CN109971818A - A method of preparing rare ginsenoside - Google Patents
A method of preparing rare ginsenoside Download PDFInfo
- Publication number
- CN109971818A CN109971818A CN201910230931.3A CN201910230931A CN109971818A CN 109971818 A CN109971818 A CN 109971818A CN 201910230931 A CN201910230931 A CN 201910230931A CN 109971818 A CN109971818 A CN 109971818A
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- CN
- China
- Prior art keywords
- streptococcus thermophilus
- ginsenoside
- culture medium
- culture
- notoginsenoside
- Prior art date
- Legal status (The legal status 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 status listed.)
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- JUJWROOIHBZHMG-RALIUCGRSA-N pyridine-d5 Chemical compound [2H]C1=NC([2H])=C([2H])C([2H])=C1[2H] JUJWROOIHBZHMG-RALIUCGRSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
- UOJAEODBOCLNBU-UHFFFAOYSA-N vinaginsenoside R4 Natural products C1CC(C2(CC(O)C3C(C)(C)C(OC4C(C(O)C(O)C(CO)O4)OC4C(C(O)C(O)C(CO)O4)O)CCC3(C)C2CC2O)C)(C)C2C1C(C)(CCC=C(C)C)OC1OC(CO)C(O)C(O)C1O UOJAEODBOCLNBU-UHFFFAOYSA-N 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
- C12P33/20—Preparation of steroids containing heterocyclic rings
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- 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
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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Abstract
The invention belongs to biologies, pharmaceutical technology field, are related to a kind of using the various ginsenosides of streptococcus thermophilus (Streptococcus thermophiles) microbe conversion, notoginsenoside, the method for preparing ginsenoside Compound K.Include the following steps: 1) to take the streptococcus thermophilus (Streptococcus thermophiles) of activation to be inoculated in culture medium, cultivated under the conditions of 30-38 DEG C of temperature, prepares transformation fermentation liquid；2) ginsenoside or notoginsenoside, microbe conversion are added in the resulting transformation fermentation liquid of step 1)；3) collection step 2) gained reaction solution, is filtered, filtrate is separated with chromatography, is collected the flow point containing ginsenoside compound K, is drying to obtain ginsenoside compound K.Compound K is prepared with the method for the present invention, production cost is low, impurity is few, and efficiency of pcr product is high, green safe pollution-free.
Description
Technical field
The invention belongs to biomedicine technical fields.It is related to a kind of preparation of rare ginsenoside Compound K (CK)
Method, and in particular to ginsenoside is converted by microbial fermentation or notoginsenoside obtains the preparation method of Ginsenoside compound K.
Background technique
Ginseng is the rare drug of East Asia Region tradition, and medicinal history is long.Modern pharmacy is studies have shown that ginsenoside is people
The main pharmacodynamics ingredient of ginseng has significant pharmacological activity, is the important effective substance of ginseng, widely acts on the mind of human body
Through system, immune system, in the treatment of the difficult and complicated cases such as cardiovascular disease and tumour, distinguished unique effect is shown
Fruit.There are about tens kinds for specific ginsenoside monomer, such as Rb1、Rb2、 Rb3、Re、Rd、Rg1、Rg2、Rg3、Rh1、Rh2And Re
Etc..Studies have shown that these ginsenosides are metabolized into containing sugar is less, molecular weight is smaller, can under the action of biological enzyme in vivo
With the Rg being directly absorbed by the body3、Rg5、Rh2、Rh3, CK, protopanoxadiol, Rg2、Rh1, the rare active soap such as protopanaxatriol
Glycosides, absorbed into serum play therapeutic effect.Therefore, Rg3、Rg5、Rh2、Rh3, CK, protopanoxadiol, Rg2、Rh1, protopanaxatriol
It is the main active that ginseng plays medical health care function etc. rare saponin(e.In these rare ginsenosides, ginsenoside
CK is not present in natural ginseng saponin(e, be selectively broken after convert in enteron aisle for ginsenoside ginsenoside 3 with
The primary product of the performance ginseng pharmacological activity formed after 20 glycosidic bonds.Generally believe that Ginsenoside compound K is natural glycol at present
The active metabolite of group ginsenoside, and Rb, Re, Rd etc. may be natural precursor drug.Internal in vitro test in recent years
The study found that proliferation, infiltration and transfer of the CK in addition to inhibiting tumour cell, inducing apoptosis of tumour cell inhibit chemical carcinogens
The chromosomal gene of induction be mutated and the multidrug resistant of reversing tumor cell outside, also have anti-inflammatory, antiallergy, it is anti-nervous and
Many biological activities such as immunological regulation.
Since CK in natural ginseng, Radix Notoginseng and American Ginseng and is not present, previous domestic and foreign scholars mainly pass through chemistry side
It is prepared by method or human enteric bacteria metabolism, other microorganisms or enzymatic conversion glycol group ginsenoside monomer.Ginsenoside parent nucleus knot
3 and 20 glycosidic bonds have disequilibrium on structure, and lower 3 glycosidic bonds of neutrallty condition are easily broken off, 20 under acid or alkaline condition
Position glycosidic bond is easily broken off, therefore is difficult effectively to be precisely controlled the fracture of glycosidic bond using chemical method, is unfavorable for rare ginseng soap
The production of glycosides.Bioconversion has because its regioselectivity is good and reaction is mild single-minded, thus in terms of preparing rare ginsenoside
There is unique advantage.But there are significant individual difference, different races, physical condition, diet for human body intestinal canal flora metabolism
The factors such as mode, pressure and habit directly affect enterobacteriaceae metabolic activity.Commodity in use enzyme preparation, although easy to operate, valence
Lattice are expensive, it is inconvenient, at high cost to there is a problem of enzyme source, therefore application is restricted.Pass through fermentation method and enzyme process both at home and abroad at present
In the report for obtaining CK, higher conversion ratio can be obtained by substrate of glycol group ginsenoside monomer, but cost of material is very high,
Industrialization value is not had；And sanchi leaf total saposins, general ginsenoside and American ginseng total saponins are converted with fungi fermentation to make
Standby CK, cost of material is low, and industrialization prospect is preferable, but has not yet to see industrialized production, and there are industrial biological pollutions
Risk.Therefore, for a long time the preparation method of CK be always this field research hotspot.
Probiotics is a kind of microorganism addition for applying beneficial effect to host by improving intestinal microbial balance
Object.The probiotics often applied is mainly the bacterium for generating lactic acid, such as lactobacillus and Bifidobacterium, and to body, there are many it for they
Its incomparable physiological action of normal physiological flora, comprising: adjustment intestinal flora；Improve the metabolism of protein and vitamin；
Prevent constipation；Generate antibiotic；Alleviate lactose intolerance；Treat hepatic injury；Reduce the disease incidence of colon cancer；It is antitumor；Enhancing
Immune system；Reduce cholesterol；Treat urinary system infection contamination etc..Therefore it can not only be obtained using probiotics conversion ginsenoside
The rare ginsenoside of high activity, while can directly be used in its fermentation liquid there is no to human body and the harmful substance of nature
In the production of drug or food, the generation of biological pollution is reduced.
Summary of the invention
Technical problem solved by the invention is to provide a kind of method for preparing rare ginsenoside CK, and this method has letter
Just, efficiently, can scale fermentation, pollution-free and the advantages of can be directly used for drug and food production.
The present invention is achieved through the following technical solutions:
1) streptococcus thermophilus (Streptococcus thermophiles) of activation is taken to be inoculated in culture medium, in temperature
It is cultivated under the conditions of 30-38 DEG C, prepares transformation fermentation liquid；
2) ginsenoside or notoginsenoside, microbe conversion are added in the resulting transformation fermentation liquid of step 1)；
3) collection step 2) gained reaction solution, is filtered, filtrate is separated with chromatography, collects the stream containing Ginsenoside compound K
Point, Ginsenoside compound K is obtained after dry.
Wherein,
Culture medium described in step 1) is MRS culture medium, MC culture medium, M17 culture medium, tomato juice culture medium, yeast
Cream-dextrose culture-medium, BCG cow's milk culture medium, cow's milk culture medium, 10% skimmed milk culture medium and based on above-mentioned culture medium
Improved culture medium；
The OD of the streptococcus thermophilus of activation described in step 1)600=0.6~0.8；
The inoculative proportion of streptococcus thermophilus described in step 1) is 1~30% (v/v), preferably are as follows: 5~15%；
The revolving speed 100-600r/min of culture described in step 1)；
The OD of transformation fermentation liquid described in step 1)600>0.8；
The streptococcus thermophilus of activation described in step 1), which refers to, is transferred to streptococcus thermophilus accordingly from solid medium
Fluid nutrient medium or accordingly in the fluid nutrient medium containing ginsenoside；
Transformation fermentation liquid described in step 1) is the culture solution containing streptococcus thermophilus, the culture for filtering out streptococcus thermophilus
What is obtained after liquid or streptococcus thermophilus are broken contains enzyme solutions；
The culture solution containing streptococcus thermophilus is the OD containing streptococcus thermophilus600> 0.8 culture solution；
The culture solution for filtering out streptococcus thermophilus is the OD containing streptococcus thermophilus600> 0.8 culture solution is 3000
The resulting supernatant of centrifugation medium under the conditions of~10000r/min；
Obtaining after resulting streptococcus thermophilus is broken is the OD containing streptococcus thermophilus containing enzyme solutions600> 0.8 culture
Liquid resulting thallus of centrifugation medium under the conditions of 3000~10000r/min, is resuspended in the PBS buffer solution of 50~1000mL
In, the crude enzyme liquid that is obtained after ultrasonication on ice-water bath, centrifugation；
The final concentration of 0.01-15% (w/v) of ginsenoside in step 2)；
Ventilation ratio is 0-10 (v/v) in step 2)；It is preferred that are as follows: 0.5-5 (v/v)
Ginsenoside described in step 2) or notoginsenoside are ginsenoside Rb1、Rb2, Rd, Rc, notoginsenoside D, Fa,
K, R4, T, L, O, P, Q, S, Fc, Fe or contain the ginseng extract of glycol saponins, the West in ginsenoside or notoginsenoside
Conopsea extraction, Notogineng Extract, ring conopsea extraction, extract of Radix Ginseng stem and leaf, stem and leaf of Radix Panacis Quinquefolii extrac, notoginseng haulm extract
Object, panax japonicus stem-leaf extract.
The temperature of microbe conversion described in step 2) is 25-45 DEG C, pH 4.0-7.5, stirring rate 0-600r/min,
Transformation time is 24-120h, preferably are as follows: 24-72h；When with streptococcus thermophilus obtained after broken when being converted containing enzyme solutions,
Microbe conversion can carry out under static state.
Chromatography described in step 3) is normal phase chromatography, RP chromatography or molecular exclusion chromatography, preferably silica gel
Column chromatography, ODS column chromatography and Sephadex LH-20 gel column chromatography.
Specifically,
Technical scheme is as follows:
1) it takes streptococcus thermophilus to be inoculated in culture in 50~2000mL culture medium to activate to OD600After=0.6~0.8, by 1
~30% ratio (v/v) is inoculated in 10L fermentor, cultivated under the conditions of 30-38 DEG C of temperature, revolving speed 100-600r/min to
OD600> 0.8, that is, it can be directly used for subsequent fermentation conversion；Or the centrifugation medium at 3000~10000r/min, gained supernatant
It can also be used for subsequent fermentation conversion；Gained thallus after centrifugation can be also resuspended in the PBS buffer solution of 50~1000mL, Yu Bing
Ultrasonication in water-bath obtains crude enzyme liquid after centrifugation, which can also be used for subsequent fermentation conversion；
2) ginsenoside or Radix Notoginseng soap of final concentration of 0.01-15% (w/v) are added in the transformation fermentation liquid of step 1)
After glycosides, start microbe conversion.It feeds intake after 12-24h, adjusts ventilation ratio 0-10 (v/v)；Microbe conversion temperature is 25-45 DEG C, and pH is
4.0-7.5, stirring rate 0-600r/min, transformation time 24-120h.
3) collection step 2) gained reaction solution, is filtered, filtrate is separated with chromatography, is enriched with, and collection contains Ginsenoside compound K
Flow point, it is dry after Ginsenoside compound K, can further improve CK purity through recrystallization.
The present invention by the various ginsenosides of streptococcus thermophilus (Streptococcus thermophiles) microbe conversion,
Notoginsenoside prepares CK, using obtaining the monocrystalline of CK after pillar layer separation and recrystallization.
It is as follows to measure its spectral data: HR-ESI-MS:m/z 645.4340 [M+Na]+(calcd for C36H62NaO8,
645.4342)；1H-NMR (400MHz, pyridine-d5)δH: 5.19 (1H, d, J=7.6Hz, Glc-H-1), 1.63 (3H,
S, H21), 1.60 (6H, s, H-26,27), 1.23 (3H, s, H-28), 1.00 (3H, s, H-29), 1.00 (3H, s, H-30),
0.95 (3H, s, H-18), 0.89 (3H, s, H-19)；13C-NMRδC: 39.4 (C1), 28.3 (C-2), 78.1 (C-3), 39.6
(C-4), 56.4 (C5), 18.8 (C-6), 35.2 (C-7), 40.1 (C-8), 50.3 (C9), 37.4 (C-10), 30.8 (C-11),
70.2 (C-12), 49.7 (C-13), 51.5 (C-14), 31.0 (C-15), 26.7 (C-16), 51.7 (C-17), 16.4 (C-
18), 16.1 (C-19), 83.3 (C-20), 22.4 (C-21), 36.2 (C-22), 23.3 (C-23), 126.0 (C-24), 131.0
(C-25), 25. 8 (C-26), 17.8 (C-27), 28.7 (C-28), 16.4 (C-29), 17.4 (C-30), 98.3 (C-20-Glc-
1), 75.2 (C-20-Glc-2), 79.3 (C-20-Glc-3), 71.6 (C-20-Glc-4), 78.3 (C-20-Glc-5), 62.9
(C-20-Glc-6).It is consistent with the structure of document report CK to confirm its structure.
Method of the invention has the advantage that streptococcus thermophilus is common probiotics, it is used not only may be used as zymophyte
To realize high CK conversion ratio, can realize up to nearly 85% conversion ratio using ginsenoside monomer as substrate, and its fermentation material without
It need to be further processed and can be used to drug and food production, and is environmentally friendly, inanimate object pollution is suitable for that large-scale industry is raw
It produces.
Specific embodiment
Influence of the embodiment l fermentation temperature to conversion ratio
Streptococcus thermophilus (Streptococcus thermophiles) is transferred to 100mL from MRS solid medium
In MRS fluid nutrient medium, 36 DEG C of culture 12h.20mL bacterium solution is taken to be inoculated into the 200mL MRS liquid containing 10mg ginsenoside Rd
In body culture medium, ferment at 22,27,32,37,42 DEG C respectively, stirring rate 300r/min, pH 6.5.48h post-fermentation
Terminate.Fermentation liquid is centrifuged under 5000r/min, gained supernatant is concentrated to dryness after removing thallus, is dissolved and is determined with 20mL methanol
Hold to 50mL, analyze measurement CK content through HPLC, as the result is shown at 37 DEG C CK conversion ratio highest, conversion ratio 76.2%.
| Temperature (DEG C) | 22 | 27 | 32 | 37 | 42 |
| Conversion ratio (%) | 10.7 | 41.9 | 71.6 | 76.2 | 63.2 |
The result shows that conversion ratio can achieve 60% or more when temperature is 32~42 DEG C of ranges, further, when
When temperature is 32~37 DEG C of ranges, conversion ratio can reach 70% or more.
Influence of 2 fermentation time of embodiment to conversion ratio
Streptococcus thermophilus (Streptococcus thermophiles) is transferred to 100mL from MRS solid medium
In MRS fluid nutrient medium, 37 DEG C of culture 12h.20mL bacterium solution is taken to be inoculated into containing 10mg ginsenoside Rb1200mL MRS liquid
In body culture medium, ferment 12,24,48,72,96 and 120h respectively at 37C, stirring rate 300r/min, pH 6.5.Hair
After ferment, fermentation liquid is centrifuged under 5000r/min, gained supernatant is concentrated to dryness after removing thallus, is dissolved with 20mL methanol
And it is settled to 50mL, and measurement CK content is analyzed through HPLC, the conversion ratio of CK reaches plateau after the 72h that ferments as the result is shown, turn
Rate is 82.0%.
| Time (h) | 12 | 24 | 48 | 72 | 96 |
| Conversion ratio (%) | 15.2 | 44.8 | 75.1 | 82.0 | 83.6 |
Test result show when fermenting between within the scope of 48~96h when, conversion ratio is up to 70% or more, further
Ground, when fermenting between within the scope of 72~96h when, conversion ratio is up to 80% or more.
Embodiment 3
By streptococcus thermophilus (Streptococcus thermophiles) from MRS solid medium be transferred to containing
0.01% (w/v) ginsenoside Rb1100mL MRS fluid nutrient medium in, 30 DEG C of culture 12h；Be then seeded into containing
1000mg ginsenoside Rb16L MRS fluid nutrient medium in, ferment for 37 DEG C in 10L automatic fermenter.It is adjusted after 12h
Whole ventilation ratio is 0.5 (v/v), stirring rate 400r/min, pH 6.5.72h post-fermentation terminates.Fermentation liquid is after filter-cloth filtering
5.8L filtrate is obtained, filtrate obtains crude extract 293.6g after being concentrated to dryness, by ODS pillar layer separation, with methanol-water
Gradient elution (40% methanol~100% methanol) is carried out for mobile phase, collects the flow point containing CK, after being concentrated to dryness, with
95% ethyl alcohol recrystallization obtains CK 367mg, yield 65.3%, and it is 91.3% that HPLC, which detects its purity,.
Streptococcus thermophilus (Streptococcus thermophiles) is transferred to by embodiment 4 from M17 solid medium
In 100mL M17 fluid nutrient medium, 37 DEG C of culture 12h；It is then seeded into 6L M17 fluid nutrient medium, is automatically sent out in 10L
37 DEG C of culture 36h in fermentation tank, culture solution obtain 5.7L filtrate after filter-cloth filtering, general ginsenoside 5g are added in filtrate, in 10L
It ferments for 40 DEG C in automatic fermenter, stirring rate is adjusted to 400r/min.96h post-fermentation terminates.Fermentation liquid is by subtracting
Crude extract 285.0g is obtained after pressure concentration is dry, is separated by silica gel column chromatography, is with the methylene chloride-methanol of the water containing one thousandth
Mobile phase carries out gradient elution (CH2Cl2: MeOH=19:1~1:19), the flow point containing CK is collected, after being concentrated to dryness, with
95% ethyl alcohol recrystallization obtains CK 362mg, yield 7.24%, and it is 87.2% that HPLC, which detects its purity,.
Embodiment 5 is by streptococcus thermophilus (Streptococcus thermophiles) from 10% skimmed milk solid culture
Base is transferred in 10% skimmed milk fluid nutrient medium of 100mL, 37 DEG C of culture 12h；It is then seeded into 10% skimmed milk of 6L
In fluid nutrient medium, for 24 hours, culture solution obtains thallus after filter-cloth filtering for 37 DEG C of cultures in 10L automatic fermenter.Gained thallus
It is resuspended in 100mL PBS, is centrifuged 10min in 5000rpm after ultrasonication under condition of ice bath, supernatant is taken to be diluted with PBS
To 500mL as reaction solution, is converted, 40 DEG C, be incubated for for 24 hours under the conditions of pH=5.0 using 1.0g arasaponin as substrate.
Solids 905mg is obtained after reaction solution freeze-drying, by Sephadex LH-20 column chromatography, using 30% methanol as mobile phase, is received
Collect the flow point containing CK, after being concentrated to dryness, then analyses and separate through silica gel column chromatography, with the methylene chloride-of the water containing one thousandth
Methanol is that mobile phase carries out gradient elution (CH2Cl2: MeOH=75:25~45:55), the flow point containing CK is collected, is concentrated under reduced pressure
To after dry, with 95% ethyl alcohol recrystallization, CK 50.3mg, yield 5.03% are obtained, it is 93.1% that HPLC, which detects its purity,.
Claims (10)
1. a kind of method for preparing rare ginsenoside Compound K, which is characterized in that converted using streptococcus thermophilus fermentation
Ginsenoside or notoginsenoside produce Compound K.
2. the method as described in claim 1, which is characterized in that comprise the following steps:
1) streptococcus thermophilus (Streptococcus thermophiles) of activation is taken to be inoculated in culture medium, in temperature 30-
It is cultivated under the conditions of 38 DEG C, prepares transformation fermentation liquid；
2) ginsenoside or notoginsenoside, microbe conversion are added in the resulting transformation fermentation liquid of step 1)；
3) collection step 2) gained reaction solution, is filtered, filtrate is separated with chromatography, and collection contains ginsenoside Compound K's
Flow point obtains ginsenoside Compound K after dry.
3. method according to claim 2, which is characterized in that culture medium described in step 1) is MRS culture medium, MC culture
Base, M17 culture medium, tomato juice culture medium, yeast extract-dextrose culture-medium, BCG cow's milk culture medium, cow's milk culture medium, 10% take off
Rouge cow's milk culture medium or improved culture medium based on above-mentioned culture medium.
4. method according to claim 2, which is characterized in that step 1) the transformation fermentation liquid is to contain streptococcus thermophilus
What is obtained after culture solution, the culture solution for filtering out streptococcus thermophilus or streptococcus thermophilus are broken contains enzyme solutions.
5. method according to claim 2, which is characterized in that the inoculative proportion of streptococcus thermophilus described in step 1) is 1
~30% (v/v), preferably are as follows: 5~15%.
6. method as claimed in claim 4, which is characterized in that
The culture solution containing streptococcus thermophilus is the OD containing streptococcus thermophilus600> 0.8 culture solution；
The culture solution for filtering out streptococcus thermophilus is the OD containing streptococcus thermophilus600> 0.8 culture solution 3000~
The resulting supernatant of centrifugation medium under the conditions of 10000r/min；
Obtaining after resulting streptococcus thermophilus is broken is the OD containing streptococcus thermophilus containing enzyme solutions600> 0.8 culture solution exists
The resulting thallus of centrifugation medium, is resuspended in the PBS buffer solution of 50~1000mL under the conditions of 3000~10000r/min, Yu Bing
Ultrasonication in water-bath, the crude enzyme liquid obtained after centrifugation.
7. method according to claim 2, which is characterized in that ginsenoside described in step 2) or notoginsenoside are ginseng soap
Glycosides Rb1、Rb2, Rd, Rc, notoginsenoside D, Fa, K, R4, T, L, O, P, Q, S, Fc, Fe or containing ginsenoside or notoginsenoside
Ginseng extract, American ginseng extract, Notogineng Extract, ring conopsea extraction, extract of Radix Ginseng stem and leaf, stem and leaves of American ginseng extract
Object, Panax Notoginseng Folium Saponins, panax japonicus stem-leaf extract.
8. method according to claim 2, which is characterized in that microbe conversion temperature is 25-45 DEG C, pH 4.0-7.5, ventilation
Than for 0-10 (v/v), stirring rate 0-600r/min, transformation time 24-120h.
9. method according to claim 2, which is characterized in that ginsenoside or notoginsenoside is final concentration of in step 2)
0.01-15% (w/v).
10. method according to claim 2, which is characterized in that the chromatography includes normal phase chromatography, RP chromatography or
Molecular exclusion chromatography, preferably silica gel column chromatography, ODS column chromatography and Sephadex LH-20 gel column chromatography.
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| CN111118095A (en) * | 2019-12-10 | 2020-05-08 | 武汉克鲁金生物科技有限公司 | Method for producing ginsenoside CK extract by hydrolyzing ginsenoside with Kluyveromyces lactis |
| CN111118095B (en) * | 2019-12-10 | 2023-10-24 | 武汉克鲁金生物科技有限公司 | Method for producing ginsenoside CK extract by hydrolyzing ginsenoside by kluyveromyces lactis |
| CN113244148A (en) * | 2021-04-30 | 2021-08-13 | 云南莱因生物科技有限公司 | A skin care composition |
| CN115068490A (en) * | 2022-06-23 | 2022-09-20 | 四川大学 | Application of ginsenoside CK in preparation of bacteriostatic agent for mycobacterium abscessus or/and mycobacterium tuberculosis |
| CN116731880A (en) * | 2023-06-16 | 2023-09-12 | 昆明理工大学 | Endophytic fungus Mucor multicastus and application thereof |
| CN116731880B (en) * | 2023-06-16 | 2024-04-26 | 昆明理工大学 | Endophytic fungus Mucor multicastus and application thereof |
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