The present embodiment relates to a process for producing a foam-like grown material, and more particularly to a process for producing a foam-like packaging or multipurpose grown material, from assisted growth of multiple fungal species on flowers, agricultural residue, spawn etc and/or their extracts/parts.
SUMMARY OF THE PRIOR ART
In the present day, a range of material, for a range of purposes is being grown or manufactured today using several starting materials. The most prominent materials especially for more prominent purposes are usually manufactured using several chemicals, minerals, ores, fossils etc. The materials have also been prepared using trees, plants and animals for several purposes and such materials are called grown materials. The production of many materials and composites produces significant hazardous environmental effects in form of pollution that is caused by their production and processing, as well as their non-biodegradability or recalcitrance.
The conventional materials such as petroleum-based foams are not biodegradable and require significant energy for their production. The grown materials such as trees, crops, and plants require sunlight, fertilizers and large farmable land, and are time intensive. Both of these strategies of producing material lead to a significant ‘waste stream or waste streams’ that can be both aqueous or non-aqueous in nature.
Fungi are one of the faster growing organisms having a cell wall getting extended at the tips of the hyphae. While the cell wall of a plant is made up of cellulose, the structural components of animal cells mainly include collagen, whereas fungal cell walls largely comprise Chitin, which is a hard and strong fibrous substance. A few prior art references disclose use of fungal species to grow material based on Chitin; however, the processes make use of defined media or substrates.
Simultaneously, there is a problem pertaining to flower waste. Flowers are universally used for several such occasions such as marriage, celebrations, funerals, religious rites etc. However, the problem pertaining to their waste has not been addressed extensively apart from their use as biofertilizer or plant growth materials.
As mentioned, there remains a need for producing foam-like materials from natural substrates.
SUMMARY OF THE INVENTION
In view of the foregoing, a process for obtaining foam-like grown material from flowers is provided.
In an aspect, a process for preparing a grown foam-like material is provided. The process includes inoculating grain spawn, a mixture of grains and CaCO3 with a culture consisting a plurality of fungal species to obtain colonized grain spawns, in step (104); inoculating a flower substrate with colonized grain spawns, in step (106) to obtain a myceliated mixture; incubating the myceliated mixture inside moulds of required shape or size to obtain a moulded myceliated material in step (108); baking the moulded myceliated material to reduce moisture and kill microbes to obtain harvested moulded myceliated material (110); and drying the harvested moulded myceliated material to obtain the grown foam-like material.
The process further includes a method of preparation of the flower substrate; wherein the method of preparation comprises soaking sawdust and lignocellulosic additives in water and mixing with flower or flower petals and CaCO3. The grain spawns are prepared by boiling grains in water and mixing CaCO3. The flower substrate has 40-80% w/w flower or flower petals, 1-3% w/w CaCO3, 10-40% w/w straw, 5-20% w/w sawdust and 5-20% w/w lignocellulosic material/additives. The flower substrate includes flower or flower petals that are fresh or dried, and segregated or non-segregated. The grain spawns and the flower substrates are sealed inside a bag or jar or bottle after inoculation. The flower substrate is inoculated with 5-15% w/w of the colonized grain spawns. The process further includes crushing myceliated material obtained after inoculation of flower substrate to break clumps.
The process further includes adding wheat flour, corn flour, any grain flour, guar gum, starch or barley to the myceliated mixture from the flower substrate before incubating the myceliated mixture inside moulds of required shape or size to obtain a moulded myceliated material in step (108). The myceliated mixture is filled inside the moulds under pressure.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
The embodiments herein and the various features and advantageous details thereof are explained more fully with reference to the non-limiting embodiments that are illustrated in the accompanying drawings and detailed in the following description. Descriptions of well-known components and processing techniques are omitted so as to not unnecessarily obscure the embodiments herein. The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the examples should not be construed as limiting the scope of the embodiments herein.
The source and geographical origin of the biological resources used herein is India.
As mentioned, there remains a need for sustainable processes using natural substrates/resources to manufacture materials for multiple purposes. The embodiment herein provides a process of producing a foam-like material or a composite foam-like material by growing one or more fungal species on a flower substrate or flower-based substrate or media. The process includes growing fungi by first inoculating a grain spawn with a fungal colony and then using inoculated grain spawn to inoculate a flower substrate or a flower-based substrate to obtain a myceliated material to which additives such as wheat flour and/or corn flour may be added. The myceliated material obtained, after mixing with additives, is filled inside a polycarbonate mould of a desired shape and the moulds are then incubated for a period of a few days so that the material attains the shape of the mould.
Figure 1 illustrates a process (100) for obtaining a grown foam-like material, according to an embodiment herein. The process includes preparing agar culture plates, preparing of grain spawn, preparing of flower substrate, moulding the grown material by primary and secondary growth, and harvesting and drying the material obtained. More specifically, the process (100) includes inoculating a mixture of grains and CaCO3 with a culture consisting of a number of fungal species to obtain colonized grain spawns, in step (104); inoculating a flower substrate with colonized grain spawns, in step (106) to obtain a myceliated mixture; incubating the myceliated mixture inside moulds of required shape or size to obtain a moulded mycelium in step (108); baking the moulded mycelium to reduce moisture and kill microbes to obtain harvested moulded mycelium in step (110); and drying the harvested moulded mycelium to obtain the grown foam-like material.
In step (102), the agar culture plates are formed/prepared to obtain fungal colonies. In a preferred embodiment, Potato Dextrose Agar (PDA) solution is used to prepare agar plates, which are then inoculated with a fungal colony or an existing colonised agar, and the freshly prepared agar plates are thus incubated. In an embodiment, a Malt Extract Agar (MEA) solution is used to prepare agar plates. In another embodiment, a Czapek Dox Agar or Wort Agar solution is used to prepare agar plates. The agar plates are sterilized and cooled before they are inoculated with an existing culture of desired fungal strains or species. The inoculated agar plates are then incubated for complete colonization of mycelium.
In the optional step (104), the grain spawn is prepared by boiling the grains in water and mixing them with CaCO3 (Calcium Carbonate) and inoculating the mixture with a number of fungal species or fungal cultures having a number of fungal species or desired fungal species. In a preferred embodiment, wheat grains are boiled. The boiling grains may be continuously stirred before mixing CaCO3. In an embodiment, Ca(OH)2, CaSO4, Lime or Gypsum are mixed with the boiling grains to obtain grain spawn. In an embodiment, the amount of CaCO3 to be added to boiling grains ranges from 1-3% w/w.
In an embodiment, the fungal species/strains are selected from Pleurotus ostreatus (Oyster), Trametes versicolor (Turkey Tail), Hericium erinaceus (Lions Mane), Lentinula edodes (Shiitake), Grifola frondosa (Maitake), Ganoderma lucidum (Reishi).
The grain spawn is then filled inside bags, preferably PPE bags, and then inoculated with the colonized agar plates. In an embodiment, the agar plates are colonized with mushroom or fungal strains/species. In a preferred embodiment, mushrooms are medicinal mushrooms. In an embodiment, PPE bags are replaced with plastic bottles or glass jars to contain the grain spawn. In a preferred embodiment, the grain spawn bags, jar or bottles are sterilized before inoculation. After inoculation, the inoculated grain spawn bags are sealed and incubated. During incubation, the bags may be given horizontal and vertical strokes to uniformly spread the inoculum across the grain spawn. The prepared grain spawn may be frozen for later use.
The step (104) is optional since the floral substrate may directly be inoculated from fungal strains/species colonies obtained from PDA, of step (102). However, the step (104) may be preferred.
In step (106), a flower substrate is prepared. In a preferred embodiment, the flower substrate includes 40-80% w/w flower petals, 1-3% w/w CaCO3, 10-40% w/w straw, 5-20% w/w sawdust and 5-20% w/w other lignocellulosic material/additives. In an embodiment, any flower waste is used. In a preferred embodiment, dried or fresh flower petals which may be segregated or non-segregated. In an embodiment, the whole flower is used. The sawdust and lignocellulosic additives are soaked in water. They may be soaked for 3-5 hours, preferably 3-4 hours. Thereafter, flower or flower petals and CaCO3 are mixed with the soaked ingredients and the mixture is transferred to PPE filter bags. The filled in PPE bags may be sterilized.
In an embodiment, Ca(OH)2, CaSO4, Lime or Gypsum are mixed with soaked ingredients and flower/flower petals instead of CaCO3. Similarly, the PPE bags may be replaced with glass/PPE bottles/jars.
In a preferred embodiment, the sawdust is of any long leaf tree. In another preferred embodiment, wood chips, shavings and bark pieces are used in lieu or after mixing with sawdust.
In an embodiment, the straw or lignocellulosic alternatives include agricultural residues such as grain straw, rice husk, corn stalk, hemp, kenaf, coconut coir, sugarcane bagasse.
The colonized grain spawn is then used to inoculate the flower substrate bags and are allowed to incubate at room temperature of a fixed duration. The bags are incubated until a uniform growth of mycelium is obtained in the bags.
The PPE bags containing the flower substrate having 40-80% w/w flower petals, 1-3% w/w CaCO3, 10-40% w/w straw, 5-20% w/w sawdust and 5-20% w/w other lignocellulosic material/additives is inoculated with the colonized grains spawns as described earlier. In an embodiment, 5-15% w/w of the grain spawns are added to the PPE bags having flower substrate. The PPE bags or glass/plastic bottles/jars are sealed and incubated at room temperature. In an embodiment, they are incubated for 7-15 days. The PPE bags or glass/plastic bottles/jars may be shaken horizontally and vertically to obtain uniform growth of mycelium.
In step (108), moulding by primary and secondary growth of the mycelium obtained in step (106) is carried out. The myceliated material from flower substrate bags is crushed to break clumps. The wheat flour and corn flour are then added as additives. In an embodiment, 1%-5% w/w wheat flour and 1%-5% w/w corn flour is added. In an embodiment, the wheat flour or corn flour are replaced by or are added with any high nutritional support additive such as any grain flour, guar gum, starch, barley. Alternatively, more flower substrate mixture may also be added to increase the substrate amount to fill in the moulds.
This mixture is then filled inside polycarbonate moulds of various shapes and sizes as desired. In an embodiment, the moulds are sterilized. The moulds may be sterilized by cleaning the mould with 5% acetic acid solution followed by treatment with 70% ethanol solution. In practice, any established method of sterilizing surfaces may be used. The myceliated material from the flower substrate bags is then filled inside the sterilized moulds under pressure. In an embodiment, the pressure is 1-5 psi (0.069 bar to 0.35 bar approximately). The filled-up moulds are then incubated for a fixed duration, preferably 3-5 days, until the material is colonized on a primary level and attains shape of the mould.
The moulded mycelium obtained after incubation of the moulds is then pushed out from moulds using pressure and is incubated inside a humid chamber. In a preferred embodiment, the relative humidity of the chamber is 70-90%, and the incubation period is 3-5 days.
The material so grown, in step (108), or obtained after incubation under humid conditions is then harvested and baked, in step (110), so as to kill microbes and to remove moisture. The dried product so obtained is then used for multiple purposes and specifically for packaging purposes. The grown material is harvested and baked at, preferably, 1000C to 1100C. A hot air dryer may be used. Alternatively, sun drying is used. The harvesting step removed 60-70% moisture content of the grown material obtained from step (108). The foam-like material is then obtained after drying.
EXAMPLES:
Example 1: Preparation of agar culture plates
Potato Dextrose Agar (PDA) or Malt Extract Agar (MEA) solution was prepared by addition of appropriate amounts of nutrient in 100 ml water and was then sterilized at 1210C in an autoclave. The agar solution was then poured in 90 mm diameter petri-plates and was allowed to cool inside the Laminar Flow Hood for 40-45 minutes, which solidified the solution to form gel in the plate. A colonized agar piece of size 5mm x 5mm obtained from an old culture plate was introduced inside the freshly prepared plate as inoculum. The inoculated plates were incubated at 27 °C for 8 days in BOD (Biological oxygen Demand) incubator for complete colonization of mycelium. The fully colonized plates were stored at 4°C or -20°C for future use.
Example 2: Preparation of grain spawn
The grain spawns were prepared by boiling the wheat grains in water for approximately 20 minutes to 2 hrs to achieve an adequate amount of moisture in the grains. The mixture was continuously stirred to assure the homogeneous moisture in the grains. The boiled grains were then mixed with 1-3 % w/w CaCO3 (Calcium carbonate) and transferred to PPE filter bags. The bags were then sterilized at 121 0C and 15 psi pressure for more than 30 minutes in an autoclave. The autoclaved grains were allowed to cool to room temperature inside a laminar flow hood. Thereafter, fungal culture from well colonized agar plate that was prepared was added to the grain bag and sealed. The sealed bags were then incubated at room temperature for more than 2 weeks to allow the growth of fungal culture. The prepared grain spawn was stored at 4 °C for later use.
Example 3: Preparation of grain spawn
In another method, the wheat grains were washed in tap water and soaked overnight After soaking, the excess water was drained out from the grains. The water soaked grains were then mixed with appropriate amount of CaCO3 (Calcium carbonate) and transferred to PPE bags. Thereafter, 40-80% w/w water was added to grain mixture. The bags prepared were then sterilized at 1210C and 15 psi pressure for 60-90 minutes in an autoclave. The autoclaved grains were allowed to cool inside a laminar flow hood. Thereafter, approximately 30-40% of well colonized agar plate that was prepared was added to the grain bag and sealed. The sealed bags were then incubated at room temperature for around 2 weeks. The prepared grain spawn was stored at 4°C for later use.
Example 4: Preparation of flower substrate
A dry substrate containing flower petals, CaCO3, straw, sawdust, agricultural residues and other lignocellulosic materials was prepared. The dried mix of flower petals and other constituent were added with appropriate amounts of limestone powder (2-4% preferable) and water to achieve the 60-80%RH moisture level and transferred to PPE filter bags, which are then heat sterilized at 121°C and 15 psi pressure for more than 30 minutes in an autoclave. The autoclaved bags were allowed to cool inside a laminar flow hood. The substrate bags were then inoculated using any well grown fungal culture source (preferably fresh growing flower substrates bags) and sealed. The sealed bags were incubated at room temperature for 15 days. .
Example 5: Moulding, primary and secondary growth of material
The myceliated material from fully colonized substrate bags was thoroughly crushed to break all the clumps. Thereafter, high nutritional additives were added uniformly to the myceliated material. The polycarbonate molds of various shapes as per the requirement were sterilized. The well mixed myceliated material was filled inside the sterilized polycarbonate mold under a pressure of 1-5 psi. The filled-up molds were incubated at room temperature for few days. In this period, the material got colonized on a primary level and attained the shape of the mold. The colonized material was then pushed out from moulds using pressure and incubated inside a humidity chamber having 70-90% RH for few more days.
Example 6: Harvesting and drying
The grown colonized material was harvested and baked at higher temp (preferebly above 40°C) in a hot air dryer for hours to heat kill the microorganism present and also to remove the 60-70% moisture content present in it. The foam-like grown material was then obtained after drying.
As will be readily apparent to those skilled in the art, the present embodiment may easily be produced in other specific forms without departing from its essential characteristics. The present embodiment are, therefore, to be considered as merely illustrative and not restrictive, the scope being indicated by the claims rather than the foregoing description, and all changes which come within therefore intended to be embraced therein.

We claim:
1. A process for preparing a grown foam-like composite material using floral or agricultural substrates comprises:
inoculating a flower substrate with a plurality of fungal species/strains, in step (106) to obtain a myceliated mixture;
incubating the myceliated mixture inside moulds of required shape or size to obtain a moulded myceliated material in step (108);
baking the moulded myceliated material to reduce moisture and kill microbes to obtain harvested moulded myceliated material in step (110); and
drying the harvested moulded myceliated material to obtain the grown foam-like material.
2. The process as claimed in claim further comprising inoculating grain spawn, a mixture of grains and CaCO3 with a culture consisting a plurality of fungal species to obtain colonized grain spawns, in step (104), wherein the step (104) is performed before inoculating the flower substrate.
3. The process as claimed in claim 1 further comprising a method of preparation of the flower substrate; wherein the method of preparation comprises soaking sawdust and lignocellulosic additives in water and mixing with flower or flower petals and CaCO3.
4. The process as claimed in claim 1, wherein the grain spawns are prepared by boiling grains in water and mixing CaCO3.
5. The process as claimed in claim 2, wherein the flower substrate has 40-80% w/w flower or flower petals, 1-3% w/w CaCO3, 10-40% w/w straw, 5-20% w/w sawdust and 5-20% w/w lignocellulosic material/additives.
6. The process as claimed in claim 4, wherein the flower substrate includes flower or flower petals that are fresh or dried, and segregated or unsegregated.
7. The process as claimed in claim 1, wherein the grain spawns and the flower substrates are sealed inside a bag or jar or bottle after inoculation.
8. The process as claimed in claim 1, wherein the flower substrate is inoculated with 5-15% w/w of the colonized grain spawns.
9. The process as claimed in claim 1 further comprising adding wheat flour, corn flour, any grain flour, guar gum, starch or barley to the myceliated mixture from the flower substrate before incubating the myceliated mixture inside moulds of required shape or size to obtain a moulded myceliated material in step (108).
10. The process as claimed in claim 1, wherein the myceliated mixture is filled inside the moulds under pressure.