MAAVi Innovation Center

Research Areas

SAR

How can the child know in advance the shape that fits every mold? What if we changed the shape of the different pieces?

This is an exagerated simplification of the existing relationship between Chemistry and Biology.

“Turning a problem into an opportunity”

Social awareness has been followed by a series of official policies aimed at reducing, reusing and recycling said by-products, creating a niche market for companies that are capable of leveraging on the value provided by that waste. 

The use of this waste may be quite straightforward, with not much added value, such as: Compost for the development of fertilizers or production of biofuels, biomethane, etc. But also highly technological, with a higher added value: Extraction of active biomolecules for the production of nutraceuticals; pectins for the food industry; production of nanocellulose for the cosmetics industry, textile applications, adhesives, etc.; extraction of hemicellulose for drug encapsulation, among others.

“MAAVi Lab Model”

Here at MAAVi we work hand in hand with large agricultural producers and the agroindustry. We evaluate their waste, both in terms of quantity and quality: Type of waste, annual volume, seasonality, origin of that waste, chemical composition, biological stability, potential proven applications, etc. 

For applications in the field of agriculture, we organize agronomic trials; however, for other applications unrelated to agriculture, we involve third companies with a potential interest in the products. 

All of it after studying the technical and economical viability of the project.

Research Areas

Phytochemistry

Plants are the largest biofactory of molecules known to date. According to estimates, over 400,000 plant species have been identified so far, which accounts for only 3% of all plant biodiversity. Any of those species is capable of producing from 5,000 up to tens of thousands of different compounds by itself. If we consider all the possible combinations between those compounds, we are literally speaking of a nearly infinite chemical space (thousands of millions of molecules). This chemical diversity is the key to plant survival throughout millions of years of evolution, and it is considered the largest source of active substances to which all kinds of biological properties are attributed.

In our Botany Department, we study and decode the chemical diversity of plants, to understand the interaction between different molecules. Thus, we can design new combinations targeting a broad spectrum of biological activities against numerous challenges.

The chemical space

The natural chemical space encompasses all the possible combinations of molecular structures found in nature. This is nearly infinite (thousands of millions of compounds), 95% unknown, and its largest known source of compounds are plants (over 70%). We develop a collaborative network with the largest centers dedicated to plant biodiversity in the world. 

We can extract molecules by applying an innovative amplification node (hipervínculo a apartado Nodo de amplificación de otra sección) to the plant matrix, which consists of a unique combination of technological and biotechnological sequential processes. We proceed to their identification and structural elucidation through combined chromatographic and spectrographic techniques (hipervínculo a apartado Analítica) and chemometric analysis. 

We define and isolate the marker compounds of the biological activity through advanced techniques based on bioassay-guided chemical fractionation, and analyze the results using chemometrics and artificial intelligence (hipervínculo apartado IA diferente al anterior)

We boost the power of nature through in vitro culture techniques, genetic enhancement and selective breeding of wild chemotypes.  

We increase the diversity and functionality by selecting the leading natural molecules, using SAR studies and artificial intelligence algorithms.

We optimize the biological and toxicological properties of the molecules resulting from the targeted design of multifunctional synergies.

The chemical principles supporting our product and the responsible use of natural resources are important to us.

Natural chemistry principles supporting the product

Our chemical processes are based on molecules that are, or at least may be found in nature.

Responsible Use

We use natural resources responsibly. And we do it on three different levels

How do we understand multifunctional synergies

A synergistic effect consists in finding a molecular combination that offers a superior biological effect compared to the one achieved by the simple addition of the different components. This is a solid approach applied in numerous areas of the Medicine and Pharmacology domains. We are applying the same approach to other areas, such as Agriculture, Aquaculture, Livestock Farming, the Food Industry and Cosmetics. 

We look for synergies to: Increase efficacy, lower application rates and active ingredient exposure levels, expand the “multitarget” spectrum of action, amplify action mechanisms, improve physicochemical properties, promote systemic activity and residuality, and meet any other requirements.

We are experts in the design of synergistic combinations of natural molecules. We are different in that (1) we study natural synergies and (2) we design exclusive combinations using an interdisciplinary approach comprising Botany, Microbiology, Chemistry, Biology, Bioinformatics, and Mathematics.

We begin by determining all of the chemical space contained within the plant. We use guided bioassays to perform chemical fractionation, in which each chemically characterized fraction is subjected to different biological challenges. In this manner, we can determine the function (synergistic, additive, cooperative, and/or antagonistic) of each chemical entity with regard to the crude extract’s activity levels. 

The key to our success is that we understand how to extrapolate natural synergies into new combinatorial matrices. We use the Gibbs Triangle to construct a matrix with over 100 secondary and tertiary interactions between the chemical entities of the chosen marker molecules based on sublethal concentrations (less than LD50 or EC50). Using a phenotyping analysis, the efficacy levels of each combination are compared to those of each individual molecule, taking into consideration the respective 50% and 90% efficacy rates. The synergistic effects are validated on three levels: isobolographic analysis, the Limpel equation, and the Chou and Talalay combinatory index.

Research Areas

Plant Physiology

Plants are just as important as any other living organism on the planet. Plants are essentially the reason that most organisms survive on Earth. They provide us with the oxygen we breathe and the food we eat. 

Plant Physiology is a field of Science that studies the structure and function of plants and their components.

Plant physiology is the study of all aspects of plant biology at the structural, molecular, and systemic levels, as well as their interactions with the environment (ecophysiology).

It combines information from a variety of scientific disciplines, including physics, chemistry, biochemistry, genetics, and molecular biology, to better understand how plants develop and survive within different environments.

It relies on a number of techniques and tools to assess a plant’s health and performance under a variety of cultivation or development conditions.

In agriculture, it is essential to understand how the plants of a specific crop function in order to analyze, recover, and enhance their performance and yield. 

Throughout their life cycle, plants are exposed to many kinds of stressors that jeopardize their ability to survive and, in the case of crops, result in yield losses. These stressors can be biotic (a pest or disease) or abiotic (a drought, a frost, for example).

We can more accurately design products targeted at avoiding, overcoming, and recovering from these stressors if we understand plant physiology. It is a component of what we do at Maavi.

How can we determine if a plant is stressed and what changes take place?

Environmental stressors such as high temperatures, frost, drought, changes in light intensity, salinity, nutritional deficiencies, heavy metals, herbicides, ozone, and so on affect plant function directly or indirectly, influencing fluorescence emission. Multiple techniques can be used to reveal response mechanisms, evaluate stress responses, and pinpoint specific pollutants and their source. 

Some of these techniques are:

• Chlorophyll fluorescence

• IRGA: infrared gas analyzer.

• Biochemical techniques: MDA, quantification

Research Areas

Microalgal biotechnology

Microalgae are unicellular plants that we turn into tiny natural factories, from which we extract millions of compounds with high biotechnological value.

Unicellular plants

Microalgae belong to the plant kingdom and, like higher plants, they can be autotrophic, cosmopolitan organisms that accumulate similar secondary metabolites in response to specific environmental conditions. But what characteristics have made them appealing in the area of biotechnology?

• Not being obligate autotrophs, because these microbes, in addition to photosynthesis, can use organic carbon sources, resulting in heterotrophic nutrition, or combine both methods to obtain energy, resulting in mixotrophic nutrition.
• They can tolerate extremely harsh conditions in terms of pH, salinity, and/or temperature due to certain physiological characteristics that enable them to thrive in extreme environments such as glaciers, deserts, or even unusual locations such as burial sites. 

Given the aforementioned, it is possible to direct the accumulation of secondary metabolites in microalgae with specific physiological properties, turning them into tiny biotechnological factories.

Secondary metabolites

Microalgae have different physiological reactions to different exogenous conditions.

The phenotypic behavior  of microalgae in response to external factors changes their phytohormone, pigment, and fatty acid content, as well as their biopesticide activity.

At MAAVi, we study how organisms behave in their natural environments by identifying and replicating under controlled settings the conditions that result in the activation of metabolic pathways for compounds of biotechnological interest.

Natural factories

At MAAVi, we value natural diversity; as a result, we prospect, characterize, and optimize the accumulation of bioactive compounds in order to find the best production system on a larger scale that balances the amount of biomass generated and the accumulation of metabolites of interest, at the lowest possible production cost. 

• Identifying bioactives in microalgae species 

We take into account the wide variety of microalgae that exist (over 30,000 species), which is why we favor strains of microalgae that have been largely unexplored, bearing in mind that they are not all the same and taking advantage of their phenotypic plasticity.

• High resolution characterization 

We can extract molecules by applying an innovative amplification node to the plant matrix, which consists of a unique combination of technological and biotechnological sequential processes. We proceed to their identification and structural elucidation through combined chromatographic and spectrographic techniques and chemometric analysis.  

We increase the diversity and functionality by selecting the leading natural molecules, using SAR studies and artificial intelligence algorithms.  

We optimize the biological and toxicological properties of the molecules resulting from the targeted design of multifunctional synergies.  

• Optimization with non-invasive strategies  

We seek to add value to what is natural in a sustainable manner. A strain must be able to maintain an equilibrium between its ability to generate biomass and accumulate metabolites of interest, while remaining scalable and profitable in terms of production processes. To do this, we focus on adaptive evolution strategies in the laboratory.

• Testing and scalability systems 

We have several 2L controlled culture systems (mixing, aeration, temperature, pH, and lighting), which enable us to produce enough biomass for later analysis, as well as 20L reactors that allow us to begin the production of cultures with which to inoculate our semi-industrial unit. 

The semi-industrial unit, on the other hand, includes different processes that allow us to increase the quantity of biomass in the cultures. These include open culture (775 L raceway) and closed culture, with tubular (700L), cylindrical and flat (500 L) photobioreactors, and 4 cylindrical systems (150 L each). Each PBR has sensors that monitor the pH and temperature of the culture as well as other control and monitoring parameters. This information can be electronically analyzed through a computer program, allowing for remote access to the data in real time for quick decision-making and regulation of growth conditions. 

It should be noted that our pilot plant consists of a system of independent feeding and harvesting pumps, which allow operation at different flow rates or operating regimes (continuous, semi-continuous or batch). Therefore, the structure of this design allows us to compare each system and look for the one that is most appropriate for each strain and target metabolite.  

Biotechnological value

Las microalgas, junto a las demás fuentes naturales con las que trabajamos en Maavi (4HEALTH) representan un patrimonio biológico importante. Desde ellas obtenemos moléculas con las que desarrollamos nuevos e innovadores productos para los distintos mercados a los que ayudamos en su transición hacia lo natural.

Y es que las microalgas presentan un sinfín de posibilidades y en Maavi estamos alertas a las nuevas tendencias investigativas para abordarlos y ensayarlos. Y es que uno de nuestros aspectos que nos diferencian de cualquier otro centro de investigación microalgal es que contamos con gran cantidad de plataformas de ensayo, donde realizamos desde ensayos in vitro hasta pruebas en campo. Además, nos apoyamos en nuestras redes de colaboración nacionales e internacionales, académicas, investigativas y comerciales, logrando así un escenario multidisciplinario a partir del cual generar nuestros desarrollos biotecnológicos.

Microalgae, together with the other natural sources with which we work at MAAVi (4HEALTH)constitute a significant biological legacy. They provide us with the molecules we need to create new, cutting-edge products for various markets as we support their transition toward natural alternatives.

This is due to the fact that microalgae offer infinite possibilities and at MAAVi, we are continuously on the lookout for new research trends to address and test. We also have a number of testing platforms where we can carry out everything from in vitro tests to field trials, which distinguishes us from other microalgal research facilities. Additionally, we rely on our academic, research, and business national and foreign cooperation networks to create a multidisciplinary framework from which to develop our biotechnological innovations.

Products

Among the products developed by MAAVi using microalgae, Seawex stands out as the only all-natural product based on the joint biostimulant action of seaweed and microalgae. It is an active source of natural phytohormones, such as cytokinins, auxins and gibberellins, but also of other components, such as free amino acids, vitamins, carbohydrates and antioxidants, which promote stress resistance and prepare the crop for periods of high energy demand or phenological stage changes. Its perfect combination optimizes all metabolic and physiological processes, maximizing crop yield.

Research Areas

Priming

Plants have developed their own defense mechanisms, similar to how humans have developed an immune system against pathogens. 

Do you agree that it is exciting to try to understand how an organism can spend its entire life in one location without having the choice of moving elsewhere to escape adverse weather conditions or pathogens?

Can we argue that plants have an immune system? Do plants have an immunological memory capable of recalling attacks that they have previously overcome? 

The Evolution of Plant Defense Systems

Priming is an evolution of a plant’s defense mechanism. 

According to Martínez-Medina and collaborators (2016), it is the physiological state in which plants are prepared or pre-conditioned for a more efficient activation of their defenses against environmental challenges, both biotic and abiotic.

How do plants train or develop their defense mechanisms?

In the natural world, when a plant senses a threat, it activates its defense mechanisms in response to the threat it detected. These stimuli can include a pathogen infecting plant cells, a beneficial organism interacting with the plant, mechanical friction with a human or animal, and so on. 

MAAVi conducts research to prepare plants in advance, before they detect a threat or attack, using natural molecules known as “elicitors”. We are talking about “elicited” or “primed” plants.

These elicitors will boost plant cell alertness, enabling them to trigger defense mechanisms more effectively when the real threat emerges. 

Would you like to know which kind of elicitors we research at MAAVi?

Plants can recognize certain molecular patterns and respond accordingly. These substances can be categorized as MAMPs, PAMPs, or DAMPs depending on whether they are derived from microbes, pathogens, or plant cell damage. 

Plants, in addition to these molecules, can react to certain chemical or environmental stimuli.  

Some natural compounds, such as oligosaccharides, glycosides, amides, vitamins, carboxylic acids, and aromatic compounds, have already been linked to the Priming phenomenon as growth boosters, activators, or inducers of plant defenses. 

In general, plants handle natural compounds better than most synthetic compounds analyzed. (Iriti et al., 2010; Noutoshi et al., 2012; Aranega-Bou et al., 2014).

As a result, at MAAVi Innovation Center, we look for these compounds within the primary natural sources that our solutions are based on: 

– from Botany, for example: Milsana (extracted from Reynoutria sachalinensis), oligogalacturonides, burdock fructooligosaccharides…

– from Microalgae, carrageenans, fucans, ulvans, laminarin…

– from Microbiology, flagellin, harpin-proteins, lipopeptides, β-glucans, chitosan, chitin, ergosterol…

Would you like to know how we uncover novel Priming elicitors?

Research Areas

Enthomology

Insect Breeding and Targeted Insect Testing Area.

MAAVi has pest insect breeding systems in place that allow us to undertake biopesticide trials throughout the year.

Discover where we do research with our “allies” to develop our solutions..

Enthomology is the scientific study of insects, which account for two-thirds of all documented species worldwide (1.3 million in total). 

This area of MAAVi is in charge of breeding the insect pests that are the target of our biocontrol solutions. 

Once we provide optimal conditions, insect populations become established in our breeding boxes, with some species reaching densities of up to 10,000 individuals.

Then, as part of our Hit 2 Lead process, we subject them to several testing phases.

Candidate Primary Trials:

• Contact mortality on the pest’s stage of development that is most aggressive for the crop.

• We set efficacy standards of 75% or above for candidates to advance to the next round of trials.

• Trials performed in vitro on plant material.

Secondary trials of Hits, Leads, Synergies and Preformulates or pest behavior trials:

• Settlement inhibition

• Oviposition inhibition

• Feeding inhibition

• Performend “in planta”

• Dose–response assessment

Therefore, we conduct studies at all phases of pest life cycles in order to define modes of action and identify population-reduction mechanisms. 

En MAAVI contamos con las siguientes instalaciones de cría y ensayos:

• We have the following breeding and testing facilities at MAAVi:

• 200 m2 of greenhouses divided into:

  • Ten 20m2 sub-units with overhead ventilation, air extractors, and temperature control.
  • Two 20m2 growth chambers with temperature, humidity, and photoperiod control.

• 80 m2 of in vitro and in planta testing area.

We conduct over 600 trials per week

Research Areas

Phenotyping

The term phenotype refers to any observable characteristic of a plant, such as its height, number and size of leaves, level of chlorophyll, etc. This characteristic or attribute is the result of the interaction between the genotype of the plant and its environment throughout development. Plant phenotyping analyzes how a plant’s genetic make-up interacts with its environment during development to produce specific traits or  characteristics. Our platform LINNA, enables precise mass phenotyping of plants through the use of non-invasive approaches based on digital images and sensors. Digital images obtained with various sensors (hyperlink to click 2) are processed at the pixel level and transformed into binary code by our artificial vision system, translating nature into data understandable by LINNA® prediction models. This allows us to evaluate, for example, the response of plants under different types of environmental stress, as well as the influence of biostimulants and bioherbicides on their development and growth. The evaluation of individual qualitative and quantitative attributes serves as a basis for assessing complex characteristics.

Hierarchical cluster generated from chlorophyll fluorescence images. The most important photosynthetic response metrics in plants treated with a herbicide (glil) or not treated (ctrl) are shown.

Would you want to learn more about MAAVi’s phenotyping platform?

We have a phenotyping platform at LINNA which offers automated measurements using multiple sensors. This high-performance integrative phenotyping integrates several automated, simultaneous, and non-destructive plant evaluation approaches. This provides information about the growth, morphology and physiology of plants.  The high performance allows us to analyze plants on a broad scale and use statistics to identify subtle yet significant variations between different treatments or genotypes.

Images obtained with visible red-green-blue (RGB) sensors are used to measure visual parameters in the LINNA platform; hyperspectral sensors are used to obtain information on the internal structure of the plant and its content of compounds of interest (pigments, proteins, carbohydrates, water, nitrogen, lignin, and so on); and chlorophyll fluorescence sensors are used to determine photosynthetic efficiency. Given that phenotyping entails a cascade of events affecting gene expression (transcriptome), data from the LINNA transcriptomics platform are integrated into the phenotyping process. Through data analytics and artificial intelligence-based algorithms , LINNA is able to collect key data for the development of natural solutions at MAAVi.

This data integration enables us to predict or identify the variables causing variations in crop physiology and morphology. For instance, it helps in the identification of a herbicide’s mode of action, changes that lead to an improvement in crop yield, and the subsequent development of new products.

Simultaneously, the system reduces research time, increases accumulated knowledge, and, most importantly, reduces failure by making a faster and more accurate transition to natural molecules and compounds to replace pesticides with biopesticides, artificial preservatives with natural ones, or find new protein sources for food.

Would you want to learn more about MAAVi’s phenotyping platform?

RGB Images 

It is used to quickly and accurately assess growth rate, leaf area, projected area, leaf greenness, leaf damage, senescence, among others. It is also possible to create growth curves using models and analytics.  

Chlorophyll fluorescence images.

PAM fluorometry and high-sensitivity CCD cameras used for chlorophyll fluorescence imaging provide quantitative information on the quantum yield of photosynthetic energy conversion. This technique also allows for the separation of photochemical and non-photochemical events in leaves. This sensor offers 17 essential chlorophyll fluorescence metrics in addition to chlorophyll fluorescence imaging. This method may be used to collect images of photosynthetic activity and associated spatiotemporal oscillations. The fluorescence parameter Ft is continuously monitored. Fo and Fm are evaluated following adaptation to darkness. Using the saturation pulse approach, various characteristics relating to fluorescence quenching are derived from these. Fv / Fm (maximum quantum yield of PS II following darkness acclimation), Y(II) (quantum yield of PS II during illumination), regulated energy dissipation quantum yields Y(NPQ) and unregulated Y(NO), apparent electron transport rate (ETR), non-photochemical quenching (NPQ), and photochemical quenching (qP) images are obtained. 

Hyperspectral images.

We have two cameras.  The Specim FX10 operates in a line scan mode in the 400-1000 nm visible and near infrared (VNIR) range, and color optimized in the 400-780 nm region. Specim FX17 gathers hyperspectral data in the near infrared NIR range (900 to 1700 nm) while operating in line scan mode. Our analytical program allows us to calculate around 150 metrics such as NDVI, GDVI, NDLI, WI, NDWI, Crt, and LSI.

We may examine biochemical properties in plants using hyperspectral technologies, such as measuring water content, changes in pigment composition, moisture content, proteins, carbohydrates, sugars, and many more. We can also determine the extent of damage caused by insects and bacteria.

Research Areas

Formulation and systemicity

We combine ingredients and insights to create novel solutions from natural molecules and microorganisms.

We combine molecules with complex mixtures of multiple chemical identities derived from natural sources. We also design solutions that contain helpful microorganisms including bacteria, fungi, microalgae, and viruses. In this area, we develop exclusive formulas that are stable, compatible, persistent, and with improved efficacy for application in Agriculture, Livestock, Food Industry, Aquaculture, and Cosmetics.

We combine molecules with complex mixtures of multiple chemical identities derived from natural sources. We also design solutions that contain helpful microorganisms including bacteria, fungi, microalgae, and viruses. In this area, we develop exclusive formulas that are stable, compatible, persistent, and with improved efficacy for application in Agriculture, Livestock, Food Industry, Aquaculture, and Cosmetics.

Natural product formulation has the significant problem of integrating a broad and complex chemical diversity with substantial variances in their physicochemical qualities.  It is an important step in the product development process that may make-or-break a project. 

We begin the formulation development process with an in-depth understanding of the chemistry underlying each extract, synergistic combination, specific molecule, or microorganism. Each project and product requires its own formulation and comprehensive analysis. As a result, formulations may be developed employing a variety of technologies, including drying methods (spray-drying, freeze-drying), emulsions, emulsifiable concentrates, and concentrated suspensions, among others. 

The challenge in microbiological formulations is to keep cells viable at high concentrations while also preserving the production capacity of molecules of interest, because cells that are not properly formulated can be exposed to unfavorable conditions during product production, distribution, and storage.

Our formulation technology innovation focuses on the following areas:

• Research of new multifunctional adjuvants and/or co-formulants of natural origin.
• Development of encapsulation and/or protective technologies for microorganisms and sensitive molecules. 
• Development of controlled release matrices.
• Development of natural molecules that act as selective carriers of active substances and which allow: (1) Penetrability, (2) Translaminarity, (3) Translocation and (4) Systemy.

Our formulations are developed with the desired characteristics of a product in mind. For instance, we can offer multiple strategies/solutions such as adhesive capacity, rain resistance, UV light protection, and phagostimulants, among others. 

Our main objective is to maximize efficacy while also ensuring formulation stability. Among our processes, we implement:

• Stability tests
• Accelerated stability tests (CIPAC MT 46.4). Their objective to determine if the shelf life of the formulations is met under normal storage conditions.
• Real-time stability tests (storage stability-shelf life). Performed under predetermined storage conditions, for the duration of the product’s lifespan and in its original packaging.
• Low temperature stability tests. The objective is to determine how storage at low temperatures affects the formulation.
• Packaging compatibility tests.  To verify that no unfavorable interactions occur between the formulation and its primary container.
• Homogeneity tests (5-batch analysis)
• Chemical compatibility tests with other products

Assessment testing of the formulations’ physicochemical behavior (solubility, dispersion, suspensibility, wettability, etc.).

• Effect residuality tests.

All of this allows us to guarantee that the products’ active and inert components satisfy technical standards and may be registered in line with the sector’s and current legislation.

Research Areas

Analytical Chemistry

We look for molecules, not extracts

Natural extracts are comprised of a huge number of molecules, the concentration of which can greatly vary depending on the type of plant, age, season of harvest, climate, etc. In this area, we monitor the active molecules to guarantee that the efficacy of our products is homogeneous.

Research Areas

Toxicology

At MAAVi Innovation Center, we are particularly mindful of the safety of the products that we manufacture. Toxicity is a dose-dependent phenomenon even in nature. Isn’t it fascinating? Keep reading

As a result, studying the toxicity of the products that we develop is a critical step in the manufacturing process. We employ cell cultures to determine the toxicity of the compounds we work with, as well as the levels at which they may be toxic. 

MAAVi has a laboratory devoted to toxicological testing at the cellular level. The trials are conducted using a liver cell line, namely HepG2, cultivated in multiwell plates. This cell line was chosen since the liver is the first organ to be affected by the toxicity of any foreign substance to the body, and it has the greatest detoxifying ability. These tests are performed by evaluating a number of escalating dosages of the compound and using the XTT technique to measure the cytotoxicity the compound causes in the cells that receive each dose. The algorithms suggested by the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) Acute Toxicity Working Group (ATWG) are then used to create a correlation between the components’ IC50x cytotoxicity and acute oral toxicity LD50 without the need of animal testing.

Research Areas

Scalability

The MAAVi Innovation Center is renowned for its scale-up infrastructure that enables the industrial replication of laboratory-tested processes. This potential for realistic scalability allows us to make much safer and more accurate decisions about a process’s exploitability, and takes us even closer to meeting the needs of the industry. 

We have pilot plants with extraction tanks up to 3,000 liters, vacuum concentrators with 300 liters/hour of water and 800 liters/hour of ethanol concentration, fermenters that can hold 100 and 3,000 liters, centrifuges with a capacity of 250 liters/hour, membrane plants that can process up to 100 liters/hour (depending on pore size), and mixing and formulation tanks with a 500 liter capacity and high shear and emulsion systems.

Every process must be designed and tested in the lab, but if it cannot later be replicated at an industrial scale, it is absolutely useless. This is why we, at MAAVi, place such a high importance on our pilot plants. Based on the knowledge gained during the laboratory design of a process, we may execute it at the industrial level, providing us with vital information that will allow us to make investment decisions for the industrialization of a process. Parameters like temperature, viscosity, calorimetric properties, shear rate, gas exchange capacity, or the design of the facilities that will allow the laboratory-developed products to be manufactured, considering the geometry of the deposits, the heat exchange surfaces, the torque of the agitation motors, or the design of the agitators allow us to determine whether a process is technically feasible on an industrial scale, estimate the volume of investment that said process may require, how much energy it will consume, what volume and type of waste will be generated, and what automation capacity could be implemented. In addition to determining the workforce and its characteristics related to the process, what services will be required (steam, compressed air, gases, sterility level, hot water, chiller water, osmosis water, etc.), the required space or footprint for its implementation, the carbon footprint generated, and so on. In short, any information and experience that is as near to reality as possible can aid in estimating the ROI of a new process, as well as other environmental factors that are currently having a major impact.

Research Areas

Pyrolysis

Human observations of natural phenomena, such as wildfires, have led to the identification of mechanisms involved in the synthesis of molecules with high value in various sectors, such as molecules with biopesticide activity, biostimulant activity, or other qualities. These processes can be integrated under the umbrella of pyrolysis.  For more information.

Our pyrolysis technique causes the thermal breakdown of a natural matrix under partial absence of oxygen, so that the components present are degraded by heat rather than combustion processes. The process is carried out in a system designed by Kimitec, which yields two types of products: on the one hand, a solid residue that remains after the procedure, known as biochar, and which has interesting characteristics that contribute to healthier soils. The volatile components created in the process, on the other hand, are collected, and it is in this fraction that molecules with strong activity are obtained at extremely low dosages.

This is done at relatively moderate temperatures (below 300 °C) in comparison to standard pyrolysis processes, which are conditions that most closely resemble natural fires. At these temperatures, the most common reactions are depolymerization, oxidation, dehydration, and decarboxylation. Additionally, the majority of the affected components are all those with low molecular weight, with amorphous cellulose and hemicellulose standing out among those with high molecular weight. Sound interesting? Keep reading.

Research Areas

Microbiology

Microorganisms and their entire metabolic diversity have evolved over the last 3.8-4.3 billion years, to perform very different functions in all terrestrial ecosystems. 

In the area of microbiology, we study the genetic and metabolic potential of all these microorganisms in order to guide them to produce or perform the action of interest. 

Fungi, bacteria, or viruses (such as bacteriophages) are used to produce compounds with plant biostimulation or biocontrol activity, as well as food additives, antibiotics, and microbial cells. As a consequence, the microorganisms we work with at MAAVi could be considered metabolic specialists, capable of flourishing in a wide range of substrates, synthesizing molecules of interest with high output, or biostimulating the plant when used as probiotics.

Microorganisms, prolific producers of bioactive metabolites (or molecules). We identify and optimize the metabolic capabilities of microorganisms. Our goal is to produce molecules of interest.

Fermentation

Fermentation is used to obtain microbial cells to develop probiotics or to produce specific molecules of interest. In this specific case, the fermentation process applies to any large-scale microbial process, whether or not it is fermentation in the biochemical sense. In fact, many of the fermentations we perform at MAAVi are aerobic. Fermentors vary in capacity from 1 to 5 liters to 3,000 liters. Fungi, bacteria, and bacteriophages are the most common microbes used in our fermentations. We perform both liquid and solid fermentations with various plant materials as a matrix. 

Microorganisms must exhibit certain traits in order to be used in industrial-scale fermentations: 1) It must first be able to develop rapidly and synthesize the product in either a liquid or solid culture, and 2) it must be able to grow in a cost-effective culture medium. We use waste by-products from other industries in many of our fermentations, in collaboration with our revaluation vertical.

Our work in the area of fermentations

Our work in the area of fermentations

We integrate information from microbial genomics, culturomics, and bioinformatics to perform microbial fermentations with the goal of getting metabolites of interest. 

To select a suitable microorganism for fermentation, an in silico search is carried out within the more than 10,000 genomes of bacteria that we have in the database. The microorganism with the ideal genetic potential for producing the desired molecule can be predicted using this search. 

Once the microorganism has been defined in silico, we either acquire it from our own collection of more than XY microbial strains (which can generate different compounds with the potential for biocontrol or biostimulation) or from research institutions with which we frequently collaborate. 

Following that, small-scale microfermentations are carried out to maximize the production of the desired metabolite. We can perform more than 1,000 microfermentations per week thanks to the system developed at MAAVi. 

Chromatographs are used to assess these microfermentations by tracking the metabolite and its concentration in different settings. The bioreactors with the largest capacity are used once the production conditions for the metabolite have been determined. We can perform more than 5,000 fermentations annually in our 20 bioreactors, which range in capacity from 1 to 5 liters.

Probiotics

The importance of understanding and maintaining the microbiological homeostasis of plants. Is it a plant, an animal or a holobiont? In nature, plants are never alone, they coexist in close proximity to bacteria, fungi, viruses, and other microorganisms (what is known as the plant microbiota). Because of this, at MAAVi, we regard plants as members of holobionts, which is made up of a multicellular organism (the plant) and all of its ancillary microbes. We restore the microbiological equilibrium that plant systems have drastically deviated from as a consequence of extensive farming.

More info on how do we operate in this area

It is important to understand and maintain the plant’s microbiological homeostasis, in which the plant is in harmony not only with its surroundings but also with its microbiota, (what is known as eubiosis). Today, this equilibrium has been disrupted as a result of the unrestricted use of chemicals. This vertical’s objective is to restore plant systems to the microbiological equilibrium from which they have greatly deviated due to extensive farming. In order to achieve this, we choose, cultivate, and develop  communities of probiotic microbes that biostimulate plant development, aid in mineral nutrition, and mitigate biotic and abiotic stress. We rely on resources like ecology, microbial genomics, and metagenomics to accomplish this.

Genomics and Artificial Intelligence. Our understanding of it at MAAVi.

Microbial genomics, in conjunction with artificial intelligence, allows us to understand the full metabolic potential of the microorganisms we have at MAAVi, as well as the full metabolic potential of microorganisms whose genomes are available in open access collections. This tool is very useful in three areas:

1. When developing a probiotic or a microbial consortium with biopesticide activity, because it will identify microorganisms with a potential for the desired activity and will also create synergies in a consortium. 
2. To find microbes that produce bioactive molecules.
3. It is also very helpful when setting up precision fermentations where the culture medium and fermentation parameters are pre-determined. 

Many of the microbial inoculants available today yield highly inconsistent results, based on the plant and environment in which they are used. This is due to the microbial inoculant’s failure to colonize and establish itself in the growth medium in order to serve its purpose. The combination of metagenomics and artificial intelligence enables us to determine whether the applied microbial product was successful in establishing itself and changing the microbial community. Furthermore, by examining the microbial communities of more and less productive crops, we can find important species for microbial community structure and function, thereby improving plant productivity. This helps us in developing prebiotic products capable of promoting crop beneficial communities.

Use of metagenomics and microbial ecology to develop pre- and probiotics for crops and soil-directed products.

In nature, microorganisms never survive in isolation; rather, they exist in groups of microbial cells known as microbial communities that interact with one another, with other organisms, and with their surroundings. And we need to take that into consideration if we want to study them and understand their mechanisms. Additionally, the soil is the terrestrial ecosystem that presents the greatest biodiversity. In a typical edaphic bacterial population, there are between 107 and 109 cells per gram of soil, and anywhere from 20,000 and 40,000 distinct bacterial species may be present. These microbial communities play an important role in the carbon and nitrogen biogeochemical cycles, in the preservation of natural ecosystems, and in agricultural production.  As a result, studying these edaphic microbial communities is critical when developing an effective product that works in the rhizosphere (edaphic section under the influence of the root, where numerous processes promoted by microorganisms take place). Many rhizosphere-targeted products incorporating microbial consortia, for example, generate very inconsistent outcomes, depending on the plant, the habitat, and the soil in which they are used. This is due to the microbial inoculant’s failure to colonize and establish itself in the growth medium in order to serve its purpose.

Because of its enormous diversity, soil appears to be a perfect ecosystem for researching microbial ecology; however, it is also likely the medium that poses the most technical challenges to microbiologists. This is due not only to the magnitude of their microbial community and the vast number of species present, but also to the medium’s multiphasic character and the fact that only 99% of the microorganisms in these microbial communities can be isolated and cultivated on standard cultivation mediums. This is why molecular approaches to microbial community research are so important. Environmental genomics, also known as metagenomics, entails the extraction and sequencing of all DNA from the microbial community being studied. By studying these genes we can learn how these microbial communities operate as well as their phylogenetic composition. Massive sequencing techniques enable us to analyze measurable changes in response to environmental variations, both naturally occurring and induced by the application of a microbiological product, allowing us to gain insight into all of the soil’s hidden diversity. In this way, at the MAAVi Innovation Center (in collaboration with Biome-makers), we conduct microbial ecology studies in order to develop microbial consortia for crop- and soil-directed applications, as well as analyze whether the microbial consortium is capable of colonizing the soil once introduced, and what physicochemical or microbiological characteristics of the soil have been decisive for this colonization.

The 16S gene

Universal phylogenetic marker

The sequencing of the 16S rRNA, a tiny subunit ribosomal gene that provides information about the kinds of microorganisms present in a specific habitat, is a major factor in determining the extent of this hidden diversity. One of the key areas of research over the past 30 years has been the study of this gene, which has changed how microbial ecology is conducted. We are able to determine evolutionary relationships from the ribosomal gene sequences because:

• They are essential for the synthesis of proteins and are found in all organisms, frequently with multiple copies.
• The protein’s primary and secondary structures, which contain both variable and other highly conserved regions, make it possible it to design both broad-spectrum and specialized primers.
• Because their sequences have stayed remarkably consistent throughout evolution and because changes to their sequence appear to happen very slowly, they can be thought of as molecular clocks.

These traits help us to recognize phylogenetic variants and identify shared evolutionary origins and developmental patterns. Conceptually, random tree structures are simple. 16S rRNA sequences from various species are compared to one another, and any variations are evaluated and used as a gauge of how far apart they evolved from one another. 

*Tamames et al. BMC MICROBIOLOGÍA 2010, 10:85. Environmental distribution of prokaryotic taxa

Phages

Bacteriophages, also known as phages for short (from the Greek phagein, which means “to eat”), are the most prevalent type of life on Earth and bacteria’s primary enemies. They are 100 times smaller than bacteria and are in charge of controlling harmful microorganisms. Every 48 hours, they replace half of the world’s bacterial population; the human gut alone contains one trillion phages, but they can also be found on the skin, in food, and crops.

Bacteriophages are naturally occurring microorganisms that are capable of infecting and killing bacteria. Discover how they behave.

The most common mode of action of phages against bacteria is bacteriolysis, which occurs at the end of the phage lytic cycle. (Figure). 

In order to reproduce, phages must first enter the host bacterial cell. Cells with a cell wall, like most bacteria, are infected differently than animal cells without a cell wall. As a result, the phages adhere to particular receptors on the exterior of the bacterial cell, creating a hole in the bacterial membrane through which they inject their genetic material. The phage DNA self-replicates inside the infected bacterium and is then integrated into bacterial infectious particles produced from chemicals isolated from the bacterial host. The formation and release of phage particles allows the phage to attack more bacteria in an exponential pattern, resulting in a faster killing impact than other alternatives.

There are several advantages to using phages as bacterial biocontrol agents. They are bacterium-specific, so other strains similar to the pathogenic strain are unaffected, and they have fast mutation rates, enabling them to adapt quickly to any resistance. MAAVi develops and formulates phages on a big scale to combat various bacterial diseases such as those produced by the pathogen Erwinia amylovora.

Curiosities: Did viruses invent DNA? Read all about it

In the area of symbionts, we co-culture symbiotic endophytic microorganisms with their host plant species, both in vitro and on a large scale. This one-of-a-kind system mimics how symbionts interact in nature and has two advantages: i) it enables us to mass-produce symbionts that support plant development; and ii) it allows us to study the molecules produced by the molecular interaction established between the two species. Many of these molecules have varied effects on the plant, such as biostimulation and defense activation, and may be useful in different projects.

Curiosities: How can fungi aid in the battle against climate change? Read all about it

Research Areas

The natural way to overcome diseases

Plant diseases have traditionally been tackled by the widespread use of biocidal agents, which have been shown to have negative impacts on both the environment and the health of humans and many other living species, such as insects. Their population has been significantly diminished on a global scale owing to the misuse of these hazardous compounds, particularly bees, who are required to pollinate thousands of crops throughout the world. 

At Kimitec, we have an area in charge of identifying and evaluating different candidates obtained from nature against plant pathogens such as fungi or nematodes, in order to find natural solutions with the same efficiency as synthetic chemistry, to combat the diseases that these cause on crops for farmers all over the world.

Would you like to learn more about phytopathology?

Phytopathology is the study of plant diseases. It analyzes the infectious organisms that can cause them, such as fungi, viruses, bacteria, and nematodes, as well as the abiotic factors, with the exception of pest insects researched in entomology.

We fine-tune and conduct bioassays with multiple pathogens, mostly fungi and nematodes, and analyze them using our Hit 2 Lead approach.

We also carry out diagnostic procedures to identify diseases observed in the field during trials or affecting producers with whom we collaborate.

Would you like to learn more about bioassays and diagnostic procedures?

Research Areas

Molecular Biology

Molecular biology studies the structure, function, and composition of biologically important molecules in order to understand the processes that occur in organisms. These molecules are mostly nucleic acids (DNA and RNA), which contain an organism’s genetic information, and proteins, which perform vital functions for that organism (structural, transport, defense, and so on).

The study of these molecules has made significant progress with the introduction of techniques like massive sequencing, which allows us to evaluate the nucleic acids contained in a cell, tissue, organ, or complete organism in depth and on a large scale. 

The advancement of molecular biology has also enabled the development of disruptive technologies such as RNA interference, which is based on the discovery that earned Andrew Z. Fire, Craig and C. Mello the Nobel Prize in Medicine and Physiology in 2006.

Would you like to know more about it?

All living beings’ genetic information is stored in nucleic acids, which are formed by the union of distinct “pieces” known as nucleotides (equivalent to the letters of an alphabet). Sequencing allows us to decode these letters in order to understand the genetic information of every living organism.

Massive sequencing, or NGS for its English acronym “Nest Generation Sequencing,” comprises all types of large-scale and parallel nucleic acid sequencing, considered the most advanced genetic research technique available today.

Do you want to know more about the sequencing techniques that we carry out at MAAVi?

With massive sequencing, we may discover something as basic about an organism as its genome (the whole of its DNA), or its transcriptome (the collection of all RNA molecules). Additionally, the distribution of bacteria and fungi in an environmental sample can also be precisely mapped through metabarcoding, and their genomes can be reconstructed through metagenomics.

At MAAVi, we gather all of this molecular data using the Illumina NovaSeq 6000 system, the most reliable, scalable, and comprehensive sequencer on the market, which produces up to 6 TB of data per study and enables us to obtain accurate and comprehensive results. 

A significant computational advancement is needed to analyze the vast quantity of data generated by the massive sequencing studies. The use of bioinformatics is essential for this purpose.

RNA interference natural

NA interference (or RNAi) or post-transcriptional gene silencing is a universal biological response to double-stranded RNA (dsRNA) molecules that affects gene expression and protein coding. RNAi is currently employed as a tool determine the function of any gene, but it has made significant progress as an emerging technology for the design of new drugs. MAAVi collaborates with Sylentis, a PharmaMar group company that is a pioneer in the development of innovative medicines based on RNA interference (RNAi) technology, to carry out the rational development of new plant protection products.

Would you like to learn more about RNA interference?

RNAi pathway triggers include viral-derived double-stranded RNA, abnormal transcription of repetitive regions in the genome such as transposons, and pre-microRNAs. A simple model of the RNAi pathway consists of two phases: the first, in which the triggering double-stranded RNA is processed by RNase II Dicer enzymes into short double-stranded small interfering RNA (siRNA). In the second phase, the generated siRNAs are loaded into a molecular complex called RISC (RNA-induced silencing complex) and a single-stranded RNA is created that hybridizes with the target messenger RNA, inducing gene silencing. This gene silencing is the result of nucleotide degradation of messenger RNA by the enzyme RNase H Argonaut. 

This natural mechanism can be experimentally triggered by the exogenous introduction of long double-stranded RNAs (dsRNAs) or siRNAs, typically between 20 and 24 nucleotides, with the goal of not only conducting functional genomics studies, but also developing therapies for the pharmacological treatment of viral infections and even many types of cancer. At MAAVi, we use this technology to develop biopesticides and bioherbicides.

Research Areas

Bioinformatics

The field of bioinformatics focuses on the research and development of statistical and computational tools for the analysis and use of biological data. These tools use computational resources to solve biological problems that extend beyond the limits of human understanding, such as the extraction of information from biological data produced on a large scale by massive sequencing. Bioinformatics is especially helpful for comparing DNA, RNA, and protein sequences both within and across species in order to study the evolutionary connections between organisms and try to ascribe functions to particular genes and proteins. In that sense, bioinformatics is the linguistics of molecular biology, analyzing patterns in DNA, RNA, and protein sequences in the same manner that linguistics examines patterns in language.

Research Areas

Artificial Intelligence

The objective of Artificial Intelligence is to develop intelligent machines, particularly computer programs, capable of performing complicated tasks akin to those performed by humans, such as reasoning or learning. Nowadays, artificial intelligence is able to play chess, communicate concepts through drawings, operate self-driving vehicles, and even write stories.

Artificial Intelligence is capable of detecting patterns that are not evident to humans with remarkable precision, thanks to the performance of routine tasks in which large amounts of data are analyzed reliably and without getting tired. MAAVi’s artificial intelligence platform incorporates machine learning and deep learning.

More info on machine learning and deep learning

Both machine learning and deep learning are Artificial Intelligence subfields that differ in how they do automated learning. In this way, machine learning allows the computer to learn on its own by providing a set of rules to an algorithm, which it applies when relevant data are found. As a result, it is capable of adjusting existing rules and developing new ones in order to increase its success rate. 

Deep learning, on the other hand, varies from machine learning in that it incorporates more complicated and detailed automated learning. Deep learning algorithms are built on artificial neural networks, also known as simulated neural networks, which imitate how the human brain operates.

At MAAVi, we use both machine learning and deep learning, integrating regression, random forests, dimensionality reduction, and unsupervised clustering models for tasks like characterizing an organism’s transcriptomic response to the application of a candidate, or exploring the chemical space of any given candidate. In addition, graph neural networks and encoders-decoders are coupled with these machine learning models.

Research Areas

Phytotrons LINNA

We have 6 growth chambers, or phytotrons, in our LINNA platform where plants can be grown under controlled conditions. They are used for both seed-grown and in vitro plants. We set up each of these growth chambers with the ideal conditions for the target crop under study. We may also recreate varying or extreme weather conditions that are characteristic of specific regions in order to understand how diverse weather conditions around the world impact crop growth and development.  For instance, heat stress, water stress, low light, etc. All of this is dependent on the specifics of each trial, which are critical in the development of biostimulants, bioherbicides, and biopesticides.

Would you like to learn more about our controlled conditions growth chambers?

Under controlled growth conditions, several factors are configured, monitored, and recorded in real time and historical data. We may adjust the photoperiod length, humidity, CO2 concentration, and temperature. Each phytotron is 24 m2 in size and can hold more than 2,000 plants. They are fully adaptable, where the height between shelves may be adjusted to different layouts to carry out varied tests, ranging from herbaceous to taller plants.

Would you like to know which conditions we can simulate in our phytotrons?

The temperature range is 5 °C to above 40 °C. The range of humidity is 10% to 95%. It is possible to maintain CO2 levels between 100 ppm and 3,000 ppm.

Research Areas

Trial area

Our solutions are tested in the “lungs” of the MAAVi Innovation Center.

The analysis of the behavior of our solutions under greenhouse testing conditions is one of the key elements in our research process. MAAVi has green lungs, check it out

At MAAVi, we conduct “in planta” testing in boxes and growth chambers where we have complete control over the growth conditions. This not only enables us to organize testing throughout the year, but also to know how our solutions will perform under realistic climate conditions.

• We germinate and grow healthy plants in two seedbed sections and a healthy plant chamber.
• We have 30 boxes for testing biopesticides, probiotics, and biostimulants in addition to 5 boxes for disease assessment. 
• 12 growth chambers or phytotrons. 

With all the facilities combined, we can accommodate more than 20,000 plants in total. Do you wish to learn how they are distributed? Keep reading!

Healthy plants Chamber

This 86 m2 unit has temperature, humidity, and photoperiod control systems, allowing for year-round plant growth. LED lighting produce red and blue wavelengths that are essential for photosynthesis and plant development. These facilities allow us to produce a large number of healthy, high-quality plants that are free of pathogens and bacteria. The chamber includes 60 shelves spread at four levels of height, allowing for a capacity of more than 3,000 plants and a rotation of more than 1,200 plants every two weeks. 

Plants are sent from this unit to the Pests, Phytopathology, and Semi-field departments for formulation validation testing, as well as the Entomology department for use as insect meal. This chamber provides the main horticultural crops to the various units, which include beans, tomatoes, peppers, eggplants, cucumbers, and zucchini. However, we also grow agronomically important plants from other regions, such as tobacco, cotton, soybeans, and sunflowers. 

Semi-field area

MAAVi has more than 3,100 m2 of facilities organized into several units to respond to the rising demand for testing and thereby be able to respond to the present agricultural difficulties.

• Seedbeds for the year-round planting and development of healthy, high-quality plants. 
• Greenhouses for testing biostimulants and probiotics. 
• 35 boxes (20–25 m2) for research on biopesticides and the assessment of formulations’ efficacy against various pests of agronomic importance. They all have special features that allow working in environments free of external or cross contamination. 

We have state-of-the-art facilities equipped with cutting-edge technological equipment that allow us to get the most out of our research. As a result, the greenhouses are outfitted with shading meshes that allow us to reduce the temperature inside, evaporative cooling panel systems that allow us to regulate the temperature and humidity of the compartments, greenhouses with lateral and overhead ventilation, irrigation by nebulizers or floodable tables, and anti-insect meshes that provide advanced control of thrips and insects of similar size. 

Phytotrons

We have 12 growth chambers, or phytotrons, where plants can be grown under controlled conditions. They are used for both seed-grown and in vitro plants. We set up each of these growth chambers with the ideal conditions for the target crop under study. We may also recreate varying or extreme weather conditions that are characteristic of specific regions in order to understand how diverse weather conditions around the world impact crop growth and development.  For instance, heat stress, water stress, low light, etc. All of this is dependent on the specifics of each trial, which are critical in the development of biostimulants, bioherbicides, and biopesticides. 

Under controlled growth conditions, several factors are configured, monitored, and recorded in real time and historical data. We may adjust the photoperiod length, humidity, CO2 concentration, and temperature.

Our AI or LINNA area comprises six full growth chambers, each of which is 24 m2 in size and can hold more than 2,000 plants. They are fully adaptable, where the height between shelves may be adjusted to different layouts to carry out varied tests, ranging from herbaceous to taller plants. The temperature range is 5 °C to above 40 °C. The range of humidity is 10% to 95%. It is possible to maintain CO2 levels between 100 ppm and 3,000 ppm.

We have six phytotrons more distributed by work areas:

• A cutting-edge walk-in phytotron that allows for exact control of radiation, temperature, relative humidity, and CO2 levels, which supports growth of both small and tall plants. 
• A walk-in chamber equipped to operate with high radiation and a large number of plants, allowing us to work under varying conditions to achieve specific goals. 
• Three PLH-LED phytotrons with specialized LED tubes for plant growth, AR12 spectrum. 
• A PLH-LED phytotron powered by red and blue LEDs. T

All of these chambers are software-controlled, allowing us to monitor a wide variety of parameters needed for scientific trials.

The combined capacity of these six units is 1,000 plants.

Research Areas

Hydrolysis

At Kimitec, hydrolysis technology has become one of the most important sources of biostimulants for crops

Hydrolysis is the process of breaking down large molecules into smaller components that may then be metabolized by other organisms. Protein hydrolysis is widely employed in agriculture to extract free amino acids or short chains of these, known as oligopeptides, which have powerful biostimulant properties.

There are three forms of hydrolysis: chemical hydrolysis, which utilizes strong acids to aggressively break down proteins, generating compounds not present in nature and yielding substantial concentrations of salts due to the use of powerful alkalis to neutralize the acids. The products that are obtained in this manner are not permitted in organic agriculture.

There are two approaches to carry out hydrolysis in a natural and respectful manner: those based on guided enzymatic attacks and those based on microbial fermentations.

Enzymatic hydrolysis produces amino acid and oligopeptide combinations known as protein hydrolysates, which have several uses in agriculture, aquaculture, livestock farming, the food industry, and cosmetics. Their composition will be determined mostly by the origin of the protein and the enzyme or enzyme cocktail utilized.

Microbial hydrolysis produces more complex compounds because microorganisms have an extensive arsenal of enzymes capable of hydrolyzing many more components of the natural matrix as well as secreting new molecules.

Enzymatic processes are chemical reactions catalyzed by an enzyme that uses water to break down a molecule into two or more fractions. When the enzyme is applied to a solution containing proteins, these are broken down into smaller chains of amino acids known as hydrolyzed proteins. Due to the specificity and selectivity of enzymes, the process variables may be adjusted to produce the appropriate type of hydrolysate. The size of the peptide chains generated, as well as the quality and uniformity of the final product, depend on the process variables (temperature, pH), identification of the enzyme/substrate combination, and the appropriate ratio. To achieve a good reaction rate and maximize the enzymatic efficiency, it is critical to understand all the reaction variables in an enzyme hydrolysis process.

One of the most important benefits of non-chemical hydrolytic processes is the low frequency of amino acids in the racemic form D, which cannot be taken by plants; yet, this form is far more common in chemical processes.

The microbial hydrolysis or fermentation process is slower and more expensive, but it produces a higher quality product because the presence of L-form amino acids is the highest of the three options, and during the microbial growth process, in addition to hydrolysis reactions, biosynthesis processes give rise to molecules with higher biostimulant activity, such as polyamines. Microbial fermentation processes are often carried out with bacteria of the Bacillus or Aspergillus species, which have a diverse set of enzymes capable of breaking down a large number of plant compounds while also creating molecules with specific biostimulant activity.