Platform for the production of microbial triglyceride mixtures with adjustable composition

Abstract

The present invention lies in the field of microbial fermentation and provides a process for producing triglyceride mixtures with an adjustable fatty acid profile. Furthermore, the present invention provides the use of suitable microorganisms for the production of such triglyceride mixtures, as well as the triglyceride mixtures produced in this way.

Description

[0001] The present invention lies in the field of microbial fermentation and provides a process for producing triglyceride mixtures with an adjustable fatty acid profile. Furthermore, the present invention provides the use of suitable microorganisms for the production of such triglyceride mixtures, as well as the triglyceride mixtures produced in this way.

[0002] Vegetable oils and triglyceride mixtures are indispensable in industry. As flavor carriers in food, as additives in cosmetics, or in the form of emulsions such as varnishes or detergents, vegetable oils are used in almost all consumer products. The many applications are driving an ever-increasing demand for vegetable oils, each with its own specific requirements. Many vegetable oils are facing growing criticism because their production is associated with significant environmental damage in the producing countries and long transport routes.

[0003] Furthermore, conventional vegetable oils often have highly variable fatty acid compositions and therefore different physical properties, which can complicate the processing process, e.g. in the case of a solid triglyceride mixture.

[0004] Triglycerides are the main form of fat in food and in the body, where they are stored as an energy reserve. The breakdown of triglycerides releases the fatty acids they contain. Vegetable oils always consist of a mixture of triglycerides, which are released as fatty acids.

[0005] Microorganisms can also produce such triglyceride mixtures. The microbial metabolic pathway for triglyceride production begins with the synthesis of fatty acids via the acetyl-CoA pathway. Microorganisms such as yeast convert acetyl-CoA, an intermediate product of carbohydrate breakdown, into fatty acids through a series of enzymatic reactions. These fatty acids are then sequentially bound to a glycerol molecule by the enzyme acyltransferase. First, monoacylglycerol is formed, then diacylglycerol, and finally, the addition of a third fatty acid forms the triglyceride, which is stored in lipid droplets.

[0006] The microbial production of fatty acids is widespread. The production of a specific fatty acid length using the production organism Yarrowia lipolytica, a yeast, is particularly favored.

[0007] European patent application EP 2 649 887 A2 describes the production of eicosapentaenoic acid (EPA) using Yarrowia ipolytica. EPA is a polyunsaturated fatty acid with a chain length of 20 carbon atoms. This fatty acid belongs to the class of omega-3 fatty acids and has a positive effect on health as part of a human diet.

[0008] The publication “Yarrowia lypolytica as a model for bio-oil production” by Beopoulos et al. (Prog Lipid Res. 2009 Nov;48(6):375-87. doi: 10.1016) describes the elucidation of the metabolic pathways of the yeast Yarrowia lypolytica for the targeted production of one or more desired fatty acids. The molecular basis, including potential genetic modifications, is described in particular.

[0009] The international application WO 2004 / 101757 A2 describes the production of Omega3 and Omega6 fatty acids using Yarrowia lypolytica.

[0010] Independently of the organism Yarrowia lypolytica, other organisms are also used for the production of fatty acids, such as a microalga of the genus Prototheca in publication WO 2012 / 106560 A1 or a modified fungal cell in publication WO 2011 / 161317 A2.

[0011] All these publications have in common that they do not offer a system for easily and specifically adjusting a fatty acid profile, nor for replicating natural, plant-based oils.

[0012] Therefore, the primary objective of the present invention was to provide a method for producing a microbial oil or a microbial triglyceride mixture, which is characterized by simple adaptation to a desired fatty acid profile.

[0013] This task was accomplished by a fermentative process for the production of a triglyceride mixture with an adjustable fatty acid profile, consisting of or comprising: i. Providing at least one microorganism that exhibits optimized triglyceride production, ii. Providing at least one carbon source as a substrate, preferably selected from the group consisting of carbon-containing waste streams from the food, forestry, paper and agricultural industries, preferably lignocellulose, hemicellulose, molasses, starch, pectin, glucose, xylose, fructose, sucrose, galactose, galacturonic acid and arabinose; iii. Cultivate the microorganism under conditions that allow the microorganism to grow; iv. Subsequent cultivation of the microorganism under conditions that do not allow or only allow limited growth of the microorganism and result in the production of triglycerides; v. Obtaining a triglyceride mixture with a fatty acid profile defined by the conditions in step iv. the resulting triglyceride mixture has a fatty acid profile with

[0014] Lauric acid in a proportion of 0 to 5 wt.%, preferably 0 to 2 wt.%, Myristic acid in a proportion of 0 to 5 wt.%, preferably 0.1 to 3 wt.%, particularly preferably 0.5 to 2.5 wt.%,

[0015] Palmitic acid in a proportion of 0 to 70 wt.%, preferably 0.1 to 60 wt.%, particularly preferably 0.5 to 50 wt.%,

[0016] Palmitoleic acid in a proportion of 0.1 to 20%, preferably 1 to 15%, particularly preferably 1.5 to 10% by weight,

[0017] Stearic acid in a proportion of 0.1 to 30%, preferably 0.5 wt.% to 25 wt.%, particularly preferably 0.5 to 20 wt.%,

[0018] Oleic acid in a proportion of 10 to 70 wt.%, preferably 12 to 60 wt.%, particularly preferably 15 to 55 wt.%, and optionally Arachidic acid in a proportion of 0.1 to 10 wt.%, preferably 0.2 to 7.5 wt.%, behenic acid in a proportion of 0.1 to 10 wt.%, preferably 0.2 to 7.5 wt.%, lignoceric acid in a proportion of 0.1 to 5 wt.%, preferably 0.2 to 4 wt.%, nervonic acid in a proportion of 0 to 7.5 wt.%, preferably 0.5 to 5 wt.%, each in relation to the total amount of the triglyceride mixture, exhibits.

[0019] Surprisingly, the invention demonstrated that adjusting the cultivation conditions after the growth phase significantly influences the composition of the resulting triglyceride mixture. This allows for the targeted production of triglyceride mixtures tailored to specific requirements or the replication of naturally occurring triglyceride mixtures. Unlike the methods described in the prior art, this does not require targeted intervention at the metabolic level. Any microorganism capable of triglyceride production can be used in the process according to the invention.

[0020] The invention is particularly advantageous in the provision of fermentatively produced oils or triglyceride mixtures that have a corresponding fatty acid profile to naturally occurring oils or triglyceride mixtures. This allows oils, such as palm oil, which is used in a wide variety of products and has a poor ecological footprint, to be produced cost-effectively and sustainably in large quantities. The resulting fermentative product has a corresponding fatty acid profile to the natural oil. Fermentatively produced oil offers a location-independent alternative with a significantly smaller land requirement than the production of natural oils. With the oil-producing microorganism used and precise process control, it is possible to control the fatty acid profile of the produced oil and thus tailor the oil to specific requirements.

[0021] Examples of natural oils that can be produced using the process according to the invention are avocado oil, palm oil, peanut oil, hazelnut oil, cocoa butter, argan oil, rapeseed oil, olive oil or sunflower oil.

[0022] A “triglyceride mixture” is also commonly known as an oil and consists of a mixture of triglyceride molecules. Triglycerides have the general formula (I) where the R groups 1 , R 2 and R 3 These are fatty acid residues that are bound in oils or triglyceride mixtures and are released in the body, for example, after ingestion via food.

[0023] When reference is made to "triglyceride mixtures with short fatty acid residues" within the scope of the present invention, these refer to triglycerides with fatty acid residues having a length of no more than four carbon atoms. The term "short-chain fatty acid residues" can also be used synonymously.

[0024] When reference is made to "triglyceride mixtures with medium-length fatty acid residues" within the scope of the present invention, these refer to triglycerides with fatty acid residues having a length of five to fourteen carbon atoms. The term "medium-chain fatty acid residues" can also be used synonymously.

[0025] When reference is made to "triglyceride mixtures with long fatty acid residues" within the scope of the present invention, this refers to triglycerides with fatty acid residues having a length of more than fourteen carbon atoms. The term "long-chain fatty acid residues" can also be used synonymously.

[0026] Within the scope of the present invention, triglyceride mixtures with a variety of fatty acid residues can be produced, such as, for example,

[0027] Fatty acids are classified as saturated or unsaturated. Saturated fatty acids have no double bonds in their carbon chain, meaning that all carbon atoms are completely saturated with hydrogen atoms. Unsaturated fatty acids, on the other hand, possess one or more double bonds, resulting in fewer hydrogen atoms being bonded. The process of the present invention can be used to produce both saturated and unsaturated fatty acids.

[0028] Within the scope of the present invention, a "microorganism exhibiting optimized triglyceride formation" is one that either naturally exhibits increased triglyceride production or in which the metabolic pathway for triglyceride formation is promoted by genetic modification through the inhibition of other metabolic pathways for byproducts. Surprisingly, such genetic modification does not require targeted intervention in metabolic pathways to obtain individual desired triglycerides. Rather, it was recognized that optimizing triglyceride formation by eliminating or regulating byproduct formation enables the process according to the invention. In this process, a single production organism is used for a variety of triglyceride mixtures.

[0029] The present invention is not limited to a specific microorganism, but can be carried out with all oil-producing microorganisms. Examples of such microorganisms are yeasts, fungi, algae, bacteria, or archaea. Advantageously, conventional biotechnological production organisms can also be used within the scope of the present invention.

[0030] Within the framework of the process of the present invention, a variety of carbon sources can be used. These can be obtained, for example, from waste streams of the food, agricultural, and forestry industries, and are preferably lignocellulose, molasses, starch, or pectin-containing waste streams from the sugar industry. The monosaccharides glucose, xylose, fructose, sucrose, galactose, and arabinose or galacturonic acid can also be used as carbon sources.

[0031] The process according to the invention comprises two main phases. First, the oil-producing microorganism is cultivated to enable its growth (step iii). This is followed by the production phase (step iv), in which triglyceride formation takes place. By precisely adjusting the cultivation conditions in this phase, a desired triglyceride mixture with a defined fatty acid profile is obtained. Within the scope of the present invention, the relevant cultivation conditions have been identified. The cultivation conditions defined in step iv determine the desired fatty acid profile of the triglyceride mixture.

[0032] These are preferably selected within the scope of the present invention from the parameters temperature, pH value, oxygen availability, osmotic pressure, secondary substrate availability, cultivation time and / or the addition of additives, preferably wherein organic acids are added as an additive.

[0033] The “osmotic pressure” in the context of the present invention refers to the pressure created by the difference in the concentration of solutes (e.g., salts, sugars) between the interior of a microorganism and the surrounding culture medium. If the concentration of solutes in the environment is higher than inside the cell (hypertonic environment), the osmotic pressure draws water out of the cells. Conversely, if the concentration outside is lower (hypotonic environment), too much water can enter the cells. By adjusting the osmotic pressure, the fatty acid profile of the triglyceride mixture can be adjusted.

[0034] The availability of the secondary substrate, or "secondary substrate availability," is also referred to as "secondary substrate limitation." In fermentation technology, secondary substrate limitation refers to a situation in which the growth of microorganisms or the production of a desired substance is restricted by the availability of a secondary substrate present at a concentration below the microorganism's requirements. Even if the primary substrate, such as glucose, is sufficiently available, the organism's growth is slowed or stopped by the limited secondary substrate, such as oxygen or an essential nutrient. In the context of the present invention, the fatty acid profile of the triglyceride mixture can be adjusted via secondary substrate limitation.

[0035] Oxygen availability refers to the presence of oxygen during the cultivation period. Some microorganisms require oxygen for growth and product formation (obligate aerobes), some microorganisms can utilize oxygen (facultative aerobes / anaerobes), and others grow in the absence of oxygen (obligate anaerobes). Therefore, controlling oxygen availability can also influence the product formation of the triglyceride mixture.

[0036] In addition to the parameters defined above, the cultivation temperature influences the appearance of the triglyceride mixture. A cultivation temperature of ≤ 25 °C preferably prevents the formation of palmitic acid, while a cultivation temperature of > 25 °C promotes the formation of palmitic acid. Preferably, a cultivation temperature of ≥ 30 °C results in the formation of more than 20 wt% palmitic acid, based on the total amount of fatty acid residues.

[0037] Furthermore, an increase in pH value leads to an increased formation of linoleic acid up to 35 wt% and a decrease in the formation of steraric acid to less than 30 wt%.

[0038] Another way to adjust the fatty acid profile is by adding various additives, preferably organic acids. The addition of citric acid leads to an increased formation of palmitic acid, up to 58% by weight.

[0039] Furthermore, the product formation of triglyceride mixtures can preferably also be achieved via auxotrophy of the microorganism used. Auxotrophy refers to the state of an organism that, due to a genetic mutation or defect, is unable to synthesize a specific essential compound, such as an amino acid, a vitamin, or a nucleotide building block. This organism depends on the missing compound being supplied from its environment, such as the culture medium. If the supply of this missing amino acid is insufficient, growth is inhibited, and only product formation in step iv. takes place. Therefore, within the scope of the present invention, an oil-forming auxotrophic microorganism can be provided in step i. By limiting the essential compound in the culture medium, the triglyceride formation can then be influenced.

[0040] Preferably, in step iv., the organism is cultivated under secondary substrate limitation. This prevents further growth of the organism and promotes triglyceride formation. Preferably, in step iv., the microorganism is cultivated under secondary substrate limitation of the phosphorus, sulfur, and / or nitrogen source.

[0041] Preferably the microorganism is selected from the family Ustilaginaceae or the division of Basidiomycetes, preferably selected from the genus Ustilago, and the oil-producing microorganism Ustilago maydis is particularly preferred.

[0042] It is preferred within the scope of the present invention that the formation of by-products in the microorganism Ustilago maydis of ustilaginic acid, itaconic acid, mannosylerythritol lipids and / or malate is prevented or reduced.

[0043] Another aspect of the present invention relates to the use of at least one microorganism, in which the microorganism is optimized for a metabolic pathway for the production of triglycerides, for the production of a triglyceride mixture with a desired fatty acid profile.

[0044] Preferably, the metabolic pathway is optimized to avoid the formation of by-products; particularly preferably, the glycolipid biosynthesis pathway is interrupted.

[0045] Another aspect of the present invention is a triglyceride mixture produced or producible according to the inventive method, wherein the produced triglyceride mixture has a desired fatty acid profile and wherein long-chain triglycerides with a fatty acid chain length of 20 to 24 carbon atoms are present in such a mixture in an amount of over 2 wt.%, preferably over 5 wt.%, particularly preferably over 10 wt.%, based on the total weight of the mixture.

[0046] Comparing the desired fatty acid profile of a natural oil with a corresponding oil produced according to the present invention, it is noticeable that an oil produced according to the invention has a higher proportion of long-chain fatty acids compared to a natural oil. Therefore, naturally occurring oils and oils produced according to the invention can be distinguished from one another, but they fulfill the same requirements in their application.

[0047] Another aspect of the inventive method relates to a device for carrying out the inventive method, wherein the device has a controller which, when a triglyceride mixture to be produced with a predetermined fatty acid profile is entered, sets the corresponding parameters in the device after comparison with a stored database.

[0048] First, the desired fatty acid profile of a triglyceride mixture is entered via an input mask. The controller in the device, which could be a computer, compares the target value for the composition of the desired triglyceride mixture with the values ​​stored in a database and thus determines the fermentation conditions that must be used to produce the desired triglyceride mixture and sets these conditions in the device.

[0049] The temperature is measured via a temperature controller familiar to experts, and the pH value via a pH probe. By controlling connected pumps, the pH value can be adjusted during fermentation by adding acidic or basic solutions. The carbon source can also be controlled via pump control, and the oxygen concentration can be controlled by controlling the gas inflows. Suitable control methods for the individual parameters are known to the expert.

[0050] Furthermore, the input device and the fermentation device, including the control devices, can be in one design, in several separate designs, or parts of the individual devices can be combined.

[0051] The invention is characterized below by illustrative, non-limiting examples: Brief description of the illustrations Fig. This shows the influence of organic acids and low and high osmolality on the fatty acid composition in a triglyceride mixture. The total amount of triglycerides and their fatty acid residues was defined as the reference value for determining the percentage (in wt%). Fig. This study demonstrates the influence of secondary substrate limitation (phosphorus, sulfur, and nitrogen sources) on the fatty acid composition of a triglyceride mixture. The total amount of triglycerides and their fatty acid residues was defined as the reference value for determining the percentage (in wt%). Fig. This shows the influence of pH on the fatty acid composition in a triglyceride mixture. The total amount of triglycerides and their fatty acid residues was defined as the reference value for determining the percentage (in wt%). Fig. This figure shows the influence of oxygen concentration, measured against the maximum oxygen transfer capacity, on the fatty acid composition in a triglyceride mixture. The total amount of triglycerides and their fatty acid residues was defined as the reference value for determining the percentage (in wt%). Fig. This shows the influence of the cultivation temperature on the fatty acid composition in a triglyceride mixture. The total amount of triglycerides and their fatty acid residues was defined as the reference value for determining the percentage (in wt%). Fig. This shows the influence of cultivation time on the fatty acid composition in a triglyceride mixture. To determine the percentage (in wt%), the total amount of triglycerides and their fatty acid residues was defined as the reference value. The in Fig. The arrows shown are for illustrative purposes only and serve to clarify the axis scaling. Fig. This figure shows the influence of the mono- and disaccharides galactose, arabinose, xylose, fructose, sucrose, and glucose on the fatty acid composition of a triglyceride mixture. The total amount of triglycerides and their fatty acid residues was defined as the reference value for determining the percentage (in wt%). Examples Example 1: Fermentation to produce a triglyceride mixture using the organism Ustilago maydis

[0052] First, an unlimited-growth preculture of Ustilago maydis MB215Δcyp1Δemt1 is prepared from a glycerol stock stored at -80 °C. This preculture serves to improve the reproducibility of the subsequent main experiments. The cultivation conditions for the preculture are defined as follows: 250 mL shake flasks with a filling volume of 20 mL are used. The culture is cultivated at a temperature of 30 °C and a shaking speed of 350 revolutions per minute (rpm) with a shaking diameter of 50 mm. The preculture medium contains a carbon source, e.g., glucose, at a concentration of 20 g / L, as well as diammonium sulfate at a concentration of 5 g / L. Once the culture has entered the exponential growth phase, the preculture is stopped, washed with a 0.9% sodium chloride (NaCl) solution, and adjusted to an optical density (OD) of 10.

[0053] In the next step, the main culture is inoculated with the washed pre-culture to a final OD of 0.1. The cultivation conditions for the main culture are defined as follows: A constant temperature (reference = 30 °C) and a constant pH (reference = 6.5) are maintained. The carbon-to-nitrogen ratio can be adjusted between 0.008 and 0.04 g. N / G C If oxygen limitation is not desired, a cascade system is used to ensure a dissolved oxygen concentration above 30%. This includes regulating the agitator speed and the aeration rate. Additionally, for specific fermenters, the fermenter head pressure can be adjusted from 1 to a maximum of 8 bar. For specific fatty acid profiles, the individual parameters must be adjusted.

[0054] The main culture for oil production lasts between 72 and 168 hours, depending on the chosen process parameters such as the nitrogen to carbon (N / C) ratio, temperature and other relevant factors.

[0055] The fermentation process of the main culture is divided into two cultivation phases: 1. Growth phase: In this phase, the U. maydis culture grows without limitation until the nitrogen source is completely depleted. The amount of available nitrogen is adjusted to utilize as much of the maximum oxygen capacity of the respective cultivation system as possible without causing oxygen limitation. 2. Triglyceride Production Phase: After the nitrogen source is consumed, the triglyceride production phase begins. This phase is characterized by reduced and slowly decreasing respiration in the U. maydis culture. During this phase, the cells accumulate triglycerides intracellularly until either the cells are completely filled with lipid droplets or the carbon source is depleted. Once the carbon source is exhausted, the cultivation is terminated.

[0056] After cultivation is complete, the triglyceride produced can be obtained by cell disintegration and extraction. In this process, the cell structures are broken down mechanically or chemically to release the intracellular triglyceride, which is then extracted or separated by centrifugation.

[0057] This two-phase cultivation process ensures efficient triglyceride production, allowing for optimal control of both growth and product formation. Example 2: Investigations into product formation

[0058] In the Fig. This study demonstrates which parameters influence the fatty acid profile of the resulting triglyceride mixture. One parameter was changed per fermentation run, and the fatty acid profile of the resulting triglyceride mixture was analyzed.

[0059] The data obtained can be used, for example, to train an algorithm and to determine the required fermentation conditions based on an input of a desired fatty acid profile. Example 3: Production of a specific triglyceride mixture (avocado oil)

[0060] A fermentation process was carried out as described in Example 1, with the following parameters set for the production phase (step iv): Temperature: 30°C pH: 6.5 Carbon source: Glucose Secondary substrate limitation: nitrogen Oxygen administration: No limitation Cultivation time: 144 hours

[0061] The resulting triglyceride mixture has the following composition: 22% by weight palmitic acid, 48% by weight oleic acid, and 18% by weight linoleic acid. This composition is comparable to avocado oil. QUOTES INCLUDED IN THE DESCRIPTION

[0000] This list of documents cited by the applicant was automatically generated and is included solely for the reader's convenience. The list is not part of the German patent or utility model application. The DPMA accepts no liability for any errors or omissions. Cited patent literature

[0000] EP 2 649 887 A2

[0007] WO 2004 / 101757 A2

[0009] WO 2012 / 106560 A1

[0010] WO 2011 / 161317 A2

[0010] Cited non-patent literature

[0000] Prog Lipid Res. 2009 Nov;48(6):375-87. doi: 10.1016

[0008]

Claims

[1] Fermentative process for producing a triglyceride mixture with an adjustable fatty acid profile, consisting of or comprising: i. Providing at least one microorganism optimized for the production of triglycerides, f ii. Providing at least one carbon source as a substrate, preferably selected from the group consisting of carbon-containing waste streams from the food, forestry, paper and agricultural industries, preferably lignocellulose, hemicellulose, molasses, starch, pectin, glucose, xylose, fructose, sucrose, galactose, galacturonic acid and arabinose; iii. Cultivate the microorganism under conditions that allow the microorganism to grow; iv. Subsequent cultivation of the microorganism under conditions that do not allow or only allow limited growth of the microorganism and result in the production of triglycerides; v. Obtaining a triglyceride mixture with a fatty acid profile defined by the conditions in step iv. the resulting triglyceride mixture has a fatty acid profile with Lauric acid in a proportion of 0 to 5 wt.%, preferably 0 to 2 wt.%, Myristic acid in a proportion of 0 to 5 wt.%, preferably 0.1 to 3 wt.%, particularly preferably 0.5 to 2.5 wt.% Palmitic acid in a proportion of 0 to 70 wt.%, preferably 0.1 to 60 wt.%, particularly preferably 0.5 to 50 wt.%, Palmitoleic acid in a proportion of 0.1 to 20%, preferably 1 to 15%, particularly preferably 1.5 to 10% by weight, Stearic acid in a proportion of 0.1 to 30%, preferably 0.5 wt.% to 25 wt.%, particularly preferably 0.5 to 20 wt.%, Oleic acid in a proportion of 10 to 70 wt.%, preferably 12 to 60 wt.%, particularly preferably 15 to 55 wt.%, and optionally Arachidic acid in a proportion of 0.1 to 10 wt.%, preferably 0.2 to 7.5 wt.%, Behenic acid in a proportion of 0.1 to 10 wt.%, preferably 0.2 to 7.5 wt.%, Lignoceric acid in a proportion of 0.1 to 5 wt.%, preferably 0.2 to 4 wt.%, Nervonic acid in a proportion of 0 to 7.5 wt.%, preferably 0.5 to 5 wt.%, each in relation to the total amount of the triglyceride mixture, exhibits. [2] Method according to claim 1, wherein the conditions which require the production of triglycerides are selected from the parameters temperature, pH value, oxygen availability, osmotic pressure, secondary substrate availability, cultivation time and / or the addition of additives, preferably wherein organic acids are added as an additive. [3] Method according to claim 1 or 2, wherein An increase in pH value leads to increased formation of linoleic acid up to 35 wt% and a decrease in the formation of stearic acid down to less than 30 wt% and / or The addition of organic acids leads to the increased formation of palmitic acid up to 60 wt. %, under certain conditions. [4] Method according to one of the preceding claims, wherein A cultivation temperature of ≤ 25 °C prevents the formation of palmitic acid and A cultivation temperature of > 25 °C leads to the formation of palmitic acid. preferably wherein a cultivation temperature of ≥ 30°C results in the formation of more than 20 wt% palmitic acid, based on the total amount of fatty acids produced. [5] Method according to one of the preceding claims, wherein in step iv. the organism is cultivated under a second substrate limitation. [6] Method according to any of the preceding claims, wherein the secondary substrate limitation is the limitation of the phosphorus, sulfur and / or nitrogen source. [7] Method according to any of the preceding claims, wherein the oil-producing microorganism is selected from the division of Basidiomycetes or the family of Ustiliganaceae, particularly preferably selected from the genus Ustilago, preferably wherein the oil-producing microorganism is Ustilago maydis. [8] Use of at least one microorganism in which the microorganism is optimized for the metabolic pathway for the production of triglycerides, preferably wherein the metabolic pathway for glycolipid biosynthesis is interrupted, for the production of an oil with a desired fatty acid profile. [9] Use according to claim 8, wherein the microorganism is selected from the group consisting of the family Ustilaginaceae or the division Basidiomycetes, preferably from the genus Ustilago and particularly preferably wherein the microorganism is Ustilago maydis. [10] Triglyceride mixture, produced or producible according to any one of claims 1 to 7, wherein the produced triglyceride mixture has a desired fatty acid profile and wherein long-chain triglycerides with a fatty acid chain length of 20 to 24 carbon atoms are present in a mixture produced in such a manner in an amount of more than 2 wt.%, preferably more than 5 wt.%, particularly preferably more than 10 wt.%, based on the total weight of the mixture. [11] Device for carrying out the method according to any one of claims 1 to 7, wherein the device has a controller which, upon input of a triglyceride mixture to be produced with a predetermined fatty acid profile, sets the corresponding parameters in the device after comparison with a stored database.

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