Method for aging meat using mucor aligarensis

Abstract

The present invention relates to a method for aging meat using a Mucor aligarensis strain deposited under accession number KACC 83102BP, and more specifically, to a meat-aging composition comprising the strain, meat aged with the strain, and a method for aging meat using the strain. The strain for aging meat according to the present invention enhances nutty, blue cheese, and buttery flavors in meat, thereby satisfying consumer preference for meat products.

Description

Meat Curing Method Using Mucor Aligarensis

[0001] The present invention provides a method for aging meat using Mucor aligarensis.

[0002] Recently, to meet the diversification of processed foods and consumer preferences for meat, there is a growing demand for flavors derived from natural ingredients, in addition to artificial flavors such as fermented seasonings (MSG), nucleic acid seasonings, and artificial meat flavors. Livestock undergo rigor mortis over a period of time immediately after slaughter. Rigor mortis refers to the stiffening of muscles and joints after death. Rigor mortis causes meat to become hard, tasteless, and leak moisture, reducing its marketability. However, after maximum stiffness, muscles begin to undergo autodigestion through proteolytic enzymes, softening the meat. Autodigestion slightly restores the muscle pH and increases water-holding capacity, leading to enzymes that produce amino acids and unsaturated fatty acids such as oleic acid, enhancing flavor. The above process is collectively referred to as "aging," and two primary methods for aging meat are wet aging and dry aging. First, wet aging is a method of aging meat while it contains moisture. It is a method of aging meat while sealed with a tool such as vinyl. This wet aging method is known to require aging for at least 14 days and as much as 30 days or more at a temperature of 0 to 4℃. Not only does it take a long time to mature, but it also has a high risk of microbial contamination from the external environment, internal organs, and blood during the slaughtering, processing, and aging processes, posing a safety risk due to the proliferation of microbial contamination. There are problems such as the meat being distributed without undergoing a sufficient aging process, or the meat spoiling before the aging date or having a shortened shelf life after the aging date depending on the initial contamination level. Next, dry aging is a method of aging meat by storing it in a well-ventilated aging room without sealing it. Unlike wet aging, which is a common aging method for meat, this method naturally evaporates moisture and matures the meat using traditional meat aging methods that predate technological advancements.The conventional dry aging method has the advantage of being able to produce a unique flavor not only through the action of protein autolytic enzymes inside the meat but also through the action of microorganisms such as bacteria and molds on the surface. However, because the surface of the meat is excessively dried and a hard shell is formed, the microorganisms that affected the early stage of aging can no longer affect it, so even if the aging period is extended, aging does not progress deep into the meat. In addition, since the dried part must be removed when used, there is a problem in that not only is the loss rate high, but it cannot provide excellent sensory characteristics that satisfy consumers.

[0003] The Rural Development Administration has developed a rapid meat aging technology using radio waves (Title of the invention: Rapid meat aging device using radio waves and rapid meat aging method using the same; Application number: 10-2020-0176857; Registration number: 10-2654965). Ji-Hye Moon et al. have developed a meat flavoring agent using supercritical extracted lard (Ji-Hye Moon, In-Wook Choi, Hee-Don Choi, Yun-Sook Kim, "Aroma pattern and sensory characteristics of meat flavoring agent derived from Maillard reaction product using supercritical extracted lard," Korean J. Food Sci. An. Vol. 32, No. 5, pp. 644-651 (2012)). The flavor components of meat are produced by the thermal decomposition of amino acids and peptides, caramelization of carbohydrates, decomposition of nucleotides, decomposition of thiamine, and thermal decomposition of fat, and it is generally known that volatile components containing sulfur compounds or carbonyl groups play a key role. The first study to investigate the aging effect using mold growing on the surface of aged meat was conducted by Kotula et al. in 1988, but reported that there was no difference in preference and cooking quality between meat aged for 14 days with three types of mold (Thamnidium elegans, Mucor mucedo, Chaetostylum fresenii) and meat aged after mold inoculation (Ineffectiveness of three added mold species to enhance the rapid aging of beef (Kotula AW, Campano SG, Kinsman DM. Ineffectiveness of Three Added Mold Species to Enhance the Rapid Aging of Beef. J Food Prot. 1988 Feb;51(2):126-129. doi: 10.4315 / 0362-028X-51.2.126. PMID: 30978777).As a method to increase the flavor of meat using fungi, Hanagasaki et al. proposed beef aging using Mucor flavus, and reported that the functional amino acids GABA (gamma-aminobutyric acid), proline, and aspartic acid increased by dry aging (Hanagasaki T, Asato N. Changes in free amino acid content and hardness of beef while dry-aging with Mucor flavus: Changes in the quality of beef while dry-aging with Mucor flavus. J Anim Sci Technol. 2018 Jul 26;60:19. doi: 10.1186 / s40781-018-0176-6. Erratum in: J Anim Sci Technol. 2018 Sep 14;60:21. PMID: 30065846; PMCID: PMC6062914.). The above Hanagasaki et al. reported that dry aging increases the nutty flavor of Mucor flavus, but did not mention increasing the nutty, blue cheese, and buttery flavors. The present inventors discovered a novel strain that increases the nutty, blue cheese, and buttery flavors of meat when dry aging, and completed the present invention.

[0004] The purpose of the present invention is to provide a new strain that enhances the flavor of meat when inoculated into meat and cultured.

[0005] Another object of the present invention is to provide a meat aging composition comprising the strain.

[0006] Another object of the present invention is to provide aged meat containing the strain.

[0007] Another object of the present invention is to provide a method for aging meat using the strain.

[0008] For the above purpose, the first aspect of the present invention provides a Mucor aligarensis strain having a microbial deposit number of KACC 83102BP. The strain is characterized by increasing the nutty flavor, blue cheese flavor, and butter flavor of meat when inoculated and cultured on meat.

[0009] A second aspect of the present invention provides a meat maturing composition comprising a Mucor aligarensis strain of microbial deposit number KACC 83102BP. The maturing composition can be used to inoculate meat to increase nutty flavor, blue cheese flavor, and butter flavor. The microbial strain can be included in the maturing composition alone or in combination with other microorganisms. As microorganisms that can be used in combination to increase the flavor of meat, kimchi lactic acid bacteria and yeast were reported in Korean Patent Application No. 10-2018-0077868 (filed on July 4, 2018), and Lactiplantibacillus plantarum KM2 strain was reported in Korean Patent Application No. 10-2021-0124768 (filed on September 17, 2021). Therefore, the microorganisms that can be used in combination can be selected from any microorganisms that are known to increase the flavor of meat.

[0010] The above-mentioned maturing composition can be prepared and administered in various formulations and methods according to methods known in the art. For example, the Mucor aligarensis strain of the present invention, the culture solution thereof, the concentrate of the culture solution, or the dried product thereof, may be mixed with a carrier commonly used in food science and prepared in the form of powders, liquids and solutions, tablets, capsules, syrups, suspensions, or granules, and administered. The carrier may be, for example, a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, a coloring agent, a flavoring agent, etc., but is not limited thereto. In addition, the administration dosage may be selected differently depending on the meat, as long as the amount is such that the active ingredient in the meat can be fixed and fermented.

[0011] The maturing composition according to the present invention may include other conventional food additives, and its suitability as the "food additive" is determined by the specifications and standards for the relevant item according to the general provisions and general test methods of the Food Additive Code approved by the Ministry of Food and Drug Safety, unless otherwise specified.

[0012] Items listed in the above "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, high-molecular-weight pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents for noodles, preservative preparations, and tar color preparations.

[0013] The meat may be, but is not limited to, beef, pork, lamb, chicken, goat, fish, or a mixture thereof. Preferably, it may be beef and pork, and more preferably, beef. The microorganism of the present invention may be provided in the form of mycelia or spores. Mycelia have the characteristic of growing rapidly after inoculation, which is advantageous for rapid maturation, but have the disadvantage of making inoculation and storage of mycelia difficult. The spore form has the problem of growing after a considerable period of time after inoculation, which prolongs the maturation time, but is advantageous in terms of storage and transportation of spores.

[0014] A third aspect of the present invention provides meat having a nutty, blue cheese, and buttery flavor, comprising mycelia cultured with the strain Mucor aligarensis, Microbial Accession No. KACC 83102BP. The meat may be, but is not limited to, beef, pork, lamb, chicken, goat, fish, or a mixture thereof, as described above. Beef and pork are preferred, and beef is more preferred.

[0015] The fourth aspect of the present invention provides a method for maturing meat, comprising the steps of inoculating the surface of meat with spores of Mucor aligarensis strain of microbial deposit number KACC 83102BP; and the step of low-temperature culturing the meat inoculated with the spores at 2 to 6°C. More preferably, the method may additionally include a step of maturing the meat using radio waves. For the method for maturing meat using radio waves, the method of "Rapid meat maturation device using radio waves and rapid meat maturation method using the same" (Application number: 10-2020-0176857; Registration number: 10-2654965), which is another patent of the present inventor, can be used. More specifically, 20 to 45 W of meat is treated per kg using radio waves of 27.12 MHz or 13.56 MHz in a meat maturation chamber. In the step of maturing meat using the above radio waves, the temperature at the center of the meat can be maintained at 20 to 30°C. The meat, as described above, may be, but is not limited to, beef, pork, lamb, chicken, goat, fish, or a mixture thereof. Beef and pork are preferred, and beef is more preferred.

[0016] When meat is dry-aged with Mucor aligarensisBC01 according to the present invention, flavors such as nutty flavor, blue cheese flavor, and butter flavor are increased, thereby increasing consumer satisfaction with aged meat.

[0017] Figure 1 shows (a) a photograph of mycelia of Mucor aligarensisBC01 according to the present invention, and (b) a state in which Mucor aligarensisBC01 according to the present invention is cultured on meat during radio wave maturation.

[0018] Figure 2 shows the results of phylogenetic analysis performed using the ITS portion of Mucor aligarensisBC01 according to the present invention, which was purely isolated.

[0019] Figure 3 shows a phylogenetic tree of Mucor aligarensisBC01 according to the present invention with other microorganisms, created using the maximum likelihood phylogenetic tree analysis method.

[0020] Figure 4 shows (a) a photograph of fungal sporangia of Mucor aligarensisBC01 according to the present invention, and (b) a photograph of the 5th day of culture at 20°C on PDA medium.

[0021] Figure 5 shows the results of a sensory evaluation of aged meat according to the present invention.

[0022] Figure 6 shows beef aged after treatment with a strain according to the present invention. (a) Beef treated with Mucor aligarensisBC01 and then radio-treated for 2 days, (b) Beef treated with Mucor aligarensisBC01 and then radio-treated for 2 days and then cultured at low temperature at 4°C for 3 days, (b) Beef treated with Mucor aligarensisBC01 and then radio-treated for 2 days and then cultured at low temperature at 4°C for 6 days.

[0023] [Example 1] Pure isolation and identification of strains

[0024] Among dry-aged meats, dry-aged meat with a better flavor than general dry-aged meat was selected, and the resulting meat was pulverized to isolate and culture microorganisms. The pulverized solution was diluted and cultured on PDA (Potato Dextrose Agar) medium, and the culture was repeated several times by colony re-culturing to isolate yeast and molds, respectively. When smelling the state in which it was cultured on PDA medium, if there was a difference from other microorganisms, it was thought that it would have a great influence on the ripening flavor, so the smell of each isolated and cultured mold was confirmed. Among these, Mucor aligarensisBC01 showed a clearly different PDA smell, developing a new flavor such as a buttery or nutty scent. Mucor racemosus, which grows quickly, showed a sour scent in the medium smell and showed a yeast-like smell change. There was no difference in the medium smell for Penicillium mold.

[0025] Each of the isolated and cultured molds and yeasts was used for beef aging, but the yeast only added sourness to the meat, the Penicillium mold had no effect on improving flavor, and the isolated Mucor racemosus had the fastest growth rate among the Mucor molds, but had a minimal effect on increasing flavor. On the other hand, when Mucor aligarensisBC01 according to the present invention was treated, an increase in nutty flavor, blue cheese flavor, and butter flavor was clearly observed.

[0026] When the Mucor aligarensisBC01 fungus according to the present invention was cultured on a PDA medium, the sour smell of the medium disappeared and a strong nutty, blue cheese, and buttery smell was expressed. Accordingly, Mucor aligarensisBC01 was deposited as a microorganism with the National Institute of Agricultural Sciences on July 1, 2024, and was assigned the microorganism accession number KACC 83102BP. When the Mucor aligarensisBC01 fungus according to the present invention was cultured on a PDA medium at three temperatures: refrigeration (4°C), room temperature (20-25°C), and bacterial culture temperature (37°C), mycelia were confirmed after 2 days at room temperature, and spores began to form after 3 days. Mycelia were confirmed after 2 weeks under refrigeration (4°C), and at 37°C, spores did not germinate even after 4 weeks, and did not germinate even when transferred back to room temperature. Therefore, the appropriate culture temperature for Mucor aligarensisBC01 could be estimated to be in the temperature range of 20-25℃.

[0027]

[0028] [Example 2] Confirmation of increase in nutty flavor, blue cheese flavor, and butter flavor of beef during dry aging after inoculation with strain according to the present invention

[0029] (Comparative Example 1-1)

[0030] The sensory results for the grade 1 Korean beef sirloin cuts without inoculation with the fungal spores of Mucor aligarensisBC01 according to the present invention [comparative example] are as shown in Table 1 below. A total of 14 panelists participated in the sensory experiment. Of these, 5 were male and 9 were female.

[0031]

[0032] (Comparative Example 1-2)

[0033] The sensory results for grade 1 Korean beef sirloin cuts, when radio-wave aging was performed at 25°C for 48 hours without inoculating with the Mucor aligarensisBC01 fungal spores of the present invention and then at 4°C for 3 days [Comparative Example 1-2], are as shown in Table 2 below. A total of 14 panelists participated in the sensory experiment. Of these, 5 were male and 9 were female.

[0034]

[0035] (Example 2-1)

[0036] When distilled water containing Mucor aligarensisBC01 fungal spores at a level of 3 log CFU / mL or higher was inoculated onto the surface of grade 1 Korean beef sirloin cuts, radio-wave aging was performed at 25°C for 48 hours, and then low-temperature incubation was performed at 4°C for 3 days (Example 2-1), the sensory results for grade 1 Korean beef sirloin cuts are as shown in Table 3 below. A total of 14 panelists participated in the sensory experiment. Of these, 5 were male and 9 were female.

[0037]

[0038] When the beef was cultured at a low temperature of 4℃ for 3 days, mold grew on the surface of the beef and the flavor was also good.

[0039]

[0040] (Example 2-2)

[0041] After inoculating with the fungal spores of Mucor aligarensisBC01 according to the present invention, the sensory results for grade 1 Korean beef sirloin cuts were as shown in Table 4 below, after radio wave aging at 25°C for 48 hours and then at 4°C for 6 days (Example 2-2). A total of 14 panelists participated in the sensory experiment. Of these, 5 were male and 9 were female.

[0042]

[0043] [Example 3] Effect of radiofrequency treatment and dry aging on free amino acid content

[0044] Flavor changes in meat aging are driven by the increased production of free amino acids by protein-degrading enzymes, a decrease in neutral and polar fats, and an increase in free fatty acids. The effects of radiofrequency treatment and dry aging on free amino acids were analyzed, and the results are presented in Table 5.

[0045] Amino acid name Free amino acid (mg / 100g) Comparative example 1-1 Comparative example 1-2 Example 2-1 Example 2-2 Alanine 13.605a ± 0.3742 0.734b ± 0.8312 2.785c ± 1.2732 4.189c ± 0.499 Ammonia 1.200a ± 0.0182.502b ± 0.1192.579b ± 0.421 3.287b ± 0.604 Anserine 21.034a ± 0.5112 4.496b ± 3.4302 8.825c ± 0.5232 5.036b ± 2.416 Arginine 3.579a ± 0.2110b ± 00b ± 0.097b ± 0.169 Aspaltate 1.530a ± 0.0613.791b ± 0.3163.738b ± 0.8253.886b ± 0.188 Carnosine 81.542a ± 1.419101.990b ± 4.623104.893b ± 4.362109.479b ± 5.122 Citrulline 0.670a ± 0.1250.726a ± 0.2000.902a ± 0.31.096a ± 0.205 Cystathionine 0.217a ± 0.1880.420a ± 0.1490.530a ± 0.150.606a ± 0.235Cystine0.596a ± 0.1120.725a ± 0.0330.827a ± 0.2241.016a ± 0.37Ethanolamine0a ± 00.036ab ± 0.0630.046a ± 0.0790.1a ± 0.087Glutamic acid4.461a ± 0.7328.113b ± 0.9259.573b ± 1.79611.985b ± 3.02Glycine2.843a ± 0.1454.401b ± 0.1394.898b ± 0.0785.475b ± 0.541Histidine1.615a ± 0.0893.397b ± 0.8583.488b ± 0.2484.221c ± 0.346Hydroxylysine0a ± 00.252b ± 0.0790.294b ± 0.0920.363b ± 0.095Isoleucine3.240a ± 0.3354.282a ± 0.4377.124b ± 0.9888.549b ± 1.074Leucine5.876a ± 0.53810.365b ± 0.65512.975b ± 1.68115.602b ± 2.094Lysine3.795a ± 0.2976.446b ± 0.6269.165c ± 0.39710.703c ± 1.802Methionine3.188a ± 0.2635.828b ± 0.3476.138b ± 0.7577.283b ± 0.966Ornithine1.143a ± 0.145.878b ± 0.1846.509b ± 0.2396.805b ± 0.207Phenylalanine4.277a ± 0.4725.971b ± 0.4588.638c± 1.23210.341c± 1.582Phosphoethanolamine0a ± 00a ± 00.411b ± 0.0320.423b ± 0.025Phosphoserine0.082a ± 0.1420.482b ± 0.0260.545b ± 0.0770.618b ± 0.108Sarcosine0.333a ± 0.2880.419ab ± 0.7780.732a ± 0.2070.864a ± 0.331Serine3.495a ± 0.2174.149b ± 0.3976.595c± 0.1977.376c± 0.699 Taurine 7.303a ± 0.165 8.115ab ± 1.346 8.465ab ± 0.534 8.12b ± 0.713 Threonine 2.554a ± 0.157 5.206b ± 0.397 5.923b ± 0.314 7.03c ± 0.575 Tyrosine 0.431a ± 0.139 0.383a ± 0.012 0.39a ± 0.14 0.593a ± 0.183 Urea 5.282a ± 0.14 5.717a ± 1.243 6.591a ± 0.77 5.113a ± 1.958 Valine 4.004a ± 0.29 35.645b ± 0.5329.834c± 0.7311.571d± 0.993β-Alanine0.296a ± 0.0340.389ab ± 0.2020.482ab ± 0.1340.606b ± 0.172β-Amino Butyric Acid0.730a ± 0.1511.042a ± 1.2031.739a ± 0.5401.605a ± 1.300γ-Amino-n-Butyric Acid0.345a ± 0.090.345a ± 0.1520.396a ± 0.1720.623a ± 0.266Total178.067 ± 3.704242.245 ± 20.76273.452 ± 4.208291.375 ± 20.722.

[0046] * Numerical values ​​represent the mean ± standard deviation (mg / 100g) for three replicates. If the letters following the mean differ for the same amino acid, a significant difference of p<0.05 is indicated by Duncan's multiple range test.

[0047] Compared to the comparative example, in the example, protein was decomposed and most free amino acids increased. In particular, alanine, glycine, lysine, proline, serine, and threonine, which exhibit sweetness, and aspartic acid and glutamic acid, which exhibit umami, increased more in the example than in the comparative example. In particular, since Example 2-1 differed only in the inoculation of Mucor aligarensisBC01 mold spores compared to Comparative Example 1-2, it could be confirmed that the microorganism had a significant effect in increasing free amino acids. This result is also consistent with the occurrence of flavor differences shown in the sensory evaluation.

[0048]

[0049]

Claims

1. Mucor aligarensis strain with microbial deposit number KACC 83102BP.

2. A Mucor aligarensis strain characterized in that the strain in paragraph 1 increases the nutty flavor, blue cheese flavor, and butter flavor of meat.

3. A meat aging composition comprising a strain according to paragraph 1.

4. A meat aging composition according to claim 3, characterized in that the strain is in the form of mycelia.

5. A meat aging composition according to claim 3, characterized in that the strain is in the form of a spore.

6. A meat aging composition according to any one of claims 3 to 5, characterized in that the composition increases nutty flavor, blue cheese flavor, and butter flavor.

7. Meat having a nutty, blue cheese and buttery flavor containing mycelia cultured with the strain according to paragraph 1.

8. Meat according to paragraph 7, characterized in that the meat is beef.

9. A step of inoculating the surface of meat with spores of the strain according to paragraph 1 diluted in distilled water; and A meat aging method comprising a step of culturing meat inoculated with the above spores at a low temperature of 2 to 6°C.

10. A meat aging method characterized in that, in paragraph 9, it additionally includes a step of aging meat using radio waves.

11. A meat aging method characterized in that, in the step of aging meat using radio waves in the 10th paragraph, the temperature of the center of the meat is maintained at 20 to 30°C.

12. A meat aging method according to any one of claims 9 to 11, characterized in that the meat is beef.

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