WO2019093543A1 - Procédé de production d'une composition améliorant l'immunité comprenant un extrait de noni traité par enzyme - Google Patents

Procédé de production d'une composition améliorant l'immunité comprenant un extrait de noni traité par enzyme Download PDF

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Publication number
WO2019093543A1
WO2019093543A1 PCT/KR2017/012679 KR2017012679W WO2019093543A1 WO 2019093543 A1 WO2019093543 A1 WO 2019093543A1 KR 2017012679 W KR2017012679 W KR 2017012679W WO 2019093543 A1 WO2019093543 A1 WO 2019093543A1
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WIPO (PCT)
Prior art keywords
enzyme
extract
treated
noni
immunity
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PCT/KR2017/012679
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English (en)
Korean (ko)
Inventor
조용준
백광수
심재석
최수영
박영민
김현지
김상우
이진우
윤만석
권진혁
Original Assignee
주식회사 뉴트리사이언스
(주)뉴트리바이오텍
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Publication of WO2019093543A1 publication Critical patent/WO2019093543A1/fr

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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
    • A23L29/00—Foods or foodstuffs containing additives; 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/06—Enzymes
    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system

Definitions

  • the present invention relates to a noni ( Morinda citrifolia ) extract and a preparation method thereof, and relates to a noni extract and a preparation method thereof capable of improving the immunity enhancing effect.
  • Morinda citrifolia is called Indian mulberry in India, Pajitian in China, painkiller tree in Caribbean, cheese fruit in Australia, and Nono in Tahiti island. Jiu) or the Pharaoh ( ⁇ ⁇ ⁇ ) is called.
  • Noni leaves, stems, flowers, fruits and seeds have been used in folk remedies and are recorded as the best natural remedy according to the ancient literature in the South Pacific. Indeed, the anthraquinone and serotonin in noni have been shown to help digestion, relieve pain, and help with hypertension and cancer.
  • the present invention also provides a method for producing an immunity enhancing composition comprising an enzyme-treated noni extract.
  • the antagonist may include one or more selected from the group consisting of peclyve, rohament, viscozyme, and cytolase.
  • the above-mentioned enzyme-degrading enzyme may be added at a concentration of 0.01 to 0.5 parts by weight based on 100 parts by weight of the raw material.
  • the treatment temperature of the enzyme is 20 to 80 ° C.
  • the present invention may further comprise a step of extracting, filtering and drying the product after the treatment with the enzyme.
  • a noni extract which is produced according to the above-described method, can be provided.
  • a health functional food containing the extract may be provided.
  • the production of an immune-related factor can be improved, and thus an extract having an immunity-enhancing effect can be provided.
  • an extract having an immunity-enhancing effect can be provided.
  • it can be applied to various fields such as food as a health functional composition.
  • FIG. 1 is a schematic view of a method for producing an extract according to an embodiment of the present invention.
  • FIG. 3 is a graph showing changes in macrophage-derived TNF-a according to the concentration of the extract.
  • FIG. 4 is a graph showing changes in TNF-a from rat serum according to the concentration of the extract.
  • FIG. 5 is a graph showing changes in mouse spleen-derived IFN-? -Related mRNA according to the concentration of the extract.
  • FIG. 6 is a graph showing changes in mouse spleen-derived TNF-.alpha. MRNA according to the concentration of extract.
  • FIG. 7 is a graph showing changes in IL-12-related mRNA from mouse spleen according to the concentration of the extract.
  • the immunity enhancing composition comprising the enzyme-treated noni extract according to the embodiment of the present invention and a method for producing the same will be described in detail.
  • &quot raw material " refers to a material that has been subjected to washing, hydrolysis, pulverization, and drying after removing the foreign matter after harvest, without being subjected to any other chemical treatment, extraction or filtration.
  • the noni matter can be obtained by hydrolysis, pulverization and lyophilization of noni.
  • the raw material can be obtained, for example, by pulverizing water without adding water to the noni, followed by extraction and filtration, and freeze-drying.
  • freeze-drying can be carried out by preliminarily freezing and then drying to a moisture content of 5% or less. For example, after freezing at -40 ° C, 30 to 50 ° C, for example, at 40 ° C for 30 to 60 hours, For example, such that the moisture content is not more than 5%, for example, 3 to 5% for 48 hours.
  • a pin mill, a disc mill, or the like may be used as the step of crushing the raw material, and the crushing method is not particularly limited as long as it is a general crushing method.
  • the amount of water in the hydrolysis step may be 2 to 50 times, for example, 5 to 20 times, based on the weight of noni.
  • the enzyme may include one or more selected from the group consisting of peclyve, rohament, viscozyme, and cytolase, but is not limited as long as it is a commonly used protease.
  • Peclyve can be generated by Aspergillus niger and can include peclyve CP, peclyve PR, or a combination thereof.
  • peclyve CP may contain a pectin-lyase, a polygalacturonase, and a pectin-methyl-esterase.
  • peclyve PR may contain pectin-lyase, pectin-methyl-esterase.
  • Peclyve may be provided in liquid form and has an optimal temperature range of 40 to 55 ⁇ ⁇ . It can also be used in diets and adjuvants, specifically for anti-oxidant and vitamin extraction.
  • Rohament can be produced by Trichoderma reesei and is a liquid fungal cellulase enzyme for the hydrolysis of starch-free polysaccharides. It is a cellulase enzyme that is used to hydrolyze starch-free polysaccharides. Cellulase, beta-glucan, And other enzymes that degrade plant cell wall polysaccharides such as silanes (xylan, xylan).
  • the rohament can be processed in a distillation process and a mashing process.
  • Viscozyme can be produced by Aspergillus sp and can be produced by a variety of enzymes such as arabanase, cellulose, beta-glucanase, hemicellulose and xylanase , xylanase).
  • the major enzyme activity of viscozyme is the activity of endo- ⁇ -glucanase which hydrolyzes 1,3 or 1,4-mers in ⁇ -D-glucan. Viscozyme is known to be an effective enzyme for extracting polyphenols from apples and the like.
  • Cytolase can be produced by Aspergillus niger and is an acid resistance pectinase and can be used as a pressing enzyme for depectinization and compaction in the processing of various fruit juices.
  • the enzyme of the present invention may be added in an amount of 0.01 to 0.5 parts by weight, for example, 0.1 to 0.3 parts by weight, based on 100 parts by weight of the raw material.
  • the treatment temperature of the said enzyme is 20 to 80 ° C, for example 40 to 60 ° C.
  • a powdered enzyme it is necessary to warm and stir the enzyme so that the powder does not aggregate when the enzyme is melted, and it can be maintained at a temperature range that does not affect the activity of the enzyme.
  • the enzyme treatment can be carried out with stirring for 30 minutes to 10 hours, for example 1 to 5 hours.
  • the method may further include a step of extracting, filtering and drying the product after the treatment with the enzyme-degrading enzyme.
  • the conditions for extracting the product after the enzyme treatment can be 50 to 120 ° C, for example, 80 to 110 ° C for 1 to 10 hours, for example, 2 to 8 hours.
  • the extraction method may include a solvent extraction method, a supercritical extraction method, an ultrasonic extraction method, a hot water extraction method, and the like.
  • the filtration and drying step may include, for example, a freeze-drying method, an evaporation concentration method, a reverse osmosis concentration method, a hot air drying method, a spray drying method, a membrane separation concentration method, and the like.
  • the drying step of the present invention may comprise a spray drying step.
  • additives such as excipients and the like can be added.
  • dextrin can be added in an amount of 10 to 80%, for example, 30 to 60% based on the solid content.
  • the additive is not particularly limited as long as it is a general food additive, and its content can be appropriately selected within a range that does not affect the activity and function of the composition.
  • the present invention also provides an immunoenhancing composition comprising an enzyme-treated noni extract as an active ingredient, which is produced according to the above-described method.
  • an immunoenhancing composition comprising an enzyme-treated noni extract as an active ingredient, which is produced according to the above-described method.
  • a health functional food containing the composition can be provided.
  • a composition was prepared by varying the type of the enzyme.
  • Noni was hydrolyzed, ground and lyophilized to obtain a raw material.
  • the obtained raw material was pulverized using a pin mill, a disc mill or the like, and water 10 times as much as the raw material was added.
  • the enzyme-degrading enzyme as shown in Table 1 was added in an amount of 0.2 part by weight based on 100 parts by weight of the raw material, and the mixture was reacted at 50 DEG C for 2 hours.
  • the mixture was extracted by hot water extraction at 95 ° C. for 4 hours, concentrated under reduced pressure at a temperature of 50 to 60 ° C., and hot air temperature was 220 ° C.
  • mouse macrophage cell line RAW264.7 cells were cultured in 100 mm cell culture dishes at a density of 70-80% using RPMI 1640 medium containing 100 IU / ml penicillin and 100 ug / ml streptomycin and 10% FBS.
  • the cultured mouse macrophage cell line RAW264.7 was adjusted to a concentration of 1 ⁇ 10 6 cells / ml using RPMI 1640 medium, inoculated into a 96-well plate, and preincubated for 18 hours at 5% CO 2 and 37 ° C. .
  • the medium was then removed and the extracts were treated with media containing the concentrations of 200, 400, and 800 ug / mL, respectively.
  • 10 ⁇ l of MTT solution stock concentration: 5 mg / ml
  • 100 ⁇ l of DMSO was added to each well for reaction termination and formazan crystal dissolution.
  • the cell viability was calculated from the OD value obtained by measuring the absorbance at 570 nm of the amount of MTT reduced to formazan, and the result is shown in Fig.
  • Comparative Example 1 an extract according to the same method as in Example 1 was used, except that the enzyme-degrading enzyme was not treated.
  • TNF macrophage-derived TNF- ⁇
  • TNF is a cytokine that mediates and regulates innate immunity. TNF is secreted by antigen-stimulated T lymphocytes, NK cells, and mast cells, but since activated macrophages are the main producing cells, TNF production is measured as a factor for analyzing the immuno-promoting effect.
  • the mouse macrophage cell line RAW264.7 cultured in the same manner as in Experimental Example 1 was adjusted to a concentration of 1 ⁇ 10 6 cells / ml using RPMI 1640 medium, inoculated into a 96-well plate, and cultured in 5% CO 2 and 37% Lt; 0 > C for 18 hours. The medium was then removed and the extracts were treated with medium containing 200, 400 and 800 ug / mL of each extract. Twenty-four hours later, the supernatant was transferred to another 96-well plate in an amount of 100 ⁇ l, and the concentration of TNF- ⁇ was measured according to manufacturer's instructions using an enzyme-linked immunosorbent assay (ELISA) kit of R & D Systems. .
  • ELISA enzyme-linked immunosorbent assay
  • TNF- ⁇ production was the highest in the extract prepared by treating Peclyve CP.
  • TNF- ⁇ TNF- ⁇ from rat serum was measured.
  • the C57BL / 6 mice were sacrificed and the spleen was taken out.
  • the splenocytes were picked and adjusted to a concentration of 2 ⁇ 10 6 cells / ml using RPMI 1640 medium, and then inoculated into a 6-well plate.
  • IFN-y-related mRNA In order to analyze the effect of the extract on the immunity enhancement, the production of IFN-y-related mRNA from rat spleen was measured.
  • IFN-y is a cytokine that mediates and modulates adaptive immunity. It activates macrophages to remove microorganisms, promote the differentiation of sub-sensitized CD4 + T cells into TH1 cells, inhibit the differentiation into TH2 cells, and is produced in T lymphocytes and NK cells.
  • the C57BL / 6 mice were sacrificed and the spleen was taken out. The splenocytes were picked and adjusted to a concentration of 2 ⁇ 10 6 cells / ml using RPMI 1640 medium, and then inoculated into a 6-well plate.
  • the sense and antisense primer sequences of the target proteins used herein were prepared by reference to the existing literature, and GAPDH was used as a control gene. Specific sequences are shown in Table 2.
  • PCR amplification was performed using SYBR Premix Ex Taq (Takara Bio) and real-time thermal cycler (Bio-Rad) according to the manufacturer's manual. The results are shown in Fig.
  • TNF- ⁇ -related mRNA from rat spleen was measured.
  • the TNF-a nucleotide sequences used are shown in Table 3, and the sequence of GAPDH is shown in Table 2.
  • the measurement was carried out in the same manner as in Experimental Example 4, and the results are shown in FIG.
  • IL-12 is a cytokine that mediates and regulates innate immunity. It plays an important role in initiating a series of reactions involving macrophages, NK cells, and T lymphocytes, which are involved in the removal of intracellular microorganisms. do.
  • the used IL-12 base sequences are shown in Table 4, and the sequence of GAPDH is shown in Table 2. The measurement was carried out in the same manner as in Experimental Example 4, and the results are shown in FIG.
  • immunity factors such as TNF, IL and IFN can be enhanced, and as a result, it can be expected that the immunity enhancing effect can be expected.

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Abstract

La présente invention concerne un procédé de production d'une composition d'amélioration de l'immunité comprenant un extrait de noni traité par enzyme en tant que principe actif. La présente invention permet d'obtenir un extrait qui peut améliorer la production de facteurs liés à l'immunité et a ainsi un effet d'amélioration de l'immunité. De plus, en tant que composition fonctionnelle de santé, la composition peut être appliquée dans divers domaines tels que des produits alimentaires et analogues.
PCT/KR2017/012679 2017-11-08 2017-11-09 Procédé de production d'une composition améliorant l'immunité comprenant un extrait de noni traité par enzyme WO2019093543A1 (fr)

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KR10-2017-0147696 2017-11-08
KR1020170147696A KR102009662B1 (ko) 2017-11-08 2017-11-08 효소 처리된 노니 추출물을 포함하는 면역 증진 조성물의 제조방법

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KR102160689B1 (ko) * 2020-03-19 2020-09-28 농업회사법인 주식회사 시우 식물세포 분해효소를 이용한 천연 첨가물 쥬스 제조 방법, 이에 의해 제조된 식물세포 분해효소를 이용한 천연 첨가물 쥬스

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6440410B1 (en) * 2001-01-19 2002-08-27 A. Glenn Braswell Reducing oxysterols with extracts of Morinda citrifolia, red wine, prune, blueberry, pomegranate, apple and enzymes
US20090068204A1 (en) * 2007-09-06 2009-03-12 Darien Benjamin J Morinda citrifolia Based Formulations for Regulating T cell Immunomodulation in Neonatal Stock Animals
US20090196944A1 (en) * 2008-02-01 2009-08-06 Brad Rawson Methods of Manufacture of Morinda Citrifolia Based Compositions for Treatment of Anti-Inflammatory Diseases through Inhibition of Cox-1, Cox-2, Interleukin -1beta, Interleukin-6, TNF-alpha, HLE, and iNOS
US20120238506A1 (en) * 2009-11-30 2012-09-20 Laboratoires Expanscience Acacia macrostachya seed extract and compositions containing same

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KR101701895B1 (ko) * 2014-08-20 2017-02-02 주식회사 비케이바이오 고미가 제거된 면역 증강용 감귤류 추출물, 이의 제조방법 및 이를 포함하는 건강기능식품

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6440410B1 (en) * 2001-01-19 2002-08-27 A. Glenn Braswell Reducing oxysterols with extracts of Morinda citrifolia, red wine, prune, blueberry, pomegranate, apple and enzymes
US20090068204A1 (en) * 2007-09-06 2009-03-12 Darien Benjamin J Morinda citrifolia Based Formulations for Regulating T cell Immunomodulation in Neonatal Stock Animals
US20090196944A1 (en) * 2008-02-01 2009-08-06 Brad Rawson Methods of Manufacture of Morinda Citrifolia Based Compositions for Treatment of Anti-Inflammatory Diseases through Inhibition of Cox-1, Cox-2, Interleukin -1beta, Interleukin-6, TNF-alpha, HLE, and iNOS
US20120238506A1 (en) * 2009-11-30 2012-09-20 Laboratoires Expanscience Acacia macrostachya seed extract and compositions containing same

Non-Patent Citations (1)

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Title
DE SOUSA: "Effects of short-term consumption of Morinda citrifolia (Noni) fruit juice on mice intestine, liver and kidney immune modulation", FOOD AND AGRICULTURAL IMMUNOLOGY, vol. 28, no. 3, 2017, pages 528 - 542, XP055607683 *

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