Rimosilactobacillus fermentum LRCC 5387 strain for improving the flavor and texture of bread and its uses

Abstract

This invention provides the Rimosilactobacillus fermentum LRCC 5387 strain, which improves the flavor and texture of bread, and its uses. [Solution] Rimosilactobacillus fermentum LRCC 5387, deposited as KCCM13523P derived from milk. Limosilactobacillus fermentum It constitutes the LRCC 5387) strain.

Description

Technical Field

[0001] The present invention relates to the strain Lactobacillus fermentum LRCC 5387 which improves the flavor and texture of bread and its uses.

Background Art

[0002] Bread refers to a product made by mixing various materials mainly with wheat flour, fermenting them, and then baking or steaming. At this time, it can be divided into many types according to the manufacturing method, raw materials used, blending ratios, etc. In the case of the manufacturing method, it can be divided into the sponge method, the tangzhong method, the direct method, etc., and the types of bread have been developed very diversely, such as sandwich bread, butter rolls, sweet buns, bagels, etc. In the past, each natural yeast derived from raw materials was utilized to make bread by long-term fermentation, but with the development of technology, the commercial bread market has been able to grow significantly due to the development of bread-making yeast.

[0003] According to Global Bread Market (2024 - 2033) by Custom Market Insights, the global bread market in 2023 was at a level of approximately 230 million US dollars and was analyzed to show an average annual growth rate of approximately 3.66%. Among these, the market with the most rapid growth is the Asia-Pacific region, which was found to be in line with the domestic trend of pursuing quick and convenient meal substitute products. As the consumption of domestic bread increases, the preferences of consumers diversify, and there is a need for diversification of bread products to meet this.

[0004] Lactic acid bacteria produce lactic acid during their growth process. Fermented foods using lactic acid bacteria acquire various advantages due to the organic acids and metabolites produced by the bacteria during their metabolic process. These advantages include improved shelf life by lowering the pH of the product, improved digestion and absorption through the breakdown of protein, and enhanced flavor and palatability. According to references, there is a growing trend in the baking industry for consumers to prefer natural substances to improve quality, and there are examples of using lactic acid bacteria to enhance flavor or improve the physical properties of the dough. Furthermore, Korean Patent No. 10-01823088 describes how fermenting bread with kimchi-derived lactic acid bacteria resulted in superior sensory properties and delayed staling. Based on this, the present inventors sought to find lactic acid bacteria that could improve the flavor of bread, and after understanding the flavor characteristics of lactic acid bacteria, attempted to develop bread and a method for producing it using them. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] Korean Patent No. 10-1823088 Specification [Non-patent literature]

[0006] [Non-Patent Document 1] Korean J.Food Cookery SCI.Vol.22,No.3(P270~281) June,2006 [Overview of the Initiative] [Problems that the invention aims to solve]

[0007] The object of the present invention is to provide the Rimosilactobacillus fermentum LRCC 5387 strain and its uses for improving the flavor and texture of bread.

[0008] The objects of the present invention are not limited to those mentioned above. The objects of the present invention will become more apparent in the following description and will be realized by the means and combinations thereof described in the claims. [Means for solving the problem]

[0009] To achieve the above objective, the present invention relates to Rimosilactobacillus fermentum LRCC 5387, which was deposited as KCCM13523P derived from milk. Limosilactobacillus fermentum This provides the LRCC 5387) strain.

[0010] The aforementioned strain may use, but is not limited to, one or more of the following as a carbon source: arabinose (L-Arabinose), ribose (D-Ribose), galactose (D-Galactose), glucose (D-Glucose), fructose (D-Fructose), mannose (D-Mannose), maltose (D-Maltose), lactose (D-Lactose), melibiose (D-Melibiose), sucrose (Sucrose), and raffinose (D-Raffinose).

[0011] The aforementioned bacterial strain may have 16S rRNA containing the base sequence of Sequence ID No. 1.

[0012] The aforementioned strain can improve the flavor of bread.

[0013] The aforementioned flavors may include Maillard notes and sour notes.

[0014] The aforementioned bacterial strain can improve the texture of bread.

[0015] Another object of the present invention is to provide a lactic acid bacteria fermentation liquid comprising one or more of the following: the bacterial strain, its crushed product; its culture; its fermented product; and extracts of the bacterial strain, its crushed product, culture solution, or fermented product.

[0016] Still another object of the present invention is to provide a method for producing bread including a step of using the lactic acid bacteria fermentation broth.

[0017] Still another object of the present invention is to provide bread containing one or more of the strain, its disrupted product; its culture; its fermented product; and an extract of the strain, disrupted product, culture solution or fermented product.

Advantages of the Invention

[0018] The present invention relates to the Lactobacillus fermentum LRCC 5387 strain for improving the flavor and texture of bread and its uses. More specifically, when making bread by producing dough using the strain, it was confirmed that compared to the case where the strain is not used, the Maillard aroma and sour aroma increase and the flavor is improved.

[0019] The effects of the present invention are not limited to the effects mentioned above. It should be understood that the effects of the present invention include all effects inferable from the following description.

Brief Description of the Drawings

[0020] [Figure 1] It is a diagram showing the result of confirming the flavor using an electronic nose when producing a wheat flour fermentation broth using the strain of the present invention. [Figure 2] It is a diagram showing the result of confirming the flavor using an electronic nose when producing white bread using the strain of the present invention. [Figure 3] It is a diagram showing the result of confirming the change in the aroma pattern using an electronic nose when producing white bread using the strain of the present invention.

Modes for Carrying Out the Invention

[0022] While explaining each drawing, like reference numerals have been used for like components. In the accompanying drawings, the dimensions of each structure are shown enlarged for the clarity of the present invention.

[0023] In this specification, terms such as "comprising" or "having" are intended to specify that the features, numbers, steps, operations, components, parts or combinations thereof described in the specification exist, and it should not be understood as precluding the existence or addition possibility of one or more other features, numbers, steps, operations, components, parts or combinations thereof. Also, when a part such as a layer, film, region, plate, etc. is "on" another part, this includes not only the case where it is "directly on" the other part but also the case where there are further other parts in between. Conversely, when a part such as a layer, film, region, plate, etc. is "under" another part, this includes not only the case where it is "directly under" the other part but also the case where there are further other parts in between.

[0024] Unless otherwise explicitly stated, all numbers, values, and / or expressions used herein to express quantities of components, reaction conditions, polymer compositions, and formulations should be understood to be, in all cases, modified by the term “approximately,” as each such number is an approximation that reflects the various uncertainties of measurement that arise when obtaining such values ​​from essentially different sources. Furthermore, where a range of numbers is disclosed herein, such range is continuous and, unless otherwise noted, includes all values ​​from the minimum to the maximum value of such range. Additionally, where such range represents integers, it includes all integers from the minimum to the maximum value of such range, unless otherwise noted.

[0025] This invention aims to enhance the flavor of fermented bread by increasing the Maillard aroma and sour aroma, which are caramelization aromas, using the Rimosilactobacillus fermentum LRCC 5387 strain. More specifically, when applied to bread making, the Rimosilactobacillus fermentum LRCC 5387 strain, which has excellent functionality and palatability, can reduce the cheese-like fermentation odor characteristic of lactic acid bacteria and increase the savory aroma.

[0026] This invention relates to Rimosilactobacillus fermentum LRCC 5387, which was deposited as KCCM13523P derived from milk. Limosilactobacillus fermentum We provide the LRCC 5387) strain.

[0027] The aforementioned "market milk" refers to crude oil that has been processed and sterilized.

[0028] The aforementioned strain can use one or more of the following as a carbon source: arabinose (L-Arabinose), ribose (D-Ribose), galactose (D-Galactose), glucose (D-Glucose), fructose (D-Fructose), mannose (D-Mannose), maltose (D-Maltose), lactose (D-Lactose), melibiose (D-Melibiose), sucrose (Sucrose), and raffinose (D-Raffinose).

[0029] The aforementioned bacterial strain may have 16S rRNA containing the base sequence of Sequence ID No. 1.

[0030] The aforementioned strain can improve the flavor of bread.

[0031] The aforementioned flavors may be Maillard aroma and sour aroma.

[0032] The aforementioned bacterial strain can improve the texture of bread.

[0033] Furthermore, the present invention provides a lactic acid bacteria fermentation liquid comprising one or more of the above-mentioned bacterial strain, its crushed product; its culture; its fermented product; and extracts of the above-mentioned bacterial strain, crushed product, culture solution, or fermented product.

[0034] Furthermore, the present invention provides a method for producing bread that includes a step of using the lactic acid bacteria fermentation liquid.

[0035] In the above manufacturing method, the weight ratio of wheat flour to the lactic acid bacteria fermentation liquid may be 10:0.5 to 20, but preferably 10:1.

[0036] In the above manufacturing method, the lactic acid bacteria fermentation liquid may be 10 Baker's % relative to 100 Baker's % of wheat flour.

[0037] Baker's % is a method of expressing the amounts of all ingredients as percentages based on the amount of flour, where the weight of the flour (g) is set as 100%, and all remaining ingredients are calculated as percentages of the weight of the flour.

[0038] Furthermore, the present invention provides bread containing one or more of the above-mentioned bacterial strain, its crushed product; its culture; its fermented product; and extracts of the above-mentioned bacterial strain, crushed product, culture solution, or fermented product.

[0039] The bread may be fermented bread. [Examples]

[0040] The present invention will be described in detail below with reference to examples and comparative examples. However, the following examples and comparative examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0041] Example 1. Isolation of Rimosilactobacillus fermentum LRCC 5387 strain. 1. Sample collection and isolation of lactic acid bacteria To select lactic acid bacteria suitable for improving the flavor of fermented bread, approximately 150 kimchi samples were purchased from traditional markets throughout the country. 44 samples were then collected from the collected milk at the Lotte Wellfood Pasteur Factory (Hoengseong, Gangwon-do, South Korea), based on the collection point. The kimchi and milk samples were diluted 10-fold with sterile distilled water and then homogenized using a pulverizer (Stomacher, Pro-Media SH-001, ELMEX). The diluted solution was then multi-diluted with sterile saline, and 0.1 ml was taken. This was then spread onto MRS solid medium containing 0.002% by weight BCP (Bromocresol purple) and 1.5% by weight Agar, and cultured in a 37°C incubator for 48 hours.

[0042] After culturing, colonies exhibiting a yellow ring were selected and individually subcultured 2-3 times on the BCP-MRS solid medium described above to isolate each strain as a pure culture.

[0043] The aforementioned bacterial strains were individually inoculated into MRS liquid medium and re-cultured at 37°C for 48 hours. The cells were then placed in 1 ml of a freezing solution (a mixture of MRS liquid medium and glycerol in a 4:1 ratio, pre-dispensed into cryo-tubes) and stored frozen at -70°C. The frozen strains were used as starters for other experiments of the present invention.

[0044] 2. Measurement of bacterial strain growth and pH reduction (1) Confirmation of bacterial strain growth and pH changes in wheat flour To improve the flavor of fermented bread, it is important that lactic acid bacteria grow within the flour and produce many metabolites. Furthermore, it is necessary to prevent the generation of off-odors during flour fermentation while enhancing the flavor of the flour. Therefore, the growth rate within the flour and the flavor of the fermented flour product of each strain isolated above were compared, and superior strains were selected.

[0045] Of the lactic acid bacteria isolated as described above, those isolated from fresh milk were inoculated onto MRS agar plates and cultured at 37°C for 24 hours. A portion of the cultured colonies was taken, inoculated into MRS liquid medium, and then cultured again at 37°C for 24 hours to produce a starter culture solution.

[0046] To test the growth and pH reduction of isolated lactic acid bacteria in wheat flour, the starter culture solution was centrifuged at 8000g for 10 minutes, the supernatant was discarded, and the solution was diluted with the same volume of distilled water and then mixed uniformly again. One part by weight of the resulting starter wash solution was inoculated into 100 parts by weight of a mixture of wheat flour and distilled water in a 1:2 weight ratio. A lactic acid bacteria fermentation solution was then produced by culturing at 37°C for 18 hours, and the number of viable lactic acid bacteria was measured using the serial dilution method and the agar streaking method. One g of the strain wheat flour culture solution was taken, diluted to the appropriate dilution ratio with 0.85% sterile physiological saline, and 100 μL of the diluted solution was streaked onto MRS agar medium. After culturing for 48 hours, the number of colonies was measured. pH changes were also confirmed using a pH meter. The analytical results are shown in Table 1 below.

[0047] [Table 1]

[0048] 3. Flavor measurement of lactic acid bacteria fermented liquid To evaluate the flavor of the lactic acid bacteria fermented liquid, a lactic acid bacteria fermented liquid was prepared using the same method as in Example 1.2.(1), and the changes in taste and aroma patterns were analyzed using gas chromatography / mass spectrometer (GC-MS) (GC: Aglient Technologies 7890B, MS: Aglient Technologies 5977A) and an electronic nose Hercles II (Alpha MOS, Toulouse, France). The results are shown in Figure 1, and the experimental values ​​are shown in Table 2.

[0049] [Table 2]

[0050] The electronic nose is an electronic device that mimics the human olfactory system, designed to complement the limitations of the human nose, which cannot continuously detect different odors and has limitations in the range of odors it can detect. The electronic nose can detect volatile fragrance components released from a sample. The electronic nose separates complex volatile organic compounds into single components through two columns, a non-polar column and a weakly polar column, and uses a flame ionization detector (FID) to output electrical signals for each single component, which can then be detected. In other words, the electronic nose can detect specific components contained in a sample through the detection of fragrance components.

[0051] Using a statistical program, we were able to perform discriminant function analysis on information obtained through an electronic nose (Alpha Mos).

[0052] As a result, it was confirmed that the sour and fruity aromas were stronger and the green aroma was weaker compared to the control group (Figure 1).

[0053] Analysis of the aroma components revealed that the main components for each aroma characteristic were acetic acid for sour aromas, and ethyl acetate and ethyl lactate for fruity aromas. Ethyl acetate is known to have fruity and sweet sensory characteristics, while ethyl lactate is known to have sharp and buttery sensory characteristics. Candidate bacterial strains with fruity aromas were selected compared to the control group. In this case, the control group was Lactic acid bacteria Lacticaseibacillus paracasei, as specified in Korean Patent No. 10-2320816 and No. 10-2466220. Lacticaseibacillus paracasei )LRCC5240 and Lactiplantibacillus plantarum ( Lactiplantibacillus plantarum LRCC5318 was used in a mixture.

[0054] Example 2. Bread production and analysis of physical properties and flavor 1. Bread production To confirm the flavor-improving effect of applying lactic acid bacteria fermentation liquid to fermented bread, we manufactured white bread and evaluated its flavor. The lactic acid bacteria fermentation liquid was manufactured using the method described in Example 1, and the sponge and dough method was used for manufacturing the white bread. The mixing ratios used in the test are shown in Table 3 below. Baker's % used as the unit for the mixing ratio is a method of expressing the amount of all ingredients as a percentage based on the amount of flour, with the weight of flour (g) set as 100%, and all remaining ingredients calculated as a percentage of the weight of flour.

[0055] As shown in Table 3, the ingredients were weighed in Baker's % units so that the lactic acid bacteria fermentation solution was 10% of 100% of strong flour. The ingredients for the starter dough were mixed for 6 minutes, and then fermented in a constant temperature and humidity chamber at 27°C and 75% humidity for 4 hours. The starter dough, with the shortening removed, was added to the fermented starter dough and mixed for 8 minutes, after which the shortening was added. This dough was mixed for an additional 8 minutes, given a resting period of 15 minutes, then divided and rounded into equal portions, and given an intermediate fermentation for 15 minutes. After the fermentation was complete, the dough was shaped, placed in a loaf pan, and fermented in a constant temperature and humidity chamber at 38°C and 85% humidity for 1 hour. After fermentation was complete, it was baked in an oven with top heating at 180°C and bottom heating at 210°C for 25 minutes. At this time, the control group was the lactic acid bacterium *Lacticacea paracasei* described in Korean Patent No. 10-2320816 and No. 10-2466220. Lacticaseibacillus paracasei )LRCC5240 and Lactiplantibacillus plantarum ( Lactiplantibacillus plantarum LRCC5318 was used in a mixture, and the other manufacturing methods were the same as those for the bread manufacturing method described above.

[0056] [Table 3]

[0057] 2. Physical property analysis - volume and hardness When evaluating the quality of bread, flavor is important, but volume and texture are also crucial factors. Therefore, we attempted to evaluate the physical properties of the bread prepared as in Example 2. To measure the volume of the bread, we cut a suitable portion from the middle, measured its weight, and then measured its volume using a volume measuring instrument (Volcan Profiler 300). Subsequently, we calculated the specific volume (mL / g) of the bread using the measured weight and volume.

[0058] Furthermore, the hardness of the bread was measured using a texture analyzer (TA-XT plus). The hardness of the bread is expressed as a TA value, and a lower value indicates a softer texture. The moisture content of the bread was measured using an infrared moisture meter (FD-720), and the results are shown in Table 4 below.

[0059] [Table 4]

[0060] As a result, compared to the control group, we confirmed that the volume increased, the hardness decreased, a softer texture was imparted, and the degree of moisture loss was also reduced.

[0061] 3. Flavor and sensory analysis To evaluate the flavor of bread treated with flavor-enhancing lactic acid bacteria, changes in taste and aroma patterns were analyzed for the bread prepared in Example 2 using gas chromatography-mass spectrometry (GC-MS) (GC: Aglient Technologies 7890B, MS: Aglient Technologies 5977A) and an electronic nose Hercles II (Alpha MOS, Toulouse, France). The results are shown in Figure 2, and the experimental values ​​for Figure 2 are shown in Table 5. Furthermore, based on the experimental values, the change in the aroma pattern of the LRCC 5387-treated bread compared to the control group was represented in a spider chart as shown in Figure 3.

[0062] [Table 5]

[0063] Analysis of the fragrance components revealed that the main components for each fragrance characteristic are diacetyl and acetoin for creamy fragrance, acetic acid for sour fragrance, and furfuryl alcohol, furfural, and 2-pentylfuran for Maillard fragrance. Diacetyl is known to have a strong buttery and creamy sensory characteristic, while acetoin is known to have a sweet, buttery, and milky sensory characteristic. Furfuryl alcohol is known to have a sweet and caramel-like sensory characteristic, furfural is known to have a sweet, almond-like, and baked bread-like sensory characteristic, and 2-pentylfuran is known to have a bean-like and earthy sensory characteristic.

[0064] A panel sensory evaluation was conducted on bread treated with flavor-enhancing lactic acid bacteria to investigate its flavor, texture, taste, and palatability. The panel consisted of 18 people who blindly evaluated bread made using the method described in Example 2, and the results are shown in Table 6.

[0065] [Table 6]

[0066] Flavor, texture, and taste were investigated using a 9-point scale. The preference for the control group and the bread treated with the LRCC 5387 strain was assessed by selecting the product with the higher preference score. As a result, the bread treated with the LRCC 5387 strain received high scores in flavor, texture, and taste, and was selected as the final preference by 13 out of 18 panelists. Based on these results, the LRCC 5387 strain was ultimately selected as the flavor-improving lactic acid bacterium.

[0067] Example 3. Identification of Rimosiractobacillus fermentum LRCC 5387 strain 1. Morphological and biochemical identification The morphological and biochemical characteristics of the selected strains were analyzed. The analysis results are shown in Table 7 below.

[0068] [Table 7]

[0069] As can be seen from Table 7 above, the bacterial strain of the present invention was confirmed to be Gram-positive, rod-shaped, spore-forming, and catalase-negative.

[0070] 2. Confirmation of sugar utilization To easily identify the aforementioned strains, we attempted to measure their sugar utilization using the API 50 CHL kit (bioMerieux, France). More specifically, colonies cultured on MRS solid medium were suspended in 10 ml of API 50 CHL medium, inoculated into tubes with API 50 CH strips, and cultured in a 37°C incubator for 48 hours. The results of each strain's fermentation of the sugar substrate were then interpreted and are shown in Table 8 below.

[0071] [Table 8]

[0072] As can be seen in Table 7, the LRCC 5387 strain was able to use arabinose (L-Arabinose), ribose (D-Ribose), galactose (D-Galactose), glucose (D-Glucose), fructose (D-Fructose), mannose (D-Mannose), maltose (D-Maltose), lactose (D-Lactose), melibiose (D-Melibiose), sucrose (Sucrose), and raffinose (D-Raffinose) as carbon sources.

[0073] 3. Genetic Identification For accurate identification, the 16s rRNA sequences of the aforementioned strains were amplified using the general-purpose primers 785F (GGATTAGATACCCTGGTA) and 907R (CCGTCAATTCMTTTRAGTTT), and identified as Rimosiractobacillus fermentum using the NCBI GenBank database. Limosilactobacillus fermentum The nucleotide sequence similarity of the strain was analyzed. As a result, the nucleotide sequence of the strain's 16s rRNA gene was as shown in Sequence ID No. 1, confirming that it has up to 99% homology with Rimosilactobacillus fermentum.

[0074] Therefore, the inventors identified the aforementioned strain as a new Rimosiractobacillus fermentum strain, named it Rimosiractobacillus fermentum LRCC 5387, and deposited it with the Korea Microbial Conservation Center on October 23, 2024 (deposit number KCCM13523P).

[0075] In summary, when the wheat flour fermentation liquid produced using the Rimosilactobacillus fermentum LRCC 5387 strain according to the present invention was applied to bread, it was confirmed that Maillard aroma and sour aroma increased compared to the control group. This was not merely a simple physicochemical measurement; it was also selected as a preferred product in panel analysis, confirming that the flavor characteristics were enhanced.

[0076] Therefore, it is expected that using the strain of the present invention will enable the production of bread with enhanced Maillard aroma, which is known as the savory flavor of bread, and that this will be useful later when combining it with lactic acid bacteria that suit the characteristics of the product to impart a variety of flavors.

[0077] The above description of the present invention is illustrative, and a person with ordinary skill in the art to which the present invention pertains will understand that it can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. Therefore, it should be understood that each embodiment described above is illustrative in all respects and not limiting.

[0078] The scope of the present invention is defined by the claims described below, and all modifications or alterations derived from the meaning, scope, and equivalent concepts of the claims shall be construed as being included within the scope of the present invention. [Accession Number]

[0079] Depository name: Korea Center for Microbial Conservation (KCCM) Accession number: KCCM13523P Date of acceptance: 20241023

Claims

1. The Limosilactobacillus fermentum LRCC 5387 strain, deposited as KCCM13523P, originated from city milk.

2. The rimosylactobacillus fermentum LRCC 5387 strain according to claim 1, characterized in that the strain uses one or more selected from the group consisting of arabinose (L-Arabinose), ribose (D-Ribose), galactose (D-Galactose), glucose (D-Glucose), fructose (D-Fructose), mannose (D-Mannose), maltose (D-Maltose), lactose (D-Lactose), melibiose (D-Melibiose), sucrose (Sucrose), and raffinose (D-Raffinose) as a carbon source.

3. The rimosilactobacillus fermentum LRCC 5387 strain according to claim 1, characterized in that the strain has 16S rRNA containing the base sequence of Sequence ID No.

1.

4. The aforementioned strain is characterized by improving the flavor of bread, as described in claim 1, for the rimosilactobacillus fermentum LRCC 5387 strain.

5. The limosilactobacillus fermentum LRCC 5387 strain according to claim 4, characterized in that the aforementioned flavor is a Maillard aroma and a sour aroma.

6. The aforementioned strain is characterized by improving the texture of bread, as described in claim 1, for the rimosilactobacillus fermentum LRCC 5387 strain.

7. A lactic acid bacteria fermentation liquid comprising one or more of the following: a bacterial strain according to any one of claims 1 to 6; a crushed product thereof; a culture thereof; a fermented product thereof; and an extract of the bacterial strain, crushed product, culture solution, or fermented product.

8. A method for producing bread, comprising the step of using the lactic acid bacteria fermentation liquid of claim 7.

9. Bread comprising one or more of the following: a bacterial strain according to any one of claims 1 to 6, its crushed product; its culture; its fermented product; and an extract of the bacterial strain, its crushed product, its culture solution, or its fermented product.

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