Method for producing biotechnological products comprising fermented algae

Abstract

The invention relates to a method for obtaining a biotechnological product containing fermented algae, intended for use as a raw material component or as a final product in the agriculture, food, and cosmetics sectors. The method enhances the agricultural activity of algae, improves its nutritional content and bioavailability, enabling the obtain functional components and eliminates sensory issues associated with algae.

Description

[0001] METHOD FOR PRODUCING BIOTECHNOLOGICAL PRODUCTS COMPRISING

[0002] FERMENTED ALGAE

[0003] TECHNICAL FIELD

[0004] The invention relates to a method for obtaining a biotechnological product containing fermented algae, intended for use as a raw material component or as a final product in the agriculture, food, and cosmetics sectors. The method enhances the agricultural activity of algae, improves its nutritional content and bioavailability, enabling the obtain functional components and eliminates sensory issues associated with algae.

[0005] PRIOR ART

[0006] Algae, also known as seaweeds, are photosynthetic organisms that inhabit aquatic and terrestrial ecosystems, and their sizes can range from microscopic microalgae to large macroalgae. Algae produce energy through photosynthesis, like plants, and contribute significantly to the production of atmospheric oxygen. Thanks to their rich nutritional content and biotechnological potential, they are utilized in the technical fields of agriculture, food, and cosmetics.

[0007] The strong taste, color, and odor profiles of algae limit their use in many technical fields. In products with a high algae content, the characteristic color, strong taste, and odor associated with the added algae components pose significant challenges. Although methods such as genetic modification have been employed to improve these unfavorable sensory characteristics, such approaches are subject to strict regulatory procedures. Moreover, these modification processes lead to a reduction in the nutritional content of the products, a decrease in growth rate, and a loss of genetic stability.

[0008] The rich nutritional content of algae is located within the cells and is protected by the cell membrane and cell wall. These structures hinder the extraction of the desired components and reduce their bioavailability in the end consumer. Mechanical or chemical disruption of the cell wall or membrane requires high energy and cost, and often results in the degradation of sensitive biological and biochemical compounds.

[0009] As a result, it has become essential to develop a method that enables the production of biotechnological products containing algae, by improving extraction efficiency, reducing production costs, enhancing the sensory properties of algae, preserving or increasing their nutritional value, and significantly improving bioavailability.

[0010] BRIEF DESCRIPTION OF THE INVENTION

[0011] The present invention relates to a method for producing biotechnological products that contain fermented algae as a component.

[0012] The primary objective of the invention is to develop a cost-effective and efficient method for producing biotechnological products incorporating fermented algae as a component, with enhanced functional and economic value. In this method, algae are processed using microbial systems without involving downstream processes that would result in chemical pollution or high energy costs. During this process, biological and biochemical components are extracted, while sensory properties are improved and the nutritional and functional composition is enriched. With these characteristics, the method distinguishes itself from those used in existing techniques.

[0013] Another objective of the invention is to improve the strong odor and taste profile of algae through the methods described in the invention. In this way, the biotechnological products obtained by the disclosed method can be made suitable for use in a wide range of technical fields.

[0014] Another objective of the invention is to provide a method for obtaining biotechnological products containing fermented algae as a component, which have high nutritional content and increased bioavailability of these components. Another objective of the invention is to provide a method specific to the species and / or the bioactive molecule by using different microbial system combinations, process parameters, and / or fermentation specifications depending on the targeted bioactive molecule to be extracted. In this way, an optimized and specific production process is provided for each bioactive component.

[0015] Another objective of the invention is to include pre-treatment steps that are adaptable to the type of algae to be processed and the type of algal component to be obtained, and that enhance fermentation efficiency.

[0016] DETAILED DESCRIPTION OF THE INVENTION

[0017] In this detailed description, the subject of the invention relates to a method for obtaining biotechnological products containing fermented algae-based components, and is explained with examples that are provided solely for a better understanding of the subject and that do not have any limiting effect.

[0018] As known in the related art, algae are used as raw materials in many different technical fields, primarily in agriculture, food, and cosmetics, due to their rich nutritional content. The method described in the invention aims to eliminate the disadvantages or improve the shortcomings present in the existing techniques.

[0019] Fundamentally, in the present invention, it becomes possible to weaken or break down the cell walls or membranes present in algae with high efficiency by using microbial systems through the fermentation environment to be created. The biotechnological products obtained in this way possess high nutritional content and allow for the extraction and enrichment of macromolecules such as proteins, fats, and carbohydrates, as well as bioactive and biological / chemical micromolecules such as pigments, antioxidants, and various secondary metabolites contained within the algae.

[0020] In this invention, algae are essentially included as a partial nutrient source for the microorganisms in the fermentation environment. The microorganisms interact with the cell walls and / or membranes directly or indirectly, enabling the release of intracellular components. To achieve this, it is necessary to configure suitable fermentative microorganisms and fermentation conditions.

[0021] The provision of a suitable fermentation environment is maintained under control through various parameters such as culture medium, pH, temperature, mixing, and aeration.

[0022] At the same time, the fermentative microorganisms metabolize and modify certain amino acid groups and volatile compounds that cause strong taste and odor profiles in the structure of algae, thereby enabling the improvement of the sensory performance of algae.

[0023] In the preferred embodiment, the fermentation environment is established in a manner that allows various microorganisms, such as bacteria or yeast, to carry out the fermentation process. In this environment, at least one microorganism is used to perform the fermentation.

[0024] The microorganisms and algae species mentioned here are preferred because they are capable of enabling the aforementioned interactions. The microorganisms described in the invention are able to carry out the necessary interactions to break down or weaken the cell membranes / walls of the specified algae.

[0025] The fermentation process steps described in this invention are as follows:

[0026] - Supplying algae in dry form as the fermentation raw material and adding it to the fermentation medium at specified ratios,

[0027] - Adding at least one fermentative microorganism to the fermentation medium to carry out the fermentation, and performing the fermentation process according to defined specifications to obtain a biotechnological product in the form of a solid-liquid phase mixture.

[0028] In the preferred embodiment, at least one of the following algae species is used for fermentation: Chlorella (Chlorella vulgaris, Chlorella protothecoides, Chlorella sorokiniana, Auxenachlorella pyrenoidosa), Nannochloropsis, Dunaliella, Haematococcus, Scenedesmus, Euglena (Euglena sp., Euglena gracilis), Schizochytrium, Spirulina (Arthrospira platensis, Spirulina maxima), Aphanizomenon flos-aquae (AFA), Tetraselmis, Eustigmatophytes, Euglenophytes, Rhodophytes, Porphyridium, Bacillariophyceae, Phaeodactylum, Odontella, Thraustochytriaceae, Ascophyllum nodosum, Asparagopsis taxiformis, Fucus vesiculosus, Sargassum spp., Ulva spp., Laminaria spp., Gracilaria spp., Chondrus crispus, Porphyra spp., Gelidium spp., Enteromorpha prolifera, Agarophyton vermiculophyllum, Saccharina latissima.

[0029] In a preferred embodiment, the algae to be used as raw material are provided in powder, granule, or flake form. In this way, the surface area of the algae is increased for fermentation processes.

[0030] In a preferred embodiment, the algae ground into powder for fermentation processes are mixed with water at a concentration ranging from 10 to 125 grams per liter.

[0031] In the method of the invention, pre-treatment is preferably applied to the algae before the fermentation step. The mentioned pre-treatments are carried out to enhance fermentation efficiency. These process steps comprise at least one of homogenization, sonication, bead milling, or enzymatic pre-treatment. The main objective here is to physically or chemically weaken the cell walls and / or membranes of the algae prior to fermentation in order to increase fermentation efficiency.

[0032] The sonication process, which can be applied as a pre-treatment step, is carried out at a frequency range of 10 to 100 kHz and a power range of 50 to 2000 W. Preferably, the sonication process is performed for a duration of 1 to 240 minutes.

[0033] The homogenization process, which can be applied as a pre-treatment step, is carried out at a pressure range of 50 to 2000 bar, in single or multiple cycles, and at a temperature range of 30 to 60 °C. Preferably, the homogenization process is performed for a duration ranging from 1 second to 30 minutes. In the bead milling process, which can be applied as a pre-treatment step, the diameter of the beads used ranges from 0.2 to 10 mm. In these processes, the rotation speed is in the range of 500 to 10,000 rpm. The duration of these processes ranges from 1 to 240 minutes.

[0034] In the enzymatic hydrolysis process, which can be applied as a pre-treatment step, at least one enzyme from the cellulase, hemicellulase, protease, lipase, glucanase, or chitinase group is used. The concentration of these enzymes is in the range of 0.1% to 10% (w / v). The pH value in the mentioned enzymatic hydrolysis processes ranges from 3 to 9. The temperature for carrying out the processes is in the range of 30 to 70 °C. Preferably, the duration of these processes ranges from 1 to 48 hours.

[0035] In the method of the invention, at least one fermentative microorganism from the group consisting of Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus acidophilus, Bifidobacterium lactis, Saccharomyces cerevisiae, Bifidobacterium bifidum, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus paracasei, Lactococcus lactis, Lactobacillus bifidobacteria sp., Lactobacillus plantarum, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus helveticus, Bacillus belgii, Bacillus subtilis, Bacillus megaterium, Bacillus licheniformis, and Azotobacter vinelandii is used for the fermentation processes. In a preferred embodiment, a mixture of these microorganisms may also be used.

[0036] In a preferred method, at least one component selected from carbon sources such as sugars, nitrogen, vitamins, and salts, or mixtures thereof, is added to the fermentation medium as a culture medium component to support the fermentative microorganisms.

[0037] The fermentation medium described in the invention is configured to allow or prevent contact with air depending on the oxygen requirement of the microorganisms used. The reactor material may be plastic, metal, alloy, or glass. In cases where aeration is required, the aeration rate is in the range of 0.1 to 5 vvm. In the mentioned fermentation medium, the algae-to-fermentative microorganism ratio is preferably in the range of 1 :1 to 1 :100 by weight.

[0038] The pH value of the fermentation medium is in the range of 3 to 10. This range is determined to ensure the optimal growth and activity of the microorganisms used in the fermentation. The pH of the fermentation medium can be regulated using acid / base chemicals and / or their dilute solutions such as HCI, H2SO4, CH3COOH, C6H8O7, NaOH, KOH, Na2HPO4, and Ca(OH)2.

[0039] The temperature of the fermentation medium is in the range of 20 to 70 °C. These temperature values determined for the fermentation medium represent the optimal growth and activity conditions for the microorganisms used.

[0040] In a preferred embodiment, the fermentation processes are carried out for a duration ranging from 6 to 120 hours, depending on the components used, the product to be obtained, the microorganism employed, and the fermentation parameters.

[0041] In a preferred embodiment, the final product obtained as a result of fermentation contains at least one emulsifier. As the mentioned emulsifier, at least one from the group consisting of lecithin, carrageenan, xanthan gum, and guar gum is included; however, it is not limited to these compounds and also encompasses other emulsifier compounds known in the relevant technical field. The emulsifier is present in the final product in an amount ranging from 0.1% to 1% by weight. The emulsifier is included to ensure the stability of the final product and to emulsify components that are not naturally miscible.

[0042] In a preferred embodiment, the final product contains at least one preservative compound. As the mentioned preservative compound, at least one from the group consisting of citric acid, potassium sorbate, sodium benzoate, sorbic acid, and trehalose is included; however, it is not limited to these compounds and also encompasses other preservative compounds known in the relevant technical field. The preservative compound is present in the final product at a concentration ranging from 0.2 to 0.6 g / L. The preservative compound is included to extend the shelf life of the biotechnological product.

[0043] In a preferred embodiment, mixing processes are carried out during or after the fermentation processes. The mentioned mixing operations are performed at a speed in the range of 50 to 1500 rpm.

[0044] The product obtained after fermentation is a biotechnological product in liquid-solid phase form and is collected from the medium. In a preferred embodiment, at least one of the following process steps is applied to the obtained product: clarification, salting, homogenization, and emulsification. By applying at least one of these process steps, it becomes possible to eliminate process-related impurities and to obtain a product with increased purity.

[0045] The solid-liquid phase biotechnological product obtained through the fermentation processes in the method of the invention can be used in various technical fields. The present inventors are able to separate the solid and liquid phases from the solid-liquid mixture and obtain biotechnological products in both phases for use in technical applications. The following sections will provide characterization data of the obtained products.

[0046] - Obtaining a biotechnological product in solid-liquid phase

[0047] The product obtained through the fermentation process is a biotechnological product in a dual liquid-solid phase. In a preferred embodiment, at least one of the following process steps may be applied to this product: pasteurization, filtration, UHT, or UV sterilization. In this way, microbial activity within the liquid-solid phase biotechnological product can be terminated.

[0048] The solid-liquid phase biotechnological product obtained in the invention is packaged and stored. The product obtained in this manner can be used in the technical fields of agriculture, food, and cosmetics.

[0049] Obtaining the liquid phase biotechnological product The inventors are able to obtain two separate biotechnological products, as solid phase and liquid phase, by subjecting the fermentation-derived products to separation processes for use in various technical fields.

[0050] As the said separation process, one of filtration, flocculation, or centrifugation is applied. If preferred, these processes may be applied more than once.

[0051] As the said filtration process, at least one of the following methods may be applied: porous membrane, paper filter, mesh filtration, or membrane filtration. In membrane applications, microfiltration or ultrafiltration membranes may be used depending on the desired output profile.

[0052] The said centrifugation process can be carried out continuously or intermittently at a speed ranging from 4500 to 18000 rpm.

[0053] With the application of these processes, the liquid portion is collected. In a preferred embodiment, at least one of the following process steps may be applied to this product: pasteurization, filtration, UHT, or UV sterilization. In this way, microbial activity within the liquid phase biotechnological product can be terminated.

[0054] The liquid phase biotechnological product obtained in the invention is packaged and stored. The resulting liquid phase biotechnological product is rich in bioactive and macro / micro biological and chemical molecules such as antioxidants, flavonoids, pigments, proteins, fats, and carbohydrates. The product obtained in this way, without being limited thereto, can be used in various applications such as biofertilizers, biostimulants, microbial fertilizers, natural food colorants, food raw materials, and fermented cosmeceutical extracts. It can thus be utilized in the technical fields of agriculture, food, and cosmetics.

[0055] Obtaining the solid phase biotechnological product By subjecting the solid-liquid phase biotechnological product to separation processes, two distinct biotechnological products solid phase and liquid phase are obtained.

[0056] As the said separation process, one of filtration, flocculation, or centrifugation is applied. If preferred, these processes may be applied more than once.

[0057] As the said filtration process, at least one of the following methods may be applied: porous membrane, paper filter, mesh filtration, or membrane filtration. In membrane applications, microfiltration or ultrafiltration membranes may be used depending on the desired output profile.

[0058] The said centrifugation process can be carried out continuously or intermittently at a speed ranging from 4500 to 18000 rpm.

[0059] With the application of these processes, the solid portion is collected. In a preferred embodiment, at least one of the following process steps may be applied to this product: pasteurization, UHT, or UV sterilization. In this way, microbial activity within the solid phase biotechnological product can be terminated.

[0060] In a preferred embodiment, the solid phase product is subjected to at least one drying process. The said drying process may be carried out using various drying methods such as an oven, drying cabinet, spray dryer, freeze dryer, or drum dryer. The solid phase product obtained after drying has a moisture content of 30% or less. The process temperature for drying is in the range of 60 to 200 °C for ovens and drying cabinets, 100 to 300 °C for spray dryers, 80 to 200 °C for drum dryers, and -20 to -100 °C for freeze dryers.

[0061] The solid phase biotechnological product produced in the invention may contain high levels of macro-biological components such as proteins, fats, and carbohydrates depending on the type of algae used, and may also be rich in biological / chemical molecules such as vitamins, minerals, antioxidants, and flavonoids. The obtained product is packaged and stored. In this form, the product can be used in the technical fields of agriculture, food, and cosmetics. With the method described in the invention, it is possible to obtain biotechnological products containing fermented algae as a component, which are suitable for use in various technical fields, have high added value and efficiency, and are cost- effective. In the method described in the present invention, processed algal biomass is obtained by extracting bioactive molecules through microbial systems using selective biotechnological applications, without involving sub-processes that would cause chemical pollution or high energy costs. While carrying out this process, sensory performance and content are also enhanced, which distinguishes the invention from existing methods in this respect.

[0062] The method subject to the invention enables the production of biotechnological products containing algae with improved odor and taste profiles. In this way, the biotechnological products obtained by the method described in the invention can be utilized in many different technical fields.

[0063] The method subject to the invention enables the production of biotechnological products from algae that have high nutritional content and improved bioavailability of these components.

[0064] The scope of protection of the invention is specified in the appended claims and cannot be limited to what is described for illustrative purposes in this detailed description. It is clear that a person skilled in the art can produce similar embodiments in the light of what is explained above, without deviating from the main theme of the invention.

Claims

CLAIMS1. A method for obtaining a biotechnological product containing components present in algae, characterized in that it comprises the following process steps:- providing algae in dry form as fermentation raw material and adding them to the fermentation medium,- adding at least one fermentative microorganism to the fermentation medium,- obtaining a biotechnological product in a solid-liquid phase mixture by performing fermentation processes in the fermentation medium.

2. A method according to claim 1 , characterized in that at least one algae species selected from the group consisting of Chlorella (Chlorella vulgaris, Chlorella protothecoides, Chlorella sorokiniana, Auxenachlorella pyrenoidosa), Nannochloropsis, Dunaliella, Haematococcus, Scenedesmus, Euglena (Euglena sp., Euglena gracilis), Schizochytrium, Spirulina (Arthrospira platensis, Spirulina maxima), Aphanizomenon flos-aquae (AFA), Tetraselmis, Eustigmatophytes, Euglenophytes, Rhodophytes, Porphyridium, Bacillariophyceae, Phaeodactylum, Odontella, Thraustochytriaceae, Ascophyllum nodosum, Asparagopsis taxiformis, Fucus vesiculosus, Sargassum spp., Ulva spp., Laminaria spp., Gracilaria spp., Chondrus crispus, Porphyra spp., Gelidium spp., Enteromorpha prolifera, Agarophyton vermiculophyllum, and Saccharina latissima is added to the fermentation medium.

3. A method according to any one of the preceding claims, characterized in that at least one pre-treatment step selected from homogenization, sonication, heating or freezing, and bead milling is applied to the algae before being added to the fermentation medium.

4. A method according to claim 3, characterized in that the said sonication process is carried out at a frequency in the range of 10 to 100 kHz and at a power in the range of 50 to 2000 W.

5. A method according to claim 3 or 4, characterized in that the said homogenization process is carried out at a pressure in the range of 50 to 2000 bar and at a temperature in the range of 30 to 60 °C.

6. A method according to any one of claims 3 to 5, characterized in that the said bead milling process is carried out at a rotation speed in the range of 500 to 10,000 rpm.

7. A method according to any one of claims 3 to 6, characterized in that at least one enzyme selected from the group consisting of cellulase, hemicellulase, protease, lipase, glucanase, and chitinase is used in the enzymatic hydrolysis process.

8. A method according to any one of claims 3 to 7, characterized in that the enzymatic hydrolysis process is carried out at a temperature in the range of 30 to 70 °C and at a pH value in the range of 3 to 9.

9. A method according to any one of the preceding claims, characterized in that an aqueous mixture containing algae in an amount ranging from 10 to 125 grams per liter is used as the raw material.

10. A method according to any one of the preceding claims, characterized in that at least one fermentative microorganism selected from the group consisting of Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus acidophilus, Bifidobacterium lactis, Saccharomyces cerevisiae, Bifidobacterium bifidum, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus paracasei, Lactococcus lactis, Lactobacillus bifidobacteria sp., Lactobacillus plantarum, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus helveticus, Bacillus belgii, Bacillus subtilis, Bacillus megaterium, Bacillus licheniformis, and Azotobacter vinelandii is added to the fermentation medium.11 .A method according to any one of the preceding claims, characterized in that at least one component selected from different carbon sources such as sugars, nitrogen, and vitamins is added as a culture medium component.

12. A method according to any one of the preceding claims, characterized in that the product obtained after fermentation contains a preservative compound comprising at least one selected from the group consisting of potassium sorbate, sodium benzoate, and sorbic acid.

13. A method according to claim 12, characterized in that the product obtained after fermentation contains the preservative compound in an amount ranging from 0.2 to 0.6 g / L.

14. A method according to any one of the preceding claims, characterized in that at least one emulsifier selected from the group consisting of lecithin, carrageenan, xanthan gum, and guar gum is added to the product obtained after fermentation.

15. A method according to claim 14, characterized in that at least one emulsifier is added to the product obtained after fermentation in an amount ranging from 0.1 % to 1 % by weight.

16. A method according to any one of the preceding claims, characterized in that the fermentation medium comprises algae and fermentative microorganisms at a weight ratio in the range of 1 :1 to 1 :100.

17. A method according to any one of the preceding claims, characterized in that the pH value of the fermentation medium is in the range of 3 to 10.

18. A method according to any one of the preceding claims, characterized in that the temperature of the fermentation medium is in the range of 20 to 70 °C.

19. A method according to any one of the preceding claims, characterized in that a liquid phase biotechnological product and a solid phase biotechnologicalproduct are obtained by applying at least one filtration and / or at least one centrifugation process, respectively, to the obtained solid-liquid phase biotechnological product.

20. A method according to claim 19, characterized in that a filtration method using a membrane structure with microfiltration or ultrafiltration properties is applied.

21. A method according to claim 19 or 20, characterized in that the said centrifugation process is carried out at a speed in the range of 4500 to 18000 rpm.

22. A method according to any one of claims 19 to 21 , characterized in that at least one purification process is applied to the said liquid phase biotechnological product.

23. A method according to any one of claims 19 to 22, characterized in that a drying process is applied to the said solid phase biotechnological product: o at a temperature in the range of 60 to 200 °C for oven and drying cabinet, or o at a temperature in the range of 100 to 300 °C for spray dryer, or o at a temperature in the range of 80 to 200 °C for drum dryer, or o at a temperature in the range of -20 to -100 °C for freeze dryer24. A method according to any one of claims 19 to 23, characterized in that at least one method selected from filtration, pasteurization, UHT sterilization, and / or UV sterilization is applied to terminate the viable microbial activity within the liquid phase or solid phase biotechnological product.