CN1229086A - Method for preparing rare ginsengoside using enzymatic method to modify ginsenoside glycoside - Google Patents

Method for preparing rare ginsengoside using enzymatic method to modify ginsenoside glycoside Download PDF

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CN1229086A
CN1229086A CN99112764A CN99112764A CN1229086A CN 1229086 A CN1229086 A CN 1229086A CN 99112764 A CN99112764 A CN 99112764A CN 99112764 A CN99112764 A CN 99112764A CN 1229086 A CN1229086 A CN 1229086A
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saponin
ginsenoside
glycosyl
scarce
enzyme
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CN1105781C (en
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金凤燮
鱼红闪
保永明
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/25Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
    • A61K36/258Panax (ginseng)
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/56Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00Function of food ingredients
    • A23V2200/30Foods, ingredients or supplements having a functional effect on health
    • A23V2200/308Foods, ingredients or supplements having a functional effect on health having an effect on cancer prevention
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10Preparation or pretreatment of starting material
    • A61K2236/19Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/10General cosmetic use

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  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
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Abstract

An enzymolysis process for preparing rare ginseng saponin by modifying the glycosyl of ginseng saponin hydrolyzing the ginseng saponins with the saponinases including beta-glucosidease, alpha- or beta-arabinosylase, alpha-rhamnosidease, etc to hydrolyze out part of glycosyl radicals on moleculae of saponin, resulting in new rare saponin. Its advantages are low cost and high economic value.

Description

Enzyme process changes the method that the ginsenoside glycosyl prepares scarce ginsenoside
The present invention relates to a kind of biological method that changes the ginsenoside glycosyl, particularly adopt enzyme process to change the method that the ginsenoside glycosyl prepares scarce ginsenoside.
Genseng is that the genseng (Panax) in the plant Araliaceae (araliaceae) belongs to, four not of the same race and other mutation are arranged, as genseng (P.ginseng), Radix Panacis Quinquefolii (P.quenquefolinus), pseudo-ginseng ginseng (P.notoginseng), rhizome of Japanese Ginseng (P.uaponicus) etc. and some other mutation.Genseng is the valuable tonic of the strengthening by means of tonics that has won fame both at home and abroad, and the effective constituent that mainly contains of genseng is a saponin.The ginsenoside of having found has 34 kinds; Ginsenoside can be divided into three kinds according to the difference of sapogenin, i.e. glycols saponin class (protopanaxdill-type Gisenosides), triol saponin class (Protopanaxtriol-typeGisenosides).Wherein the glycols saponin accounts for 50~60% of whole saponin, secondly is three alcohols saponins 30~35%, also has olea acids saponin.The glycols saponin Ra1, Ra2, Ra3, RB1, Rb2, Rb3, Rc, Rd and the Rg3 that account for saponin total amount 50~60% in the genseng are identical with scarce ginsenoside Rh2 sapogenin, and just sugar chain is different.Wherein saponin content height in genseng such as Rb, Rc, Rd obtain easily.But the Rh2 saponin that antitumour activity is very high exists in white ginseng hardly, only contain 100,000 at red ginseng/.On glucoside unit the 3rd carbon of Rh2 and R6, Rc, the same saponin molecule of Rd a β-glucosyl group is arranged, therefore, can obtain being worth high anticancer saponin Rh2 as long as the part glycosyl on the saponin molecules such as Rb, Rc, Rd is removed.Equally,, remove the part glycosyl, can obtain being worth high saponin Rh1 for accounting for saponin total amount 40~50% panoxatriol class saponin Re, Rf, Rg1 and Rg2 saponin in the genseng.
The present invention seeks to: utilize biotechnology, provide a kind of enzyme process to change the method that the ginsenoside glycosyl prepares scarce ginsenoside.
The present invention is achieved in that enzyme process changes the method that the ginsenoside glycosyl prepares scarce ginsenoside, it is characterized in that: adopt saponin beta-glucosidase, α-, saponin enzymes such as β-arabinofuranosidase/xylosidase, α-mouse season Glycosylase, β-glycuronide enzyme or beta-galactosidase enzymes, for the processing that is hydrolyzed of the ginsenoside of all kinds of Panax ginseng, can make the part glycosyl in the saponin molecule glucoside unit be hydrolyzed, become new rare saponin.
Realize when of the present invention that with such scheme described various saponin enzymes can be extracted by microorganism such as bacterium, streptomycete, mould, yeast, basidiomycetes or plant, plant seed processing.
Use the present invention, can β-glucose saponin ginsenoside enzymatically hydrolyzing Rg 3, obtain scarce ginsenoside Rh 2Also can α-Ji sugar saponin ginsenoside enzymatically hydrolyzing Rg 2, obtain scarce ginsenoside Rh 1Also can
Can produce the saponin enzyme and directly be inoculated on ginseng powder or the sheet and cultivate, obtain high ginseng powder of scarce ginsenoside component content or sheet.
Core of the present invention is to utilize the part glycosyl of the higher ginsenoside of saponin enzyme hydrolysis content in each seed ginseng, thereby obtains scarce ginsenoside.In fact, the general cellulase or the excision enzyme of hemicellulase are difficult to hydrolysis saponin glycosyl, because saponin is different with Mierocrystalline cellulose, the glucoside unit ring with big has only saponin enzyme could realize hydrolysis saponin glycosyl.
The invention has the advantages that: it can allow people in simple and economic mode, and some ginsenoside compositions " transformation " that occurring in nature is extensively existed become Rh1, Rh2 and so on ginsenoside composition rare, that economic worth is very high again.
Below in conjunction with embodiment the present invention is done further narration:
Embodiment 1:
Bacillus sp.JF bacterium with high-temperature aerobic, in the 0.01MgSO4 substratum that contains 3% Semen Maydis powder, 1% genseng extract, temperature is ventilated down for 60 ℃ and was cultivated 30~40 hours, then bactofugation, 70% ethanol sedimentation zymoprotein, collect albumen, freeze-drying, obtain can hydrolysis the beta-glucoside saponin enzyme of β-glucose key.
Get the 3 above-mentioned saponin enzymes of gram and 140 milliliters of physiological waters mixing; 10 gram panaxoside Rg 3,20 ml physiological salines and 40 milliliters of ethanol mix.Two kinds of above-mentioned mixed solution remix were 60 ℃ of following stirring reactions of temperature 12 hours.The reaction back adds 800 milliliters of ethanol, removes by filter albumen precipitation, and excess ethyl alcohol is washed precipitation, filtrate decompression evaporate to dryness, and dry product distilled water flush away salt three times deducts evaporate to dryness, obtains 7 grams and contains 40~95%Rh2 saponin.
Embodiment 2:
Degerming after aspergillus tubigensis is cultivated, nutrient solution ethanol sedimentation zymoprotein with the 0.02M sodium-acetate buffer dissolving of pH5, separates sugared saponin enzyme of α-mouse season, freeze-drying with DEAE-Cullulose ion exchange resin salt gradient elution method.
10 grams, three alcohols ginsenoside Res are dissolved in 200 milliliters of 0.02M sodium-acetate buffer dissolvings with PH5, add 0.05 gram α-mouse season sugar saponin enzyme 40 ℃ of stirring reactions of temperature 12 hours, be heated to 100 ℃ 30 minutes, filter out the Deproteinization precipitation, macroporous resin 200 gram AB-8 absorption, 500 milliliter of 70% ethanol elution, its ethanol liquid evaporated under reduced pressure gets thick Rg1 saponin.Its thick Rg1 saponin separates through the silicagel column partition method, and underpressure distillation gets 5.5 gram Rg1.
Same quadrat method, above-mentioned enzyme are handled 10 gram triol panaxoside Rgs 2, can obtain valuable pH1 saponin 4.5~6 grams.
Embodiment 3:
Cultivate the back degerming after getting genus bacillus, use the ethanol sedimentation zymoprotein, freeze-drying.10***** is restrained genseng mixing saponin be dissolved in 200 milliliters, add the above-mentioned mixing saponin enzyme of 0.3 gram, 40 ℃ of stirring reactions of temperature 12 hours with the dissolving of the 0.02M sodium-acetate buffers of pH5.Through efficient liquid phase chromatographic analysis, 80% above saponin loses a glycosyl at least.Rare anticancer saponin Rh2, RH1 saponin content have improved hundred times.Reaction solution is through adding the heat extraction zymoprotein, and through 200 gram AB-8 porous resin absorption, 500 millis, 90% ethanol elution, evaporated under reduced pressure can obtain the high mixing saponin of the 8 rare saponin contents of gram.
Embodiment 4
The bacterium aspergillus tubigensis (not producing poison or harmful thing) of producing the saponin enzyme is inoculated on the ginseng tablet of moisture content 25~150%, cultivated 2~7 days, be drying to obtain the genseng that height contains rare saponin at 28~38 ℃.Through efficient liquid phase chromatographic analysis, 80% above saponin loses a glycosyl at least.Rare anticancer saponin RH2, RH1 saponin content have improved hundred times.

Claims (5)

1, enzyme process changes the method that the ginsenoside glycosyl prepares scarce ginsenoside, it is characterized in that: adopt saponin beta-glucosidase, α-, saponin enzymes such as β-arabinofuranosidase/xylosidase, α-mouse season Glycosylase, β-glycuronide enzyme or beta-galactosidase enzymes, for the processing that is hydrolyzed of the ginsenoside of all kinds of Panax ginseng, can make the part glycosyl in the saponin molecule glucoside unit be hydrolyzed, become new rare saponin.
2, method according to claim 1 is characterized in that: described various saponin enzymes can be extracted by microorganism such as bacterium, streptomycete, mould, yeast, basidiomycetes or plant, plant seed processing.
3, method according to claim 1 is characterized in that: with β-glucose saponin ginsenoside enzymatically hydrolyzing Rg 3, get scarce ginsenoside Rh 2
4, method according to claim 1 is characterized in that: with α-Ji sugar saponin ginsenoside enzymatically hydrolyzing Rg 2, get scarce ginsenoside Rh 1
5, method according to claim 2 is characterized in that: the mushroom that can produce the saponin enzyme directly is inoculated on ginseng powder or the sheet to be cultivated, and obtains high ginseng powder of scarce ginsenoside component content or sheet.
CN99112764A 1999-03-17 1999-03-17 Method for preparing rare ginsengoside using enzymatic method to modify ginsenoside glycoside Expired - Fee Related CN1105781C (en)

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KR1019990045180A KR100329259B1 (en) 1999-03-17 1999-10-14 manufacturing process of ginseng saponins

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WO2002053722A1 (en) * 2000-12-29 2002-07-11 Fengxie Jin Ginsenoside glycosidase which hydrolyzes ginsenoside glycosyl and the use thereof
CN1305479C (en) * 2003-09-28 2007-03-21 中国科学院大连化学物理研究所 Low polarity ginsenoside combination possessing anti-tumor activity
WO2007035723A3 (en) * 2005-09-19 2007-07-12 Herbalscience Singapore Pte Ltd Compositions and methods comprising panax species
CN102154428A (en) * 2011-01-13 2011-08-17 佟心 Method for preparing ginsenoside Rh2
CN102805750A (en) * 2011-05-31 2012-12-05 天津药物研究院 Composition of aescine derivative and salt thereof, preparation method and medical uses thereof
CN101570746B (en) * 2008-04-29 2013-07-10 天士力制药集团股份有限公司 Novel saikosaponin glycosidase, preparation method and application thereof
CN103571809A (en) * 2012-07-24 2014-02-12 中国科学院上海生命科学研究院 Novel beta-glucosidase, coding gene thereof and applications thereof
CN105695552A (en) * 2016-04-08 2016-06-22 南京林业大学 Method using enzymatic method to prepare rare ginsenoside Rh1
CN107384896A (en) * 2017-08-11 2017-11-24 南京林业大学 A kind of enzymatic compositions for preparing rare ginsenoside and its application
CN108138211A (en) * 2015-10-22 2018-06-08 株式会社爱茉莉太平洋 The method for selectively preparing Ginsenoside F2, Mc compounds and O compounds from the saponin of ginseng using enzyme process
CN108138210A (en) * 2015-10-22 2018-06-08 株式会社爱茉莉太平洋 The method for selectively preparing ginsenoside Rd from the saponin of ginseng using enzyme process
CN111118096A (en) * 2020-01-07 2020-05-08 陕西岳达德馨生物制药有限公司 Enzymatic preparation method of saponin
CN112273655A (en) * 2020-10-29 2021-01-29 光亚生物科技(广州)有限公司 Small-molecule ginseng cream with anti-aging effect and preparation method thereof
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KR101776005B1 (en) 2015-09-01 2017-09-12 셀루스원 주식회사 Production of ginsenoside Rh2, Rk2 and Rh3 through combinative usage of enzymes and organic acids
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US7759101B2 (en) 2000-12-29 2010-07-20 Fengxie Jin Ginsenoside glycosidases hydrolyzing ginsenoside sugar moieties and uses thereof
WO2002053722A1 (en) * 2000-12-29 2002-07-11 Fengxie Jin Ginsenoside glycosidase which hydrolyzes ginsenoside glycosyl and the use thereof
CN1305479C (en) * 2003-09-28 2007-03-21 中国科学院大连化学物理研究所 Low polarity ginsenoside combination possessing anti-tumor activity
WO2007035723A3 (en) * 2005-09-19 2007-07-12 Herbalscience Singapore Pte Ltd Compositions and methods comprising panax species
CN101570746B (en) * 2008-04-29 2013-07-10 天士力制药集团股份有限公司 Novel saikosaponin glycosidase, preparation method and application thereof
CN102154428B (en) * 2011-01-13 2014-04-30 佟心 Method for preparing ginsenoside Rh2
CN102154428A (en) * 2011-01-13 2011-08-17 佟心 Method for preparing ginsenoside Rh2
CN102805750A (en) * 2011-05-31 2012-12-05 天津药物研究院 Composition of aescine derivative and salt thereof, preparation method and medical uses thereof
CN103571809A (en) * 2012-07-24 2014-02-12 中国科学院上海生命科学研究院 Novel beta-glucosidase, coding gene thereof and applications thereof
CN103571809B (en) * 2012-07-24 2015-06-03 中国科学院上海生命科学研究院 Novel beta-glucosidase, coding gene thereof and applications thereof
CN108138211A (en) * 2015-10-22 2018-06-08 株式会社爱茉莉太平洋 The method for selectively preparing Ginsenoside F2, Mc compounds and O compounds from the saponin of ginseng using enzyme process
CN108138210A (en) * 2015-10-22 2018-06-08 株式会社爱茉莉太平洋 The method for selectively preparing ginsenoside Rd from the saponin of ginseng using enzyme process
CN108138211B (en) * 2015-10-22 2021-08-10 株式会社爱茉莉太平洋 Method for selectively preparing ginsenoside F2, Mc compound and O compound from ginsenoside of ginseng by enzyme method
CN105695552A (en) * 2016-04-08 2016-06-22 南京林业大学 Method using enzymatic method to prepare rare ginsenoside Rh1
CN107384896A (en) * 2017-08-11 2017-11-24 南京林业大学 A kind of enzymatic compositions for preparing rare ginsenoside and its application
CN107384896B (en) * 2017-08-11 2020-07-31 南京林业大学 Enzyme composition for preparing rare ginsenoside and application thereof
CN111118096A (en) * 2020-01-07 2020-05-08 陕西岳达德馨生物制药有限公司 Enzymatic preparation method of saponin
CN112273655A (en) * 2020-10-29 2021-01-29 光亚生物科技(广州)有限公司 Small-molecule ginseng cream with anti-aging effect and preparation method thereof
WO2023066002A1 (en) * 2021-10-19 2023-04-27 耿胜利 Method for preparing ginsenoside preparation by adopting bioengineering technology

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