WO2006082264A1 - Method of obtaining biostimulants from agro-industrial residues - Google Patents

Method of obtaining biostimulants from agro-industrial residues Download PDF

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Publication number
WO2006082264A1
WO2006082264A1 PCT/ES2006/000034 ES2006000034W WO2006082264A1 WO 2006082264 A1 WO2006082264 A1 WO 2006082264A1 ES 2006000034 W ES2006000034 W ES 2006000034W WO 2006082264 A1 WO2006082264 A1 WO 2006082264A1
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Prior art keywords
agroindustrial
biostimulants
obtaining
phase
procedure
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PCT/ES2006/000034
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Spanish (es)
French (fr)
Inventor
Juan Parrado Rubio
Juan Dionisio Bautista Palomas
Eduardo Jesús ROMERO RAMÍREZ
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Universidad De Sevilla
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Publication of WO2006082264A1 publication Critical patent/WO2006082264A1/en

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    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • C05F5/006—Waste from chemical processing of material, e.g. diestillation, roasting, cooking
    • C05F5/008—Waste from biochemical processing of material, e.g. fermentation, breweries
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/10—Fertilisers containing plant vitamins or hormones
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/50—Isolated enzymes; Isolated proteins
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/141—Feedstock
    • Y02P20/145—Feedstock the feedstock being materials of biological origin
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
    • Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Definitions

  • the subject of the present invention is a process for obtaining plant biostimulants from agroindustrial wastes.
  • the process includes: a) A fermentative phase with microorganisms producing hydrolytic enzymes using as a base of the culture medium, agroindustrial residues and CaCI 2 in concentrations that favor the excretion of enzymes and b) a second phase in which the fermentation broths will be used as a hydrolytic tool on new waste that will be modified by the enzymes produced in the first step.
  • Agroindustrial wastes are characterized by being organic matter of high protein content and with a high susceptibility to be hydrolyzed due to the denaturation of the proteins produced in the agroindustrial processes from which they originate.
  • the residues of the bioalcohol industry whose origins are sorghum, corn, wheat and oats seeds, residues of the oil industry (from sunflower seeds, rapeseed, etc.), residues of the wine industry stand out.
  • the final products obtained are natural substances with biostimulant function (stimulates growth, flowering, rooting, ...) and also fertilizer (provides organic nutrients to plants).
  • the current technique makes the low cost (composting) incompatible with the high quality and product control (hydrolysis).
  • the present invention contemplates the use of agro-industrial waste with defined compositions so that the final product has an ideal proportion of nutritional elements for The crop to be treated. It is a process of production of biostimulants through enzymes produced in the same substrates that give rise to biostimulants, reducing process costs.
  • the present invention therefore consists of a two-phase process for obtaining a liquid product rich in soluble phytonutrients and of high absorption, of comparable quality to those obtained by enzymatic hydrolysis but of much lower cost. Description of the invention
  • the present invention aims at a process for obtaining a liquid product rich in soluble phytonutrients and high absorption for use as fertilizers and germination stimulants, rooting, growth, flowering, fruit set and maturation of plants and its fruits It can be applied in its different variants to any type of plant, time of plant development, soil and form of cultivation, and potentially usable in organic farming.
  • the process object of this invention comprises two phases that we will call the fermentation and hydrolytic phase respectively and two types of products are obtained: Soluble substances in liquid form with biostimulant and fertilizer capacity and solid by-product with fertilizer capacity.
  • the raw material used in the process of the present invention as a source of nitrogen and energy for the generation of enzymes are insoluble and defined plant substrates and, generally, of high insolubility. All have as a common characteristic its high content in Protein Nitrogen and a high susceptibility to be hydrolyzed due to the denaturation of the proteins produced in the agro-industrial processes from which they originate.
  • the substrates used in this process will be of vegetable origin and with high protein content as residues of the carob industry, the bioalcohol industry, (with different origins such as corn, sorghum, wheat, or barley), oil industry (sunflower degreased flours , rapeseed, etc.) wine industry (grape pomace) etc.
  • the process of the present invention comprises two phases:
  • the Fermentative Phase it is based on a vegetable substrate with high nitrogen content and high protein denaturation from agro-industrial processes, being the optimal culture medium for Bacillus licheniformis, an enzyme excretory microorganism.
  • a production of a liquid phase enriched in hydrolytic activity of microbial origin must be achieved.
  • a concentration of 0.5% - 2% of plant substrate (the most optimal conditions around 1%) and an inoculum of B. licheniformis together with CaCI 2 in quantities starting from 5 are introduced into a biorector mmolar to favor the synthesis and stability of the enzymes to be produced.
  • the bioreactor must be at neutral pH, T a optimum for the development of the microorganism (35-37 0 C) and aeration for 40-50 hours, obtaining a solution of the modified substrate with a high content of proteases excreted by the microorganism, which is used in Ia Hydrolytic phase of the process as an enzymatic base to increase the productivity of the biostimulant object of the present invention.
  • the Hydrolytic Phase of the process consists in taking the fermentation broth of the fermentation phase and diluting it 20-30 times until reaching an optimum concentration to carry out the hydrolysis. This process requires several controls among which the pH (optimal for enzymatic activity, between 7-8), the temperature (55-6O 0 C) and time (100min-150min) as fundamental factors could be cited.
  • This phase can be carried out in the same container as the previous phase simply by adding water and the agro-industrial waste with high biostimulant potential (10-20% w / v), obtaining the solubility of the introduced plant material with biostimulant capacity.
  • the solid-liquid separation is carried out by centrifugation, at the concentration by evaporation and by acidifying the medium to lower the pH, facilitating storage, transport and, above all, because at that concentration it is totally stable.
  • the resulting solids not solubilized can be used as conventional organic fertilizer and the resulting liquid is the product with biostimulant capacity for use as fertilizers and stimulants of germination, rooting, growth, flowering, fruit set and maturation of your plants and fruits
  • the first phase is the fermentation phase, common in the production of both types of biostimulants, with the difference in the substrate used in the second phase.
  • the fermentation medium includes the following components:
  • microorganisms used are bacteria of the genus Bacillus enzyme superproducers, in the example a strain of Bacillus licheniformis ATCC 21415 has been used.
  • the fermentation phase is centrifuged and the separation is carried out in a liquid and a solid phase.
  • the cell-free liquid phase will be used as a hydrolytic tool because of its high protease content, being cold stable for a month.
  • the second phase is the hydrolysis phase in which the conversion of agroindustrial wastes into biostimulants occurs:
  • the difference lies in the raw material that is hydrolyzed being the process itself in both cases:
  • First Biostimulant called BM From residues of the bioalcohol industry from corn seeds.
  • Second Biostimulant called BA From residues of the food industry of the carob bean.
  • the biostimulant product of corn residues (BM) has the following composition: Physical-chemical characteristics
  • the biostimulant product of carob (BA) has the following composition Physicochemical characteristics
  • biostimulant capacity by shortening flowering times and increase in the number of fruits Using as raw materials corn residues of the bioalcohol industry in the fermentation phase and the carob residues as a substrate in the hydrolytic phase (BA), the biostimulant power It has been studied in tomatoes, in a greenhouse with light and temperature control (Licopericon pimpinellifolium) and a parameter such as the ability to increase the number of total flowers and the total number of fruits has been evaluated.
  • the product has been used in a diluted liquid form for fertirrigation, resulting in its optimal dose ranging from 1/500 dilution of BA.

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Botany (AREA)
  • Plant Pathology (AREA)
  • Virology (AREA)
  • Environmental Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Agronomy & Crop Science (AREA)
  • Dentistry (AREA)
  • Pest Control & Pesticides (AREA)
  • Biochemistry (AREA)
  • Environmental & Geological Engineering (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Fertilizers (AREA)
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Abstract

The invention relates to a method of obtaining plant biostimulants from agro-industrial residues. The inventive method comprises the following phases, namely: a) a fermentation phase with hydrolytic-enzyme-producing micro-organisms, in which the culture medium base comprises agro-industrial residues and CaCl2 in concentrations that encourage enzyme excretion; and b) a second phase in which the fermentation broths are used as a hydrolytic instrument on novel residues modified by the enzymes produced in the first step. The end products obtained are essentially natural and have a biostimulating function (stimulates growth, flowering, root development, etc.) and a fertilising function (supplies plants with organic nutrients).

Description

TítuloTitle
Procedimiento para Ia obtención de bioestimulantes a partir de residuos agroindustrialesProcedure for obtaining biostimulants from agroindustrial waste
ObjetoObject
La presente invención tiene por objeto un procedimiento para Ia obtención de bioestimulantes vegetales a partir de residuos agroindustriales. El proceso comprende: a) Una fase fermentativa con microorganismos productores de enzimas hidrolíticas usando como base del medio de cultivo, residuos agroindustriales y CaCI2 en concentraciones que favorecen Ia excreción de enzimas y b) una segunda fase en Ia que los caldos de fermentación serán utilizados como una herramienta hidrolítica sobre nuevos residuos que serán modificadas por las enzimas producidas en el paso primero.The subject of the present invention is a process for obtaining plant biostimulants from agroindustrial wastes. The process includes: a) A fermentative phase with microorganisms producing hydrolytic enzymes using as a base of the culture medium, agroindustrial residues and CaCI 2 in concentrations that favor the excretion of enzymes and b) a second phase in which the fermentation broths will be used as a hydrolytic tool on new waste that will be modified by the enzymes produced in the first step.
Los residuos agroindustriales se caracterizan por ser materia orgánica de alto contenido proteico y con una alta susceptibilidad a ser hidrolizado debido a Ia desnaturalización de las proteínas producido en los procesos agroindustriales de los cuales proceden. Destacan los residuos de Ia industria del bioalcohol cuyos orígenes son semillas de sorgo, maíz, trigo y avena, residuos de Ia industria del aceite (procedentes de semillas de girasol, colza, etc.), residuos de Ia industria vitivinícola..Agroindustrial wastes are characterized by being organic matter of high protein content and with a high susceptibility to be hydrolyzed due to the denaturation of the proteins produced in the agroindustrial processes from which they originate. The residues of the bioalcohol industry whose origins are sorghum, corn, wheat and oats seeds, residues of the oil industry (from sunflower seeds, rapeseed, etc.), residues of the wine industry stand out.
Los productos finales obtenidos son sustancias naturales con función bioestimulante (estimula el crecimiento, Ia floración, el enraizamiento,...) y también fertilizante (provee de nutrientes orgánicos a las plantas).The final products obtained are natural substances with biostimulant function (stimulates growth, flowering, rooting, ...) and also fertilizer (provides organic nutrients to plants).
Estado de Ia técnicaState of the art
Actualmente, los fertilizantes orgánicos parten de sustratos orgánicos y sufren una serie de tratamientos que los transforman en mayor o menor grado en sustancias asimilables por Ia planta. Los procesos productivos se dividen en fermentativos y enzimáticos: Los procesos fermentativos utilizados en Ia actualidad de forma mayoritaria, son tratamientos microbianos, procesos de fermentación en estado sólido (compostaje). Hay numerosas patentes que describen dichos procesos (Véase como ejemplo de ello, el "Procedimiento para Ia fabricación de un fertilizante orgánico a partir de excreta ganadera y residuos agroforestales", patente española N0 2 151 845; "Procedimiento para Ia obtención de un fertilizante orgánico", patente española N0 518.595).Currently, organic fertilizers are based on organic substrates and undergo a series of treatments that transform them to a greater or lesser extent into substances assimilable by the plant. The production processes are divided into fermentative and enzymatic: The fermentation processes currently used in the majority are microbial treatments, solid state fermentation processes (composting). There are numerous patents that describe these processes (See as an example of this, the "Procedure for the manufacture of an organic fertilizer from livestock excreta and agroforestry waste", Spanish patent No. 0 151 151 845; "Procedure for obtaining a fertilizer organic ", Spanish patent No. 0 518,595).
En estos procesos, debido al poco control que se ejerce sobre el crecimiento microbiano que es inespecífico, hace que el producto final no sea homogéneo de una partida a otra, por Io que las producciones que se obtienen con ellos tampoco son siempre comparables.In these processes, due to the little control that is exerted on the microbial growth that is nonspecific, it makes the final product not homogeneous from one item to another, so that the productions that are obtained with them are also not always comparable.
.Los procesos de producción mediante hidrólisis enzimática directa, son mucho más eficientes y dan lugar a productos líquidos que presentan una mayor capacidad bioestimulante, mayor biodisponibilidad y funcionalidad a Ia hora de su aplicación. Estos procesos son mucho más eficientes, el tiempo de producción es de horas con respecto a semanas y meses de los procesos fermentativos. Sin embargo, las hidrólisis enzimáticas de diversos orígenes tienen el problema de coste de Ia enzima (o enzimas) empleadas en el proceso, Io cual suele hacer prohibitivo su uso. Otros mecanismos como Ia hidrólisis química suele producir, como efectos colaterales, una gran cantidad de sustancias no absorbibles por Ia planta y, potencialmente tóxicas.The production processes through direct enzymatic hydrolysis are much more efficient and give rise to liquid products that have a greater biostimulant capacity, greater bioavailability and functionality at the time of application. These processes are much more efficient, the production time is hours with respect to weeks and months of the fermentation processes. However, enzymatic hydrolysis of various origins have the problem of cost of the enzyme (or enzymes) used in the process, which usually makes its use prohibitive. Other mechanisms such as chemical hydrolysis usually produce, as collateral effects, a large number of substances that are not absorbable by the plant and, potentially toxic.
La técnica actual, hace incompatible el bajo coste (compostaje) con Ia alta calidad y control de producto (hidrólisis) La presente invención contempla el uso de residuos agroindustriales con composiciones definidas a fin de que el producto final posea una proporción ideal de elementos nutritivos para el cultivo a tratar. Se trata de un proceso de producción de bioestimulantes mediante enzimas producidas en los mismos sustratos que dan lugar a los bioestimulantes, reduciéndose los costes del proceso. La presente invención consiste por tanto, en un proceso en dos fases para Ia obtención de un producto líquido rico en fitonutrientes solubles y de alta absorción, de calidad comparable a los obtenidos por hidrólisis enzimática pero de coste muy inferior. Descripción de Ia invenciónThe current technique makes the low cost (composting) incompatible with the high quality and product control (hydrolysis). The present invention contemplates the use of agro-industrial waste with defined compositions so that the final product has an ideal proportion of nutritional elements for The crop to be treated. It is a process of production of biostimulants through enzymes produced in the same substrates that give rise to biostimulants, reducing process costs. The present invention therefore consists of a two-phase process for obtaining a liquid product rich in soluble phytonutrients and of high absorption, of comparable quality to those obtained by enzymatic hydrolysis but of much lower cost. Description of the invention
La presente invención tiene por objeto un procedimiento para Ia obtención de un producto líquido rico en fitonutrientes solubles y de alta absorción para su uso como fertilizantes y estimulantes de Ia germinación, el enraizamiento, el crecimiento, Ia floración, el cuajado y Ia maduración de plantas y sus frutos. Puede aplicarse en sus diferentes variantes a cualquier tipo de planta, momento del desarrollo vegetal, suelo y forma de cultivo, y potencialmente utilizable en agricultura ecológica.The present invention aims at a process for obtaining a liquid product rich in soluble phytonutrients and high absorption for use as fertilizers and germination stimulants, rooting, growth, flowering, fruit set and maturation of plants and its fruits It can be applied in its different variants to any type of plant, time of plant development, soil and form of cultivation, and potentially usable in organic farming.
El procedimiento objeto de esta invención comprende dos fases que llamaremos Fase fermentativa e hidrolítica respectivamente y se obtienen dos tipos de productos: Sustancias solubles en forma líquida con capacidad bioestimulante y fertilizante y subproducto sólido con capacidad fertilizante.The process object of this invention comprises two phases that we will call the fermentation and hydrolytic phase respectively and two types of products are obtained: Soluble substances in liquid form with biostimulant and fertilizer capacity and solid by-product with fertilizer capacity.
La materia prima utilizada en el procedimiento de Ia presente invención como fuente de nitrógeno y energía para Ia generación de enzimas son sustratos vegetales insolubles y definidos y, generalmente, de una elevada insolubilidad. Todos tienen como característica común su alto contenido en Nitrógeno proteico y una alta susceptibilidad a ser hidrolizado debido a Ia desnaturalización de las proteínas producido en los procesos agroindustriales de los cuales proceden. Los sustratos utilizados en este proceso serán de origen vegetal y con altos contenidos proteicos como residuos de Ia industria de Ia algarroba, industria del bioalcohol, (con diferentes orígenes como maíz, sorgo, trigo, o cebada), industria aceitera (harinas desengrasadas de girasol, colza, etc) industria vitivinícola (orujo de uva) etc.The raw material used in the process of the present invention as a source of nitrogen and energy for the generation of enzymes are insoluble and defined plant substrates and, generally, of high insolubility. All have as a common characteristic its high content in Protein Nitrogen and a high susceptibility to be hydrolyzed due to the denaturation of the proteins produced in the agro-industrial processes from which they originate. The substrates used in this process will be of vegetable origin and with high protein content as residues of the carob industry, the bioalcohol industry, (with different origins such as corn, sorghum, wheat, or barley), oil industry (sunflower degreased flours , rapeseed, etc.) wine industry (grape pomace) etc.
El procedimiento de Ia presente invención comprende dos fases:The process of the present invention comprises two phases:
La Fase Fermentativa: tiene como base un sustrato vegetal de alto contenido en nitrógeno y elevada desnaturalización proteica procedente de procesos agroindustriales siendo el medio de cultivo óptimo de Bacillus licheniformis, un microorganismo excretor de enzimas. En Ia fase fermentativa se ha de conseguir una producción de una fase líquida enriquecida en actividad hidrolítica de origen microbiano. Para ello, se introducen en un biorrector una concentración de 0,5%- 2% de sustrato vegetal (siendo las condiciones más óptimas en torno al 1 %) y un inoculo de B. licheniformis junto con CaCI2 en cantidades a partir de 5 mmolar para favorecer Ia síntesis y estabilidad de las enzimas a producir. El biorreactor debe estar a pH neutro, Ta óptima para el desarrollo del microorganismo (35-370C) y aireación durante 40-50 horas, obteniéndose una disolución del sustrato modificado con un elevado contenido en proteasas excretadas por el microorganismo, que se emplea en Ia fase hidrolítica del procedimiento como base enzimática para aumentar Ia productividad del bioestimulante objeto de Ia presente invención.The Fermentative Phase: it is based on a vegetable substrate with high nitrogen content and high protein denaturation from agro-industrial processes, being the optimal culture medium for Bacillus licheniformis, an enzyme excretory microorganism. In the fermentation phase, a production of a liquid phase enriched in hydrolytic activity of microbial origin must be achieved. For this, a concentration of 0.5% - 2% of plant substrate (the most optimal conditions around 1%) and an inoculum of B. licheniformis together with CaCI 2 in quantities starting from 5 are introduced into a biorector mmolar to favor the synthesis and stability of the enzymes to be produced. The bioreactor must be at neutral pH, T a optimum for the development of the microorganism (35-37 0 C) and aeration for 40-50 hours, obtaining a solution of the modified substrate with a high content of proteases excreted by the microorganism, which is used in Ia Hydrolytic phase of the process as an enzymatic base to increase the productivity of the biostimulant object of the present invention.
La Fase Hidrolítica del proceso consiste en tomar el caldo de fermentación de Ia fase fermentativa y diluirlo 20-30 veces hasta llegar a una concentración óptima para llevar a cabo Ia hidrólisis. Éste proceso requiere diversos controles entre los que podría citarse el pH (óptimo para Ia actividad enzimática, entre 7-8), Ia temperatura (55-6O0C) y el tiempo (100min-150min) como factores fundamentales.The Hydrolytic Phase of the process consists in taking the fermentation broth of the fermentation phase and diluting it 20-30 times until reaching an optimum concentration to carry out the hydrolysis. This process requires several controls among which the pH (optimal for enzymatic activity, between 7-8), the temperature (55-6O 0 C) and time (100min-150min) as fundamental factors could be cited.
Esta fase puede llevarse a cabo en el mismo recipiente que Ia fase anterior simplemente añadiendo agua y el residuo agroindustrial con elevado potencial bioestimulante (10-20% p/v), obteniéndose Ia solubilidad del material vegetal introducido con capacidad bioestimulante.This phase can be carried out in the same container as the previous phase simply by adding water and the agro-industrial waste with high biostimulant potential (10-20% w / v), obtaining the solubility of the introduced plant material with biostimulant capacity.
Por último se procede a Ia separación sólido - líquido mediante centrifugación, a Ia concentración por evaporación y acidificando el medio para disminuir el pH facilitando el almacenaje, el transporte y, sobre todo, porque a esa concentración es totalmente estable. Los sólidos resultantes no solubilizados, pueden emplearse como abono orgánico convencional y el líquido resultante es el producto con capacidad bioestimulante para su uso como fertilizantes y estimulantes de Ia germinación, el enraizamiento, el crecimiento, Ia floración, el cuajado y Ia maduración de sus plantas y frutos.Finally, the solid-liquid separation is carried out by centrifugation, at the concentration by evaporation and by acidifying the medium to lower the pH, facilitating storage, transport and, above all, because at that concentration it is totally stable. The resulting solids not solubilized, can be used as conventional organic fertilizer and the resulting liquid is the product with biostimulant capacity for use as fertilizers and stimulants of germination, rooting, growth, flowering, fruit set and maturation of your plants and fruits
Modo de realizaciónEmbodiment
A continuación se describe el proceso de obtención de bioestimulantes para dos casos concretos:The process of obtaining biostimulants for two specific cases is described below:
A) Residuos de Ia industria del bioalcohol procedentes de semillas de maíz.A) Waste from the bioalcohol industry from corn seeds.
B) Residuos de Ia industria de Ia algarroba. La primera fase es Ia fase fermentativa, común en Ia producción de ambos tipos de bioestimulantes, radicando Ia diferencia en el substrato utilizado en Ia segunda fase. El medio de fermentación incluye los siguientes componentes:B) Waste from the carob industry. The first phase is the fermentation phase, common in the production of both types of biostimulants, with the difference in the substrate used in the second phase. The fermentation medium includes the following components:
Residuos de Ia industria del bioalcohol 10 gramos (base de medio de cultivo) Cloruro de calcio 0.55 gramos (Activador de Ia producción de proteasas) Cloruro de sodio 3 gramos Agua hasta 1 litroWaste from the bioalcohol industry 10 grams (culture medium base) Calcium chloride 0.55 grams (Activator of protease production) Sodium chloride 3 grams Water up to 1 liter
Los microorganismos utilizados son bacterias del género Bacillus superproductoras de enzimas, en el ejemplo se ha utilizado una cepa de Bacillus licheniformis ATCC 21415.The microorganisms used are bacteria of the genus Bacillus enzyme superproducers, in the example a strain of Bacillus licheniformis ATCC 21415 has been used.
Las condiciones de cultivo fueron los siguientes: pH 7The culture conditions were as follows: pH 7
Temperatura 37 grados centígrados Tiempo de cultivo 53 horas. El resultado de Ia fase anterior es una suspensión de partículas y bacterias en un líquido que lleva disuelto una cantidad considerable de actividad hidrolítica de Ia bacteria B. licheniformis empleada en el proceso.Temperature 37 degrees Celsius Cultivation time 53 hours. The result of the previous phase is a suspension of particles and bacteria in a liquid that has dissolved a considerable amount of hydrolytic activity of the B. licheniformis bacteria used in the process.
La fase de fermentación es centrifugada y se procede a Ia separación en una fase líquida y otra sólida. La fase líquida libre de células se empleara como herramienta hidrolítica por su gran contenido en proteasas, siendo estable en frío durante un mes.The fermentation phase is centrifuged and the separation is carried out in a liquid and a solid phase. The cell-free liquid phase will be used as a hydrolytic tool because of its high protease content, being cold stable for a month.
La segunda fase es Ia fase de hidrólisis en Ia cual se produce Ia conversión de los residuos agroindustriales en bioestimulantes: En nuestro ejemplos se muestra Ia producción de dos tipos de bioestimulantes, Ia diferencia radica en Ia materia prima que se hidroliza siendo el proceso el mismo en ambos casos:The second phase is the hydrolysis phase in which the conversion of agroindustrial wastes into biostimulants occurs: In our examples the production of two types of biostimulants is shown, the difference lies in the raw material that is hydrolyzed being the process itself in both cases:
Primer Bioestimulante denominado BM: Procedente de residuos de Ia industria del bioalcohol procedentes de semillas de maíz. Segundo Bioestimulante denominado BA: Procedente de residuos de Ia industria alimentaria de Ia algarroba.First Biostimulant called BM: From residues of the bioalcohol industry from corn seeds. Second Biostimulant called BA: From residues of the food industry of the carob bean.
En un reactor hidrolítico se introducen los siguientes materiales: 200 gramos de Residuo de Ia industria del bioalcohol procedentes de semillas de maíz para Ia producción BM ó 200 gramos de Residuo procedentes de Ia industria de Ia algarroba para Ia producción de BAThe following materials are introduced into a hydrolytic reactor: 200 grams of Waste from the bioalcohol industry from corn seeds for BM production or 200 grams of Waste from the carob industry for the production of BA
50 mi de Ia Fase liquida producida en Ia etapa anterior50 ml of the liquid phase produced in the previous stage
Agua hasta 1000 mi En el reactor se mantendrá constante dos parámetros:Water up to 1000 ml Two parameters will remain constant in the reactor:
Temperatura 55 0C pH 7Temperature 55 0 C pH 7
Tiempo de reacción: 120 minutosReaction time: 120 minutes
Una vez finalizado, el producto es centrifugado, y evaporado siendo su composición para cada caso:Once finished, the product is centrifuged, and evaporated being its composition for each case:
El producto bioestimulante de residuos de maíz (BM) tiene Ia siguiente composición: Características físico-químicasThe biostimulant product of corn residues (BM) has the following composition: Physical-chemical characteristics
%p/p% p / p
Humedad 65-60 pH 5.0-5.5 Densidad (Kg/I) 1.10-1.12Humidity 65-60 pH 5.0-5.5 Density (Kg / I) 1.10-1.12
Cenizas 1.04-1.29Ashes 1.04-1.29
Nitrógeno total 4.44-5.08Total Nitrogen 4.44-5.08
Nitrógeno amoniacal 1.22-1.40 Nitrógeno proteico 3.22-3.68Ammoniacal nitrogen 1.22-1.40 Protein nitrogen 3.22-3.68
Proteína Bruta 28-32Crude Protein 28-32
Aminoácidos libres 7-9Free Amino Acids 7-9
El producto bioestimulante de algarroba (BA) tiene Ia siguiente composición Características físico-químicasThe biostimulant product of carob (BA) has the following composition Physicochemical characteristics
%p/p% p / p
Nitrógeno total 2,6-3,2Total Nitrogen 2.6-3.2
Nitrógeno amoniacal 0,26-0,33Ammoniacal nitrogen 0.26-0.33
Nitrógeno proteico 2,30-2,87Protein nitrogen 2.30-2.87
Contenido Proteico 14,0-18,0 Aminoácidos libres 7-9Protein Content 14.0-18.0 Free Amino Acids 7-9
Humedad 70-74 pH 5,4Humidity 70-74 pH 5.4
Densidad (Kg/I) 1 ,08-1 ,1 1 Materia orgánica 24,3-28,2Density (Kg / I) 1, 08-1, 1 1 Organic matter 24.3-28.2
Cenizas 1 ,6-2,0Ashes 1, 6-2.0
Ejemplo de capacidad bioestimulante acortando tiempos de floración e incremento en el número de frutos Utilizando como materias primas residuos de maíz de Ia industria del bioalcohol en Ia fase de fermentación y los residuos de algarroba como substrato en Ia fase hidrolítica (BA), Ia potencia bioestimulante se ha estudiado en tomates, en invernadero con control de luz y temperatura (Licopericon pimpinellifolium) y se ha evaluado un parámetro como es Ia capacidad de incrementar el número de flores totales y el número total de frutos.Example of biostimulant capacity by shortening flowering times and increase in the number of fruits Using as raw materials corn residues of the bioalcohol industry in the fermentation phase and the carob residues as a substrate in the hydrolytic phase (BA), the biostimulant power It has been studied in tomatoes, in a greenhouse with light and temperature control (Licopericon pimpinellifolium) and a parameter such as the ability to increase the number of total flowers and the total number of fruits has been evaluated.
Se han utilizado dos controles, uno es Ia no bioestimulación (se utiliza sólo agua), y otro es el uso de un bioestimulante comercial de origen animal denominado Isabion y comercializado por Syngenta.Two controls have been used, one is non-biostimulation (only water is used), and another is the use of a commercial biostimulant of animal origin called Isabion and marketed by Syngenta.
El producto, se ha empleado en forma líquida diluida para fertirrigación resultando que su dosis óptima va de una dilución 1/500 del BA.The product has been used in a diluted liquid form for fertirrigation, resulting in its optimal dose ranging from 1/500 dilution of BA.
Los resultados muestran una gran bioestimulación de las plantas las cuales producen más flores y más frutos que en los controles sin bioestimulación (sólo con agua) y con Isabion. Parámetros en el cultivo del tomateThe results show a great biostimulation of the plants which produce more flowers and more fruits than in the controls without biostimulation (only with water) and with Isabion. Parameters in tomato cultivation
Tiempo Altura de Ia planta (cm) Número de flores por plantaTime Height of the plant (cm) Number of flowers per plant
(semanas)(weeks)
Agua Isabion BA/BM Agua Isabion BA/BWater Isabion BA / BM Water Isabion BA / B
Nd Nd Nd 1a - 1aNd Nd Nd 1st - 1st
10 Nd Nd Nd 3a - 4a10 Nd Nd Nd 3a - 4a
12 Nd Nd Nd 8a - 7a12 Nd Nd Nd 8a - 7a
14 Nd Nd Nd 9ab - 1514 Nd Nd Nd 9ab - 15
16 Nd Nd Nd 9ab 2a 32b16 Nd Nd Nd 9ab 2a 32b
18 53.7af 69.5ab 83.8b 9ab 16ab 33b18 53.7a f 69.5ab 83.8b 9ab 16ab 33b
Tiempo Número de frutos por plantaTime Number of fruits per plant
(semanas)(weeks)
Agua Isabion BA/MWater Isabion BA / M
88
1010
12 1a 1a12 1st 1st
14 1a 1a14 1st 1st
16 2a 4a16 2nd 4th
18 3a 1a 15ab18 3rd 1st 15ab
Nd: No determinadaNd: Not determined
^Diferentes letras indican que son significativamente diferentes. ^ Different letters indicate that they are significantly different.

Claims

Reivindicaciones Claims
1. Procedimiento para Ia obtención de bioestimulantes a partir de residuos agroindustriales caracterizado porque comprende dos fases: a) una fase fermentativa con microorganismos productores de enzimas hidrolíticas, usando como base del medio de cultivo residuos agroindustriales en agua1. Procedure for obtaining biostimulants from agroindustrial wastes characterized in that it comprises two phases: a) a fermentative phase with microorganisms producing hydrolytic enzymes, using agroindustrial residues in water as the basis of the culture medium
(0.5-2% p/v) y CaCI2 en condiciones de pH neutro, Ta óptima para el desarrollo de microorganismos (35-370C) y aireación durante un tiempo de 40-50 horas y b) una segunda fase en Ia que los caldos de fermentación serán utilizados como una herramienta hidrolítica sobre residuos agroindustriales que serán modificados por las enzimas producidas en el paso primero.(0.5-2% w / v) and CaCI2 under conditions of neutral pH, T a optimal for the development of microorganisms (35-37 0 C) and aeration for a time of 40-50 hours and b) a second phase in which The fermentation broths will be used as a hydrolytic tool on agroindustrial waste that will be modified by the enzymes produced in the first step.
2. Procedimiento para Ia obtención de bioestimulantes a partir de residuos agroindustriales según reivindicación 1 caracterizado por el empleo de Bacillus licheniformis como microorganismo productor de enzimas hidrolíticas.2. Procedure for obtaining biostimulants from agroindustrial waste according to claim 1 characterized by the use of Bacillus licheniformis as a microorganism producing hydrolytic enzymes.
3. Procedimiento para Ia obtención de bioestimulantes a partir de residuos agroindustriales según reivindicaciones 1 y 2 caracterizado porque los residuos agroindustriales se componen de materia orgánica de alto contenido en nitrógeno proteico y con una alta susceptibilidad a ser hidrolizado.3. Procedure for obtaining biostimulants from agroindustrial waste according to claims 1 and 2 characterized in that agroindustrial waste is composed of organic matter with high protein nitrogen content and with a high susceptibility to be hydrolyzed.
4. Procedimiento para Ia obtención de bioestimulantes a partir de residuos agroindustriales según reivindicaciones 1-3 caracterizado por concentraciones del 0,5-2% de residuos agroindustriales y cantidades de CaCI2 a partir de 5 milimolar para favorecer Ia síntesis y estabilidad de las enzimas a producir.4. Procedure for obtaining biostimulants from agroindustrial wastes according to claims 1-3 characterized by concentrations of 0.5-2% of agroindustrial wastes and amounts of CaCI 2 from 5 millimolar to favor the synthesis and stability of enzymes to produce
5. Procedimiento para Ia obtención de bioestimulantes a partir de residuos agroindustriales según reivindicaciones 1-4 caracterizado porque en Ia segunda fase las condiciones óptimas de temperatura y tiempo oscilan entre 55-600C y 100-150 minutos, respectivamente. 5. Procedure for obtaining biostimulants from agroindustrial waste according to claims 1-4, characterized in that in the second phase the optimum temperature and time conditions range between 55-60 0 C and 100-150 minutes, respectively.
PCT/ES2006/000034 2005-01-28 2006-01-26 Method of obtaining biostimulants from agro-industrial residues WO2006082264A1 (en)

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WO2016005706A1 (en) 2014-07-11 2016-01-14 Prp Holding Use of an organo-mineral composition to increase assimilation of nutrients from the ground by the plant
CN110999920A (en) * 2019-12-16 2020-04-14 渭源县农业技术推广中心 Compound preparation for preventing and treating diseases and insect pests of angelica sinensis and reducing bolting rate of angelica sinensis as well as preparation method and application of compound preparation
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WO2013030422A1 (en) 2011-08-29 2013-03-07 Heineken España, S.A. Method for producing biofertilisers and biostimulants for agriculture and animal feeding
WO2016005706A1 (en) 2014-07-11 2016-01-14 Prp Holding Use of an organo-mineral composition to increase assimilation of nutrients from the ground by the plant
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CN110999920A (en) * 2019-12-16 2020-04-14 渭源县农业技术推广中心 Compound preparation for preventing and treating diseases and insect pests of angelica sinensis and reducing bolting rate of angelica sinensis as well as preparation method and application of compound preparation

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