WO2022030493A1 - Agent de formation de protoporphyrine ix de zinc contenant des bactéries lactiques, et procédé d'amélioration de ton de couleur et procédé de fabrication d'aliments traités utilisant celui-ci - Google Patents

Agent de formation de protoporphyrine ix de zinc contenant des bactéries lactiques, et procédé d'amélioration de ton de couleur et procédé de fabrication d'aliments traités utilisant celui-ci Download PDF

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WO2022030493A1
WO2022030493A1 PCT/JP2021/028777 JP2021028777W WO2022030493A1 WO 2022030493 A1 WO2022030493 A1 WO 2022030493A1 JP 2021028777 W JP2021028777 W JP 2021028777W WO 2022030493 A1 WO2022030493 A1 WO 2022030493A1
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lactic acid
meat
lactococcus
bacterium
color tone
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PCT/JP2021/028777
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Japanese (ja)
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純一 若松
カウサー ウル アラム ムド
徹 早川
朗人 玖村
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国立大学法人北海道大学
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
    • A23L13/60—Comminuted or emulsified meat products, e.g. sausages; Reformed meat from comminuted meat product
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/40—Colouring or decolouring of foods
    • A23L5/42—Addition of dyes or pigments, e.g. in combination with optical brighteners
    • A23L5/46—Addition of dyes or pigments, e.g. in combination with optical brighteners using dyes or pigments of microbial or algal origin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/10—Nitrogen as only ring hetero atom

Definitions

  • the present invention comprises an agent and method for forming zinc protoporphyrin IX under aerobic conditions using Lactococcus lactic acid bacteria, an agent and method for improving the color tone of a meat product, and a meat product. Regarding the method.
  • the color tone of meat is affected by the state of myoglobin, which is the main pigment protein in meat.
  • Myoglobin has one heme, which is an iron complex of protoporphyrin IX, in its molecule, and its color tone changes greatly depending on the type of molecule coordinate-bonded to the sixth ligand of this heme iron and the charge state of iron. do.
  • the color tone of bright red meat seen in fresh meat is due to a derivative called oxymyoglobin in which an oxygen molecule is bound to the 6th ligand and the charge of heme iron is divalent.
  • oxymyoglobin in which an oxygen molecule is bound to the 6th ligand and the charge of heme iron is divalent.
  • the color tone becomes brown and it looks bad, but this is because the heme iron in myoglobin is oxidized to trivalent, and the oxygen of the 6th ligand is removed and replaced with water molecules. This is because the absorption spectrum of light changes by changing to a derivative called.
  • the meat when the meat is heated, it turns grayish brown because the protein portion (globin) of myoglobin in the meat is denatured by heating to become denatured globin hemicromium. All heme iron in the modified globin hemichrome is trivalent in the oxidized state.
  • nitrite derived from a color former produces a carcinogenic N-nitroso compound when it reacts with amines and amides under acidic conditions.
  • processed meat products are classified into Group 1 which is said to have high grounds for carcinogenicity to humans. ..
  • the amount added is extremely small, and in many countries including Japan, the acceptable daily intake (ADI) is scientifically calculated and the usage standard is set, but there are consumers who are concerned about the danger.
  • ADI acceptable daily intake
  • Non-Patent Document 1 ZnPP is more stable to heat and light than heme due to the difference in the coordinated metal. Therefore, elucidation of the formation mechanism and applied technology have been studied, and a patent application has been filed as a color tone improvement technology using ZnPP (Patent Document 1 and Patent Document 2).
  • Non-Patent Document 2 Non-Patent Document 3
  • ZnPP is gradually formed during manufacturing and accumulated in the final product, so it is considered to be particularly suitable for improving the color tone of long-term aged meat products.
  • Long-term aged prosciutto ham and dry sausages such as salami which are examples of such long-term aged meat products, have established a solid position as high-class foodstuffs and continue to be widely produced even today. There is.
  • Non-Patent Document 4 This has been reported in both the system using only the intrinsic factor of meat excluding the influence of microorganisms and the experimental system considering external factors such as microorganisms.
  • large bone-in hams made of thigh meat such as palma ham that does not use a coloring agent, most of them are covered with skin and subcutaneous fat, and the red meat inside is not exposed to oxygen, so ZnPP is used. It is formed and exhibits a vivid color tone.
  • the excised part near the hip joint is significantly dried from the surface layer and has an effect of oxidation, the surface is generally excised and the inside is sliced and eaten.
  • An object of the present invention is to provide a new means capable of forming ZnPP in a meat product whose influence of oxygen cannot be eliminated because it is produced under aerobic conditions.
  • the present inventors have found that lactic acid bacteria belonging to the genus Lactococcus have the ability to form ZnPP even under aerobic conditions in which oxygen, which is a ZnPP formation inhibitor, is present. Was completed.
  • An agent for forming zinc protoporphyrin IX under aerobic conditions in a meat-containing material containing Lactococcus lactic acid bacteria (2) The agent according to (1), wherein the lactic acid bacterium is a bacterium belonging to Lactococcus lactis. (3) The agent according to (1) or (2), wherein the lactic acid bacterium is a bacterium belonging to the Lactococcus lactis subsp. Cremoris. (4) A method for forming zinc protoporphyrin IX in a meat-containing material under aerobic conditions, which comprises adding Lactococcus lactic acid bacteria to the meat-containing material.
  • (10) A method for improving the color tone of a whole meat product produced under aerobic conditions, which comprises adding Lactococcus lactic acid bacteria to a meat product raw material.
  • (11) The method according to (10), wherein the lactic acid bacterium is a bacterium belonging to Lactococcus lactis.
  • (12) The method according to (10) or (11), wherein the lactic acid bacterium is a bacterium belonging to the Lactococcus lactis subspecies Cremoris.
  • (13) A meat product having an improved color tone as a whole, including the addition of Lactococcus lactic acid bacteria to the raw material of the meat product and the production of the meat product from the raw material of the meat product after the addition of the lactic acid bacteria under aerobic conditions. How to manufacture.
  • ZnPP in meat products manufactured under aerobic conditions, not only in the central portion thereof, but also on the surface where it is conventionally difficult to form ZnPP due to the influence of oxygen.
  • ZnPP can also be formed on the surface layer portion.
  • the present invention provides an agent for forming ZnPP under aerobic conditions in a meat-containing material containing Lactococcus lactic acid bacteria.
  • the present invention also provides a method for forming zinc protoporphyrin IX in a meat-containing material under aerobic conditions, which comprises adding Lactococcus lactic acid bacteria to the meat-containing material.
  • ZnPP is Dihydrogen [3,8,13,17-tetramethyl-7,12-divinyl-21H, 23H-porphine-2,18-dipropionato (4-)-N21, N22, N23, N24] zincate (4-) Also represented (CAS No. 15442-64-5), zinc (Zn) is added to porphyrin (protoporphyrin IX), which has a structure in which four methyl groups, two vinyl groups, and two propionic acid groups are bonded to the porphyrin ring. It is a coordinated complex compound.
  • the lactic acid bacterium used in the present invention is a bacterium belonging to the genus Lactococcus, for example, Lactococcus allomyrinae, Lactococcus chungangensis, Lactococcus formosensis, Lactococcus fujiensis, Lactococcus garvieae, Lactococcus hircilactis, Lactococcus kimchi, Lactococcus kimchi, Lactococcus lactis Lactococcus laudensis, Lactococcus nasutitermitis, Lactococcus petauri, Lactococcus piscium, Lactococcus piscium, Lactococcus planta It can be a bacterium belonging to (Lactococcus raffinolactis), Lactococcus reticulitermitis, Lactococcus taiwanensis or Lactococcus termiticola.
  • the lactic acid bacterium is a bacterium belonging to Lactococcus lactis or Lactococcus rafinolactis.
  • Bacteria belonging to Lactococcus lactis are, for example, Lactococcus lactis subsp. Tructae, Lactococcus lactis subsp. Lacttis, and Lactococcus lactis subsp. Lactococcus. It can be a bacterium belonging to lactis subsp. Cremoris) or Lactococcus lactis subsp. Hordniae.
  • the lactic acid bacterium is a bacterium belonging to Lactococcus lactis subspecies Turkutae, Lactococcus lactis subspecies Lactis or Lactococcus lactis subspecies Cremoris.
  • the lactic acid bacterium is a bacterium belonging to the Lactococcus lactis subspecies Cremoris.
  • Lactococcus lactic acid bacteria stored or commercially available in various microbial strain storage institutions can be used.
  • Examples of such lactic acid bacteria are, for example, Lactococcus lactis subspecies Lactis NBRC12007 strain, Lactococcus lactis subspecies Lactis NBRC100933 strain, and lacto deposited at the Bioresource Center (NBRC), Incorporated Administrative Agency.
  • GB (A) -1 strain was issued to the Patent Microorganisms Depositary Center, National Institute of Technology and Evaluation, 2-5-8 Kazusakamatari, Kisarazu City, Chiba Prefecture, Japan, on June 10, 2020. It has been deposited internationally as 03229 (Indication of identification: GB (A) -1).
  • Table 1 shows the mycological properties of the GB (A) -1 strain.
  • the lactic acid bacteria are Lactococcus lactis subspecies Cremoris GB (A) -1, Lactococcus lactis subspecies Lactis NBRC12007 strain, Lactococcus lactis subspecies Lactis NBRC100933 strain, Lactococcus lactis. Subspecies Turkutae NBRC110453 strain or Lactococcus lactis CH-1 strain. Particularly preferred is the Lactococcus lactis subspecies Cremoris GB (A) -1 strain.
  • Lactococcus lactic acid bacteria are prepared by culturing under commonly used culture conditions, for example, using MRS medium at a temperature of 25 to 37 ° C. for 8 to 48 hours under aerobic or anaerobic conditions. can do.
  • Lactococcus lactic acid bacteria can produce ZnPP under aerobic conditions in a material containing meat.
  • the aerobic condition means that the meat-containing material is placed in the air or in a gas containing oxygen having a concentration similar to that in the air.
  • Lactococcus lactic acid bacteria can produce ZnPP by coexisting with a meat-containing material and leaving it under aerobic conditions, for example, at a temperature of 4 to 37 ° C. for several days to several weeks.
  • Lactococcus lactic acid bacteria can produce ZnPP even under anaerobic conditions in a material containing meat.
  • the anaerobic condition means that the gas is placed in a gas having almost no oxygen, for example, an oxygen concentration of less than 0.1 vol%, which is generally used for culturing anaerobic bacteria.
  • the meat-containing material is not limited as long as it contains meat, and is a test sample for an experiment, for example, pork homogenate or minced pork described in J. Wakamatsu et al., Meat Science, 2020, 165, 107989. It may be a raw material for meat products. Examples of meat include beef, pork, mutton, goat meat, horse meat, deer meat, chicken meat, rabbit meat, whale meat and other meats of mammals having a bright red, pinkish red or reddish color. , Not limited to these. When a meat product raw material is used as a material containing meat, the meat product raw material may contain, in addition to the meat, ingredients necessary for the production of the meat product, such as salt and optionally spices.
  • ZnPP can be extracted from the meat-containing material after coexisting with Lactococcus lactic acid bacteria using 75% acetone or the like, and can be detected or measured by HPLC or other methods.
  • ZnPP is detected and measured by measuring the fluorescence intensity at 590 nm when irradiated with excitation fluorescence at 420 nm according to the method described in J. Wakamatsu et al., Meat Science, 2007, 77, 580-586. You can also do it.
  • Color tone improving agent for meat products provides an agent for improving the color tone of all meat products produced under aerobic conditions, including Lactococcus lactic acid bacteria.
  • the present invention also provides a method for improving the overall color tone of a meat product produced under aerobic conditions, which comprises adding Lactococcus lactic acid bacteria to the raw material of the meat product.
  • the present invention includes the addition of Lactococcus lactic acid bacteria to the raw material of the meat product and the production of the meat product from the raw material of the meat product after the addition of the lactic acid bacteria under aerobic conditions, and the color tone is improved as a whole.
  • Provide a method for producing a meat product is provided.
  • color tone improvement means to change the color tone of a meat product, which causes browning or fading under the influence of oxygen when manufactured and stored by a conventional method, to a preferable color tone, specifically, freshness. It means to change to a highly palatable color tone such as red, pink red or reddish color, or to maintain the color tone.
  • the color tone improvement is performed by measuring the lightness L *, the chromaticity a * and b * by a known method for evaluating the color tone of the meat product, for example, by visual observation or by using a color difference meter, and the saturation C * and the hue. It can be evaluated by a method of calculating the angle h or the like.
  • Lactococcus lactic acid bacteria can produce ZnPP in meat-containing materials under aerobic conditions. Therefore, by adding Lactococcus lactic acid bacteria to the raw material of meat products and subjecting them to manufacturing processes such as aging under aerobic conditions, the surface that comes into contact with oxygen and the surface through which oxygen can permeate to a depth of about 5 mm. It is possible to produce a meat product in which ZnPP is formed in a portion (referred to as a surface layer portion). Furthermore, since Lactococcus lactic acid bacteria can produce ZnPP even under anaerobic conditions, the above-mentioned meat products produced using Lactococcus lactic acid bacteria can be produced in any of the central, surface and surface layers thereof. It is also possible that ZnPP is formed in.
  • ZnPP which is a bright red pigment
  • Lactococcus lactic acid bacteria to form a sufficient amount of ZnPP in the whole meat product for improving the color tone, it is possible to produce a meat product having an improved color tone as a whole.
  • Meat products are required to have more remarkable color deterioration due to oxygen, that is, products that require long-term aging or drying under aerobic conditions in the manufacturing process, or long-term storage under aerobic conditions. Those are preferable.
  • the meat product is preferably a non-heated meat product or a dried meat product.
  • meat products include lux ham, prosciutto, semi-dry sausage, and dry sausage.
  • a particularly preferable meat product is semi-dry sausage or dry sausage.
  • oxygen blocking means such as vacuum packaging and oxygen scavengers do not need to be used in particular, but the use is not excluded.
  • conventional color formers such as nitrite do not need to be used in particular, but rather nitric oxide produced by the color former inhibits the formation of ZnPP (J. Wakamatsu et al., Meat Science, 2010, 84 (J. Wakamatsu et al., Meat Science, 2010, 84). 1), 125-128),
  • the meat product produced in this way has an excellent color tone not only in the central portion but also on the surface and the surface layer portion, even though the coloring agent and the oxygen removing means are not used. Therefore, it is possible to avoid product disposal due to deterioration of appearance, and it is expected that it will lead to saving of animal protein resources and reduction of food waste. Furthermore, by not using a color former, there is no risk of nitrosamine formation, and safety is high.
  • the brightness L *, redness a * and yellowness b * of each sample are measured using a color difference meter (CM-700d, Konica Minolta), and the saturation C * and hue angle h are calculated from a * and b *. (Fig. 2).
  • the redness of the bacterial inoculated group was higher than that of the control group and showed a value close to that of the nitrite group.
  • the bacterial inoculation group had a lower yellowness but the same hue angle as the nitrite group. This means that the shade of the bacterial inoculation plot is roughly equal to that of the nitrite plot.
  • the color tone of the bacterial inoculation area is slightly inferior in brightness and vividness, but the color tone is similar to that of the nitrite area.
  • the LbC and LLC plots showed a bright red color and were as bright as the nitrite plot.
  • strong red fluorescence derived from ZnPP was also observed.
  • strong red fluorescence was observed not only inside the cross section but also in the entire cross section including the surface layer portion, and the entire cross section including the surface layer portion had a preferable vivid color tone.
  • the LbC group red fluorescence was not observed in the surface layer portion, and the colors in the surface layer portion and the inside were significantly different.
  • FIG. 6 shows a graph in which the color tone of the surface of the dry sausage is quantified by a color difference meter.
  • the LLC group showed the same degree of redness as the nitrite group, and the hue angle and saturation were also the same.
  • Example 3 ZnPP formation with GB (A) -1 strain (pork homogenate, aerobic and anaerobic conditions) Sterile minced pork was added to physiological saline to a concentration of 30% by mass, and homogenized at 10,000 rpm for 90 seconds using a homogenizer (CELL MASTER CM-100, AZ ONE Co.). Bacterial suspensions of homogenate, NaCl and LnL, LnM, LbP, LbC and LLC prepared in Example 1 were added to a sterile test tube and mixed (final concentration of minced pork 20% by mass, final concentration of NaCl 3 mass). %, Final bacterial concentration 2.0 x 10 6 CFU / mL).
  • test tubes were covered and placed in a gas-impermeable bag and incubated at 25 ° C. for 5 days under aerobic or anaerobic conditions. Anaerobic conditions were maintained with an oxygen absorber (A-500HS, I.S.O. Inc.).
  • an oxygen absorber A-500HS, I.S.O. Inc.
  • a mixture (antibiotic group) to which the final concentration of sulfate 50 ⁇ g / mL) was added was prepared in the same manner as above and incubated. An antibiotic group was set up to confirm that there was no contamination of microorganisms capable of forming ZnPP during the operation.
  • Example 4 ZnPP formation with GB (A) -1 strain (heated or unheated pork homogenate, aerobic and anaerobic conditions) Using unheated salted pork homogenate, short-time heating (100 ° C, 20 seconds) to inactivate the enzyme, and autoclaved pressure-heating (121 ° C, 15 minutes) salted pork homogenate.
  • the ZnPP formation of the Lactococcus lactis subspecies Cremoris GB (A) -1 strain was evaluated in the same manner as in Example 3.
  • Example 5 ZnPP formation by Lactococcus lactic acid bacteria (pork homogenate, aerobic conditions) Lactococcus lactic acid bacteria shown in Table 3 below were added to the salted pork homogenate in the same manner as in Example 3, and ZnPP formation was evaluated under aerobic conditions.
  • NBRC National Institute of Technology and Evaluation Biological Resources Center

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Abstract

La présente invention concerne : un agent et un procédé de formation de protoporphyrine IX de zinc dans des conditions aérobies utilisant des bactéries lactiques appartenant au genre Lactococcus ; un agent et un procédé d'amélioration du ton de couleur d'un produit à base de viande ; et un procédé de fabrication d'un produit à base de viande. Selon la présente invention, de la ZnPP peut être formée, en particulier, non seulement au centre d'un produit à base de viande fabriqué dans un état aérobie, mais également sur la surface ou la partie de couche de surface de celle-ci, où la ZnPP se formait à peine dans l'état de la technique en raison de sa sensibilité à l'oxygène.
PCT/JP2021/028777 2020-08-03 2021-08-03 Agent de formation de protoporphyrine ix de zinc contenant des bactéries lactiques, et procédé d'amélioration de ton de couleur et procédé de fabrication d'aliments traités utilisant celui-ci WO2022030493A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013537427A (ja) * 2010-08-17 2013-10-03 セーホーエル．ハンセン アクティーゼルスカブ ビタミンｋ２高生産性ラクトコッカス・ラクティス株
WO2015111597A1 (fr) * 2014-01-22 2015-07-30 株式会社明治 Procédé de préparation de citrulline
EP3318625A2 (fr) * 2016-11-08 2018-05-09 Innolact Group Oy Souche de bactérie lactique

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2013537427A (ja) * 2010-08-17 2013-10-03 セーホーエル．ハンセン アクティーゼルスカブ ビタミンｋ２高生産性ラクトコッカス・ラクティス株
WO2015111597A1 (fr) * 2014-01-22 2015-07-30 株式会社明治 Procédé de préparation de citrulline
EP3318625A2 (fr) * 2016-11-08 2018-05-09 Innolact Group Oy Souche de bactérie lactique

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ASADUZZAMAN MD; OHYA MOMO; KUMURA HARUTO; HAYAKAWA TORU; WAKAMATSU JUN-ICHI: "Searching for high ZnPP-forming edible bacteria to improve the color of fermented meat products without nitrite/nitrate", MEAT SCIENCE., ELSEVIER SCIENCE., GB, vol. 165, 7 March 2020 (2020-03-07), GB , XP086126185, ISSN: 0309-1740, DOI: 10.1016/j.meatsci.2020.108109 *
KAUSER-UL-ALAM MD., HAYAKAWA TORU, KUMURA HARUTO, WAKAMATSU JUN-ICHI: "High ZnPP-forming food-grade lactic acid bacteria as a potential substitute for nitrite/nitrate to improve the color of meat products", MEAT SCIENCE., ELSEVIER SCIENCE., GB, vol. 176, 1 June 2021 (2021-06-01), GB , pages 108467, XP055894286, ISSN: 0309-1740, DOI: 10.1016/j.meatsci.2021.108467 *
KAUSER-UL-ALAM MD., TOBA YU, HIOKI SHOJI, HAYAKAWA TORU, KUMURA HARUTO, WAKAMATSU JUN-ICHI: "Lactococcus lactis subsp. cremoris Produces Zinc Protoporphyrin IX Both Aerobically and Anaerobically and Improves the Bright Red Color of Fermented Meat Products", FOODS, vol. 9, no. 11, pages 1583, XP055894289, DOI: 10.3390/foods9111583 *

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