DESC:FIELD OF THE INVENTION:
This invention relates to the field of textile engineering.

Particularly, this invention relates to processing of natural fibres obtained from plant sources.

Specifically, this invention relates to an eco-friendly bioprocess for separation of hemp plant fibres for various applications.

BACK GROUND OF THE INVENTION:
Hemp plant (Cannabis sativa) has been used in India for thousands of years. The Vedas, estimated to be at least 3400 years old, refer to it as one of the five most sacred plants in India. Hemp finds wide applications in the field of textile, medicines, steel industry, and as an alternative for harmful plastics.

All over the world, cotton is a preferred natural fibre and India is no exception. While cotton takes about 9 months to grow, hemp grows to 12 feet in about 3 to 4 months. Advantages of hemp cultivation include the fact that hemp consumes less water and could be a boon to non-irrigated land portions of India. Besides, it is good at suppressing weeds and also can be grown for multiple cycles in a given portion of land. Along with all these advantages, hemp fibres are being used to make clothes or textiles, as building material, to make shoes, to make paper, and to make ropes or cords. Hemp fibres have been considered as a sustainable biomaterial to replace man-made fibres in composite applications, due to their low cost and density, good mechanical properties and biodegradability. Based on environmental concerns, interest in using cellulosic fibres has increased in recent years. Applications of plant fibres, based on hemp, have potential as suitable bio-composites for fabrics and textiles industries.

To make raw hemp fibres for textile material, hemp fibres are separated by ‘retting’, which is a process of decaying pectin that binds the hemp fibres to a core of its stem. By retting, the long fibre, fibres are separated from non-fibres parts of stalks. This natural retting process takes several days to months based on climatic conditions.

PRIOR ART AND THE DRAWBACK ASSOCIATED WITH KNOWN ART:
Reference is made to US patent US8591701 dated Nov. 26, 2013; this document teaches a method of extracting hemp fibres from decorticated hemp fibre skin which involves pre-treating decorticated hemp fibre skin with an aqueous solution containing di-sodium citrate, tri-sodium citrate, or a mixture thereof, having a pH of from about 6-13 at temperature of about 90° C. or less; and, subsequently, treating recovered fibre with an enzyme. Determining the extent of completion of a plant fibre degumming process involves treating degummed fibre with a recombinant pectinase expressed in an organism that produces neither cellulose nor xylanase, to release reducing sugar from any residual pectin on the degummed fibre, and, quantifying the released reducing sugar. One of the disadvantages, of this method, is requiring pretreatment with costly chemicals and use of costly chemical such as enzyme. Harsh treatment using strong alkaline solutions like caustic soda (concentrated sodium hydroxide) is also used in this prior art process. This prior art treatment requires high temperature which is energy intensive.

Reference is made to non-patent literature ‘Hemp fibres: Enzymatic effect of microbial processing on fibre bundle structure’, authored by Thygesen, Anders and Lui, Ming and Meyer, Anne S and Daniel, Geoffrey (2013) In: Risoe-International-Symposium-on-Materials- Science-Proceedings, 2 Sept - 5 Sept 2013, Roskilde, Denmark. It was observed that, in this study, basidiomycete white-rot fungi Ceriporiopsis subvermispora and mutant strain Phlebia radiata Cel 2 were used. Disadvantages of these research enzymes are obtaining these fungi, growing, and using for degradation of pectin, hemicellulose, and lignin polymers of the fibres. This process is time consuming and is not commercially viable.

Reference is made to patent application WO2017200379 titled, methods for producing a manure-derived bioplastic and bioproducts’ which teaches separation of manure in fractions and the treatment of said fractions in order to retrieve useful components to produce manure-derived bioplastic and other manure-derived bioproducts. Here, according to this prior art, animal waste was used to produce bioplastics. This process used animal waste for production of bioplastics. This is not very much related to the current invention but it applies and uses natural bio-resources for production of industrial valuable bioplastics.

Reference is made to non-patent literature ‘Enzymatic Bioprocessing – New Tool of Extensive Natural Fibre Source Utilization’ authored by Marek Jan, Antonov Viktor, Bjelkova Marie, Smirous Prokop, Fischer Holger, Janosik Stefan. 2008 International Conference on Flax and Other Fibre Plants. This prior art teaches enzymatic retting of flax, enzymatic cottonization of fibre, enzymatic hemp separation, enzymatic processing of flax rovings before wet spinning etc. This state reviews of commercially available enzymes for fibre extraction. Disadvantage of this process is that it is expensive and time consuming.

Reference is made to non-patent literature ‘Ecofriendly Innovative Processing of Hemp Hurds Fibres for Tissue and Towel paper’ authored by Ved Naithani, Preeti Tyagi, Hasan Jameel, Lucian A. Lucia, and Lokendra Pal. This prior art teaches a method for making tissue paper from hemp hurd fibres by three pulping processes: autohydrolysis (hydrothermal), sodium carbonate-based defibration, and high yield kraft pulping, and benchmarked against hardwood pulp. To meet industrial standards, hardwood and hemp pulp fibres were mixed at a dry mass ratio of 75:25, from which tissue paper sheets were prepared. This process does not have many advantages, for separation of fibres, and suffers with use of costly processing such as hydrothermal process, and using chemicals makes this process costly and non-ecofriendly.

Reference is made to non-patent literature ‘Optimizing Hemp Fibre Production for High Performance Composite Applications’ by authors, Salvatore Musio, Jörg Müssig and Stefano Amaducci, where this prior art teaches that fibres were treated with a solution of sodium hydroxide (8 wt %) during 15 h at room temperature using a weight ratio water / biomass of 15/1 and then filter-pressed (6.8 atm =100 psi) to remove excess water. The 100 g of impregnated hemp fibres were loaded into a SE reactor and heated to 190 °C for 4 min. The sample was then exploded by sudden-drop in pressure and released in the discharge tank. The framework of the steam explosion facility has been described in a previous publication. The exploded hemp fibres (EF) were separated from the liquid fraction by vacuum filtration and dried at 40 °C for 24 h. This prior art process suffers with disadvantages as it uses costly chemicals and is energy intensive; thus, making it economically unsuitable to be used for ecofriendly hemp fibres’ separation.

Reference is made to US patent number 10415155 dated Jul 31, 2015 wherein it teaches a production method of hemp fibre for spinning, the method including: an immersion treatment process of immersing raw hemp fibre in a treatment liquid including an alkali, water, and at least one type of enzyme selected from the group consisting of cellulose-degrading enzymes and glycosidic bond hydrolyzing enzymes for an immersion time of from 30 minutes to 60 minutes under conditions of a temperature of from 60° C. to 100° C.; a water-washing process of washing the immersion treated hemp fibre with water; and a drying process of drying the water-washed hemp fibre. Disadvantages, of this method, is using cellulose degrading enzymes and glycosidic bond hydrolysing enzymes which may be costly and they degrade the cellulose which is an essential part of textile fibres. Also, they require high energy for fibre treatment.

Reference is made to non-patent literature ‘Enzymatic Bioprocessing - New Tool of Extensive Natural Fibre Source Utilization’ authored by Jan, Marek & Antonov, Viktor & Bjelková, Marie & Prokop, Smirous & Fischer, Holger & Stefan, Janosik. (2008). In this prior art, they reported that new enzymes are going to have great potential in bast fibre processing and modification for different end uses. There are several new technologies using enzymes which are able to modify fibre parameters, which achieve desired properties, and which improve processing results and ecology in the area of bast fibre processing and fabric finishing. Enzymatic retting of flax, enzymatic cottonization of bast fibre, enzymatic hemp separation, enzymatic processing of flax rovings before wet spinning etc. create a group of new technologies supported by effective mechanical treatments. A complex solution of relations between agrar and processing sphere and the utilization of new technologies of biochemical and mechanical processing enable the creation of suitable conditions to utilize traditional and perspective renewable fibre sources and to open them to ways to new qualities and to other industrial areas of use. This process suffers with disadvantages as it uses costly chemicals such pure enzymes and energy intensive thus make economical unsuitable to be used for ecofriendly hemp fibres separation. This process described is for flex fibres separation using enzymes.

Therefore, there is a need for a process which obviates all these prior art limitations.

OBJECTS OF THE INVENTION:
An object of the invention is to provide an ecofriendly bioprocess for separation of hemp plant fibres.

Another object of the invention is to provide a bioprocess, for separation of hemp plant fibres, which is eco-friendly, economical, and uses naturally available microorganisms and enzymes.

SUMMARY OF THE INVENTION:
According to this invention, there is provided an eco-friendly bioprocess for separation of hemp plant fibres for various applications.

According to one aspect of the invention, there is provided a method of separating and removing a bundle of fibres from decorticated raw hemp serving as control with water having pH 5-8 at room temperature.

In accordance with another aspect of the invention, there is provided a method for determining extent of completion of a plant fibre separation process comprising, the process comprising: treating raw fibre with biomaterial obtained from ruminant animal waste having pH 5-6 at room temperature.

In accordance with yet another aspect of the invention, there is provided a method for softening of a natural hemp fibre, said process comprising: providing wet processed fibre/s in a container that contains biomaterial obtained from ruminant animal waste and biomaterial obtained mushroom fungus. During this bioprocessing, fungus produces several enzymes which are responsible for degradation of pectin, hemicellulose, and lignin. Cellulose is most important component of fibres for industrial use which is, to some extent, protected during fermentation.

In accordance with yet another aspect of the invention, there is provided a method for separation and softening of natural hemp fibre, the method comprising treating the fibre with previously developed inoculum of consortium of microorganism and with biomaterial obtained from ruminant animal waste and mushroom fungus.

In at least an embodiment, this invention discloses a method for separation hemp fibres from decorticated hemp fibres skin, the method comprising:
- pre-treating decorticated hemp fibres’ skin with an aqueous solution containing biomaterial obtained from a bio-resource material, obtained from ruminant animal waste, keeping for natural fermentation, in order to separate its fibres, for 3-10 days;
- subsequently, washing the separated fibres washed with water; and
- treating the washed and separated fibres with one or more cleaning agents for more cleaner fibres.

In at least an embodiment, the pH of aqueous solution is 1.

In at least an embodiment, the temperature is in a range of 27-35o C.

In at least an embodiment, the decorticated hemp fibres’ skin is obtained by natural fermentation carried from 1day to 10 days.

In at least an embodiment, the biomaterial obtained from ruminant animal waste to treat fibres is used in the range of 500g/l to 1000g/l of aqueous solution which is mixed thoroughly with natural fibres to degrade pectin, hemicellulose, lignin and preventing cellulose from degradation.

In at least an embodiment, the combination of biomaterial (biomaterial obtained from ruminant animal waste as mentioned above) and mushroom fungus, as such, or extract, is used for easy separation of natural fibres.

In at least an embodiment, the decorticated hemp fibre skin is naturally fermented with biomaterial obtained from ruminant animal waste and combination of mushroom which helps in separation of fibres.

In at least an embodiment, the process and method, of this invention, has expected capability to produce combination of pectinase, hemicellulose, polyphenol oxidase / laccase and cellulase enzymes helping in fibres’ separation.

In at least an embodiment, a combination of biomaterial obtained from ruminant animal waste and mushroom (preferably, oyster mushroom), and previously grown inoculum, have been used for easy separation of natural fibres.

In at least an embodiment, the decorticated hemp fibres’ skin is naturally fermented with biomaterial obtained from ruminant animal waste and combination of mushroom and inoculum which help in separation of fibres and making smoothness of fibres.

In at least an embodiment, a combination of an aqueous solution containing 1-10 g carbon source , 0.5- 10 g nitrogen source, hemp fibres, 250 g to 300 g biomaterial obtained from ruminant animal waste, 1-50 g fungal mushroom are mixed together and kept for fermentation for 40-120 hrs. at ambient temperature.

In at least an embodiment, a combination of an aqueous solution containing hemp fibres, 250 g to 300 g biomaterial obtained from ruminant animal waste, 1-50g fungal mushroom and 7 to 10 g inoculum, 1-10 g carbon source 0.5- 10 g nitrogen source are mixed together and further kept for fermentation for 40-120 hrs. at ambient temperature.

In at least an embodiment, this invention discloses a eco-friendly bioprocess for separation of hemp plant fibres, from decorticated hemp fibres skin, said process comprising:
- pre-treating the decorticated hemp fibres skin with an aqueous solution, said aqueous solution bearing a pH in the range of 5.0 to 9.0, wherein said aqueous solution consisting of:
o 2.0 gm to 5.0 gm of a carbon source per 1 kg of hemp fibre;
o 1.0 gm to 5.0 gm of a nitrogen source per 1 kg of hemp fibre;
o biomaterial obtained from a bio-resource material;
- storing, for natural fermentation, at ambient temperature, in order to separate its fibres, for a time period ranging from 3.0 to 10 days;
- subsequently, washing the separated fibres with water having a pH in the range of 5.0 to 8.0; and
- treating the washed and separated fibres with one or more cleaning agent for more cleaner fibres.

In at least an embodiment, said ambient temperature is 20 to 37 deg. C

In at least an embodiment, said biomaterial comprising ruminant animal waste along with a combination of mushroom and inoculum, wherein the proportion of ruminant animal waste to mushroom to inoculum is 30 parts:2 parts: 2 parts.

In at least an embodiment, the pH of said aqueous solution is in a range of 5.0-9.0.

In at least an embodiment, said ambient temperature is in a range of 27 to 35o C.

In at least an embodiment, said decorticated hemp fibres skin is obtained by natural fermentation carried from 1.0 day to 10 days.

In at least an embodiment, said process comprising a step of bioprocessing by providing wet said cleaner fibres in a container that contains biomaterial obtained from ruminant animal waste and with biomaterial obtained from mushroom fungus, said fungus comprising enzyme/s which are responsible for degradation of pectin, hemicellulose, and lignin.

In at least an embodiment, said process comprising a step of bleaching said separated cleaned fibres.

In at least an embodiment, said process being performed in an environment having 70%-80% moisture.

In at least an embodiment, said separated cleaned fibres exhibit reduction of pectin in the region of 6%-20%, reduction of lignin in the region of 10%-20%, and reduction of cellulose in the region of 50%-70%.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS:
The invention will now be described in relation to the accompanying drawings, in which:

FIGURE 1a shows untreated control hemp fibres. FIGURE 1b shows treated hemp fibres;
FIGURE 2 illustrates hemp fibres that were obtained via a first method of separation;
FIGURE 3 illustrates hemp fibres that were obtained via a second method of separation;
FIGURE 4 illustrates hemp fibres that were obtained via a third method of separation;
FIGURE 5 illustrates control carried out using water;
FIGURE 6a illustrates fibres control and FIGURE 6b illustrates output of a method where fibre control, of FIGURE 6a, was treated with bio-resource material;
FIGURE 7 illustrates fibres which were processed by fungal growth and also using bio-resource material; and
FIGURE 8 illustrates fibres which were processed by fungal growth and also using bio-resource material and inoculums.

DETAILED DESCRIPTION:
According to this invention, there is provided an eco-friendly bioprocess for separation of hemp plant fibres for various applications.
In one embodiment, there is provided an eco-friendly bioprocess for separation of hemp plant fibres, from decorticated hemp fibres skin, for various applications, said process comprising the following steps:
- pre-treating the decorticated hemp fibres skin with an aqueous solution, said aqueous solution bearing a pH in the range of 5.0 to 9.0, wherein said aqueous solution consisting of: 2.0 gm to 5.0 gm of a carbon source per 1 kg of hemp fibre; 1.0 gm to 5.0 gm of a nitrogen source per 1 kg of hemp fibre; and a biomaterial obtained from a bio-resource material;
- storing, for natural fermentation, at ambient temperature, in order to separate its fibres, for a time period ranging from 3.0 to 10 days;
- subsequently, washing the separated fibres with water having a pH in the range of 5.0 to 8.0; and
- treating the washed and separated fibres with one or more cleaning agent for more cleaner fibres.

In one embodiment, the biomaterial comprises a ruminant animal waste along with a combination of mushroom and inoculum, wherein the proportion of ruminant animal waste to mushroom to inoculum is 30 parts:2 parts: 2 parts.
In one embodiment, the pH of said aqueous solution is in a range of 5.0-9.0.
In one embodiment, the ambient temperature is in a range of 27 to 35o C.
In one embodiment, the decorticated hemp fibres skin is obtained by natural fermentation which is carried from 1.0 day to 10 days.
In one embodiment, the process comprising a step of bioprocessing, the bioprocessing involves providing wet said cleaner fibres in a container that contains biomaterial obtained from ruminant animal waste and with biomaterial obtained from mushroom fungus, said fungus comprising enzyme/s which are responsible for degradation of pectin, hemicellulose, and lignin.
In one embodiment, the process comprising a step of bleaching said separated cleaned fibres.
In one embodiment, the process is performed in an environment having 70%-80% moisture.
In one embodiment, the separated cleaned fibres characterized by reduction of pectin in the region of 6% to 20%, reduction of lignin in the region of 10% to 20%, and reduction of cellulose in the region of 50% to 70.

The invention is illustrated with the help of following methods and reactions conditions.
Table 1: Reaction conditions:

Method 1:
1000 g hemp fibres was treated with 250 g to 300 g of biomaterial obtained from ruminant animal waste and kept for fermentation for 40-120 hrs.

Method 2:
1000 g hemp fibres was treated with 250 g to 300 g of biomaterial obtained from ruminant animal waste, 1-50 g of fungal mushroom, 1-10 g of carbon source, and 0.5- 10 g of nitrogen source and kept for fermentation for 40-120 hrs.

Method 3:
1000 g hemp fibres was treated with 250 g to 300 g of biomaterial obtained from ruminant animal waste, 1-50g of fungal mushroom, 1-50 g of fungal mushroom, 1-10 g of carbon source, and 0.5- 10 g of nitrogen. To this was added7 to 10 g of inoculum and kept for fermentation for 40-120 hrs.

The visual results are illustrated in the following figures:

FIGURES 1a and 1b, in general, show hemp fibres which were separated using natural fermentation bioprocess using biomaterial obtained from ruminant animal waste and in combination with Mushroom and Inoculum. FIGURE 1a shows untreated control hemp fibres. FIGURE 1b shows treated hemp fibres.

FIGURE 2 illustrates hemp fibres that were obtained via a first method of separation using natural fermentation bioprocess using ruminant animal waste for 40-120 hrs. Further, these separated fibres (22) were treated by using eco-friendly bleaching reagent to make cleaner fibres (24) .

FIGURE 3 illustrates hemp fibres that were obtained via a second method of separation using natural fermentation bioprocess using biomaterial obtained from ruminant animal waste, Mushroom fungus, carbon source and Nitrogen source. Natural fermentation was run for 40-120 hrs. Further, these separated fibres (32) were treated by using eco-friendly bleaching reagent to make cleaner fibres (34).

FIGURE 4 illustrates hemp fibres that were obtained via a third method of separation using natural fermentation bioprocess using biomaterial obtained from ruminant animal waste, Mushroom fungus, carbon source and Nitrogen source with consortium of inoculum. Natural fermentation was run for 40-120hrs. Further, these separated fibres (42) were treated by using eco-friendly bleaching reagent to make cleaner fibres (44).
The post bioprocessing results are tabulated in the following table.
Table 2: Post bioprocessing results
Control fibres Processed fibres in method 1 Processed fibres in method 2 Processed fibres in method 3
Improper segregation Successful segregation Successful segregation Successful segregation
Rough textured fibres Moderately Rough fibres Moderately smooth fibres Smooth fibres
Micron level strands of fibres aren’t formed Micron level strands of fibres are obtained Micron level strands of fibres are obtained Micron level strands of fibres are obtained

Table 2, above, illustrates, outcome of the treatment of Hemp Fibres after Bioprocessing.

In at least an embodiment of this invention, bioprocessing of hemp fibres was done by treating the hemp fibres with different fermentation methods. During natural fermentation, biodegradation of lignocellulosic materials occurs, due to secretion of several enzymes, and are responsible in separation and cleaning of complex hemp fibres. Application of lignocellulosic degrading enzymes in pure form seems to be very expensive and not economical feasible. According to prior art, several processes were being used to clean and separate the hemp fibres, from bast, which use harmful chemicals for environment and also use high temperature/s which is energy intensive. Therefore, natural ecofriendly fermentation bio-process was developed, according to this invention, by using relatively inexpensive and easily available natural bio-resource materials (biomaterial obtained from ruminant animal waste) which harbours a rich microbial diversity, containing different species of bacteria (Bacillus spp., Corynebacterium spp., and Lactobacillus spp.), protozoa, and yeast (Saccharomyces and Candida), and also containing several useful enzymes for degradation of pectin, hemicelluloses, lignin, and also cellulose.

Thus, keeping this advantage in mind, experiments were performed using bio-resource materials with different compositions and also with the combination of presence of some carbon sources natural nitrogen sources, with 70-80% moisture, room temperature, low pH, and high relative humidity.

It was observed that there was reduction of pectin by 6-20%, ignin content was reduced to 10-20%, and cellulose contents were increased by 50 to 70%, from initial fibres, in the treated fibres, with bioprocessing according to this invention.

Work was carried out to separate hemp fibres. Raw fibres were suspended in water as control and experimental raw fibre bundles were taken for bioprocessing using natural fermentation. During fermentation, several enzymes were produced during microbial growth which degraded plant polysaccharides.

In at least an embodiment, bio-resource material was used, the bio-resource material being biomaterial obtained from ruminant animal waste and / or combination of mushroom fungi and / or natural microorganisms, capable of degrading pectin, hemicellulose, and lignin.

In at least an embodiment, during fermentation, in order to facilitate microbial growth, carbon sources and nitrogen sources were supplemented with 70-80% moisture by maintaining room temperature, slightly low pH, and high relative humidity, these conditions allow the growth of natural bacteria (e.g. entities that mainly produce hydrolytic enzymes such as Bacillus, Clostridium) and fungi (e.g. Aspergillus, Penicillum). These microorganisms are basically fermenting or degrading pectic substrates (viz. pectinases, hemicellulose and lignins) from middle lamella and allow hemp’s defibrillation / separation into individual fibres and / or fibre bundles. Other microorganisms' secrete enzymes that can also degrade proteins, starches, fats, and waxes; from plant fibres cells.

Utilization of insoluble lignocellulosic substrates by fungi depends on their capability to synthesize relevant hydrolytic (cellulases and hemicellulases) and oxidative (ligninolytic) extracellular enzymes. These are responsible for degradation of substrate major components, that is, cellulose, hemicelluloses, and lignin, into low molecular weight compounds that can be assimilated for fungal nutrition. Thus, fungal selection and fermentation period are important. Thus, with this background, process steps, of this invention, comprise natural fermentation using natural bio-waste materials from animal sources and plant sources.

In at least an embodiment, natural fermentation is performed using natural microorganisms which produce pectinase, hemicellulose, lignin, and degradation enzymes along with controlled cellulose degrading enzymes. These naturally produced enzymes are capable of separating natural polymeric lignocellulosic containing materials and fibres for various industrial applications.

It is found that hemp fibres were separated efficiently using biomaterial obtained from ruminant animal waste or a combination of ruminant animal waste with mushroom fungus or a combination of ruminant animal waste with mushroom fungus and inoculum compared to the control. After bioprocessing, separation of fibres is found to be good, having good strength, when raw fibre bundle was treated with biomaterial obtained from ruminant animal waste, whereas fibres were very smooth when it was treated in a combination with biomaterial obtained from ruminant animal waste, mushroom, and inoculum having some appropriate amount nitrogen and carbon sources as a supplement for the growth of microorganisms.

The use of this novel and inventive ecofriendly bioprocess, for separation of hemp raw fibres, is for textiles, rope, clothing, paper, tissue, packaging materials, shoes and fabrics industries. This bioprocess has several advantages as it is eco-friendly, economical, and uses naturally available microorganisms and enzymes. This bioprocess takes less time to separate fibres, from each other, without damaging the length of fibres. No hazardous chemicals are used during bioprocessing of fibres in accordance with this invention; whereas, field retting for fibre separation, of the prior art process, takes anywhere from 6-8 weeks depending on weather and requires to be closely monitored.

It is to be noted that natural plant-based fibres specially hemp fibres are tightly bound with three complex biopolymers such pectine, hemillulose, cellulose, and lignin. These polymers have to be degraded to save cellulosic component of fibres. Cellulose contents, in the fibres, are very essential for industrial utilization of natural fibres. Cellulosic contents of fibres help in textile, fabric, clothing, paper making, cellulose-based polymer making, as chemical filters, and as fibre-reinforcement composites, epoxy coating for various applications, rayon etc.

Controlled biodegradation of pectine, hemicellulose and lignin were done using animal-based waste materials or in combination of fungi, bacteria for selective degradation of pectine, hemicellulose, lignin and some plant waxes. In this natural bioprocess, the process comprises a step of fermentation to release a consortium of enzymes produced by organisms and enzymes present in the animal-based waste natural product. During this bioprocessing, the step of fermentation separates the fibres without effecting the fibres’ structure; making it suitable for textile and other applications.

Thus, in the present invention a natural fermentation was used which releases desired enzymes. The bioprocess, of this invention, is suitable for industrial applications for natural fibres extraction and separation.

In accordance with a first non-limiting exemplary embodiment, control was carried out using water; this is reflected in FIGURE 5. This method is carried out by suspending hemp fibres in water for 3-10 days and it was observed that fibres were not segregated properly and had rough texture (FIGURE 5) when compare to other methods.

In accordance with a second non-limiting exemplary embodiment, bioprocess method, of this invention, was carried out by treating fibres with one or more bio-resource materials (biomaterial obtained from ruminant animal waste) which produced consortium of several useful enzymes for degradation of pectin, hemicelluloses, lignin, and also cellulose. Fermentation bioprocess was done for 3-10 days and it was observed that segregation of fibres was done properly. Further, separated fibres were treated with a bleaching agent. After treatment with bleaching agent, fibres became more segregated / separated and smoother as seen in Figure 6b when compared with Figure 5. FIGURE 6a illustrates fibres control and FIGURE 6b illustrates output of a method where fibre control, of FIGURE 6a, was treated with bio-resource material (biomaterial obtained from ruminant animal waste).

In accordance with a third non-limiting exemplary embodiment, where fibres were processed by fungal growth and also using bio-resource material, its output is seen in FIGURE 7. This method is carried out by treating fibres with Bio-resource material (biomaterial obtained from ruminant animal waste) mixed with water having appropriate amount of natural carbon and nitrogen sources. A mushroom fungus was inoculated in proper amount. This fungus grows in the presence of natural carbon and nitrogen sources. Mixture of Bio-resource material and mushroom along with nutrient medium were incubated at ambient temperature for 3-10 days for fermentation to degrade unwanted bioploymers such as pectin, lignin, hemicellulose, and protecting cellulose. After fermentation, it was observed that fibres were segregated nicely and became smoother (a). After treating with beaching agent, fibres became cleaner and whitish (b).

In accordance with a fourth non-limiting exemplary embodiment, where fibres were processed by fungal growth and also using bio-resource material, and inoculums, its output is seen in FIGURE 8. This method is carried out by treating fibres with Bio-resource material (biomaterial obtained from ruminant animal waste) mixed with water having appropriate amount of natural carbon and nitrogen sources. A mushroom fungus and a prepared inoculum of microbes were inoculated in proper amount with wetted fibres. This fungus and inoculum grow in presence of natural carbon and nitrogen sources. Mixture of Bio-resource material, mushroom, and inoculum, along with nutrient medium, were incubated at ambient temperature for 3-10 days for natural fermentation was done to degrade unwanted bio-ploymers such as pectine, lignin, hemicellulose, and protecting cellulose. After fermentation, it was observed that fibres were segregated (a) nicely and became smoother. After treating with beaching agent, fibres became more cleaner and whitish (b).

It was observed that when raw hemp fibres were treated with Bio-resource material (biomaterial obtained from ruminant animal waste) and / or with mushroom fungus and / or with inoculum showed good characteristics in terms of separation and smoothness of the fibres after fermentation, washing, and bleaching with bleaching agent.

Control fibres Processed fibres in method 1 Processed fibres in method 2 Processed fibres in method 3
Improper segregation Successful segregation Successful segregation Successful segregation
Rough textured fibres Moderately Rough fibres Moderately smooth fibres Smooth fibres
Micron level strands of fibres aren’t formed Micron level strands of fibres are obtained Micron level strands of fibres are obtained Micron level strands of fibres are obtained

Table 3, above, illustrates outcome of experiments on separation of hemp fibres using fermentation bioprocess.

Industrial Application of the present invention:
Hemp fibres have been used industrially as a source of quality fibre. Long, strong and durable, hemp fibres are about 60- 70% cellulose and contain low levels of lignin (around 8-10%). The fibre diameter ranges from 16 to 50 microns. Hemp fibre conducts heat, dyes well, resists mildew, blocks ultraviolet light and has natural anti-bacterial properties. Shorter, woody core fibres ("tow") contain higher levels of lignin. They are easy to grow without agrochemicals. Hemp is used increasingly in agrotextiles, car panels and fibreboard, and "cottonized" for clothing. Hemp stem contain fibres which can be process for textile fibres as it has very good amount of cellulose. Hemp fibres can be refined to be used for making clothes or textiles, as building materials, to make shoes, to make paper, to make ropes or cords. Hemp fibres are broadly used in industrial applications for their mechanical properties, in particular their Young’s modulus (YM) that is comparable to glass fibres, and for their low density, which makes them suitable for environmentally friendly alternatives to petrochemical compounds in applications such as in construction, bioplastics, or composite materials.
Hemp Fibres can be used for valuable composites materials which are being used for high end cars body building which revolutionising the auto industries. Hemp stalks / fibres are rich in cellulose, which makes it possible to produce a variety of bioplastics from these stalks. One of the stronger varieties of hemp bioplastic is already in use for making car parts. This bioplastic is considerably stronger than steel, as both Ford and Dietzen have demonstrated.
Hemp bioplastic is also considerably lighter than steel. The use of hemp bioplastic for production of automobile bodies can give lightweight dent-resistant vehicles. The production process would be far more carbon-neutral than the automobiles we currently make. Uses of hemp plastic is for door panels and back shelves of certain series of cars by BMW, Mercedes-Benz, and Bugatti; which are made from hemp fibres. Biodegradable hemp plastics are being used for single use for bottling of water and other materials.

Hemp fibres have been used industrially as a viable source of paper. Currently, paper is obtained from timber trees but it might as well be made from hemp. There are several advantages to hemp paper:
· Hemp grown in one acre could help produce as much paper as four acres of timber;
· Hemp has shorter yield cycles meaning we can produce more paper from it in any given time period;
· Paper made from hemp is stronger and more durable.

The TECHNICAL ADVANCEMENT of this invention lies in providing a bioprocess which does not require costly chemicals and bioprocess is performed at ambient temperature and natural enzymatic process. Advantages of the natural fermentation process, is that it can releases desired enzymes and is suitable for industrial applications for natural fibres extraction using ecofriendly bioprocess of natural fermentation. This process also does not require costly enzymes and is not energy intensive and is ecofriendly.

The bioprocess, of this invention, uses bio-resource material obtained from ruminant animal waste, mushroom, and a consortium of microbes found to be responsible for segregation of fibres without much processing; making this process ecofriendly, natural bioprocessing, relatively inexpensive, and economical. It does not involve costly chemical, enzymes, and is an efficient process to separate fibres. During bioprocessing, unwanted polymers such as pectins and lignins are degraded; but, cellulose content is increased. Thus, this process, of the invention, is very much suitable for many applications for bioprocessing of hemp, flax, coir, jute, banana, bamboo, sisal, agave, abaca, remie etc.

The bioprocess, of this invention, results in successful segregation of Hemp fibres, by disclosing different natural fermentation bioprocess methods. It was also observed that, without processing (Control), fibres were not segregated properly but, after natural fermentation processing with bio-resource materials which were obtained from ruminant animal waste along with carbon and nitrogen source and inoculums, fibres were easily segregated. The best segregation and good smoothness of fibres were observed in the second method and third method. Hence, these bio-resource materials, which include many enzymes and microorganisms, are responsible for the degradation of lignocellulosic wastes like cellulase, hemicellulases, pectinase etc. and also harbours rich microbial diversity, containing different species of bacteria and yeast hence it acts as a fundamental fermentation agent which helps fibres to segregate easily.

Advantage of the present invention is that it provides environmental benefits in terms of energy saving, economic, less labour-intensive, less time consuming to separate hemp fibres from raw hemp. Natural microorganisms present in the bio-resource material obtained ruminant animal waste which contain consortium of microorganisms which are capable to produce enzymes responsible to degrade unwanted biopolymers such as pectin, lignin and hemicellulose along with degradation of protein, carbohydrates, waxes etc. from hemp fibres. It is also advantageous to use lignin and pectin degrading mushroom such as Oyster known as Pleurotus. Inoculum was developed from natural retting containing various microorganisms capable of producing pectine , hemicellulose and lignin degrading enzymes. Thus ecofriendly bioprocess was developed which very economical and useful to separate hemp fibres efficiently in short time.

The method, of this invention, consumes less water when compared with prior art chemical processes. This bioprocess does not require any energy inputs, except for mechanical field operations. On the other hand, field retting, which is a conventional prior art method for separation of hemp fibres, has several disadvantages such as taking more time, unwanted degradation, and requiring lots of water.

Moreover, advantages, of the bioprocess, of this invention are listed as follows:
1. Ecofriendly bioprocess: Natural fermentation is carried out using biomaterial/s obtained from ruminant animal waste without any harm to animals;
2. No harmful chemicals are used which are harmful to environment;
3. No costly enzymes involved in bioprocessing of natural fibres;
4. Combination of biomaterial obtained from ruminant animal waste and easily available mushroom as such or extract has been used for easy separation of natural fibres - mushroom has capability to produce pectinase, hemicellulose, polyphenol oxidase / laccase, and some amount of cellulose; these enzymes are responsible for fibre separation;
5. Biomaterial obtained from ruminant animal waste and mushroom and consortium of microbial inoculum in combination was used and separation of the fibres were found to be efficient;
6. This bioprocess is economical and can be used for industries for large scale with cheaper infrastructure;
7. With the need of industry, this bioprocess can be modified.

While this detailed description has disclosed certain specific embodiments for illustrative purposes, various modifications will be apparent to those skilled in the art which do not constitute departures from the spirit and scope of the invention as defined in the following claims, and it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the invention and not as a limitation.
,CLAIMS:WE CLAIM,

1. An eco-friendly bioprocess for separation of hemp plant fibres, from decorticated hemp fibres skin, said process comprising:
- pre-treating the decorticated hemp fibres skin with an aqueous solution, said aqueous solution bearing a pH in the range of 5.0 to 9.0, wherein said aqueous solution consisting of:
o 2.0 gm to 5.0 gm of a carbon source per 1 kg of hemp fibre;
o 1.0 gm to 5.0 gm of a nitrogen source per 1 kg of hemp fibre;
o biomaterial obtained from a bio-resource material;
- storing, for natural fermentation, at ambient temperature, in order to separate its fibres, for a time period ranging from 3.0 to 10 days;
- subsequently, washing the separated fibres with water having a pH in the range of 5.0 to 8.0; and
- treating the washed and separated fibres with one or more cleaning agent for more cleaner fibres.

2. The process as claimed in claim 1 wherein, said ambient temperature is 20 to 37 deg. C

3. The process as claimed in claim 1 wherein, said biomaterial comprising ruminant animal waste along with a combination of mushroom and inoculum, wherein the proportion of ruminant animal waste to mushroom to inoculum is 30 parts:2 parts: 2 parts.

4. The process as claimed in claim 1 wherein, the pH of said aqueous solution is in a range of 5.0-9.0.

5. The process as claimed in claim 1 wherein, said ambient temperature is in a range of 27to 35o C.

6. The process as claimed in claim 1 wherein, said decorticated hemp fibres skin is obtained by natural fermentation carried from 1.0 day to 10 days.

7. The process as claimed in claim 1 wherein, said process comprising a step of bioprocessing by providing wet said cleaner fibres in a container that contains biomaterial obtained from ruminant animal waste and with biomaterial obtained from mushroom fungus, said fungus comprising enzyme/s which are responsible for degradation of pectin, hemicellulose, and lignin.

8. The process as claimed in claim 1 wherein, said process comprising a step of bleaching said separated cleaned fibres.

9. The process as claimed in claim 1 wherein, said process being performed in an environment having 70%-80% moisture.

10. The process as claimed in claim 1 wherein, said separated cleaned fibres exhibit reduction of pectin in the region of 6%-20%, reduction of lignin in the region of 10%-20%, and reduction of cellulose in the region of 50%-70%.

Dated this 20th day of July, 2022

CHIRAG TANNA
of INK IDÉE
APPLICANT’S PATENT AGENT
REGN. NO. IN/PA – 1785