WO2001090394A1 - Procede de production d'un antioxydant - Google Patents

Procede de production d'un antioxydant Download PDF

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Publication number
WO2001090394A1
WO2001090394A1 PCT/JP2001/004312 JP0104312W WO0190394A1 WO 2001090394 A1 WO2001090394 A1 WO 2001090394A1 JP 0104312 W JP0104312 W JP 0104312W WO 0190394 A1 WO0190394 A1 WO 0190394A1
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WO
WIPO (PCT)
Prior art keywords
antioxidant
culture
preculture
seed culture
seed
Prior art date
Application number
PCT/JP2001/004312
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English (en)
Japanese (ja)
Inventor
Ryo Kumazaki
Original Assignee
Ryo Kumazaki
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ryo Kumazaki filed Critical Ryo Kumazaki
Priority to AU2001260605A priority Critical patent/AU2001260605A1/en
Publication of WO2001090394A1 publication Critical patent/WO2001090394A1/fr

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Classifications

    • 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
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/42—Cobalamins, i.e. vitamin B12, LLD factor

Definitions

  • the present invention relates to a method for producing an antioxidant.
  • antioxidants are required to suppress sebum oxidation by UV-A and UV-B, and to prevent skin aging and damage to DNA by active oxygen generated in the process.
  • BHA butylhydroxydisole
  • BHT butylhydroxyltoluene
  • the present invention has been made in view of the above circumstances, and provides a method for extracting an antioxidant substance having excellent antioxidant activity, which is suitable for agricultural products, livestock products, food, cosmetics, pharmaceuticals, and concrete, with an organic solvent. It is an object of the present invention to provide a method for producing an antioxidant, wherein an antioxidant is obtained by a microbial reaction without using the same.
  • the inventors of the present invention have conducted research on applied technology and experiments on useful microorganisms that produce antioxidants, and as a result, have completed a production technology for producing antioxidants in large quantities.
  • the method for producing an antioxidant according to the present invention comprises:
  • the method for producing an antioxidant according to the present invention comprises:
  • FIG. 1 is a graph showing an ultraviolet absorption spectrum curve of the antioxidant liquid according to the present invention.
  • FIG. 2 is a diagram showing an oxidation-reduction potential measurement in which iron powder is added to an antioxidant solution and water according to the present invention.
  • Seed cultures are obtained by culturing the freeze-dried cells as photosynthetic bacteria in a seed medium.
  • a plurality of freeze-dried bacteria of photosynthetic bacteria can be separately seed-cultured, and a plurality of seed cultures can be used.
  • one or more species of photosynthetic bacteria that are anaerobic bacteria are used.
  • it is a bacterium belonging to Rhodopseuumonas palust ris, Rhodopseumonas spheroides, Rhodopseudomonas sphero ides, and Rhodops euaomo n "ascapsu 1 ata belonging to Rhodops euaomo n" ascapsu 1 ata. Is preferred.
  • seed culture In this specification, the terms “seed culture”, “pre-culture”, and “complex culture” are used to describe the amount. Culture medium) Expressed as the amount included. In the present invention, the “seed culture”, “preculture”, and “complex culture” are preferably prepared as a culture solution.
  • the seed culture is precultured under anaerobic conditions in the presence of an antioxidant to obtain a preculture.
  • an antioxidant is added to the seed culture, and in a container under an anaerobic environment at a temperature of 25 ° C to 30 ° C, depending on other conditions, 30 to 4 Incubate for 0 days.
  • the plurality of seed cultures are mixed and used.
  • the use of antioxidants suppresses excessive germs and suppresses oxidative effects.
  • the individual photosynthetic bacteria prepared as 1 0 8 ⁇ 1 0 1 (1 / m L photosynthetic bacteria cultures contained. It should be noted, including the number of bacteria, hereby In this document, the number of bacteria is described as the number of bacteria measured according to the plate culture method.
  • the antioxidants that can be used in this step include vitamin A, vitamin B group, and vitamin E. Among them, it is particularly preferable to use KMX of vitamin B12, a product of KORINK REA Co., Ltd. Such antioxidants are preferably used in the form of an aqueous solution.
  • a substrate As such a substrate, fish solution pull is preferable.
  • Fish Solupur is heat-treated fish residue. It consists of 16 amino acid compositions. It is preferable to use a material that has been subjected to heat treatment again or treated with lactic acid to a pH of 3.5 or less.
  • rice bran, fish cake, powdered kelp, and the like can be used or used in combination.
  • wastewater from tofu plants wastewater from starch plants, human wastewater, livestock manure wastewater, and fishery processing wastewater can also be used as substrates.
  • substrates By culturing photosynthetic bacteria using these as substrates, antioxidants can be obtained.
  • the above anaerobic condition refers to an environment in which air is shut off from the outside world, and in a sealed container, air mixed in at the time of sealing exists but light enters. Disgust When culturing photosynthetic bacteria under light conditions, the substrate can be selected so that hydrogen sulfide is generated. In this case, various bacteria are suppressed, and photosynthetic bacteria grow well.
  • the seed culture is 1 to 3 parts by weight
  • the antioxidant solution containing 0.01 to 0.05% by weight of antioxidant is 1 to 5 parts by weight
  • the substrate is 0.5 to 0.5 parts by weight. It is preferable to mix it by 5 to 5 parts by weight.
  • the substrate is added to the preculture, the main culture is performed under aerobic conditions under the presence of antioxidants until the cells are inactivated, and the culture solution is adjusted to pH 6.4 to 6. 6 to obtain an antioxidant aqueous solution.
  • the preculture can be performed without adding a substrate.
  • the preculturer and the substrate are added to the antioxidant aqueous solution, and the cells are cultured under aerobic and dark conditions while maintaining the water temperature at 25 ° C to 30 ° C.
  • This culture is performed until the cells are self-digested and inactivated.
  • Ammonia is generated by autolysis of the cells, and the culture solution becomes highly alkaline.However, under aerobic conditions, ammonia is expelled, and the culture solution has a pH of 6.4 to 6.6 as described above. Will be processed.
  • the aerobic state is maintained by passing oxygen or air through the culture solution. Thereby, an aqueous solution of an antioxidant can be obtained.
  • the antioxidant obtained according to the present invention is a vitamin; B12. This has been confirmed by a microorganism identification method using a strain of Lact obac i 11 us De lbruecki i subsp. Lact is (ATCC 7830).
  • a substrate such as fish zollible used in the preculture step.
  • This substrate may not be added.
  • the cells mainly cause autolysis without going through the growth process.
  • the subsequent process is the same as when the substrate is added.
  • Antioxidants that can be used in this process include vitamin A, vitamin B group, and vitamin E. Among them, especially, KMX of vitamin B12, a product of KOR IN K 0 REA Co., Ltd. It is preferable to use Such antioxidants are preferably used in the form of an aqueous solution.
  • the aerobic and dark conditions refer to an environment in which air is circulated from the outside world and air circulates in a closed container so that light does not enter.
  • the liquid impurities are allowed to settle naturally, and an antioxidant aqueous solution is obtained through a filter from the lower part of the tank.
  • a freeze-dried bacterium (freezedried) JCM2524 was selected from the JCM microbial strain published by the RIKEN Microorganism Preservation Facility and used.
  • a culture medium consisting of 1 L of water and having a pH of 7 was anaerobically cultured at 30 ° C. by i
  • a freeze-dried ATCC 17023 is selected from the ATCCB acteria and B acteriophages log of America Type Copper Culture Co 11ection.
  • Malic acid 2.5 g Yeast extract lg, Ammonium sulfate 1.25 g, Magnesium sulfate heptahydrate 0.2 g, potassium chloride dihydrate 0.07 g, ferric citrate 0.01 g, ethylenediaminetetraacetic acid 0.02 g, dipotassium hydrogen phosphate 0.6 g, diphosphate potassium hydrogen 0. 9 g, trace elements 1 ml (Beautique down iron (f erri cc it rat e) 0. 3g, MnSO 4 0.
  • the preculture was prepared as photosynthetic bacteria culture individual photosynthetic bacteria contain 10 8 ZML. '
  • aqueous solution of 5 parts by weight of KMX solution (product of KORIN KORE A) containing 0.01% by weight of KMX as an antioxidant, 5 parts by weight of the above preculture and 0.5 parts by weight of fish solution were added.
  • KMX solution product of KORIN KORE A
  • fish solution under aerobic and dark conditions, maintain the water temperature at 25 ° C to 30 ° C, and perform the main culture until the cells are inactivated and the cells are completely inactivated.
  • PH 6.5 After that, the impurities were allowed to settle naturally and passed through a filter from the lower part of the tank to obtain an aqueous solution of an antioxidant.
  • the antioxidant was identified as vitamin B12 by a microorganism identification method using a strain of Lact obacil lus De lbrecki is ubsp. Lact is (ATCC 7830).
  • Test substance (antioxidant obtained in Example 4 above) 20 g ZKg was orally administered to mice, and changes in general condition and body weight were observed for 7 days, and then dissected, and each organ was visually observed. As a result, there were no deaths in each group from immediately after administration to the day after the end of observation, and no piloerection, diarrhea, sweating, deep breathing or abnormal behavior was observed throughout the test period. No abnormalities were observed by gross observation at necropsy. This proved that there was no toxicity with a single dose of the test substance of 20 g / Kg (Tables 1 and 2).
  • mice Average of mice Observation period Abnormality and death
  • UV-A 400 ⁇ ! ⁇ 320 nm
  • UV-B 320 nn! ⁇ 29 Onm
  • UV-A causes suntan
  • UV-B has a property that causes sunburn on the epidermis.
  • DNA has an ultraviolet peak at 26 Onm.
  • the antioxidant solution obtained in Example 4 was tested with an ultraviolet absorption spectrum (FIG. 1, Table 3).
  • the absorbance in Table 3 when irradiated with ultraviolet light at 20 Onm, it shows an absorbance of 1.24, and then the absorbance sharply decreases as the wavelength becomes longer. At 234 ⁇ m, the absorbance is 0.086, 26 Onm At 0.042 and 30 Onm, the absorbance was 0.030 and at 40 Onm, the absorbance was 0.013. From this test, the absorbance of UV-A (400 nm to 320 nm) and UV-B (320 nm to 290 nm) ultraviolet rays was extremely low, and it was possible to protect sebum. In this test, a 100-fold diluted solution of the antioxidant substance of Example 4 was used. From this test, it was found that the absorbance was reduced by the wavelength of the ultraviolet light.
  • oxidation-reduction potentiometer ORP
  • the results are shown in Fig. 2.
  • the oxidation-reduction potential of the water into which the iron powder has been introduced shows a state of 127 mv from the third day, the iron powder turns gray, and on the fifth day the iron powder turns brown.
  • the iron powder in the natural antioxidant liquid maintained its state at the time of injection even after the 9-day test. This test demonstrated an antioxidant effect.
  • a method for extracting an antioxidant which is suitable for agricultural products, livestock products, food, cosmetics, pharmaceuticals, and concrete and has excellent antioxidant activity with an organic solvent is provided. It is possible to provide a method for producing an antioxidant in which an antioxidant is obtained by a microbial reaction without using the antioxidant. That is, the antioxidant obtained by the present invention suppresses deterioration, deterioration, discoloration, discoloration and harm caused by ultraviolet rays, and therefore has a wide field of application.

Abstract

L'invention concerne une solution aqueuse antioxydante obtenue par un procédé comprenant (1) une étape de culture de graines visant à obtenir un milieu de culture de graines d'une bactérie photosynthétique ; (2) une étape de préculture visant à obtenir un milieu de préculture par préculture du milieu de culture de graines mentionné ci-dessus en présence d'un antioxydant dans des conditions d'anaérobie et de lumière ; et (3) une étape de culture principale du milieu de préculture mentionné ci-dessus auquel a éventuellement été ajouté un substrat en présence d'un antioxydant dans des conditions d'anaérobie et d'obscurité jusqu'à ce que les cellules soient inactivées, consistant ensuite à ajuster le pH du milieu de culture liquide de 6,4 à 6,6.
PCT/JP2001/004312 2000-05-24 2001-05-23 Procede de production d'un antioxydant WO2001090394A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2001260605A AU2001260605A1 (en) 2000-05-24 2001-05-23 Process for producing antioxidant

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2000152573A JP2002306193A (ja) 2000-05-24 2000-05-24 抗酸化物質の製造方法
JP2000-152573 2000-05-24

Publications (1)

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WO2001090394A1 true WO2001090394A1 (fr) 2001-11-29

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JP (1) JP2002306193A (fr)
KR (1) KR20030036187A (fr)
AU (1) AU2001260605A1 (fr)
WO (1) WO2001090394A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007083935A1 (fr) * 2006-01-20 2007-07-26 Kyeong Yong Choi Produit cosmétique utilisant des bactéries photosynthétiques vivantes et utilisation correspondante
KR100847729B1 (ko) * 2007-01-19 2008-07-23 최경용 광합성세균 배양액의 악취제거방법 및 악취가 제거된광합성세균 배양액을 함유하는 피부 외용제

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20070075215A (ko) * 2006-01-10 2007-07-18 나광출 광합성세균을 이용한 화장료 및 그 용도
CN101455688B (zh) * 2008-12-08 2011-09-28 大连交通大学 一株兼性厌氧海洋红假单胞菌的活性提取物及其制法和用途
JP5563551B2 (ja) * 2011-12-27 2014-07-30 農業生産法人株式会社 熱帯資源植物研究所 ホモゲンチジン酸の生産方法、抗酸化剤の製造方法、および抗酸化飲料の製造方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4750397B1 (fr) * 1969-01-18 1972-12-18
DE2908769A1 (de) * 1979-03-06 1980-09-18 Biotechnolog Forschung Gmbh Verfahren zur gewinnung von metallfreien corrinoiden
JPS5678585A (en) * 1979-11-30 1981-06-27 Japan Synthetic Rubber Co Ltd Cultivation of phototrophic bacterium
EP0131456A2 (fr) * 1983-07-08 1985-01-16 Kureha Kagaku Kogyo Kabushiki Kaisha Préparation de la vitamine B12 au moyen d'un hybride de cellules fusionnées

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4750397B1 (fr) * 1969-01-18 1972-12-18
DE2908769A1 (de) * 1979-03-06 1980-09-18 Biotechnolog Forschung Gmbh Verfahren zur gewinnung von metallfreien corrinoiden
JPS5678585A (en) * 1979-11-30 1981-06-27 Japan Synthetic Rubber Co Ltd Cultivation of phototrophic bacterium
EP0131456A2 (fr) * 1983-07-08 1985-01-16 Kureha Kagaku Kogyo Kabushiki Kaisha Préparation de la vitamine B12 au moyen d'un hybride de cellules fusionnées

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
E. SIEFERT ET AL.: "Studies on the vitamin B12 auxotrophy of rhodocyclus purpureus and two other vitamin B12-requiring purple nonsulfur bacteria", ARCH. MICROBIOL., vol. 132, 1982, pages 173 - 178, XP002945450 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007083935A1 (fr) * 2006-01-20 2007-07-26 Kyeong Yong Choi Produit cosmétique utilisant des bactéries photosynthétiques vivantes et utilisation correspondante
KR100847729B1 (ko) * 2007-01-19 2008-07-23 최경용 광합성세균 배양액의 악취제거방법 및 악취가 제거된광합성세균 배양액을 함유하는 피부 외용제

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AU2001260605A1 (en) 2001-12-03
KR20030036187A (ko) 2003-05-09

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