Patent Publication Number: US-2016236996-A1

Title: A Process for Producing Highly Nutritious and Bioavailable Organic Nitrogen Fertilizer from Non GMA Organisms

Description:
TECHNICAL FIELD OF THE INVENTION 
     The invention relates to a process for producing highly nutritious and bioavailable nitrogen containing organic fertilizer with increased storage stability in liquid form, derived from natural, non-GMO sources which follow the standards of certified organic input in agriculture as per the standards required by statutory bodies of developed and developing countries. 
     BACKGROUND AND PRIOR ART 
     For a plant to grow and thrive, it requires the following different nutrient elements:
         Carbon (C), hydrogen (H) and oxygen (O) are available in air and water; therefore these are plentiful in supply;   Elements required in high concentrations are Nitrogen (N), phosphorus (P), potassium (K), sulphur (S), calcium (Ca) and magnesium (Mg)—macronutrients present in packaged fertilizers;   Micronutrients include boron (B), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo) and zinc (Zn).       

     However, nutrient input in crop production systems have come under increased scrutiny in recent years due to the potential environmental impact from nitrogen and phosphorus containing inputs. 
     Modern, intensive agriculture employing continuous and extensive use of chemical fertilizers and pesticides results in loss of natural humus, damages fertility of top soil and decreases soil porosity. Artificial chemicals destroy soil microbial flora resulting in poor soil structure, aeration and decreases nutrient availability. Consequently, roots are oxygen deprived and do not absorb salts effectively. Unabsorbed nitrates are carried away by rain water into rivers and lakes causing a threat to aquatic life. Excessive nitrates in soil elevate nitrate level in edible plants causing threat to people using them. Pests and diseases become more difficult to control as they become resistant to artificial pesticides. 
     Organic agriculture is a production system that sustains the health of soils, ecosystems and people. It relies on ecological processes, biodiversity and cycles adapted to local conditions, rather than the use of inputs with adverse effects. (International Federation of Organic Agriculture Movements) 
     The use of organic fertilizers for improving plant growth and overall development is known very well. Various industrial oil cakes unsuitable for animal consumption such as castor oil cake, neem oil cake, linseed oil cake, rapeseed cake, also edible oil cakes such as ground nut cake, are used to improve soil fertility due to their high molecular weight organic matter in the form of crude proteins, fats and complex carbohydrates. These cakes when added into soil get decomposed over a period of time by the action of bacteria present in soil. The bacteria multiply and the cakes are decomposed allowing uptake of nutrients by plants. However, complete extraction of nitrogen from the oilseed cakes is not possible by simple soil application; hence they must be processed by intensive chemical procedures for effective nitrogen utilization. 
     The procedures may introduce synthetic chemicals that are detrimental to environment. 
     In order that consumers are aware that agricultural products are actually obtained from organic ingredients without use of any chemicals and pesticides, farms are inspected as per Organic Standards. If they meet these standards they are certified organic and the produce from these farms is labeled as “Certified Organic Produce”. There are Organic standards for ‘Certified Organic Farming’ such as National Organic Program (NOP) provided by USDA; (US Department of Agriculture), National Program for Organic Production (NPOP) introduced in India, EU standards such as EC 834/2007 &amp; 889/2008 from European Union, Japanese Agricultural Standards (JAS), and Australian Input Standards. These voluntary or legislated standards regulate production methods, farm inputs and in some cases final output for organic agriculture. 
     Several organic nitrogen fertilizers have been developed by inventors by employing microbial sources and plant products, however due to intermediate processes and synthetic chemicals used these fertilizers have failed to adhere to the standards provided by the USDA. 
     NOP organic standards by USDA, (As per NOP Rule 205.105); permits the use of the following materials in organic farming fertilizer compositions:
         Compost derived animal waste or other materials.   Fresh plants material for incorporating nitrogen fixing into the soil.   Raw animal manures.   Animal &amp; Plant materials—Non Synthetic such as fish meal, fish emulsion, blood meal, feather meal, soybean meal and products of the like nature.   Non synthetic amino acids produced by plants, animals and microorganisms that have not been genetically modified and are extracted or isolated by hydrolysis or by physical or other non chemical means are considered non synthetic.       

     PCT Publication, WO2013/019933 claims organic fertilizer compositions of 1-6% nitrogen and 1-5% phosphorus comprising an effective amount of a non-genetically modified, enzymatically hydrolyzed plant protein as a nitrogen source and phytic acid as a phosphorus source for delivery to plants. However the essential element concentrations provided by the compositions are low and the source of hydrolyzed plant protein is not mentioned. 
     US 2008/0302151 relates to the manufacture of the soluble fertilizer for organic agriculture using soybean meal. The limitations of the patent are that parameters of hydrolysis such as the temperature and pH to be maintained are not mentioned. During the hydrolysis of soybean meal which contains 48% protein and 45-50% carbohydrate, the reaction mass becomes viscous and difficult to process because of the tendency of carbohydrates to swell and entrap water resulting in difficulty in filtration. Further, the process does not utilize a preservative for the liquid organic fertilizer product which being rich in amino acids and proteins is prone to microbial attack. The commercially available soymeal is solvent (uses, hexane) extracted and organic standard requires soymeal to be free from any solvent processing. 
     U.S. Pat. No. 8,308,838 relates to an organic fertilizer consisting essentially of corn steep liquor and/or oilseed extract in combination with whey and/or other protein supplements, which provide natural, nitrate free fertilizer. The process used herein comprises heating an oilseed extract, dissolving whey in the heated extract, and filtering the resultant mixture for use. The method used here does not solve the problem posed by the huge carbohydrate content provided by the corn steep liquor causing difficulty in filtration procedures. The product discussed contains the three macronutrients—nitrogen, phosphorous, potassium, however does not provide for the production of low molecular weight amino acid based nitrogen. 
     CN102584383 and JPH03237084 reveal the use of groundnut oil cakes and mustard oil cakes as the sources of element nutrition in organic fertilizer composition; however the methods employed in deriving nitrogen content and the parameters employed which follow the organic standards as per NOP of USDA are not mentioned. 
     Though, the above mentioned prior art Patents/Patent Publications teaches the production and use of an organic fertilizer that adhere to the USDA and EU standards of organic farming, there are certain limitations to the processes adopted, as mentioned above. Therefore, to overcome such limitations, the present inventors have provided an alternate process for producing an organic fertilizer containing nitrogen that meets the standards of organic farming requirements established in India and internationally, which employs ecofriendly non-GMO plant and microbial products as nitrogen sources and non-GMO plant and microbial enzyme sources, non-synthetic chemicals and natural preservatives as pH regulators. 
     Further, use of guar korma/meal as organic fertilizer is currently not in practice. The inventors have used Guar meal/Korma as the nitrogen source due to its high protein content of 33-55% and its easy availability. The process used in economically viable and the product obtained is conducive for agricultural use. 
     SUMMARY OF THE INVENTION 
     The present invention discloses a process for production of highly nutritious and bioavailable organic nitrogen fertilizer for foliar or soil application as per required organic standards established by statutory bodies of developed and developing countries. In an aspect, the invention provides an organic nitrogen fertilizer produced from products of non-Genetically Modified Organisms (N-GMO) origin including plants and microorganism as the nitrogen sources; non-synthetic chemicals derived from natural sources and processes, and natural enzymes derived from non-GMOs, each of which are employed at requisite stages of the process. 
     In another aspect, the invention provides a process for production of liquid organic nitrogen fertilizer comprising the following steps:
         a) slurrying 8-18% solid content of ground protein rich products of size of 100 mesh to 1-2 mm using demineralized water followed by heating the slurry mixture to 50°-100° C.;   b) Adding an oil absorbent to the slurry to reduce the oil content of the substrate;   c) Adding one or more carbohydrate digesting enzymes or combinations thereof and conducting digestion at pH of 5-6 and at 50° C.-55° C.;   d) Adding one or more proteolytic enzymes or combinations thereof; to carry out proteolysis at 40°-100° C., at least 3-4 hours after addition of carbohydrate digesting enzyme into the slurry followed by increasing the temperature of the reaction mass to 65°-150° C. for 2-3 hrs to inactivate the enzymes;   e) Filtering the reaction mass to separate molecules having molecular weight &lt;1000 from the mixture followed by adjusting pH of the liquid to 2.5-4 using acid buffer to obtain organic nitrogen fertilizer; and   f) Optionally concentrating and drying the liquid fertilizer.       

     Further, the invention provides guar meal/korma comprising protein content greater than 50% as the nitrogen source for preparation of the organic nitrogen fertilizer. 
     Abbreviations: 
     GMO: Genetically modified organisms 
     NOP: National Organic Program 
     USDA: US Department of Agriculture 
     NPOP: National Program for Organic Production 
     EU: European Union 
     JAS: Japanese Agricultural Standards 
     APEDA: Agriculture Produce &amp; Export Development Agency 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The present invention provides a process for production of organic nitrogen fertilizers for foliar or soil application as per organic standards established by statutory bodies of developed and developing countries. 
     The organic nitrogen fertilizer is produced from non-Genetically Modified Organisms (GMO) including plants and microorganism as the nitrogen sources; non-synthetic chemicals derived from natural processes and natural enzymes derived from non-GMOs are employed at requisite stages of the process. 
     The invention will now be described in detail in connection with certain preferred and optional embodiments, so that various aspects thereof may be more fully understood and appreciated. 
     In a preferred embodiment, the invention provides a process for production of liquid organic fertilizer comprising of the following steps:
         a) slurrying 8-18% solid content of ground protein rich products of size of 100 mesh to 1-2 mm using demineralized water followed by heating the slurry mixture to 50°-100° C.;   b) Adding an oil absorbent to the slurry to reduce the oil content of the substrate;   c) Adding one or more carbohydrate digesting enzymes or combinations thereof; and conducting digestion at pH of 5-6 and at 40° C.-60° C.;   d) Adding one or more proteolytic enzymes or combinations thereof; to carry out proteolysis at 40°-100° C., at least 3-4 hours after addition of carbohydrate digesting enzyme into the slurry followed by increasing the temperature of the reaction mass to 65°-150° C. for 2-3 hrs to inactivate the enzymes;   e) Filtering the reaction mass to separate molecules having molecular weight &lt;1000 from the, mixture followed by adjusting pH of the liquid to 2.5-4 using acid buffer to obtain organic nitrogen fertilizer; and   f) Optionally concentrating and drying the liquid fertilizer.       

     The invention is described in stage wise manner herein below: 
     Nitrogen Sources of the Organic Fertilizer 
     In an embodiment the invention provides that the nitrogen source of organic fertilizers is derived from natural, non-GMO origin, plant products selected from the group consisting of guar korma/meal, mustard seed cake, soybean, sunflower seed cake, groundnut seed cake, neem seed cake, cottonseed cake, sesame seed cake, coconut cake, palm kernel cake and castor seed cakes. A non-GMO origin, microbial product selected is baker&#39;s yeast i.e.  Saccharomyces cerevisiae.    
     These natural origin materials can be used individually and/or in combinations thereof. 
     These nitrogen sources are ground to a fine size of 100 mesh to 1-2 mm to facilitate ease in further treatment procedures. 
     The Guar or cluster bean ( Cyamopsis tetragonoloba ) is a source of guar gum. Guar meal is a by-product obtained from the downstream processing of guar seed for guar gum. It is high in protein and contains oil and albuminoids. The guar bean is principally grown in India and Pakistan, with smaller crops grown in the U.S., Australia, China and Africa. 
     Guar korma/meal is Guar korma used in the invention is sourced from the states of Rajasthan, Haryana and Gujarat. 
     Slurry Treatment 
     Water used in the treatment plant is subjected to reverse osmosis to demineralize the water and later to UV treatment to eliminate bacterial contamination. The ground, nitrogen containing material is added to demineralize water to form slurry having solid content of 8 to 18%. 
     The slurry is heated to 50°-100° C. This process is performed in a stainless steel (SS) jacketed vessel installed internally with a cooling coil. 
     1-5% natural calcite i.e. calcium carbonate is added to the slurry to absorb the oil content of the protein substrates. 
     Hydrolysis/Digestion 
     In another embodiment the invention provides a process for production of highly nutritious and bioavailable organic nitrogen fertilizer wherein carbohydrate digesting enzymes selected from the group consisting of pectinase, β-mannanase, cellulase, xylanase and amylase are used singly and/or in combinations thereof; to hydrolyze complex carbohydrate, to reduce viscosity of reaction mass 
     During hydrolysis of the protein rich substances, it is noted that fibers and complex carbohydrates tend to swell and increase the viscosity of the whole reaction mass making it difficult for filtration after the completion of hydrolysis. The swollen carbohydrate particle also entraps the hydrolyzed protein liquid into them thus the yield of organic nitrogen fertilizer based on hydrolyzed proteins is affected and reduced because some amount of hydrolysate is entrapped/hold up in the filtration cake. In order to avoid/reduce the viscosity build up of the slurry mixture and to improve the yield of hydrolyzed proteins, carbohydrate digesting enzymes are used for breakdown of complex carbohydrates present in the slurry. 
     Thick slurry/suspensions prevent the effective penetration of enzymes while the chance of interaction between the enzyme and the substrate molecule is low in very dilute slurry/suspension. Hence it is desirable to have lower viscosity of slurry for better effectiveness of the applied enzymes. Because of lower viscosity of the slurry it becomes easier for the protease enzyme to break down the proteins efficiently. 
     Hence, carbohydrate digesting enzyme selected from the group consisting of pectinase from  Aspergillus  species, β-mannanase from  Aspergillus  species,  Bacillus  species, cellulase from  Aspergillus  species,  Trichoderma  species, xylanase from  Aspergillus  species and amylase from  Bacillus  species are used singly and/or in combinations thereof; to hydrolyze complex carbohydrate, to reduce viscosity of reaction mass and reduce entrapped liquid/hold up in cake in order to increase filtration rate and yield of hydrolysate based organic N fertilizer. These enzymes are added at least 2-3 hours prior to the addition of proteolytic enzyme into the slurry. The digestion is carried out at the pH of 5-6 and at 40°-60° C. 
     In another embodiment the invention provides a process for production of highly nutritious and bioavailable organic nitrogen fertilizer wherein the proteolytic enzymes are selected from the group consisting of non-GMO plant sources bromelain and papain; non-GMO fungal proteases and bacterial proteases, and/or combinations thereof. 
     High molecular weight crude proteins from natural, non-GMO sources are hydrolyzed into smaller peptides and amino acids using proteolytic enzymes selected from the group consisting of non-GMO, natural enzymes of plant, microbial origin and/or combinations thereof. 
     The non-GMO sources of enzymes derived from plant sources are selected from the group consisting of papain and bromelain; non-GMO enzymes derived from microbial sources are selected from the group consisting of fungal proteases, bacterial proteases, neutral or alkaline protease; and/or combinations thereof The proteolytic enzymes employed in the hydrolysis are selected as per the permitted respective organic standards. The temperature of the slurry is regulated by heating or cooling processes to achieve optimum temperature required for efficient activity of proteolytic enzyme. Proteolysis of slurry mixture is carried out at 40-100° C. under constant stirring, preferably for 8-36 hrs. The reaction is carried out for several hours depending on the requirement of a-amino nitrogen concentration that provides the degree of hydrolysis in the product. 
     After achieving the desired hydrolysis level of proteins and noting the reduction in viscosity of slurry, the temperature of slurry is raised to 65°-150° C. for 2-3 hrs in order to inactivate the enzyme used in the reaction mass. 
     Filtration 
     In yet another embodiment the invention provides the process for production of highly nutritious and bioavailable organic nitrogen fertilizer, wherein the reaction mass is filtered through the plate and frame filter press, followed by filtration through micron filter and then passed through the ultrafiltration and nanofiltration unit. 
     The reaction mass is initially filtered through a plate and frame filter press in order to separate the undissolved solids from the hydrolyzed liquid. The filtrate obtained from plate and frame filter is further filtered through a micron filter to achieve a clear liquid. Further, filtration process is performed using appropriate filters to achieve separation according to the molecular weight of particles to be separated. 
     This liquid is then fed into an ultrafiltration unit for separating most of hydrolyzed proteins, peptides from carbohydrates. Then the separated peptide solution obtained from ultra filtration is fed to the nanofiltration unit which employs membrane filtration processes for separating amino acids and peptides of molecular weight &lt;1000 Daltons by applying MWCO of 1 KD. 
     The filtration involving ultra filtration and nanofiltration process yields highly nutritious and bioavailable organic nitrogen fertilizer containing hydrolyzed proteins, amino acids having low molecular weight of &lt;1000 Daltons which are quickly absorbed in plants through foliar and/or soil application and serves as a good chelating agent for minerals. The high molecular weight peptides and amino acids separated in ultra or nano filtration may be further hydrolyzed by employing the above method or recycled to produce lower molecular weight peptides and amino acids. 
     Further, in an embodiment the pH of the liquid is adjusted to 2.5-4.0 using non synthetic acetic acid derived from sugar industry or citric acid from fermentation industry and/or combinations thereof, to improve storage stability of the highly nutritious liquid organic fertilizer and reduce susceptibility of the fertilizer to microbial attack. 
     The filtrate obtained has a low concentration of hydrolyzed proteins based organic nitrogen fertilizer; hence it is concentrated in a vacuum evaporator to achieve desired concentration of the nitrogen. The concentrated liquid organic nitrogen fertilizer with concentration suitable for drying equipment is fed to suitable known dryer such as vacuum dryer, spray drying tower, drum dryer, flash dryer or freeze dryer to obtain dry, completely soluble organic fertilizer. 
     Further, the invention provides a process for production of highly nutritious and bioavailable organic nitrogen fertilizer derived from guar korma/guar meal. 
     In another preferred embodiment the invention provides a process for production of highly nutritious and bioavailable organic nitrogen fertilizer derived from guar korma/guar meal comprising the following steps:
         a) slurrying 8-18% solid content of protein rich, ground guar meal of size of 100 mesh to 1-2 mm using deionized water followed by heating the slurry mixture to 50°-100° C.;   b) Adding an oil absorbent to the slurry to reduce the oil content of the substrate;   c) Adding one or more carbohydrate digesting enzymes or combinations thereof and conducting digestion at pH of 5-6 and at 40° C.-60° C.;   d) Adding one or more proteolytic enzymes or combinations thereof; to carry out proteolysis at 40°-100° C., at least 3-4 hours after addition of carbohydrate digesting enzyme into the slurry followed by increasing the temperature of the reaction mass to 65°-150° C. for 2-3 hrs to inactivate the enzymes;   e) Filtering the reaction mass to separate molecules having molecular weight &lt;1000 from the mixture followed by adjusting pH of the liquid to 2.5-4 using acid buffer to obtain organic nitrogen fertilizer; and   f) Optionally concentrating and drying the liquid fertilizer.       

     In a representative embodiment, the invention provides a liquid and a soluble, dry, powder form of the organic nitrogen fertilizer. The liquid form of the organic fertilizer contains 2-10% organic nitrogen content and 10-65% solids; the dry, powder form contains 13.5% nitrogen content on 100% dry basis. 
     Advantageously, the invention provides production of liquid and solid organic nitrogen fertilizer from economical non-GMO protein sources by employing enzymatic sources and non-synthetic chemical that adhere to organic farming standards. 
     The organic fertilizer has enhanced functionality in terms of viscosity, bioavailability, solubility, storage stability and is endotoxin free. 
     The instant process for organic fertilizer production can be employed in India and in the international markets due to its compliance with organic farming standards. 
     The invention will now be illustrated with help of examples. The aforementioned embodiments and below mentioned examples are for illustrative purpose and are not meant to limit the scope of the invention. Various modifications of aforementioned embodiments and below mentioned examples are readily apparent to a person skilled in the art. All such modifications may be construed to fall within the scope and limit of this invention as defined by the appended claims. 
     EXAMPLES 
     Example 1 
     Nitrogen Sources of the Organic Fertilizer 
     Following raw materials from non-GMO sources were used either individually or in combination in the invention: 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                   
                 Protein content 
               
               
                   
                 Nitrogen source 
                 (%) 
               
               
                   
                   
               
             
            
               
                   
                 Guar meal/Guar Korma 
                 33-55% 
               
               
                   
                 Soybean 
                 38% 
               
               
                   
                 Mustard Seed Cake 
                 38% 
               
               
                   
                 Groundnut cake 
                 49% 
               
               
                   
                 Cottonseed cake 
                 40% 
               
               
                   
                 Sunflower oil cake 
                 33% 
               
               
                   
                 Neem cake 
                 30% 
               
               
                   
                 Castor Cake 
                 26-27% 
               
               
                   
                 Sesame oil cake 
                 35% 
               
               
                   
                 Coconut oil cake 
                 25% 
               
               
                   
                 Palm kernel cake 
                 18% 
               
               
                   
                 Baker&#39;s yeast from the species 
                 38% on dry basis 
               
               
                   
                 
                   Saccharomyces cerevisiae 
                 
               
               
                   
                 Baker&#39;s yeast compressed from the species 
                  8% 
               
               
                   
                 
                   Saccharomyces cerevisiae 
                 
               
               
                   
                   
               
            
           
         
       
     
     Example 2 
     Hydrolysis/Digestion 
     The above raw material was selected individually and/or in combination to be ground to finer size and added to demineralized water to produce slurry having solid content of 8 to 18% in a jacketed vessel. 
     Carbohydrate digesting enzymes were added at least 3-4 hrs prior to initiation of protein hydrolysis to reduce viscosity of the slurry. Hydrolysis of the non-GMO plant sources, baker&#39;s yeast and/or combinations thereof, was performed at 45-75° C. under constant stirring, preferably for 8-36 hrs using enzymes of non-GMO sources. The reaction was carried till the required concentration a amino nitrogen was detected. The enzymes were deactivated at 65°-150° C. for 2-3 hrs on completion of the protein hydrolysis and reduction in the viscosity of the liquid organic fertilizer. 
     Example 3 
     Filtration 
     The reaction slurry was filtered through a plate and frame filter press to separate undissolved particles. The resultant filtrate was further subjected to filtration through a 10 micron filter to achieve a clear liquid. This liquid was then fed into an ultrafiltration unit for separating most of hydrolyzed proteins, peptides from carbohydrates. Then the peptide solution obtained from ultra filtration process is fed to nanofiltration unit which employs membrane filtration principle by applying Molecular Weight Cut Off (MWCO) of 1 KD to separate peptides and amino acids of molecular weight &lt;1000 Daltons. The filtrate obtained from the nanofiltration unit was adjusted to pH of 2.5-4 using natural non synthetic acid buffers i.e. acetic acid and citric acid to prevent the fertilizer from microbial attack. 
     Example 4 
     Drying 
     The liquid formulation of the organic nitrogen fertilizer with concentration suitable for drying equipment was fed to suitable known dryer such as vacuum dryer, flash dryer, spray drying tower, drum dryer or freeze dryer to obtain dry, completely soluble organic fertilizer. 
     Example 5 
     The organic nitrogen fertilizer used in the liquid form has organic nitrogen content of 2-10% and 10-65% solids; the soluble, dry, powder form contains 13.5% nitrogen and 100% dry solids.