebs et al., "Lewin's Genes XI," Jones & Bartlett Learning, (2017); Berg et al., "Stryer Biochemie," Springer Verlag, 2018; and Murphy & Weaver, "Janeway's Immunobiology" (9th edition, or more recent edition), Taylor & Francis Inc, 2017.

[0033] The subject matter of the claims refers, in particular, to artificial products, which may be variants of natural (wild-type) products, or to methods of using or producing such artificial products. While there may be some degree of sequence identity with the natural structure, it is understood that the materials, methods and uses of the present invention, particularly referring to isolated nucleic acid sequences, amino acid sequences, expression constructs, transformed host cells, and modified proteins and enzymes, are “man-made” or synthetic and therefore not considered to be the result of “natural laws.”

[0034] The terms “comprise,” “contain,” “have,” and “include,” as used herein, may be used synonymously and are understood to be open definitions that allow for further members, parts, or elements. “Consists of” is considered the most closed definition, without any further elements of the characteristics of the definition of “consists of.” Thus, “includes” more broadly includes the definition of “consists of.”

[0035] As used herein, the term "approximately" refers to a value that is the same as a given value, or a value that is + / - 5% different from it. As used herein and in the claims, singular forms, such as "a," "an," and "the," include plural forms unless the context clearly indicates otherwise.

[0036] The present invention provides an isolated fungus belonging to the species Fusarium solani, which is applicable to the production of dyes applicable to substrate coloring and also applicable to the direct coloring of substrates. Dyes currently commercially used for substrate coloring are chemically synthesized. Biological dyes and staining procedures have a lower environmental impact.

[0037] The isolated fungus of this invention is deposited under the number DSM34187 and is also referred herein to as Fusarium solanii IIa (FS IIa) or Neocosmospora lichenicola. Both terms may be used interchangeably.

[0038] The fungi belonging to the species Fusarium solani are filamentous fungi in the phylum Ascomycota. Fusarium solani are a common soil fungus belonging to the classification of Ascomycetes. More than 60 species of this filamentous fungus are combined within the Fusarium solani species complex (FSSC) (Coleman, 2016). Within the Fusarium solani species complex (FSSC), many species are publicly known and described taxonomically, phylogenetically, and morphologically (Kristensen et al., 2021). According to Short et al. (2013), the FSSC is a diverse complex of many phylogenetically distinct species. There appear to be no clear morphological differences among the various Fusarium solani species (Schroers HJ et al., 2016; Chehri K et al., 2015; Matuo T and Snyder WC, 1972).

[0039] Remarkably, the isolated Fusarium solanii IIa of the present invention produces a red dye in high yield and in a short time, for example, after overnight incubation. Using this dye or the isolated fungus, a dyeing method is possible that allows for direct penetration of color and, therefore, adsorption of the pigment / dye onto textile fibers or material surfaces.

[0040] As used herein, the term "change in color" refers to a change in the color of a substrate. Therefore, the visual appearance of the substrate changes. Colors can be described using color models. The RGB color model is one way of describing colors. The RGB color model is an additive color model in which the primary colors of light—red, green, and blue—are added together in various ways to reproduce a wide range of colors. Alternative methods for describing colors are the CYMK model or the HSL color model. HSL represents hue, saturation, and lightness.

[0041] The term “substrate,” as used herein, refers to any material that may be the target of a color change. Thus, the substrate may be a textile material, but may also be a raw material that is converted into a textile material, such as yarn or thread.

[0042] As used herein, the term “textile material” refers to a material produced by creating a bundle of yarns or threads, which are generated by spinning raw fibers into long and twisted lengths. Raw fibers may be of natural or synthetic origin. Textile materials are formed by weaving, knitting, crocheting, tying, touching, felting, joining, or twisting these yarns together. The terms “textile material,” “textile,” and “woven fabric” may be used interchangeably herein.

[0043] According to a particular embodiment, the substrate is a textile material selected from the group consisting of natural textile materials, synthetic textile materials, and combinations of natural and synthetic textile materials. Natural textile materials may include, but are not limited to, cotton, silk, wool, abaca, coir, linen, flax, wood, cashmere, and mohair, or any combination thereof.

[0044] In particular, synthetic woven materials may include, but are not limited to, polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal, or any combination thereof.

[0045] The terms “dye” and “pigment” are used herein to distinguish the red dye produced by the fungus of the present invention from the dark pigment produced by adding FeCl3 to the red dye. Thereafter, the term “dye” is used for the water-soluble red dye, while the term “pigment” is used for the dark pigment, which has lower water solubility.

[0046] According to one embodiment of the present invention, a method for changing the color of a substrate is provided. The method includes the steps of providing an inoculum of an isolated fungus of the present invention, inoculating the inoculum into a culture medium, optionally pre-culturing the inoculated culture medium and optionally inactivating the fungus after pre-culturing, contacting the substrate with the culture medium from one of the preceding two steps until a desired color is obtained, and heating the substrate.

[0047] According to a particular embodiment, the method for changing the color of a substrate includes the steps of providing an inoculum of an isolated fungus of the present invention, inoculating the inoculum into a culture medium, contacting the substrate with the inoculated culture medium until a desired color is obtained, and heating the substrate.

[0048] According to a particular embodiment, the method for changing the color of a substrate includes the steps of providing an inoculum of an isolated fungus of the present invention, inoculating the inoculum into a culture medium, pre-culturing the inoculated culture medium and optionally inactivating the fungus after pre-culturing, contacting the substrate with the pre-culturised culture medium until a desired color is obtained, and heating the substrate.

[0049] According to a particular embodiment, the method for changing the color of a substrate includes the steps of providing an inoculum of an isolated fungus of the present invention, inoculating the inoculum into a culture medium, pre-culturing the inoculated culture medium and inactivating the fungus after pre-culturing, contacting the substrate with the pre-culturised culture medium until a desired color is obtained, and heating the substrate.

[0050] The term "inoculum," as used herein, refers to a population of fungi of the present invention introduced into a culture medium or any suitable medium for growing the fungi. According to certain embodiments, the inoculant may be in solid or liquid form. The inoculant may be fresh mycelium or may be taken from a frozen culture collection, such as in the form of a frozen culture. The inoculant may include a solid medium containing a culture of the fungus of the present invention, for example, a fungus grown on an agar plate. The solid fungal inoculant may be pre-cultured for several days before inoculation. In particular, the solid fungal inoculant may be pre-cultured for 1 to 30 days or longer, depending on the culture conditions of the pre-culture. Alternatively, the inoculant may include a liquid culture of the fungus of the present invention. The liquid inoculant may be pre-cultured for several days before inoculation. In particular, the liquid inoculant may be pre-cultured for 1 to 30 days or longer, depending on the culture conditions of the pre-culture. In particular, the concentration of the inoculant may vary depending on the form of the inoculant and the pre-culture of the inoculant.

[0051] As used herein, the term "inoculate into culture medium" refers to the transfer of inoculum into the culture medium. According to certain embodiments, the isolated fungi of the present invention are cultured in various ways as described herein. The terms “culture” and “pre-culture” are used herein for the growth of fungi under conditions outside of their natural environment.

[0052] According to certain embodiments, if the fungi are not inactivated before contacting the substrate with the culture medium, the fungi will continue to grow during the contact step. In other words, when the substrate comes into contact with a culture medium containing viable fungi, the color of the substrate changes simultaneously with the growth of the fungi. Therefore, the dyes produced by the fungi are used directly to change the color of the substrate.

[0053] The term "contact," as used herein, refers to the step of bringing a substrate into contact with a dye or a fungus that produces a dye. An example of contact is adding the substrate directly to a liquid culture medium in which the fungus is cultured or still present. Alternatively, if an isolated fungal dye is used to alter the color of a substrate, contact is carried out by adding the substrate to a solution of the dye.

[0054] According to certain embodiments of the present invention, fungal inactivation can be carried out by heating a fungal culture or a culture medium containing fungi to a temperature of about 121°C for at least 20 minutes. According to a particular embodiment, the substrate is removed from the culture medium after the contact step and before the heating step.

[0055] According to certain embodiments, the step of heating the substrate is applied to fix the color. According to certain embodiments, the substrate is heated at temperatures in the range of 60°C to 121°C. In particular, 121°C is used when the heating step is also used for fungal inactivation. Specifically, the substrate is heated at temperatures in the range of 60°C to 65°C, 65°C to 70°C, 70°C to 75°C, 75°C to 80°C, 80°C to 85°C, 85°C to 90°C, 90°C to 95°C, 95°C to 100°C, 100°C to 105°C, 105°C to 110°C, 110°C to 115°C, 115°C to 120°C, 100°C to 121°C, or 121°C. Specifically, the substrate is heated for at least 20 minutes and up to several hours. In particular, heating of the substrate may be carried out for 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes or longer. Specifically, heating of the substrate may be carried out for 20-25, 20-30, 20-40, 20-50, or 20-60 minutes. In particular, heating may be carried out by any method used in the art to heat the substrate. For example, the substrate may be heated in an oven or a dryer.

[0056] According to one embodiment of the present invention, a method for producing a fungal dye comprises a series of steps: providing an inoculant of the isolated fungus of the present invention; inoculating the inoculant into a culture medium; culturing the inoculated culture medium; collecting biomass and / or culture medium; and optionally, extracting a dye from the collected material.

[0057] According to certain embodiments, the culture medium may be a liquid or a solid medium. According to a particular embodiment, in a method for changing the color of a substrate using fungi, the culture medium is a liquid medium.

[0058] According to a particular embodiment, in a method for producing a fungal dye, the culture medium may be a solid medium or a liquid medium. Generally, solid culture media contain substances for solidifying the culture medium, such as agar, but may also contain other components at the same concentrations as those found in liquid culture media.

[0059] According to a particular embodiment, the culture medium includes a carbohydrate source. According to a particular embodiment, the culture medium contains glucose as a carbohydrate source. According to certain embodiments, the culture medium contains glucose in the range of 1% to 4% (m / v). In particular, the culture medium contains 1%, 2%, 3%, and 4% (m / v) glucose.

[0060] According to certain embodiments, the culture medium may include a carbohydrate source other than glucose, or a combination of glucose and a different carbohydrate source. In particular, the culture medium may include maltose instead of glucose, or a combination of maltose and glucose.

[0061] According to certain embodiments, the culture medium comprises peptone. According to a particular embodiment, the culture medium contains 1%, 5%, 10%, 15%, 20%, 1-5%, 1-10%, 5-10%, 10-15%, 5-15%, or 1-15% (m / v) peptone.

[0062] According to certain embodiments, the culture medium may contain any other components commonly used in culture media. Non-limiting examples of such components include buffers, salts, yeast extracts, malt extracts, amino acids, starch, soybean meal, potato extract, rice extract, casein, dextrin, and antibacterial agents.

[0063] According to a particular embodiment, the culture medium is Sabouraud dextrose agar. According to a particular embodiment, the culture medium is Sabouraud dextrose broth. According to a particular embodiment, the culture medium has a pH in the range of 5.0±0.2 to 6.5±0.2. In particular, the pH of the culture medium is 5.0±0.2, 5.1±0.2, 5.2±0.2, 5.3±0.2, 5.4±0.2, 5.5±0.2, 5.6±0.2, 5.7±0.2, 5.8±0.2, 5.9±0.2, 6.0±0.2, 6.1±0.2, 6.2±0.2, 6.3±0.2, 6.4±0.2, or 6.5±0.2.

[0064] According to a particular embodiment, the pH of the culture medium is maintained during fungal pre-culturing, fungal cultivation, while the substrate is in contact with a culture medium containing the inoculated culture medium, and / or while the substrate is in contact with a culture medium containing optionally pre-culturned inoculated culture medium and optionally inactivated fungi. The pH can be maintained by any known method, for example, by using a buffered culture medium and / or by adding an acid or base. The pH may be controlled by a control unit of the bioreactor.

[0065] According to certain embodiments, fungal pre-culture and culture are carried out at temperatures in the range of 10°C to 35°C. In particular, fungal pre-culture and / or culture are carried out at temperatures in the range of 15°C to 35°C. In particular, at temperatures in the range of 15°C to 30°C. In particular, at temperatures in the range of 15°C to 28°C. In particular, at temperatures in the range of 20°C to 28°C. In particular, at temperatures in the range of 25°C to 28°C. In particular, at temperatures of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35°C.

[0066] According to a particular embodiment, contact of the substrate with the culture medium containing the inoculated culture medium is carried out at a temperature in the range of 10°C to 35°C. In particular, at a temperature in the range of 15°C to 35°C. In particular, at a temperature in the range of 15°C to 30°C. In particular, at a temperature in the range of 15°C to 28°C. In particular, at a temperature in the range of 20°C to 28°C. In particular, at a temperature in the range of 25°C to 28°C. In particular, at a temperature of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35°C.

[0067] According to certain embodiments, contact of the substrate with optionally pre-cultured inoculated culture medium and optionally inactivated fungi is carried out at temperatures in the range of 10°C to 35°C. In particular, at temperatures in the range of 15°C to 35°C. In particular, at temperatures in the range of 15°C to 30°C. In particular, at temperatures in the range of 15°C to 28°C. In particular, at temperatures in the range of 20°C to 28°C. In particular, at temperatures in the range of 25°C to 28°C. In particular, at temperatures of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35°C.

[0068] According to certain embodiments, the pre-culture, culture, and contact steps are carried out under aerobic conditions. Aerobic conditions are those in which free oxygen or dissolved oxygen is present. According to certain embodiments, the collection of biomass and / or culture medium may be carried out by separating the fungal biomass from the liquid culture medium, for example, by centrifugation and / or filtration.

[0069] According to certain embodiments, sampling may also be carried out by removing the solid culture medium from its container, for example, from a Petri dish. This allows the solid culture medium containing the fungus to be fragmented.

[0070] According to certain embodiments, fungal culture, fungal pre-culture, contact of a substrate with a culture medium containing an inoculated culture medium, and / or contact of a substrate with a culture medium containing optionally pre-cultured and optionally inactivated fungi may be carried out as liquid fermentation in a bioreactor. The bioreactor may be equipped with various systems and sensors for control and automation, such as an oxygen supply system, a pH control system, and / or a mixing system.

[0071] According to another specific embodiment, for the production of dyes, fungi may be cultured on a solid medium, for example, by solid-state fermentation. Thereafter, the dyes accumulate in the medium and / or in the mycelium of the fungus.

[0072] According to a particular embodiment, in a method for producing a fungal dye, the culture step is carried out at least until red / purple color development is detected in the culture medium and / or biomass. Thus, the culture is carried out until the desired intensity of color is produced by the fungus. The duration of this step may vary depending on the specific conditions of the culture step and on the form and concentration of the fungal inoculum used.

[0073] According to another embodiment, the dye may be extracted from harvested biomass and / or from a culture medium. In the extraction method of the present invention, the final residue may be resuspended. The final residue may also be further processed for the preservation of the dye, for example, by producing a powder.

[0074] According to one embodiment, the color of the dye produced by the fungus of the present invention is in the range of RGB170±50, 10±5, and 39±10 using the RGB color model. According to one embodiment, the lightness (L) of the dye produced by the fungus is in the range of 25-45% according to the HSL color model.

[0075] According to certain embodiments, Fusarium solanii IIa produces a red pigment, which darkens to a dark red, brown-red, and burgundy-red as growth is extended. According to one embodiment of the present invention, a fungal dye may be used to alter the color of a substrate. In particular, the color of the substrate may be altered by a series of steps: contacting the substrate with the fungal dye and heating the substrate until a desired color is obtained. As previously described herein, the heating step is performed to fix the color and is carried out as described elsewhere herein. The resulting color of the fabric depends on the duration of the contact step and also on the concentration of the dye.

[0076] According to certain embodiments, in the step of contacting the substrate with the fungal dye, the dye may be in liquid or solid form. In particular, the substrate may be immersed in a solution containing the dye. More specifically, the substrate may be completely immersed in the solution containing the dye, or partially immersed in the solution. Alternatively, the dye in solution or powder form may be sprinkled onto the substrate. The substrate may be pre-treated or pre-moistened before application of the dye. The dye solution may be sprayed onto the substrate. Various additives may be added to the dye solution; for example, binders and / or thickeners may be added to the dye solution to form a paste.

[0077] According to another embodiment of the present invention, the darkened pigment is produced by adding FeCl3 to the dye of the present invention. In particular, the darkened pigment has a lightness (L) in the range of 0 to 20%. More specifically, the darkened pigment has a lightness (L) of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20%.

[0078] According to certain embodiments, the lightness (L) of the darkened pigment is reduced compared to the lightness (L) of the fungal red dye. According to another embodiment of the present invention, the color of a substrate previously stained with the fungi of the present invention described herein or with the fungal dyes described herein may be further altered by the addition of FeCl3 to the stained substrate. This alters the color of the substrate to a darker shade. The resulting color of the further treated substrate may depend on the color intensity before the staining step and may also depend on the material of the substrate.

[0079] According to certain embodiments, the lightness (L) of the substrate color decreases compared to the substrate color before contact with FeCl3. In other words, contact of the stained substrate of the present invention with FeCl3 results in a decrease in lightness (L) according to the HSL color model. The decrease in lightness (L) compared to the L value before application of FeCl3 may be 10, 20, 30, 40, 50%, or more.

[0080] According to certain embodiments, the color of the substrate can be changed by contacting the substrate with FeCl3, and the substrate was stained with the fungus of the present invention or with a fungal dye prior to contact with FeCl3.

[0081] According to certain embodiments, contact between a substrate pre-dyed with the dye of the present invention and FeCl3 results in the formation of a darkened color. According to another embodiment, the color of the substrate may be altered by a method comprising a series of steps: contacting the substrate with the darkened dye of the present invention, and heating the substrate, until a desired color is obtained. Again, heating is performed to fix the color.

[0082] According to another embodiment of the present invention, the fungal dye of the present invention has antimicrobial activity. The term "antimicrobial activity" refers to all active ingredients that can inhibit bacterial growth, prevent the formation of microbial colonies, and destroy microorganisms.

[0083] According to another embodiment, a substrate stained with the dye of the present invention has antimicrobial activity. In particular, the growth of microorganisms on the substrate is inhibited. According to certain embodiments, textile materials dyed with the dyes of the present invention have antimicrobial activity. In particular, the growth of microorganisms on or near the textile material is inhibited.

[0084] The present invention further includes the following embodiments: 1. An isolated fungus belonging to the species Fusarium solani, deposited under the number DSM34187.

[0085] 2. Use of the isolated fungi described in item 1 for staining the substrate. 3. i. Step of providing an inoculum of the isolated fungus described in item 1, ii. Step of inoculating the culture medium with the inoculum, iii. Optionally, pre-culture the inoculated culture medium, and optionally, inactivate the fungus after pre-culture. iv. The step of contacting the substrate with the culture medium of ii. or iii. until the desired color is obtained, and v. Step of heating the substrate A method for changing the color of a substrate, comprising a series of steps.

[0086] 4. The method according to item 3, wherein the culture medium contains a carbohydrate source. 5. The method according to item 4, wherein the carbohydrate source is preferably glucose in the range of 1-4% (m / v).

[0087] The method according to any one of items 3 to 5, wherein the pre-culture in 6.iii. and / or contact in iv. are carried out at a pH in the range of pH 5.0 ± 0.2 to 6.5 ± 0.2.

[0088] The method according to any one of items 3 to 6, wherein the pre-culture in 7.iii. and / or contact in iv. are carried out at a temperature in the range of 15°C to 35°C. The method according to any one of items 3 to 7, wherein the pre-culture in 8.iii. and / or contact in iv. are carried out under aerobic conditions.

[0089] 9. A method for changing the color of a substrate, comprising the method described in any one of items 3 to 8, further comprising the step of bringing the substrate into contact with FeCl3. 10. The method described in item 9, wherein contact results in a decrease in lightness (L) according to the HSL color model.

[0090] 11. i. Step of providing an inoculum of the isolated fungus described in item 1, ii. Step of inoculating the culture medium with the inoculum, iii. A step of culturing the inoculated culture medium, iv. Steps to collect biomass and / or culture medium, and v. Optionally, step iv. Extract the dye from the collected material. A method for producing a fungal dye, comprising the following sequential steps.

[0091] 12. The method according to item 11, wherein the culture medium comprises a carbohydrate source, preferably glucose. The method according to item 11 or 12, wherein the culture in 13.iii. is carried out at a pH in the range of pH 5.0±0.2 to 6.5±0.2.

[0092] The method according to any one of items 11 to 13, wherein the culture in 14.iii. is carried out at a temperature in the range of 15°C to 35°C, preferably in the range of 25°C to 28°C. The method described in any one of items 11 to 14, wherein the culture in 15.iii. is carried out under aerobic conditions.

[0093] The method according to any one of items 11 to 15, wherein the culture in 16.iii. is carried out until at least red / purple coloration is detected in the culture medium and / or biomass.

[0094] 17. The method according to any one of items 11 to 16, wherein the culture medium is a liquid or solid medium. 18. i. The step of drying the collected biomass and / or culture medium of the fungal culture described in item 1, ii.i. A step of suspending the dried material from i.i. in a solvent, preferably an alcohol, more preferably ethanol. iii.ii. The step of evaporating the solvent, iv. The residue from iii. is resuspended in a solvent different from the solvent in ii., preferably an ester, more preferably ethyl acetate. The step of evaporating the solvent in v.iv, and vi. Optionally, the step of resuspending the residue from v. A method for extracting fungal dyes, comprising the following sequential steps.

[0095] 19. Dyes produced by the method described in any one of items 11 to 17, and / or extracted by the method described in item 18, wherein the color of the dye is in the range of RGB 170±50, 10±5, and 39±10.

[0096] 20. i. The step of contacting the substrate with the dye described in item 19 until the desired color is obtained, and ii. Step of heating the substrate A method for changing the color of a substrate, comprising a series of steps.

[0097] 21. A method for producing a darkened pigment, comprising the step of adding FeCl3 to the dye described in item 19, or the step of adding FeCl3 to the production method described in any one of items 11 to 17.

[0098] 22. A darkened pigment produced by the method described in item 21, wherein the pigment has a lightness (L) in the range of 0 to 20%. twenty three. i. The step of contacting the substrate with the dye described in item 22 until the desired color is obtained, and ii. Step of heating the substrate A method for changing the color of a substrate, comprising a series of steps.

[0099] 24. The use described in item 2, or the method described in any one of items 3 to 10, 20, and 23, wherein the substrate is a textile material selected from the group consisting of natural textile materials, synthetic textile materials, and combinations of natural and synthetic textile materials.

[0100] 25. The method described in item 24, wherein the natural textile material is cotton, silk, wool, abaca, coir, linen, flax, wood, cashmere, or mohair. 26. The method according to item 24, wherein the synthetic woven material is polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, or modal.

[0101] 27. The method according to any one of items 3 to 10, item 20, and items 23 to 26, wherein the heating of the substrate is carried out at a temperature in the range of 60°C to 121°C. 28. The method according to any one of items 3 to 10, 20, and 23 to 27, wherein heating of the substrate is carried out for at least 20 minutes.

[0102] 29. The dye described in item 19, wherein the dye has antimicrobial activity. 30. A substrate having antimicrobial activity, wherein the substrate is stained with the dye described in item 19.

[0103] The examples described herein are illustrative and not intended to limit the invention. Many modifications and variations can be made to the techniques described and explained herein without departing from the scope of the invention. Therefore, it should be understood that the examples are merely illustrative and do not limit the scope of the invention. [Examples]

[0104] Example 1: Culturing of Fusarium solani (FS IIa) according to the present invention Fusarium solanii (FS) IIa was cultured in Luria bertani, Sabouraud, and malt broth, respectively. Microscopic examination revealed mycelial growth. Its body was formed by filaments or hyphae (Figures 1A, 1B, 1C), which follows the typical mycelial growth pattern of fungi belonging to the Ascomycota phylum. FS IIa is cultured aerobically at a temperature range of 25°C to 28°C.

[0105] Example 2: Dye Production Fusarium solanii IIa is grown at 28°C not only on Sabouraud 4% glucose agar (Carl Roth, X932.2) but also in Sabouraud 2% glucose broth (Carl Roth, AE23.1). Using this combined medium, after an overnight incubation (approximately 12-16 hours), a dark reddish-brown dye (see Figures 2A, 2B, and 2C) is formed, which diffuses into the agar or surrounding medium, coloring the mycelium red. In liquid medium, dye formation can take up to 48 hours.

[0106] FS IIa colonies appear burgundy red to dark purple on Sabouraud dextrose agar containing 40 g / l dextrose and 10 g / l peptone at pH 5.6. The colonies show pigmentation after 24 hours of incubation at 25°C–28°C. The pigment diffuses into the agar, resulting in a dark burgundy red colored agar that appears almost black.

[0107] 20g l at pH 5.6 -1 Dextrose and 10g l -1 Culturing in Sabouraud dextrose broth containing peptone yields a Bordeaux-red colored medium and Bordeaux-red colored, curled mycelium after 24 hours of incubation at 28°C at 150 rpm in baffled or unbaffled Erlenmeyer flasks.

[0108] Figure 2A shows Fusarium solani incubated overnight on Sabouraud agar, and Figures 2B and 2C show Fusarium solani incubated overnight in Sabouraud broth.

[0109] Example 3: Dye Extraction of the dye with ethyl acetate yields a red dye (see Figure 3, left flask). Addition of FeCl3 causes the formation of a dark, insoluble complex (see Figure 3, right flask).

[0110] Example 4: Dye Extraction The extraction of this Bordeaux red pigment was carried out according to the following protocol: - After a 10-day incubation period, the colored agar / culture medium containing the colored FS II mycelium on the surface is cut into small pieces and dried.

[0111] - Suspend the dried fragments in EtOH (+10% dH2O) and heat to a maximum of approximately 80°C for 20 minutes. Alternatively, evaporation can be carried out at room temperature over a period of 3 days. - The residue is resuspended in dH2O, and the first extraction is performed in a separatory funnel by adding ethyl acetate. After the evaporation of the ethyl acetate, a sticky, intensely red substance remains. - This red substance can be resuspended in dH2O, ethyl acetate, acetic acid, and ethanol.

[0112] Example 5: Dyeing Inoculate 3 ml of fresh medium with fresh mycelium, or inoculate directly from frozen culture using an inoculation loop. After incubation overnight at 28°C (with stirring), inoculate 200 ml of fresh medium with the starter culture from the previous day.

[0113] The blank fabric sample is added either immediately at the start of the culture or after sufficient pigment formation. Accordingly, the staining time frame ranges from 2 to 72 hours, resulting in different color intensities.

[0114] The blank fabric sample is incubated in a staining bath containing resuspended red dye. Coloration begins immediately, and the intensity increases over time. Each dyeing process is completed using a 30-minute heat treatment at 60°C to 90°C to inactivate the fungi (up to 121°C) and fix the dye to the material fibers.

[0115] Example 6: Dyeing The fungus is cultured in a liquid complex medium. After pre-culturing, the cells are inoculated into fresh liquid medium and grown in the presence of the fabric to be stained. The staining process is stopped as soon as the desired color intensity is reached. Subsequent heat treatment (20 minutes at 60°C or 20 minutes at 90°C) fixes the dye into the fabric.

[0116] Example 7: Dyeing Multifibers (MFs) were incubated in FS IIa culture medium for 72 hours at room temperature for 3 hours, followed by dye fixation at 60°C for 25 minutes. The results of the staining process are shown in column 1 of Figure 4.

[0117] Subsequent overnight incubation of the stained substrate in FeCl3*6H2O (0.1M, 0.03g ml-1) results in a color change due to complex formation between the dye and FeCl3 (see column 2 in Figure 4).

[0118] Example 8: Dyeing The fungus is grown until the desired density and dye production are reached, and then heat (121°C for 20 minutes) is used to inactivate the fungus. Only after the fungus has been inactivated is the fabric added to the culture and incubated until the desired color is reached. The heat treatment completes the dyeing procedure.

[0119] Example 9: Dyeing MF was incubated in a dye bath for 1.5 hours, followed by heat treatment at 60°C for 25 minutes. After cooling, a second incubation in FeCl3 solution resulted in a gray / black color change for cotton, cellulose, and silk. The results are shown in column 3 of Figure 4.

[0120] Example 10: Dyeing The MF was incubated in a highly concentrated dye bath for 1.5 hours and then heat-treated at 60°C for 25 minutes. The results are shown in column 4 of Figure 4.

[0121] Example 11: Dyeing A partially H2O-soluble crude extract is used for dyeing. Incubation at room temperature for approximately 3 hours already yields a dark red coloration to the fabric / material. Heat treatment at 90°C for 30 minutes increases the intensity and saturation of the color.

[0122] Example 12: Dyeing In addition to the described dyeing procedure, the red dye in FS IIa forms a complex with FeCl3, which results in a darker color change to a muted shade, almost black for dyeing silk.

[0123] Example 13: Characterization of stained substrates The stained substrates shown in Figure 4 were characterized using a color analysis device (RGB-2000, Voltcraft). The colors were analyzed using the RGB color model (see Figure 5) and the HSL color model (see Figure 6).

[0124] Example 14: Antimicrobial activity Antimicrobial activity was tested using the disk test. This tested antimicrobial activity against Escherichia coli, Staphylococcus aureus, Pseudomonas sp., and Bacillus subtilis. Fusarium solani exhibited antimicrobial activity not only against Staphylococcus aureus and Bacillus subtilis, but also, to a weaker degree, against Escherichia coli. The disk test was performed by incubating Staphylococcus aureus, Escherichia coli, and Bacillus subtilis on Mueller-Hinton agar at 37°C for 18 hours in the presence of disks prepared with 20 μL of aqueous extract of the dye of the present invention. The results of the antimicrobial activity are shown in Figure 8. The susceptibility of selected microorganisms to the Fusarium solani IIa extract was determined by measuring the diameter of the inhibition zone (ZOI) in millimeters. The results of the antimicrobial activity are also shown in Table 1 below.

[0125] [Table 1]

[0126] Example 15: Sequencing / re-identification of DSM34187 After DNA extraction, the partial gene tef (translocation elongation factor alpha-1) was sequenced using specific primers for identifying Fusarium species. The assembled DNA sequence was loaded into GenBank, MycoID, and the Fusarium ID database. Sequence determination date: March 31, 2022 DSMZ and Braunschweig (GER) sequencing >ID 22-85 Tef (Sequence ID 1) Sequence determination result:

[0127] [Table 2]

[0128] Identification: Neo-Cosmospora Likenicola (C. Massal.) Sand.-Den. & Crous, [MB#822901] Neo-cosmospora lichenicola belongs to the Fusarium solani species complex, FSSC. N. lichenicola belongs to FSSC group 16b.

[0129] Example 16: Morphology of DSM34187 Strain DSM62805 Neocosmospora solani (see references L. Lombard, NA van der Merwe, JZ Groenewald, and PWCrous Studies in Mycology 80:189-245) was used as a reference strain for isolated strain IIa. An initial investigation of potential morphological differences was carried out not only by observation of growth and aerial mycelium, but also by growth patterns in liquid culture. Both strains were cultured at 28°C on Sabouraud agar (4% glucose) and Sabouraud broth (2% glucose). See Figures 9-14.

[0130] Example 17: Molecular Identification A crude extract of the red pigment produced by isolated strain IIa was administered in 0.5 mg ml. -1 The solution was dissolved in EtOH + 25% H2O at the specified concentration and loaded onto an HPLC column (see chromatogram in Figure 15). The chromatogram shows peak A at approximately 29 mAU with a retention time of approximately 1.45–1.50 minutes, peak B at approximately 21 mAU with a retention time of approximately 1.65 minutes, peak C at approximately 3 mAU with a retention time of approximately 1.70 minutes, and peak D at approximately 4 mAU with a retention time of approximately 1.85 minutes.

[0131] Example 18: Molecular Identification The substances identified from the extract of Fusarium lichenicola IIa (DSM34187) are shown in Table 2 below:

[0132] [Table 3]

[0133] References Chehri Khosrow, Salleh Baharuddin, Zakaria Latiffah (2014) Morphological and Phylogenetic Analysis of Fusarium solani Species Complex in Malaysia, Microb Ecol (2015) 69:457-471, DOI 10.1007 / s00248-014-0494-2 Coleman JJ. The Fusarium solani species complex: ubiquitous pathogens of agricultural importance. Mol Plant Pathol. 2016;17(2):146-158. doi:10.1111 / mpp.12289 Kristensen SB, Pedersen TB, Nielsen MR, Wimmer R, Muff J, Sorensen JL. Production and Selectivity of Key Fusarubins from Fusarium solani due to Media Composition. Toxins. 2021; 13(6):376. https: / / doi.org / 10.3390 / toxins13060376 Matuo Takken, Snyder William C. (1972) Use of Morphology and Mating Populations in the Identification of Formes Speciales in Fusarium solani, Phytopathology 63:562-565 Menezes Bruna S. et al., Pigment production by Fusarium solani BRM054066 and determination of antioxidant and anti-inflammatory properties. AMB Express, 2020, 10(1), XP93014625 Molelekoa Tumisi Beiri J. et al., Production of Pigments by Filamentous Fungi Cultured on Agro-Industrial by-Products Using Submerged and Solid-State Fermentation Methods. Fermentation, 2021, 7(4), 295 Nielsen, M.R., Holzwarth, A.K.R., Brew, E. et al. A new vector system for targeted integration and overexpression of genes in the crop pathogen Fusarium solani. Fungal Biol Biotechnol 6, 25 (2019). https: / / doi.org / 10.1186 / s40694-019-0089-2 Short DP, O'Donnell K, Thrane U, et al. Phylogenetic relationships among membersof the Fusarium solani species complex in human infections and the descriptions of F. keratoplasticum sp. nov. and F. petroliphilum stat. nov. Fungal Genet Biol. 2013;53:59-70. doi:10.1016 / j.fgb.2013.01.004 Schroers Hans-Josef, Samuels Gary J., Zhang Ning, Short Dylan P.G., Juba Jean & Geiser David M. (2016) Epitypification of Fusisporium (Fusarium) solani and its assignment to a common phylogenetic species in the Fusariumsolani species complex, Mycologia, 108:4, 806-819, DOI: 10.3852 / 15-255 Slama HB, Chenari Bouket A, Pourhassan Z, Alenezi FN, Silini A, Cherif-Silini H, Oszako T, Luptakova L, Golinska P, Belbahri L. Diversity of Synthetic Dyes from Textile Industries, Discharge Impacts and Treatment Methods. Applied Sciences. 2021; 11(14):6255. https: / / doi.org / 10.3390 / app11146255 Rathna Janarthanam et al., Production of naphthoquinones and phenolics by a novel isolate Fusarium solani PSC-R of Palk Bay and their industrial applications. Bioresource Technology, 2016, 213, 289-298 Venil CK, Velmurugan P, Dufosse L, Devi PR, Ravi AV. Fungal Pigments: Potential Coloring Compounds for Wide Ranging Applications in Textile Dyeing. J Fungi (Basel). 2020;6(2):68. Published 2020 May 20. doi:10.3390 / jof6020068 In some embodiments, the present invention may be described as follows. [Aspect 1] An isolated fungus belonging to the species Fusarium solani, deposited under number DSM34187. [Aspect 2] Use of the isolated fungus described in Aspect 1 for staining a substrate. [Aspect 3] The use according to aspect 2, wherein the substrate is a textile material selected from the group consisting of natural textile materials, synthetic textile materials, and combinations of natural textile materials and synthetic textile materials, in particular, the natural textile material is selected from the group consisting of cotton, silk, wool, abaca, coir, linen, flax, wood, cashmere, and mohair, and the synthetic textile material is selected from the group consisting of polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal. [Aspect 4] i. Step of providing an inoculum of the isolated fungus described in Aspect 1, ii. Step of inoculating the culture medium with the inoculum, iii. Optionally, pre-culture the inoculated culture medium, and optionally, inactivate the fungus after pre-culture. iv. The step of contacting the substrate with the culture medium of ii. or iii. until the desired color is obtained, and v. A step of heating the substrate, wherein the heating of the substrate is carried out at a temperature in the range of 60°C to 121°C, and in particular for at least 20 minutes. A method for changing the color of a substrate, comprising a series of steps. [Aspect 5] The method according to aspect 4, wherein the pre-culture in iii. and / or contact in iv. are carried out at a pH in the range of pH 4.8 to 6.7, at a temperature in the range of 15°C to 35°C, and particularly under aerobic conditions. [Aspect 6] The method according to aspect 4 or 5, wherein the substrate is a textile material selected from the group consisting of natural textile materials, synthetic textile materials, and combinations of natural textile materials and synthetic textile materials, and in particular the natural textile material is selected from the group consisting of cotton, silk, wool, abaca, coir, linen, flax, wood, cashmere, and mohair, and the synthetic textile material is selected from the group consisting of polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal. [Aspect 7] The method according to any one of aspects 4 to 6, wherein the culture medium preferably contains a carbohydrate source, particularly glucose, in the range of 1 to 4% (m / v). [Aspect 8] The method according to any one of aspects 4 to 7, wherein the culture medium is a liquid or a solid medium. [Aspect 9] The method according to any one of aspects 4 to 8, further comprising the step of contacting a substrate with FeCl3, wherein the contact results in a decrease in lightness (L) according to the HSL color model. [Aspect 10] i. Step of providing an inoculum of an isolated fungus as described in Aspect 1, ii. Step of inoculating the culture medium with the inoculum, iii. A step of culturing the inoculated culture medium, iv. Steps to collect biomass and / or culture medium, and v. Optionally, step iv. Extract the dye from the collected material. A method for producing a fungal dye, comprising the following sequential steps. [Aspect 11] The method according to aspect 10, wherein the culture medium preferably contains a carbohydrate source, particularly glucose, in the range of 1 to 4% (m / v). [Aspect 12] The method according to aspect 10 or 11, wherein the culture in iii. is carried out at a pH in the range of pH 4.8 to 6.7, at a temperature in the range of 15°C to 35°C, preferably in the range of 25°C to 28°C, and particularly under aerobic conditions, and in particular the culture in iii. is carried out until at least red / purple coloration is detected in the culture medium and / or biomass. [Aspect 13] The method according to any one of aspects 10 to 12, wherein the culture medium is a liquid or a solid medium. [Aspect 14] The method according to any one of aspects 10 to 13, further comprising the step of adding FeCl3 to produce a darkened pigment, wherein the pigment has a lightness (L) in the range of 0 to 20%. [Aspect 15] i. A step of drying the collected biomass and / or culture medium containing the fungus described in Aspect 1, ii.i. A step of suspending the dried material from i.i. in a solvent, preferably an alcohol, more preferably ethanol. iii.ii. The step of evaporating the solvent, iv. The residue from iii. is resuspended in a solvent different from the solvent in ii., preferably an ester, more preferably ethyl acetate. The step of evaporating the solvent in v.iv, and vi. Optionally, the step of resuspending the residue from v. A method for extracting fungal dyes, comprising the following sequential steps. [Aspect 16] i. The step of contacting a substrate with a dye produced by any one of the methods of aspects 10 to 14 and / or extracted by the method of aspect 15 until a desired color is obtained, ii. A step of heating the substrate, wherein the heating of the substrate is carried out at a temperature in the range of 60°C to 121°C, and in particular for at least 20 minutes. A method for changing the color of a substrate, comprising a series of steps. [Aspect 17] The method according to aspect 16, wherein the substrate is a textile material selected from the group consisting of natural textile materials, synthetic textile materials, and combinations of natural textile materials and synthetic textile materials, and in particular the natural textile material is selected from the group consisting of cotton, silk, wool, abaca, coir, linen, flax, wood, cashmere, and mohair, and the synthetic textile material is selected from the group consisting of polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal.

[0134] [Table 4]

Claims

1. An isolated fungus belonging to the species Fusarium solani, deposited under the number DSM34187.

2. Use of the isolated fungus according to claim 1 for staining a substrate.

3. The use according to claim 2, wherein the substrate is a textile material selected from the group consisting of natural textile materials, synthetic textile materials, and combinations of natural textile materials and synthetic textile materials.

4. i. The step of providing an inoculum of an isolated fungus according to claim 1, ii. Step of inoculating the culture medium with the inoculum, iii. The step of contacting the substrate with the culture medium of ii. until the desired color is obtained, and iv. Step of heating the substrate, A method for changing the color of a substrate, comprising a series of steps.

5. The method according to claim 4, wherein the contact in iii. is carried out at a pH in the range of pH 4.8 to 6.

7.

6. The method according to claim 4, wherein the substrate is selected from the group consisting of natural textile materials, synthetic textile materials, and textile materials selected from the group consisting of combinations of natural textile materials and synthetic textile materials.

7. The method according to claim 4, wherein the culture medium contains a carbohydrate source.

8. The method according to claim 4, wherein the culture medium is a liquid or solid medium.

9. The substrate is FeCl ３ The method according to claim 4, further comprising the step of bringing into contact with

10. i. The step of providing an inoculum of an isolated fungus according to claim 1, ii. Step of inoculating the culture medium with the inoculum, iii. The step of culturing the inoculated culture medium, and iv. Step of collecting biomass and / or culture medium, A method for producing a fungal dye, comprising the following sequential steps.

11. The method according to claim 10, wherein the culture medium comprises a carbohydrate source.

12. The method according to claim 10, wherein the culture in iii. is carried out at a pH in the range of pH 4.8 to 6.

7.

13. The method according to claim 10, wherein the culture medium is a liquid or solid medium.

14. FeCl ３ The method according to claim 10, further comprising the step of adding.

15. i. A step of drying the collected biomass and / or culture medium containing the fungus described in claim 1, ii. The step of suspending the dried material from i. in a solvent, Step iii. Evaporate the solvent of iii. The steps of resuspending the residue of iv. iii. in a solvent different from the solvent of ii., and Steps to evaporate the solvent of v. iv. A method for extracting fungal dyes, comprising the following sequential steps.

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