Novel Aureobasidium pullulans strain and its use for highly efficient oligosaccharide production

Abstract

This application relates to the Aureobasidium pullulans CJFSKY2201 strain deposited under accession number KCCM13230P; a culture solution of the strain; use of the strain in the production of oligosaccharides; a composition comprising the strain, its crushed product, its culture, its concentrate, or its dried product; and a method for producing oligosaccharides, comprising the steps of culturing the strain and obtaining oligosaccharides from the culture solution.

Description

Technical Field

[0001] This application relates to Aureobasidium pullulans CJFSKY2201 strain deposited under the accession number KCCM13230P; a culture solution of the strain; use of the strain for oligosaccharide production; a composition containing the strain, its disrupted product, its culture, its concentrate, or its dried product; and an oligosaccharide production method including a step of culturing the strain and a step of obtaining oligosaccharides from the culture solution.

Background Art

[0002] Oligosaccharides play important roles in various biological processes and are used in dairy products, confectionery, beverages, etc. of oligosaccharides for the purpose of emphasizing functionality, improving sweet taste, and improving physical properties. Their functionality is known and interest in oligosaccharide synthesis is increasing. Currently, the most widely sold oligosaccharide at home and abroad is fructooligosaccharide, which is widely used as a raw material for various health functional foods and shows a tendency of increasing sales volume every year. Led by this, the demand for overall oligosaccharides is expected to continue to increase.

[0003] Oligosaccharides are also contained in natural foods such as tomatoes, bananas, onions, and mushrooms, but it is difficult to extract them directly from natural foods and in most cases, they are synthesized. Also, for research and actual application of oligosaccharides to medicine and health functional foods, their large-scale synthesis is necessary, but the synthesis of oligosaccharides is very difficult. Currently, methods using enzymes and whole-cell biocatalysis methods using microorganisms are used for the production of oligosaccharides. As conventional methods for producing high-content fructooligosaccharides, chromatography methods and crystallization methods are known, but both methods have extremely low yields, are difficult for mass production, and lack economic efficiency (Korean Registered Patent Publication KR 10-1994-0005660 B1).

[0004] Demand for oligosaccharides continues to increase, and capital investments have been made continuously, but the development of bacterial strains for their production has not yet progressed. However, further physical investment is impossible within the limited manufacturing space, and the rate of productivity improvement relative to the amount invested has been insufficient.

[0005] Against this backdrop, the applicants diligently researched and developed a strain for oligosaccharide production to enhance the activity of the strain itself. As a result, they completed this application by developing a strain that rapidly converts sugar syrup into oligosaccharides. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] Korean Registered Patent Publication KR 10-1994-0005660 B1 [Overview of the project] [Problems that the invention aims to solve]

[0007] The object of this application is to provide the Aureobasidium pullulans CJFSKY2201 strain deposited under accession number KCCM13230P and a method for producing oligosaccharides therefrom. [Means for solving the problem]

[0008] One of the purposes of this application is to provide the Aureobasidium pullulans CJFSKY2201 strain deposited under accession number KCCM13230P.

[0009] Another objective of this application is to provide a culture medium of the Aureobasidium pullulans CJFSKY2201 strain, which was deposited under accession number KCCM13230P.

[0010] Another object of this application is to provide a method for producing oligosaccharides, comprising the steps of culturing the bacterial strain and obtaining oligosaccharides from the culture medium. [Effects of the Invention]

[0011] This application confirms that the Aureobasidium pullulans CJFSKY2201 strain, deposited under accession number KCCM13230P, has high oligosaccharide synthesis ability. This allows for the effective production of oligosaccharides, and the oligosaccharides produced by the manufacturing method described in this application can be usefully used in the production of food compositions, feed compositions, and the like. [Brief explanation of the drawing]

[0012] [Figure 1] This graph shows the results of the oligosaccharide production reaction of conventional strains and selected mutant strains, illustrating the flow rate and cumulative production volume by day over 36 days. [Modes for carrying out the invention]

[0013] This can be explained in more detail as follows: On the other hand, each description and embodiment disclosed in this application also applies to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the categories of this application are not considered to be limited by the specific descriptions described below.

[0014] In one aspect of this application, this application provides the Aureobasidium pullulans CJFSKY2201 strain deposited under accession number KCCM13230P.

[0015] The "Aureobasidium pullulans" of this application is a yeast-like fungus distributed in nature. Aureobasidium pullulans is known to be found in diverse environments such as forest soil, freshwater, and air, as well as on the surfaces of various plants, and is known to possess diverse enzymatic activities, including oligosaccharide synthase, siderophores, and pullulan.

[0016] The applicants have developed a novel strain belonging to Aureobasidium pullulans, named it Aureobasidium pullulans CJFSKY2201, and deposited it under deposit number KCCM13230P.

[0017] In this application, "strain" means a collection of individuals derived from a single cell, reproducing through asexual reproduction and possessing homogeneous genetic characteristics, and within a single species, there may be multiple strains with differing genetic characteristics. In this application, "strain" means Aureobasidium pullulans, and specifically, it may be, but is not limited to, the Aureobasidium pullulans deposited under deposit number KCCM13230P.

[0018] The Aureobasidium pullulans CJFSKY2201 strain of this application may be an isolated strain or a recombinant strain, and the recombinant strain may be performed by genetic modification. It may also be a non-naturally occurring strain or a mutant strain. Furthermore, the strain may include the cells of the Aureobasidium pullulans strain, dried cells, etc. The dried cells may be, but are not limited to, spray-dried, freeze-dried, vacuum-dried, or drum-dried bacteria.

[0019] In this application, in order to store the strain stably for a long time, the cells may be dissolved in a storage solution prepared by mixing a certain amount of glycerol component with water and stored at -70°C, or suspended in sterilized 10% skim milk and freeze-dried. However, it is not limited to these, and various known methods can be used to obtain a long-term storage form.

[0020] The strain may have an increased ability to synthesize oligosaccharides compared to the wild type, or may have an increased activity of oligosaccharide synthase. The oligosaccharide synthase may be fructosyltransferase, but is not limited thereto. Fructosyltransferase is an enzyme used to produce fructooligosaccharides from sugar and raw sugar as raw materials. It is known to have a function of synthesizing oligosaccharides by binding one molecule of fructose to a raw sugar or sugar molecule after hydrolyzing the raw sugar and sugar.

[0021] Another aspect of this application provides a culture solution of the Aureobasidium pullulans CJFSKY2201 strain deposited under the accession number KCCM13230P.

[0022] The term "culture" in this application means growing the strain under appropriately artificially controlled environmental conditions. In this application, the "culture solution" of the strain may be a culture stock solution containing cells, or may be cells obtained by removing or concentrating the culture supernatant. The composition of the culture solution may further contain not only the components necessary for normal culture of Aureobasidium, but also components that synergistically act on the growth of Aureobasidium. Such a composition can be easily selected by those having ordinary skills in the art.

[0023] The medium and other culture conditions used for culturing the strain of the present application can be any medium used for culturing ordinary Aureobasidium microorganisms without special restrictions. Specifically, the strain of the present application can be cultured by adjusting temperature, pH, etc. under aerobic or anaerobic conditions in an ordinary medium containing appropriate carbon sources, nitrogen sources, phosphorus sources, inorganic compounds, inorganic salts, amino acids, vitamins and / or nucleic acids, etc.

[0024] As the medium, a natural medium or a synthetic medium can be used. As the carbon source of the medium, for example, glucose, sucrose, dextrin, glycerol, starch, etc. are used. As the nitrogen source, peptone, meat extract, yeast extract, dried yeast, soybean, ammonium salt, nitrate and other organic or inorganic nitrogen-containing compounds are used. As the phosphorus source, monopotassium phosphate, dipotassium phosphate, or the corresponding sodium-containing salt, etc. may be used, but it is not limited to such components. As the inorganic compound, sodium chloride, calcium chloride, iron chloride, magnesium sulfate, iron sulfate, manganese sulfate, calcium carbonate, etc. are used. As the inorganic salt, magnesium, manganese, calcium, iron, potassium, etc. may be used, but it is not limited thereto.

[0025] In one embodiment of the present application, the Aureobasidium pullulans strain was subjected to seed culture in a medium containing sucrose and yeast extract under the condition of pH 5-6. The main culture was carried out in a medium containing sucrose, yeast extract, dipotassium phosphate, magnesium sulfate and an antifoaming agent, etc. under the condition of about pH 7, but it is not limited thereto, and the components and the charged amounts of each material may be appropriately adjusted.

[0026] Another aspect of this application provides a bead on which the bacterial strain is immobilized. In this application, the bead is for immobilizing the bacterial strain, and there are no special restrictions on the method of its manufacture. Immobilization refers to the process of fixing bacterial cells to a space or part of an object, maintaining their biological catalytic properties, and enabling continuous reuse. Generally, the characteristics of the microorganism must be considered when selecting the immobilization carrier and immobilization method. Immobilization methods include adsorption methods and inclusion methods, and while Ca-alginate, K-carrageenan, gelatin, chitosan, and polyacrylamide gel may be used as immobilization carriers, they are not limited to these. In one embodiment of this application, an alginate solution was mixed with a cell suspension, the mixture was filtered, the mixture was filled into a bead manufacturing tank, and the mixture was deposited in a calcium chloride (CaCl2) solution to produce a bead.

[0027] Another aspect of this application provides a method for producing oligosaccharides, comprising the steps of culturing the bacterial strain and obtaining oligosaccharides from the culture medium.

[0028] The terms "culture" and "culture medium" in this application are as described above.

[0029] Oligosaccharides are sugar solutions obtained by using carbohydrate raw materials and applying enzymes to cause 10 or fewer sugar molecules to form linear or branched chains. These solutions are then filtered, purified, and concentrated to produce liquid or powdered oligosaccharides, and there are two types of food products: oligosaccharides and processed oligosaccharide products.

[0030] In this application, the term "oligosaccharides" refers to sugars with approximately 2 to 10 sugar units, and includes types such as isomaltoligosaccharides, fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, gentiooligosaccharides, maltooligosaccharides, and mixed oligosaccharides.

[0031] The "fructooligosaccharides" of this application are a mixture of sugars in which one or more fructose molecules, specifically about 1 to 5 fructose molecules, or more specifically, about 1 to 3 fructose molecules, are bound to the fructose residue of sugar, and include, but are not limited to, 1-kestose (GF2), nystose (GF3), and 1-F fructofuranosyl nystose (GF4). Fructooligosaccharides are mainly produced by enzymatic hydrolysis of raw sugar or sucrose, but are not limited to this. Fructooligosaccharides are used in a variety of processed foods and are widely used in the manufacture of mochi, castella, traditional Korean sweets, cakes, jellies, etc., but are not limited to this.

[0032] The aforementioned oligosaccharide processed products refer to products that have been processed by adding food or food additives to oligosaccharides, and each product contains 10% or more of fructooligosaccharides, isomaltoligosaccharides, galactooligosaccharides, xylooligosaccharides, and gentiooligosaccharides, and 40% or more of maltooligosaccharides.

[0033] In this application, the method for obtaining oligosaccharides using the aforementioned bacterial strain may be a method widely known in the art, or it may be a continuous process method, a semi-batch method, a batch method, etc., but is not limited thereto. Preferably, the method may be one in which sugar is mixed with the culture medium of the bacterial strain and reacted to obtain oligosaccharides from the reaction product, or one in which the fructosyltransferase enzyme is separated from the culture medium of the bacterial strain, the separated enzyme is mixed with sugar and reacted to obtain oligosaccharides from the reaction product, but is not limited thereto. In this application, "sugar (GF)" is also called sucrose or cane sugar, and refers to a disaccharide in which one molecule of fructose and one molecule of glucose are linked by a glycosidic bond.

[0034] The step of mixing and reacting the sugars may be carried out in a temperature range of 40°C to 60°C, but is not limited to this range, and the reaction time can be continued until the desired amount of the target substance is obtained.

[0035] The step of obtaining the oligosaccharide may be carried out by conventional separation methods known in the art. Such separation methods may include centrifugation, filtration, chromatography, and crystallization. For example, the reactants may be separated by ion-exchange chromatography after removing biomass by slow centrifugation of the reactants, but this is not limited to this method. Alternatively, the target substance may be recovered from the reactants by separating the bacterial strain and filtering, without a separate purification step. Alternatively, the recovery step may further include a purification step.

[0036] In this application, random mutations were induced in Aureobasidium pullulans strains used in oligosaccharide production by ultraviolet irradiation, and high-titer strains were developed even under the same equipment and culture conditions. As a result, it is possible to convert large amounts of sugar into oligosaccharides more rapidly than before. This not only leads to increased productivity but also enables responses to urgent demand. Furthermore, oligosaccharides can be produced in a shorter time than before, significantly reducing associated fixed costs such as fuel and labor, and enabling more stable oligosaccharide production.

[0037] Another aspect of this application provides a composition comprising the Aureobasidium pullulans CJFSKY2201 strain, deposited under accession number KCCM13230P, its lysates, its cultures, its concentrates, or its dried products. More detailed information regarding the strain can be found in the preceding text.

[0038] The composition containing the Aureobasidium pullulans CJFSKY2201 strain, its crushed product, its culture, its concentrate, or its dried product can be used as a composition for oligosaccharide production and can be used in the manufacture of food compositions or feed compositions.

[0039] The state of the bacterial strain may be liquid or dry, and the drying method may include, but is not limited to, forced-air drying, natural drying, spray drying, and freeze-drying.

[0040] The composition may further contain a cryoprotectant or an excipient. The cryoprotectant or excipient may be, but is not limited to, a non-naturally occurring substance or a naturally occurring substance. As another specific example, the cryoprotectant or excipient may be, but is not limited to, a substance that the Aureobasidium pullulans CJFSKY2201 strain does not naturally come into contact with or is not naturally present with the strain. As yet another specific example, the composition may further contain one or more cryoprotectants selected from the group consisting of glycerol, trehalose, maltodextrin, skim milk powder, and starch; and / or one or more excipients selected from the group consisting of glucose, dextrin, and skim milk. The cryoprotectant of this application may be included in the composition in an amount of 0.01% to 20% by weight or 0.01% to 10% by weight relative to the total weight of the composition. Specifically, the composition may contain 5% to 20% by weight of glycerol, 2% to 10% by weight of trehalose, 2% to 10% by weight of maltodextrin, 0.5% to 2% by weight of skim milk powder, and 0.1% to 1% by weight of starch. Furthermore, the excipient may be included in an amount of 75% to 95% by weight or 85% to 95% by weight relative to the total weight of the composition.

[0041] Furthermore, a method for producing a composition comprising the Aureobasidium pullulans CJFSKY2201 strain, its crushed product, its culture, its concentrate, or its dried product may include a step of mixing the Aureobasidium pullulans CJFSKY2201 strain, its crushed product, its culture, its concentrate, or its dried product with an additive. The additive may be the cryoprotective agent or excipient described above.

[0042] Another aspect of this application provides a food composition comprising oligosaccharides produced by the above manufacturing method.

[0043] At that time, the term "oligosaccharide" is as described above.

[0044] The term "food" in this application includes all foods in the ordinary sense, such as meats, sausages, bread, chocolates, candies, snacks, confectionery, pizzas, ramen noodles, other noodle products, gums, dairy products including ice cream, various soups, drinking water, tea, energy drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods.

[0045] The term "functional food" is synonymous with "food for special health use (FoSHU)" and refers to a food with high medical and therapeutic effects that has been processed to efficiently demonstrate biological regulatory functions in addition to nutritional supply. Here, "function" means obtaining effects useful for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body. The food of this application can be manufactured by methods commonly used in the industry, and can be manufactured by adding raw materials and components commonly added in the industry during said manufacturing. Furthermore, the dosage form of the food can be manufactured without restriction as long as it is a dosage form recognized as a food.

[0046] The term "health food" refers to foods that have a more active effect on maintaining or promoting health compared to general foods, while "health supplement food" refers to foods intended to supplement health. In some cases, the terms "health functional food," "health food," and "health supplement food" are used interchangeably.

[0047] Specifically, the aforementioned health functional foods are foods that are produced by adding the composition of this application to food ingredients such as beverages, teas, spices, gums, and confectionery, or by encapsulating, powdering, or suspending them. When consumed, they provide specific health benefits, but unlike general medicines, they have the advantage of not having the side effects that can occur with long-term use of medicines, as they are made from food ingredients.

[0048] At that time, the amount of oligosaccharides contained in the food is not particularly limited, but may be 0.01 to 100% by weight, specifically 1 to 80% by weight, relative to the total weight of the food composition.

[0049] The food composition may further contain a physiologically acceptable carrier, but the type of carrier is not particularly limited, and any carrier commonly used in the art may be used.

[0050] Furthermore, the food composition may contain additional ingredients commonly used in food compositions to enhance aroma, taste, appearance, etc. For example, it may contain vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. It may also contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr); and amino acids such as lysine, tryptophan, cysteine, and valine.

[0051] Furthermore, the food composition may contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder and high-grade bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color fixatives (sodium nitrite, sodium acetate, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclomate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium bitartrate, etc.), fortifiers, emulsifiers, thickeners, coating agents, gum bases, foam inhibitors, solvents, and improvers. The additives may be selected according to the type of food and used in appropriate amounts.

[0052] Another aspect of this application provides a feed composition containing oligosaccharides produced by the above manufacturing method.

[0053] At that time, the term "oligosaccharide" is as described above.

[0054] In this application, the term "feed" means any natural or artificial prescribed food, meal, or components of such meal that an animal eats, ingests, and digests.

[0055] The type of feed is not particularly limited, and feeds commonly used in the art may be used. Non-restrictive examples of the feed include plant-based feeds such as grains, root vegetables, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, meal, or grain by-products; and animal-based feeds such as proteins, inorganic substances, oils and fats, mineral substances, milk fats, single-cell proteins, zooplankton, or food. These may be used individually or in combination of two or more.

[0056] The amount of oligosaccharides contained in the feed composition of this application varies depending on the intended use and conditions of use of the feed. For example, it may be included in an amount of 0.01 to 100% by weight, more specifically 1 to 80% by weight, relative to the total weight of the livestock feed composition, but is not limited thereto.

[0057] Another aspect of this application provides the use of the Aureobasidium pullulans CJFSKY2201 strain, deposited under accession number KCCM13230P, for the production of oligosaccharides. [Examples]

[0058] The present application will be described in more detail below with reference to the following embodiments. However, the following embodiments are merely illustrative of the present application and the scope of the present application is not limited to them.

[0059] Example 1. Method for culturing bacterial strains In this application, the seed culture of the Aureobasidium pullulans strain was performed in a medium containing sucrose and yeast extract under conditions of pH 5-6, at which time the average optical density (OD) was 10.

[0060] The main culture of the bacterial strain was performed in a medium containing sucrose, yeast extract, dipotassium phosphate, magnesium sulfate, and an antifoaming agent, under conditions of approximately pH 7, with an average OD of 160.

[0061] Example 2. Selection of mutant strains Example 2-1. Primary sorting One ml of Aureobasidium pullulans culture solution was taken and diluted with 9 ml of 3 M sterile diluent (Saline), then dispensed into sterile plates (SPL Petridish) for a 10-fold dilution. The plates were then exposed to ultraviolet (UV) light from above for 20 seconds in a clean bench. After irradiation, 0.1 ml was taken from each plate and diluted 100-fold with 9.9 ml of 3 M sterile diluent (Saline). One ml of the diluted solution was dispensed into each plate and cultured in PDA (BD Difco) medium using aluminum foil to protect from light, at 28°C for 2-3 days until the bacteria had fully grown.

[0062] The cultured strains were grouped according to their characteristics, and 150 ml of culture medium was placed in a 500 ml shaking flask (Lab tech) for each group, and pre-seed culture was performed at 220 RPM. After 21 hours, the optical density (OD) and titer were measured to select superior strains.

[0063] Example 2-2. Secondary Selection The mutant strains selected in Example 1-1 were each prepared by adding 0.9 ml of 40% glycerin to 0.9 ml of culture medium, and one MCB capsule (1.8 ml of Thermo) was prepared for pure isolation. Twenty bacteria were randomly selected from these capsules, and 150 ml of the culture medium was placed in a 500 ml shaking flask for pre-seed culture at 240 RPM. After 21 hours, the OD and titer were measured to select superior strains.

[0064] After pure isolation of MCB 1 capsules produced from selected mutant strains, pure isolation was performed on vials of strains showing high titer. Ten cells were randomly selected from each vial, and 150 ml of culture medium was placed in a 500 ml shaking flask for pre-seed culture at 240 RPM. After 21 hours, OD and titer were measured to select superior strains.

[0065] Example 2-3. Final Selection After pure isolation of one capsule from each of the strains that showed high titer in Examples 1 and 2, 10 cells were selected from each according to their characteristics and subcultured in slant medium. Then, 150 ml of the culture solution was placed in a 500 ml shaking flask and pre-seed cultured at 240 RPM. After 21 hours of incubation, the OD and titer were measured to select superior strains.

[0066] [Table 1]

[0067] As shown in Table 1 above, the culture results confirmed that strain 4-1-7-1 showed an increase in titer of approximately 24% compared to the conventional strains, and this strain was ultimately selected as the superior strain.

[0068] The selected strain was named Aureobasidium pullulans CJFSKY2201 and deposited with the Korean Culture Center of Microorganisms (KCCM), an international depositary under the Budapest Convention, on August 24, 2022, receiving deposit number KCCM13230P.

[0069] Example 3. Pilot reactor experiment with selected mutant strains The final selected mutant strain (4-1-7-1) and the conventional strain were experimented with in a pilot reactor scaled down to the ratio of the in-situ reactor. The reactor flow rate and yield of the conventional strain and the mutant strain were compared. The reactor jacket temperature was set to 49°C.

[0070] A comparison of the titer of the selected final mutant strain (4-1-7-1) and the conventional strain, which directly affects oligosaccharide productivity, was performed. As shown in Table 2 below, the average titer was 370 for the conventional strain and 450 for the mutant strain, confirming an improvement of approximately 20% in average titer compared to the conventional strain.

[0071] [Table 2]

[0072] After conducting the oligosaccharide production reaction for a total of 36 days, the mutant strain produced 178L of oligosaccharides, while the conventional strain produced 144L. As shown in Table 3 below, it was confirmed that the oligosaccharide productivity of the mutant strain was approximately 23% higher than that of the conventional strain.

[0073] [Table 3]

[0074] Furthermore, as shown in Figure 1, the flow rate of the mutant strain increased by an average of +0.04 compared to the conventional strain, resulting in a cumulative oligosaccharide production of approximately 23% over 36 days.

[0075] Example 4. In-situ reactor experiment with selected mutant strains The final selected mutant strains and conventional strains were compared using an in-situ reactor. Other culture conditions were the same as in the pilot reactor experiment, and flow rate and production volume in the in-situ reactor were compared. As a result, it was confirmed that the culture patterns, such as titer, pH, DO, OD, and C / W, were almost similar to those of the pilot reactor experiment. Furthermore, the in-situ culture results showed that the mutant strains had approximately 10% higher oligosaccharide production compared to the conventional strains, as shown in Table 4 below.

[0076] [Table 4]

[0077] From the above description, a person skilled in the art will understand that this application can be implemented in other specific forms without altering its technical idea or essential features. In this regard, it should be understood that the embodiments described above are merely illustrative and not limiting. The scope of this application should be interpreted as encompassing all modified or altered forms derived from the meaning and scope of the claims, as described below, and their equivalent concepts.

Claims

1. Aureobasidium pullulans strain CJFSKY2201, deposited under accession number KCCM13230P.

2. The strain according to claim 1, wherein the strain has increased oligosaccharide synthesis ability compared to the wild type.

3. Culture medium of Aureobasidium pullulans strain CJFSKY2201, deposited under accession number KCCM13230P.

4. (a) the step of culturing the bacterial strain described in claim 1; and (b) A method for producing oligosaccharides, comprising the step of obtaining oligosaccharides from the culture medium.

5. The method for producing oligosaccharides according to claim 4, wherein step (b) involves mixing sugar with the culture medium and allowing it to react, and then obtaining oligosaccharides from the reaction product.

6. The method for producing oligosaccharides according to claim 4, wherein step (b) involves separating the fructosyltransferase enzyme from the culture medium, mixing the separated enzyme with sugar and reacting the mixture, and then obtaining oligosaccharides from the reaction product.

7. The oligosaccharide production method according to any one of claims 4 to 6, wherein the oligosaccharide is a fructooligosaccharide.

8. The method for producing oligosaccharides according to claim 7, wherein the fructooligosaccharide comprises 1-kestose, nystose, and 1-F fructofuranosyl nystose.

9. The method for producing oligosaccharides according to claim 5 or 6, wherein the reaction step is carried out in a temperature range of 40°C to 60°C.

10. A composition comprising the Aureobasidium pullulans CJFSKY2201 strain deposited under accession number KCCM13230P, its lysates, its cultures, its concentrates, or its dried products.

11. Use of Aureobasidium pullulans strain CJFSKY2201, deposited under accession number KCCM13230P, for oligosaccharide production.

Images