Isolated Fusarium solani IIa and its use for staining substrates

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 to dye 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 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 agents. [Background technology]

[0002] The textile industry is one of the largest global industrial polluters and has one of the largest water footprints. The dyeing process is one of the main sources of pollution to rivers and lakes, posing occupational hazards to textile workers and ultimately to the end consumer. Estimates reveal a water use of 79 billion cubic metres within the global textile and clothing industry in 2015, equivalent to one third of the water required for the entire EU economy in 2017.

[0003] Slama HB et al. (2021) provide an overview of synthetic dyes, their emissions impacts and treatment methods for the textile industry. Dyes are used for coloring various types of substrates, e.g. textile fibers, paper, cosmetics, but also food and medicine. The textile industry alone accounts for about 75% of the global dye market and contains approximately 10,000 different dyes. The textile industry produces fibers to form yarns, which are converted into woven fabrics. Different types of dyeing processes are used to dye textile materials, such as coating the textile evenly with dyes, printing dyes in specific areas of the textile material, bleaching, and finishing, which includes cross-linking, softening, and waterproofing the textile material. Two main categories of dyes are known (natural dyes, mainly derived from plants, and synthetic dyes, artificially synthesized from chemical compounds). Synthetic dyes are further classified into cellulosic 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 mainly derived from petrochemical compounds and are commercialized in liquid, powder, paste, or granular form. Most of these synthetic dyes cause harmful effects when discharged into the environment in untreated or partially treated form, causing multiple pollution effects on air, soil, plants, and water resources, but they also cause significant human diseases.

[0005] Furthermore, the production of synthetic dyes from petrochemical compounds has substantial impacts on the environment due to the large and widespread environmental impacts of the petrochemical industry. Microbial pigment and dye production as well as the subsequent dyeing process are promising alternatives for a "greener" and sustainable dyeing industry.

[0006] Kristensen SB et al. (2021) describe that Fusarium solani strain 77-13-4 OE:fsr6 G418R by Nielsen MR et al. (2019) produces pigments such as aurofusarin, bikaverin and fusarubin under selected culture conditions. Nielsen MR et al. (2019) reported that Zn(II) controls mycelial pigmentation. 2 Cys 6 (2019) describe a vector system for targeted integration and overexpression of genes in Fusarium solani, in which the transcription factor fsr6 was cloned and overexpressed, thereby targeting and activating the fusarubin (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 by cultured filamentous fungi from agro-industrial by-products using submerged and solid-state fermentation methods.

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

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

[0010] This object is solved by the subject matter of the present invention. Surprisingly, it has been shown that a particular isolated fungus belonging to the species Fusarium solani is capable of producing a red dye that can be used to change the color of various substrates. This isolated fungus lacks artificial targeted activation of a gene cluster involved in dye production, for example, the isolated fungus of the present invention lacks artificial activation of the PKS3:fsr gene cluster. Furthermore, it has been surprisingly shown that substrates can be colored directly by incubating the substrate with the fungus or by incubating the isolated dye with the substrate. Even more surprisingly, FeCl 3 It has been shown that when added to a fungal dye, the fungal dye is converted into a dark dye or pigment, which can also be used to color substrates.

[0011] According to the present invention, there is provided an isolated fungus belonging to the species Fusarium solani, 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 invention, the isolated Fusarium solani is particularly used for staining a substrate. According to a further embodiment of the present invention, i. providing an inoculum of the isolated fungus Fusarium solani deposited under the number DSM 34187; ii. inoculating the culture medium with an inoculum; iii. optionally pre-incubating the inoculated culture medium and, optionally, inactivating the fungus after pre-incubation; iv. contacting the substrate with the culture medium of ii. or iii. until the desired color is obtained; and v. Heating the substrate A method for changing the color of a substrate is provided, comprising the successive steps of:

[0013] According to a particular embodiment, the culture medium comprises a carbohydrate source, in particular glucose, preferably in the range of 1-4% (m / v). In particular, the pre-culture in iii. and / or the contacting in iv. is carried out at a pH in the range of pH 4.8 to 6.7, in particular in the range of pH 5.0 to 6.5.

[0014] In particular, the pre-culture in iii. and / or the contacting in iv. is carried out at a temperature in the range of 15°C to 35°C, in particular in the range of 25°C to 28°C. More specifically, the pre-culture in iii. and / or the contacting in iv. are carried out under aerobic conditions.

[0015] In particular, the method of changing the color of a substrate described herein involves treating the substrate with FeCl 3 wherein said contacting specifically results in a decrease in lightness (L) according to the HSL color model.

[0016] According to a further embodiment of the invention, the substrate is FeCl 3 Provided herein are methods of changing the color of a substrate, including the methods described herein, further comprising contacting the substrate with FeCl. 3 Contact with results in a decrease in lightness (L) according to the HSL color model.

[0017] According to a further embodiment, i. providing an inoculum of the isolated fungus Fusarium solani deposited under the number DSM 34187; ii. inoculating the culture medium with an inoculum; iii. culturing the inoculated culture medium; iv. harvesting the biomass and / or culture medium; and v. Optionally, extracting a dye from the harvested material of iv. Also provided herein is a method for producing a fungal dye, comprising the successive steps of:

[0018] According to certain embodiments, the culture medium comprises a carbohydrate source, preferably glucose, and optionally further comprises a peptone. In particular, the culture in iii. is carried out at a pH in the range of 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 culturing in iii. is carried out under aerobic conditions.

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

[0021] According to certain embodiments, the method of producing a fungal dye described herein comprises the step of adding FeCl to produce a dark pigment. 3 wherein the pigment has a lightness (L) in the range of 0 to 20%.

[0022] i. drying the harvested biomass and / or the harvested 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. Also provided herein is a method for extracting a fungal dye, comprising the successive steps of:

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

[0024] The present invention also provides i. contacting the substrate with a dye as described herein until the desired color is obtained; and ii. Heating the substrate The present invention provides a method for changing the color of a substrate, comprising the successive steps of:

[0025] In particular, heating of the substrate is carried out at a temperature in the range of from 60° C. to 121° C., in particular for at least 20 minutes. FeCl 3 to the dye described herein, or 3 Also provided herein is a method for producing a dark pigment, comprising the step of adding to the above production method.

[0026] The present invention further provides a dark pigment produced by the methods described herein, said pigment having a lightness (L) in the range of 0-20%. According to further embodiments of the present invention, there is also provided herein a method of changing the color of a substrate comprising the successive steps of contacting the substrate with a dye as described herein and heating the substrate until a desired color is obtained.

[0027] According to certain embodiments of the present invention, the substrate is a woven material selected from the group consisting of natural woven materials, synthetic woven materials, and combinations of natural and synthetic woven materials. In particular, the natural textile materials may be, but are not limited to, cotton, silk, wool, abaca, coir, linen, hemp, wood, cashmere, mohair.

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

[0029] To dye the substrate, the substrate may be heated at a temperature ranging from 60° C. to 121° C., in particular said heating lasting for at least 20 minutes. According to yet a further embodiment, the dye produced by the Fusarium solani of the present invention deposited under the number DSM 34187 has antimicrobial activity.

[0030] Specifically encompassed herein is a substrate having antimicrobial activity, said substrate being stainable with a dye produced by the Fusarium solani of the present invention, deposited under the number DSM 34187. [Brief description of the drawings]

[0031] [Figure 1A] Photographs showing phase contrast microscopy of FS IIa at 40x magnification (A), 100x magnification (B), and 40x magnification (C) showing three septate microconidia in comparison. [Figure 1B] Photographs showing phase contrast microscopy of FS IIa at 40x magnification (A), 100x magnification (B), and 40x magnification (C) showing three septate microconidia in comparison. [Figure 1C] Photographs showing phase contrast microscopy of FS IIa at 40x magnification (A), 100x magnification (B), and 40x magnification (C) showing three septate microconidia in comparison. [Figure 2A]Photographs showing incubation of Fusarium solani on Sabouraud agar (A) and in Sabouraud broth (side view: B, top view: C). [Figure 2B] Photographs showing incubation of Fusarium solani on Sabouraud agar (A) and in Sabouraud broth (side view: B, top view: C). [Figure 2C] Photographs showing incubation of Fusarium solani on Sabouraud agar (A) and in Sabouraud broth (side view: B, top view: C). [Diagram 3] Photograph showing that extraction with ethyl acetate results in a red dye / pigment (left flask) and addition of FeCl3 causes the formation of a dark, insoluble complex (right flask). [Figure 4] Photographs showing multifibers (MF) stained with FS IIa. 1 - MF is incubated in a 72-h culture of FS IIa for 3 h at room temperature followed by dye fixation at 60°C for 25 min. 2 - Subsequent overnight incubation in FeCl3*6H2O (0.1 M, 0.03 g ml-1) results in a color change due to complexation of the dye and FeCl3. 3 - MF is incubated in a dye dye bath for 1.5 h followed by heat treatment at 60°C for 25 min. After cooling, a second incubation in FeCl3 solution results in a color change to gray / black for cotton, cellulose and silk. 4 - MF is incubated in a highly concentrated dye dye bath for 1.5 h followed by heat treatment at 60°C for 25 min. [Diagram 5] FIG. 1 is a diagram showing color measurement according to the RGB system. [Figure 6] Illustrates color measurements by the HSL system. [Figure 7] FIG. 1 shows a comparison of the red dyes of the present invention (the three middle images) with commercially available red dyes / colors (the left image showing the Bordeaux color, which is located outside the figure, and the right image showing the Barbados Cherry color). [Figure 8]Photographs showing the results of disk tests on the antimicrobial activity of the dyes of the present invention. [Figure 9] Photograph showing N. lichenicola IIa (DSM 34187, left) versus N. solani DSM 62805 (right). 4 days of growth on Sabouraud agar at 28°C. Both strains produced a reddish-brownish pigment, with isolate IIa showing a higher color intensity or higher amount of pigment. Both reached a diameter of approximately 4 cm. [Figure 10] Photograph showing N. lichenicola IIa (DSM 34187, left) versus N. solani DSM 62805 (right). Growth on Sabouraud agar at +4°C for 45 days. Aerial mycelium of isolate IIa shows increased pigmentation, but both released pigment into the agar layer. [Figure 11] Photograph showing N. lichenicola IIa (DSM 34187, left) versus N. solani DSM 62805 (right). 48 hours of growth in Sabouraud broth at 28° C. Both strains released a red pigment into the surrounding medium, but again, isolate IIa shows a darker reddish color. [Figure 12] Photograph showing N. lichenicola IIa (DSM 34187, left) versus N. solani DSM 62805 (right). 72 hours of growth in Sabouraud broth at 28° C. Both strains released a red pigment into the surrounding medium, but again, isolate IIa shows a darker reddish color. [Figure 13] Photographs showing phase contrast microscopy of N. lichenicola IIa (DSM34187). [Figure 14] Photographs showing phase contrast microscopy of N. solani DSM 62805. [Figure 15] 1 is a graph showing a chromatogram of molecular identification. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] Unless otherwise indicated or defined, all terms used herein have their usual meaning in the art, which is clear to those skilled in the art.See, for example, standard manuals such as Sambrook et al., "Molecular Cloning: A Laboratory Manual" (4th edition), vols. 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 specifically to man-made products, which may be variants of natural (wild-type) products, or to methods of using or producing said man-made products. Although there may be some sequence identity with natural structures, it is well understood that the materials, methods and uses of the invention, with particular reference to, for example, isolated nucleic acid sequences, amino acid sequences, expression constructs, transformed host cells and modified proteins and enzymes, are "man-made" or synthetic and therefore are not considered the result of the "laws of nature."

[0034] The terms "comprise," "containing," "having," and "include," as used herein, may be used synonymously and are to be understood as open definitions that allow for additional members or moieties or elements. "Consisting of" is considered the most closed definition without the additional elements characteristic of the definition of "consisting of." Thus, "comprising" is broader and includes the definition of "consisting of."

[0035] The term "about" as used herein refers to a value that is the same as the given value or a value that differs by + / - 5% thereof. As used in this specification and claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0036] According to the present invention, there is provided an isolated fungus belonging to the species Fusarium solani, said isolated fungus being applicable for producing dyes applicable for substrate coloration and also for direct coloration of substrates. The dyes currently used commercially for substrate coloration are chemically synthesized. Biological dyes and dyeing procedures have a lower impact on the environment.

[0037] The isolated fungus of the present invention is deposited under the number DSM 34187 and is also referred to herein as Fusarium solani IIa (FS IIa) or Neocosmospora lichenicola, both terms may be used interchangeably.

[0038] Fungi belonging to the species Fusarium solani are filamentous fungi in the phylum Ascomycota. Fusarium solani is a common soil fungus that belongs to the classification of Ascomycota. 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 known and are taxonomically, phylogenetically, and morphologically described (Kristensen et al., 2021). According to Short et al. (2013), the FSSC is a diverse complex of many phylogenetically distinct species. No obvious differences seem to occur in the morphology of the various Fusarium solani species (Schroers HJ et al. 2016; Chehri K et al., 2015; Matuo T and Snyder WC, 1972).

[0039] Surprisingly, the isolated Fusarium solani IIa of the present invention produces a red dye in high yield and in a short time, e.g. after overnight incubation. Using this dye or the isolated fungus, a dyeing method is possible, allowing for direct penetration of color and thus adsorption of the pigment / dye on textile fibers or material surfaces.

[0040] The term "color change" as used herein refers to a change in the color of a substrate, thus changing the visual appearance of the substrate. Colors can be described using color models. The RGB color model is one way to describe color. The RGB color model is an additive color model in which the red, green, and blue primary colors of light are added together in various ways to reproduce a wide range of colors. An alternative way to describe color is the CYMK model or the HSL color model. HSL stands for hue, saturation, and lightness.

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

[0042] The term "woven material", as used herein, refers to a material made by creating interlocking bundles of yarns or threads produced by spinning raw fibers into long and twisted lengths. The raw fibers may be of natural or synthetic origin. The woven material is formed by weaving, knitting, crocheting, knotting, tatting, felting, joining, or intertwining these yarns together. The terms "woven material", "woven fabric", and "fabric" may be used interchangeably herein.

[0043] According to certain embodiments, the substrate is a woven material selected from the group consisting of natural woven materials, synthetic woven materials, and combinations of natural and synthetic woven materials. The natural textile material may be, but is not limited to, cotton, silk, wool, abaca, coir, linen, hemp, wood, cashmere, and mohair, or any combination thereof.

[0044] In particular, the synthetic textile material may be, but is not limited to, polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal, or any combination thereof.

[0045] The terms "dye" and "pigment" refer to the red dye produced by the fungus of the present invention, as FeCl 3 is used herein to distinguish it from the dark pigment produced by adding water to a red dye, whereby the term "dye" is used for the water soluble red dye, and the term "pigment" is used for the dark pigment which is less water soluble.

[0046] According to one embodiment of the present invention, there is provided a method for changing the color of a substrate, said method comprising the steps of providing an inoculum of an isolated fungus of the present invention, inoculating a culture medium with the inoculum, optionally pre-incubating the inoculated culture medium and optionally inactivating the fungus after pre-incubation, contacting the substrate with the culture medium of one of the previous two steps until a desired color is obtained, and heating the substrate.

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

[0048] According to certain embodiments, said method of changing the color of a substrate comprises the steps of providing an inoculum of the isolated fungus of the present invention, inoculating a culture medium with the inoculum, pre-incubating the inoculated culture medium and optionally inactivating the fungus after pre-incubation, contacting a substrate with the pre-incubated culture medium until a desired color is obtained, and heating the substrate.

[0049] According to certain embodiments, the method of changing the color of a substrate comprises the steps of providing an inoculum of the isolated fungus of the present invention, inoculating a culture medium with the inoculum, pre-incubating the inoculated culture medium and inactivating the fungus after pre-incubation, contacting a substrate with the pre-incubated culture medium until a desired color is obtained, and heating the substrate.

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

[0051] The term "inoculating a culture medium," as used herein, refers to the transfer of an inoculum to a culture medium. According to certain embodiments, the isolated fungi of the invention are cultured by the various methods described herein. The terms "culture" and "pre-culture" are used herein for growth under conditions outside the fungus' natural environment.

[0052] According to certain embodiments, if the fungus is not inactivated before contacting the substrate with the culture medium, the fungus is also cultivated during the contacting step. In other words, if the substrate is contacted with a culture medium containing a viable fungus, the color of the substrate changes simultaneously with the growth of the fungus. Thus, the dye produced by the fungus is directly used to change the color of the substrate.

[0053] The term "contacting" as used herein refers to the step of contacting the substrate with the dye or the dye-producing fungus. An example of contacting is adding the substrate directly to the liquid culture medium in which the fungus is cultured or still present. Alternatively, if an isolated fungal dye is used to change the color of the substrate, contacting is performed by adding the substrate to a solution of the dye.

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

[0055] According to a particular embodiment, a step of heating the substrate is applied to fix the color. According to a particular embodiment, the heating of the substrate is carried out at a temperature in the range of 60°C to 121°C. In particular, a temperature of 121°C is used if the heating step is also used for the inactivation of fungi. In particular, the heating of the substrate is carried out at a temperature 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, 115°C to 121°C, or at a temperature of 121°C. In particular, the heating of the substrate is carried out for at least 20 minutes and up to several hours. In particular, the heating of the substrate is carried out for 20, 25, 30, 35, 40, 45, 50, 55, 60 minutes or even longer. In particular, the heating of the substrate is carried out for 20-25, 20-30, 20-40, 20-50, or 20-60 minutes. In particular, the heating may be carried out by any method used in the art for heating substrates. For example, the substrate may be heated in an oven or dryer.

[0056] According to one embodiment of the invention, a method for producing a fungal dye comprises the successive steps of providing an inoculum of an isolated fungus of the invention, inoculating a culture medium with the inoculum, culturing the inoculated culture medium, harvesting the biomass and / or the culture medium, and optionally extracting the dye from the harvested material.

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

[0058] According to certain embodiments, in the method of producing a fungal dye, the culture medium may be a solid medium or a liquid medium. Generally, solid culture media contain substances for solidification of the culture medium, such as agar, but may contain other components in the same concentrations as in the respective liquid media.

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

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

[0061] According to certain embodiments, the culture medium comprises peptone. According to certain embodiments, the culture medium comprises 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 which are buffers, salts, yeast extract, malt extract, 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 certain embodiments, the culture medium has a pH in the range of pH 5.0±0.2 to 6.5±0.2. In particular, the pH of the culture medium is pH 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 certain embodiments, said pH of the culture medium is maintained during pre-cultivation of the fungus, during culturing of the fungus, during contacting the substrate with the culture medium, including the inoculated culture medium, and / or during contacting the substrate with the culture medium, including the inoculated culture medium, optionally pre-cultivated, and optionally containing the inactivated fungus. The pH can be maintained in any known manner, for example by using a buffered culture medium and / or by adding an acid or a base. The pH may be controlled by a control unit of the bioreactor.

[0065] According to a particular embodiment, the pre-culture and cultivation of the fungus is carried out at a temperature in the range of 10°C to 35°C. In particular, the pre-culture and / or cultivation of the fungus is carried out 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.

[0066] According to a particular embodiment, the contacting of the substrate with the culture medium, including 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 a particular embodiment, the contacting of the substrate with the optionally pre-cultured inoculated culture medium and the culture medium containing the optionally inactivated fungus 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.

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

[0069] According to certain embodiments, harvesting can also be performed by removing the solid medium from its container, for example from a Petri dish, whereby the solid medium containing the fungus can be disrupted.

[0070] According to certain embodiments, the cultivation of the fungus, the pre-culturing of the fungus, the contacting of the substrate with the culture medium, including the inoculated culture medium, and / or the contacting of the substrate with the culture medium, including the optionally pre-cultivated inoculated culture medium and the optionally inactivated fungus, may be carried out as submerged fermentation in a bioreactor, which 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 particular embodiment, for the production of the dye, the fungus may be cultivated on a solid medium, for example by solid state fermentation, whereby the dye accumulates in the medium and / or in the mycelium of the fungus.

[0072] According to certain embodiments, in the method for producing the fungal dye, the culturing step is carried out at least until a red / purple color development is detected in the culture medium and / or in the biomass. The culturing is thereby 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 culturing step and on the form and concentration of the inoculum of the fungus used.

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

[0074] According to one embodiment, the colour of the dye produced by the fungus of the present invention is in the range of RGB 170±50, 10±5, 39±10 using the RGB colour model. According to one embodiment, the lightness (L) of the dye produced by the fungus ranges from 25-45% according to the HSL color model.

[0075] According to certain embodiments, Fusarium solani IIa produces a red pigment that darkens to blackish red, brown-red and burgundy red with prolonged growth. According to one embodiment of the present invention, a fungal dye may be used to change the color of a substrate. In particular, the color of the substrate may be changed by successive steps of contacting the substrate with a fungal dye and heating the substrate until a desired color is obtained. As described hereinbefore, a heating step is performed to fix the color, and is performed as described elsewhere herein. The resulting color of the textile depends on the duration of the contacting step and also on the concentration of the dye.

[0076] According to a particular embodiment, in the step of contacting the substrate with the fungal dye, the dye may be present in liquid or solid form. In particular, the substrate may be immersed in a solution containing the dye. More particularly, the substrate may be completely immersed in a solution containing the dye or may be partially immersed in said solution. Alternatively, the dye solution or the dye in powder form may be sprinkled on the substrate. The substrate may be pretreated or pre-moistened before application of the dye. The dye solution may be sprayed on the substrate. Various additives may be added to the dye solution, for example, binders and / or thickeners may be added to the dye solution for the formation of a paste.

[0077] According to another embodiment of the present invention, the dark pigment is FeCl 3 to the dye of the present invention. In particular, said dark pigment has a lightness (L) in the range of 0 to 20%. More particularly, said dark 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 dark pigment is reduced compared to the lightness (L) of the red fungal dye. According to another embodiment of the present invention, the color of a substrate previously stained by the fungi of the present invention or by the fungal dyes described herein can be improved by the addition of FeCl 3 to the dyed substrate, whereby the color of the substrate changes to a darker color. The resulting color of the further processed substrate may depend on the color intensity before the dyeing step and may also depend on the material of the substrate.

[0079] According to a particular embodiment, the color lightness (L) of the substrate is determined by FeCl 3 In other words, the color of the dyed substrate of the present invention is reduced compared to the color of the substrate before contact with FeCl. 3 Contact with FeCl results in a decrease in lightness (L) according to the HSL color model. 3Said reduction in lightness (L) compared to the L value before application of may be 10, 20, 30, 40, 50%, or even more.

[0080] According to certain embodiments, the color of the substrate is determined by treating the substrate with FeCl 3 and the substrate is capable of being modified by contact with FeCl 3 The fungus was stained with a fungus of the present invention or with a fungal dye prior to contact with the fungus.

[0081] According to a particular embodiment, the FeCl of the substrate pre-stained with the dye of the present invention is 3 Contact with results in the formation of a dark color. According to another embodiment, the color of the substrate may be changed by a method comprising the successive steps of contacting the substrate with a dark pigment of the present invention and heating the substrate until the desired color is obtained. Again, heating is performed to fix the color.

[0082] According to another embodiment of the present invention, the fungal dyes of the present invention have antimicrobial activity. The term "antimicrobial activity" refers to any active ingredient that can inhibit the growth of bacteria, prevent the formation of microbial colonies, and destroy microorganisms.

[0083] According to another embodiment, the substrate stained with the dyes of the present invention has antimicrobial activity, in particular the growth of microorganisms on said substrate is inhibited. According to a particular embodiment, the textile material dyed with the dyes of the present invention has 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 fungus according to item 1 for staining a substrate. 3. i. providing an inoculum of the isolated fungus described in item 1; ii. inoculating the culture medium with an inoculum; iii. optionally pre-incubating the inoculated culture medium and, optionally, inactivating the fungus after pre-incubation; iv. contacting the substrate with the culture medium of ii. or iii. until the desired color is obtained; and v. Heating the substrate A method for changing the color of a substrate comprising the successive steps of:

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

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

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

[0089] 9. Substrate: FeCl 3 9. A method for changing the color of a substrate, comprising the method of any one of items 3 to 8, further comprising contacting said substrate with 10. The method according to item 9, wherein the contact results in a decrease in lightness (L) according to the HSL color model.

[0090] 11. i. providing an inoculum of the isolated fungus described in item 1; ii. inoculating the culture medium with an inoculum; iii. culturing the inoculated culture medium; iv. harvesting the biomass and / or culture medium; and v. Optionally, extracting a dye from the harvested material of iv. 13. A method for producing a fungal dye comprising the successive steps of:

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

[0092] 14. The method according to any one of items 11 to 13, wherein the culturing 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. The method according to any one of items 11 to 14, wherein the culturing in 15.iii. is carried out under aerobic conditions.

[0093] The method according to any one of items 11 to 15, wherein the culturing in 16.iii. is carried out at least until development of a red / purple color is detected in the culture medium and / or in the 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. drying the harvested biomass and / or the harvested culture medium of the culture of the fungus described in item 1; 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. 13. A method for extracting a fungal dye comprising the steps of:

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

[0096] 20. i. contacting the substrate with the dye according to item 19 until the desired color is obtained; and ii. Heating the substrate A method for changing the color of a substrate comprising the successive steps of:

[0097] 21.FeCl 3 or FeCl 3 to the method of production according to any one of items 11 to 17.

[0098] 22. A dark pigment produced by the method according to item 21, wherein the pigment has a lightness (L) in the range of 0 to 20%. twenty three. i. contacting the substrate with the dye according to item 22 until the desired color is obtained; and ii. Heating the substrate A method for changing the color of a substrate comprising the successive steps of:

[0099] 24. The use according to item 2 or the method according to any one of items 3 to 10, item 20 and item 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 according to item 24, wherein the natural textile material is cotton, silk, wool, abaca, coir, linen, hemp, wood, cashmere, mohair. 26. The method according to item 24, wherein the synthetic textile material is polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, modal.

[0101] 27. The method according to any one of items 3 to 10, 20, and 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, item 20, and items 23 to 27, wherein heating of the substrate is carried out for at least 20 minutes.

[0102] 29. The dye according to item 19, wherein the dye has antimicrobial activity. 30. A substrate having antimicrobial activity, said substrate being stained with the dye according to item 19.

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

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

[0105] Example 2: Dye production Fusarium solani IIa is grown at 28°C on Sabouraud 4% glucose agar (Carl Roth, X932.2) as well as in Sabouraud 2% glucose broth (Carl Roth, AE23.1). This complex medium is used to form after overnight incubation (approximately 12-16 hours) a dark red-brown dye (see Figures 2A, 2B and 2C) that diffuses into the agar or surrounding medium and colors the mycelium red. In liquid medium, dye formation can take up to 48 hours.

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

[0107] 20 g l at a pH of 5.6 -1 Dextrose and 10g l -1 Cultivation in Sabouraud-dextrose broth containing peptone results in Bordeaux red-stained medium and Bordeaux red-stained, curled mycelium after 24 h 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: Dyes Extraction of the dye with ethyl acetate gives a red dye (see Figure 3, left flask). 3 Addition of 100 mM NaCl causes the formation of a dark, insoluble complex (see FIG. 3, right flask).

[0110] Example 4: Extraction of dyes The Bordeaux Red dye extraction was carried out according to the following protocol: After an incubation period of 10 days, the stained agar / medium containing the stained mycelium of FS II on the surface is reduced into small pieces and dried.

[0111] - Dried fragments are suspended in EtOH (+10% dH2O) and heated up to approximately 80° C. for 20 minutes. Alternatively, evaporation can be performed at room temperature over a period of 3 days. - Dissolve the residue in dH2O 2 The mixture is resuspended in 200 ml of ethyl acetate and extracted a first time in a separatory funnel. After evaporation of the ethyl acetate, a sticky, intensely red material remains. - This red substance is dH 2 It can be resuspended in 0, ethyl acetate, acetic acid, and ethanol.

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

[0113] Blank fabric samples are added either immediately at the start of the incubation or after sufficient pigment formation. Accordingly, the staining time frame ranges from 2 h to 72 h, resulting in different color intensities.

[0114] Blank fabric samples are incubated in a dye bath containing resuspended red dye. Coloration begins immediately and increases in intensity with time. Each dyeing is finished using a 30 minute heat treatment at 60-90°C, which inactivates the fungus (maximum 121°C) and fixes the dye into the material fibre.

[0115] Example 6: Dyeing The fungus is cultivated in a liquid complex medium. After pre-cultivation, the cells are inoculated into fresh liquid medium and grown in the presence of the fabric to be dyed. As soon as the color intensity is reached, the dyeing process is stopped. A subsequent heat treatment (60°C for 20 minutes or 90°C for 20 minutes) fixes the dye in the fabric.

[0116] Example 7: Dyeing The multi-fibers (MF) were incubated in a 72-h culture of FS IIa for 3 h at room temperature, followed by dye fixation for 25 min at 60° C. The results of the staining process are shown in column 1 of FIG.

[0117] Subsequent staining of the substrate with FeCl 3 *Overnight incubation in 6H2O (0.1M, 0.03g ml-1) reduces the amount of dye and FeCl 3 The complex formation causes a color change (see column 2 in FIG. 4).

[0118] Example 8: Dyeing The fungus is allowed to grow until the desired density and dye production is reached, then heat (121°C for 20 minutes) is used to inactivate the fungus. Only once the fungus is 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 The MF was incubated in a dye staining bath for 1.5 h, followed by heat treatment at 60 °C for 25 min. After cooling, FeCl 3 A second incubation in the solution resulted in a color change to grey / black for cotton, cellulose and silk, the results are shown in column 3 of FIG.

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

[0121] Example 11: Dyeing H 2The crude extract, partially soluble in O, is used for dyeing. Approximately 3 hours of incubation at room temperature already results in a dark red coloration of 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 staining procedure described, the red dye for FS IIa was prepared using FeCl 3 This results in a color change to a dark color, almost black for silk coloring.

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

[0124] Example 14: Antimicrobial Activity The antimicrobial activity was tested using a disk test, whereby the antimicrobial activity was tested against Escherichia coli, Staphylococcus aureus, Pseudomonas sp. and Bacillus subtilis. Fusarium solani not only exhibits antimicrobial activity against Staphylococcus aureus and Bacillus subtilis, but also to a lesser extent against E. coli. The disk test was carried out by incubating Staphylococcus aureus, E. coli and Bacillus subtilis on Mueller-Hinton agar at 37° C. for 18 hours in the presence of a disk prepared with 20 μL of an aqueous extract of the dye of the present invention. The results of the antimicrobial activity are shown in FIG. 8. The susceptibility of the selected microorganisms to the extract of Fusarium solani IIa was determined by measuring the diameter of the zone of inhibition (ZOI) in mm. 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 the identification of Fusarium species. The assembled DNA sequences were loaded in the GenBank, MycoID and Fusarium ID databases. Sequencing date: March 31, 2022 Sequencing by DSMZ, Braunschweig (GER) >ID 22-85 Tef (SEQ ID NO:1) Sequencing results:

[0127] [Table 2]

[0128] Identification: Neocosmospora ricenicola (C. Massal.) Sand.-Den.& Crous, [MB#822901] Neocosmospora richenicola belongs to the Fusarium solani species complex, FSSC. N. richenicola belongs to the FSSC group 16b.

[0129] Example 16: Forms of DSM34187 Strain DSM 62805 Neocosmospora solani (see References L. Lombard, NA van der Merwe, J. Z. Groenewald, and P. W. Crous Studies in Mycology 80:189-245) was used as a reference strain for isolate IIa. An initial investigation of potential morphological differences was performed by observation of the growth and aerial mycelium, but also by growth patterns in liquid culture. Both strains were cultured on Sabouraud agar (4% glucose) and Sabouraud broth (2% glucose) at 28 °C. See Figures 9-14.

[0130] Example 17: Molecular Identification The crude extract of the red pigment produced by isolate IIa was diluted to 0.5 mg ml -1 EtOH+25%H 2 2H and loaded onto an HPLC column (see chromatogram in FIG. 15). The chromatogram shows peak A at about 29 mAU at a retention time of about 1.45-1.50 min, peak B at about 21 mAU at a retention time of about 1.65 min, peak C at about 3 mAU at a retention time of about 1.70 min, and peak D at about 4 mAU at a retention time of about 1.85 min.

[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 members of 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 PG, Juba Jean & Geiser David M. (2016) Epitypification of Fusisporium (Fusarium) solani and its assignment to a common phylogenetic species. 10.3852 / 15-2 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):6 https: / / doi.org / 10.3390 / app11146255 Rathna Janarthanam et al., Production of naphthoquinones and phenolics by a novel isolate of 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

[0134]

Table 4

Claims

1. An isolated fungus belonging to the species Fusarium solani deposited under the number DSM 34187.

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

3. 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 and synthetic textile materials, in particular natural textile materials selected from the group consisting of cotton, silk, wool, abaca, coir, linen, hemp, wood, cashmere, and mohair, and synthetic textile materials selected from the group consisting of polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal.

4. i. providing an inoculum of the isolated fungus of claim 1; ii. inoculating the culture medium with an inoculum; iii. Optionally, pre-incubating the inoculated culture medium and, optionally, inactivating the fungus after pre-incubation; iv. Contacting the substrate with the culture medium of ii. or iii. until the desired color is obtained; and v. Heating the substrate, in particular heating the substrate at a temperature in the range of 60°C to 121°C, in particular for at least 20 minutes. A method for changing the color of a substrate, comprising the successive steps of:

5. The method according to claim 4, wherein the pre-cultivation in step iii) and / or the contacting in step iv) is carried out at a pH in the range of 4.8 to 6.7, particularly at a temperature in the range of 15°C to 35°C, particularly under aerobic conditions.

6. 5. The method of claim 4, 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, in particular natural textile materials selected from the group consisting of cotton, silk, wool, abaca, coir, linen, hemp, wood, cashmere, and mohair, and synthetic textile materials selected from the group consisting of polyester, rayon, acrylic, polycarbonate, polyethylene, spandex, acetate, lyocell, and modal.

7. The method according to claim 4, wherein the culture medium comprises a carbohydrate source, in particular glucose, preferably in the range of 1-4% (m / v).

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

9. The substrate was FeCl ３ 5. The method of claim 4, further comprising the step of contacting the color with a color component having a predetermined color component, the color component comprising: a first color component having a predetermined color component;

10. i. providing an inoculum of the isolated fungus of claim 1; ii. inoculating the culture medium with an inoculum; iii. Incubating the inoculated culture medium; iv. Harvesting the biomass and / or culture medium, and v. Optionally, extracting the dye from the harvested material of iv.

1. A method for producing a fungal dye, comprising the successive steps of:

11. The method according to claim 10, wherein the culture medium comprises a carbohydrate source, in particular glucose, preferably in the range of 1-4% (m / v).

12. 11. The method according to claim 10, wherein the culturing in step iii) is carried out at a pH in the range of 4.8 to 6.7, in particular at a temperature in the range of 15°C to 35°C, preferably in the range of 25°C to 28°C, in particular under aerobic conditions, in particular until the development of a red / purple color is detected in the culture medium and / or biomass.

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

14. FeCl to produce a dark pigment ３ 11. The method of claim 10, further comprising the step of adding a pigment having a lightness (L) in the range of 0-20%.

15. i. drying the harvested biomass and / or the harvested culture medium comprising the fungus of claim 1; 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.

1. A method for extracting a fungal dye, comprising the successive steps of:

Images