CN109536560A - A method of improving rare saponin content in ginseng water extract - Google Patents
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Abstract
The invention belongs to fermentation arts, and in particular to a method of improve rare saponin content in ginseng water extract.Method includes the following steps: preparing ginseng aqueous extract using microwave ultrasound synergic solvent extraction, using ginseng aqueous extract as fermentation substrate, microbe conversion generates 4 kinds of rare ginsenosides.By adjusting the preparation method and conversion temperature of leavening, the rare saponin content of feature is increased substantially in the fermentation liquid made.This method simple process can compound the content that fermentation improves specific rare saponin(e, high conversion rate by strain in the case where guaranteeing while generating 4 kinds of rare saponin(es.It can be applied to different demands ginseng deep processed product.
Description
Technical field
The invention belongs to fermentation arts, and in particular to a method of improve rare saponin content in ginseng water extract.
Technical background
Ginseng (Panax ginseng C.A.Mey) is the root of Araliaceae perennial plant ginseng, is traditional rare Chinese medicine,
Ginsenoside (ginsenoside GS) is most effective medicine in the main matter basis and constituent of ginseng of ginseng physiological activity
With ingredient, wherein the less rare ginsenoside of content (rare ginsenosides) has stronger bioactivity and pharmacology
Effect has high medical value and application prospect.For example, rare ginsenoside Compound K (CK) is antitumor, anti-
Inflammation, anti-diabetic, antithrombus formation, anti-aging and liver protection etc. have good effect；There is Rg3 induction tumour cell to wither
The effects of dying, enhancing immunity of organisms.Contain rare ginsenoside in a variety of herbal mixtures, Rg3 sterling is used clinically for resisting
Tumour.However these rare saponin(e natural contents are extremely low, if content of the Rg3 in white ginseng is only 0.0003%, improve rare soap
Glycosides content is an important direction of ginseng deep processing.
Ginseng is the Chinese medicine of integration of drinking and medicinal herbs, and conventional use is to carry out decocting its aqueous extract to it, and ginseng water extracts
Common ginsenoside in liquid containing higher concentration, but rare saponin content is extremely low.Fermentation process can change ginsenoside
Sugar chain structure promotes it to convert to rare saponin(e, to change the content of saponin(e and rare saponin(e in ginseng, enhances the medicine of ginseng
Reason activity, promotes intestinal absorption.Some researches show that lactic acid bacteria can produce beta-glucosidase, which can hydrolyze a variety of β-Portugals
Polyglycoside.Using the glycosidic bond of beta-glucosidase enzyme hydrolysis ginsenoside, to generate active higher, the stronger rare soap of drug effect
Glycosides has become fermented ginseng product, for developing various ginseng deep processed products.Existing report be mostly to general ginsenoside or
The common saponin(e of monomer ferments, and the raw material of these methods fermentation must extract purifying or individually purchase from ginseng, at
This is higher；And with ginseng aqueous extract direct fermentation, though current method improves the content of rare saponin(e to a certain extent,
Transformation efficiency is not high, and obtained rare saponin content is single.
Summary of the invention
In order to solve the disadvantage that the prior art and shortcoming, the main purpose of the present invention is to provide a kind of raising ginsengs
The method of rare saponin content in water extract.
The purpose of the present invention is achieved through the following technical solutions:
A method of improving rare saponin content in ginseng water extract, comprising the following steps:
(1) sieving after ginseng crushing and grinding is obtained into Ginseng Root Powder, by Ginseng Root Powder rear progress ultrasonic-microwave soluble in water
Processing filters out ginseng pulverate's slag, obtains ginseng water extract；
(2) ginseng water extract made from step (1) is taken, Compound bacterium and sugar source is added and ferments 4~8 days at 28~40 DEG C,
The Compound bacterium is lactic acid bacteria and acetobacter；
(3) fermentation temperature is increased to 42~55 DEG C, continues fermentation 10~20 days, and after fermentation, filtering leaves fermentation liquid
?.
Preferably, the mass ratio of step (1) Ginseng Root Powder and water is 1:5~20.
Preferably, the power of step (1) ultrasound is 50~80Hz, and the power of the microwave is 400~800W.
Preferably, the time of step (1) the ultrasonic-microwave processing is 10~30min.
Preferably, the mass ratio of step (2) sugar source and ginseng water extract is 5%~20%, the inoculation of the Compound bacterium
Amount is 3%~7%.
Preferably, the inoculation quality ratio of step (2) lactic acid bacteria and acetobacter is 3~8:1.
Preferably, step (2) lactic acid bacteria is lactobacillus plantarum, Lactobacillus casei, lactobacillus paracasei, acidified milk bar
One or more of bacterium and lactobacillus acidophilus.
Preferably, step (2) acetobacter is one or both of Acetobacter pasteurianus and oxidation acetobacter.
Preferably, the granularity of step (1) described Ginseng Root Powder is 180~250 μm.
Preferably, the rare saponin(e is one or more of Rg3, CK, F2 and Rh2.
Preferably, step (3) number of days for continuing fermentation is 20 days.
Preferably, ginseng described in step (1) is white ginseng.
The principle of the present invention are as follows: microorganism grows hair using the sugar source of nutriment and addition in ginseng aqueous extract
Ferment, expression beta-glucosidase carry out bioconversion to the common saponin(e of ginseng and generate rare saponin(e.Microorganism produces beta-glucosidase
Enzyme has the wide in range property of substrate, can convert different common saponin(es and generate a variety of rare saponin(es；β-glucose that different strains generates
Glycosides enzyme is variant to different common saponin(e activity of conversion, controls, comes using the compounding fermentation of different strain and fermentation strategies
Achieve the purpose that characteristic improves certain rare saponin(e.
Compared with prior art, the invention has the advantages that and the utility model has the advantages that
(1) ginseng aqueous extract is prepared compared to traditional decocting or refluxing extraction, the present invention is assisted using microwave ultrasound
Ginseng aqueous extract is prepared with extraction, is improved extraction efficiency while shortening extraction time, is acted on by microwave ultrasound, cell
Broken complete, the dissolution of common ginsenoside is more thorough, so that total saponin content has certain promotion in aqueous extract.On the other hand
Extracting method through the invention, obtained extracting solution are conducive to the growth of fermenting microbe as fermentation substrate, are efficient
Conversion lays the foundation.
(2) present invention is fermented with compounding strain, rare in 4 kinds of microbe conversion generation by different preparation methods
On the basis of ginseng sapoglycoside Rg 3, CK, F2 and Rh2, achieve the purpose that characteristic certain rare saponin(e of increase.
(3) strain fermentation and saponin(e are converted and are separated by two step alternating temperature modes by the present invention, very effective to improve conversion effect
Rate.
(4) fermentation process is using sucrose as sugar source, while can also promote fermentation using the nutriment in ginseng aqueous extract
Any other nutriment is not added in the growth of strain, not only high conversion efficiency but also safe and non-toxic.
(5) fermentation liquid obtained in the present invention is compared with existing patent, rare saponin(e quantity and contents level have compared with
Big advantage, and without passing through concentration step, and initial solid-liquid ratio higher than 1:5, raw material utilizes abundant.Compared to patent
CN102366373B, although Rg3 and CK content improves 16.89 and 26.17 times in the patent, its raw material is red ginseng, initially
Solid-liquid ratio is 1:0.4~0.6, and there is also concentration steps in preparation process, greatly consume raw material, raw material sources ratio
White ginseng is more expensive.
(6) ginseng is mostly based on aqueous extract for drug and food, and the present invention directly carries out fermentation with aqueous extract and turns
Change, remain the various nutritional ingredients and bioactive substance of ginseng to greatest extent, be converted into obtained fermentation liquid compared with
The rare ginsenoside of high-content.
Specific embodiment
The present invention will be further described combined with specific embodiments below, but institute's protection scope of the present invention be not limited to it is described
Content.
Method of the invention has versatility to lactic acid bacteria and acetobacter, and strain used in embodiment is purchased from Guangzhou
Guangdong Province's Culture Collection of 5 building, the building of compound the 59th of city martyr Road 100 Guangdong Microbes Inst.Preservation
Number be respectively as follows: Pasteur's acetobacter (GIM1.67), acetic acid bacteria (GIM1.848), lactobacillus plantarum plant subspecies (GIM1.380),
Lactobacillus casei (GIM1.410), lactobacillus fermenti (GIM1.429) and lactobacillus acidophilus (GIM1.412).Four kinds of rare ginsengs
The standard items of saponin(e are detected using HPLC-UV, obtain the appearance time of four kinds of saponin(e standard items, as shown in table 1.Used 4 kinds
Rare ginsenoside standard items are purchased from Shanghai Yuan Ye Biotechnology Co., Ltd.If method is without specified otherwise in embodiment
Conventional method, if the reagent used is conventional commercial reagent without specified otherwise or the reagent that configures according to a conventional method.
The appearance time of 1 four kinds of saponin(e standard items of table
| 1 | F2 | 43.200 |
| 2 | Rg3 | 46.811 |
| 3 | CK | 59.825 |
| 4 | Rh2 | 65.179 |
Embodiment 1
(1) actication of culture: carrying out high pressure steam sterilization for configured MRS culture medium, be cooled to 60 DEG C or so inverted plates,
The lactobacillus plantarum plant subspecies that inclined-plane saves are seeded on MRS solid medium, 35 DEG C of inversions are cultivated 24 hours.Acetic acid bar
Bacterium (Acetobacter pasteurianus) solid medium composition: yeast extract 1%, glucose 1%, agar 2%, ethyl alcohol
2%, pH6.0, percentage are mass percent, and surplus is water.The Acetobacter pasteurianus that inclined-plane saves is seeded to acetobacter
On solid medium, 30 DEG C of inversions are cultivated 24 hours.
(2) seed culture: taking 250mL conical flask, per it is bottled enter 50mL MRS culture medium, cooling after sterilizing, picking step
(1) lactobacillus plantarum plant subspecies single colonie is inoculated in MRS fluid nutrient medium in, 35 DEG C stationary culture 24 hours.Acetic acid bar
Bacteria liquid culture medium prescription: yeast extract 1%, glucose 1%, KH2PO40.25%, ethyl alcohol 2%, pH6.0, percentage is quality
Percentage, surplus are water；250ml shaking flask is packed into culture medium 50mL, and Acetobacter pasteurianus single colonie is inoculated in picking step (1)
In acetobacter fluid nutrient medium, 30 DEG C are cultivated for 24 hours, revolving speed 150rpm.
(3) prepared by ginseng aqueous extract: taking white ginseng that 80 meshes were mechanically pulverized, obtains ginseng pulverate, takes ginseng pulverate 20g, add
Enter 200ml distilled water, is extracted after shaking up with ultrasonic-microwave synergic solvent extraction instrument, microwave power 600W, 90 DEG C of temperature, ultrasound
Power 50Hz, extraction time 15min are filtered to remove ginseng granulated slag after the completion of extracting, obtain ginseng aqueous extract.
(4) fermented and cultured: being added 150g ginseng aqueous extract in the conical flask of 250mL, and 7.5g sucrose, 65 DEG C of water are added
Bath sterilizing 45min, is inoculated with, lactobacillus plantarum plant subspecies and Acetobacter pasteurianus inoculum concentration are respectively in superclean bench
3% and 1%, 32 DEG C of standing for fermentation cultures.
(5) after fermenting 4 days, 50 DEG C is raised the temperature to, continues to ferment.
(6) after fermenting 16 days, filtering fermentating liquid removes thallus and solid impurity, obtains fermented ginseng liquid, dilute in fermentation liquid
There is saponin content to be detected with HPLC-UV.
The separation and detection of rare saponin(e: taking fermented ginseng liquid 5mL, with the water-saturated n-butanol of 3 times of volumes, extracts ginseng
Fermentation liquid 4 times, merge organic phase, vacuum rotary steam obtains ginsenoside crude product, then dissolved with 2mL Chromatographic Pure Methanol, used
HPLC-UV detects rare saponin concentrations, and testing result is as shown in table 1,
The rare saponin content of table 1 and conversion ratio yield (μ g/mL)
4 kinds of rare saponin contents improve above 200% in the present embodiment, and wherein the content of F2 improves 945.62%.
Embodiment 2
(1) actication of culture: carrying out high pressure steam sterilization for configured MRS culture medium, be cooled to 60 DEG C or so inverted plates,
Lactobacillus plantarum plant subspecies and lactobacillus fermenti that inclined-plane saves are seeded on MRS solid medium, 35 DEG C of inversion cultures
24 hours.Acetobacter (Acetobacter pasteurianus) solid medium composition: yeast extract 1%, glucose 1%,
Agar 2%, ethyl alcohol 2%, pH6.0, percentage are mass percent, and surplus is water.The Acetobacter pasteurianus that inclined-plane saves is connect
On kind to acetobacter solid medium, 30 DEG C of inversions are cultivated 24 hours.
(2) seed culture: taking 250mL conical flask, per it is bottled enter 50mL MRS culture medium, it is cooling after sterilizing, pick them separately
Lactobacillus plantarum plant subspecies and lactobacillus fermenti single colonie are inoculated in fluid nutrient medium in step (1), 35 DEG C of stationary cultures
24 hours.Acetobacter Liquid Culture based formulas: yeast extract 1%, glucose 1%, KH2PO40.25%, ethyl alcohol 2%, pH6.0,
Percentage is mass percent, and surplus is water；250ml shaking flask is packed into culture medium 50mL, Acetobacter pasteurianus in picking step (1)
Single colonie is inoculated in fluid nutrient medium, and 30 DEG C are cultivated for 24 hours, revolving speed 150rpm.
(3) prepared by ginseng aqueous extract: taking white ginseng to be mechanically pulverized, crosses 80 meshes, obtain ginseng pulverate, take ginseng pulverate 50g, add
Enter 750ml distilled water, is extracted after shaking up with ultrasonic-microwave synergic solvent extraction instrument, microwave power 800W, 90 DEG C of temperature, ultrasound
Power 50Hz, extraction time 20min.It is filtered to remove ginseng granulated slag after the completion of extracting, obtains ginseng pulverate's aqueous extract.
(4) fermented and cultured: being added 600g ginseng pulverate aqueous extract in the conical flask of 1L, and 60g sucrose, 65 DEG C of water-baths are added
Sterilize 45min, is inoculated in superclean bench, lactobacillus plantarum plant subspecies, lactobacillus fermenti and acetobacter inoculum concentration point
Not Wei 2%, 1.2% and 0.8%, the culture of 35 DEG C of standing for fermentation.
(5) after fermenting 6 days, 48 DEG C is raised the temperature to, continues to ferment.
(6) after fermenting 20 days, filtering fermentating liquid removes thallus and solid impurity, obtains fermented ginseng liquid, dilute in fermentation liquid
There is saponin content to be detected with HPLC-UV.
(7) separation and detection of rare saponin(e: with 1 step of embodiment (7), 4 kinds of rare saponin(e yield such as tables in embodiment 2
Shown in 2.
The rare saponin content of table 2 and conversion ratio yield (μ g/mL)
4 kinds of rare saponin contents improve above 300% in the present embodiment, and wherein the content of Rg3 improves 918.61%.
Embodiment 3
(1) actication of culture: carrying out high pressure steam sterilization for configured MRS culture medium, be cooled to certain temperature inverted plate,
The Lactobacillus casei of inclined-plane preservation and lactobacillus acidophilus are seeded on MRS solid medium, 35 DEG C of inversions are cultivated 24 hours.Vinegar
Acidfast bacilli (Acetobacter pasteurianus) solid medium composition: yeast extract 1%, glucose 1%, agar 2%, second
Alcohol 2%, pH6.0, percentage are mass percent, and surplus is water.The acetobacter that inclined-plane saves is seeded to acetobacter to consolidate
On body culture medium, 30 DEG C of inversions are cultivated 24 hours.
(2) seed culture: taking 250mL conical flask, per it is bottled enter 50mL MRS culture medium, cooling after sterilizing, picking step
(1) lactic acid bacterium colony is inoculated in MRS fluid nutrient medium in, 35 DEG C stationary culture 24 hours.Acetobacter Liquid Culture based formulas:
Yeast extract 1%, glucose 1%, KH2PO40.25%, ethyl alcohol 2%, pH6.0, percentage is mass percent, and surplus is water；
250ml shaking flask is packed into culture medium 50mL, and Acetobacter pasteurianus single colonie is inoculated in fluid nutrient medium in picking step (1), and 30
DEG C culture for 24 hours, revolving speed 150rpm.
(3) prepared by ginseng aqueous extract: taking white ginseng that 80 meshes were mechanically pulverized, obtains ginseng pulverate, takes ginseng pulverate 200g, add
Enter 2L distilled water, shakes ultrasonic-microwave synergic solvent extraction instrument and extract, microwave power 600W, 90 DEG C of temperature, ultrasonic power 50Hz,
Extraction time 25min.It is filtered to remove ginseng granulated slag after the completion of extracting, obtains ginseng pulverate's aqueous extract.
(4) fermented and cultured: being added 1.5kg ginseng pulverate aqueous extract in the conical flask of 2L, and 120g sucrose, 65 DEG C of water are added
Bath sterilizing 45min, is inoculated with, Lactobacillus casei, lactobacillus acidophilus and acetobacter inoculum concentration are respectively in superclean bench
3%, 1% and 0.5%, 37 DEG C of standing for fermentation cultures.
(5) after fermenting 8 days, 55 DEG C is raised the temperature to, continues to ferment.
(6) after fermenting 20 days, filtering fermentating liquid removes thallus and solid impurity, obtains fermented ginseng liquid, dilute in fermentation liquid
There is saponin content to be detected with HPLC-UV.
(7) separation and detection of rare saponin(e: with 1 step of embodiment (7), 4 kinds of rare saponin(e yield such as tables in embodiment 3
Shown in 3.
The rare saponin content of table 3 and conversion ratio yield (μ g/mL)
4 kinds of rare saponin contents improve above 300% in the present embodiment, and wherein the content of F2 and CK is respectively increased
803.39% and 813.39%.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (10)
1. a kind of method for improving rare saponin content in ginseng water extract, which comprises the following steps:
(1) sieving after ginseng crushing and grinding is obtained into Ginseng Root Powder, by Ginseng Root Powder rear progress ultrasonic-microwave processing soluble in water
Ginseng granulated slag is filtered out, ginseng water extract is obtained；
(2) ginseng water extract made from step (1) is taken, Compound bacterium and sugar source is added and ferments 4~8 days at 28~40 DEG C, it is described
Compound bacterium is lactic acid bacteria and acetobacter；
(3) fermentation temperature is increased to 42~55 DEG C, continues fermentation 10~20 days, and after fermentation, filtering leaves fermentation liquid.
2. the method according to claim 1 for improving rare saponin content in ginseng water extract, which is characterized in that step
(1) mass ratio of the Ginseng Root Powder and water is 1:5~20.
3. the method according to claim 2 for improving rare saponin content in ginseng water extract, which is characterized in that step
(2) mass ratio of sugar source and ginseng water extract is 5%~20%, and the inoculum concentration of the Compound bacterium is 3%~7%.
4. the method according to claim 3 for improving rare saponin content in ginseng water extract, which is characterized in that step
(2) inoculation quality of the lactic acid bacteria and acetobacter ratio is 3~8:1.
5. the method according to any one of claims 1 to 4 for improving rare saponin content in ginseng water extract, feature
It is, the power of step (1) ultrasound is 50~80Hz, and the power of the microwave is 400~800W.
6. the method according to any one of claims 1 to 4 for improving rare saponin content in ginseng water extract, feature
It is, the time of step (1) the ultrasonic-microwave processing is 10~30min.
7. the method according to any one of claims 1 to 4 for improving rare saponin content in ginseng water extract, feature
It is, step (2) lactic acid bacteria is lactobacillus plantarum, Lactobacillus casei, lactobacillus paracasei, lactobacillus fermenti and acidophilus cream
One or more of bacillus；Step (2) acetobacter is one in Acetobacter pasteurianus and oxidation acetobacter
Kind or two kinds.
8. the method according to any one of claims 1 to 4 for improving rare saponin content in ginseng water extract, feature
It is, the granularity of step (1) described Ginseng Root Powder is 180~250 μm.
9. the method according to claim 1 for improving rare saponin content in ginseng water extract, which is characterized in that described
Rare saponin(e is one or more of Rg3, CK, F2 and Rh2.
10. the method according to claim 1 for improving rare saponin content in ginseng water extract, which is characterized in that step
Suddenly ginseng described in (1) is white ginseng.
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| CN110591952A (en) * | 2019-09-24 | 2019-12-20 | 扬州大学 | Lactobacillus paracasei with capability of decomposing oil and fat and application thereof |
| CN110771875A (en) * | 2019-11-05 | 2020-02-11 | 质每(中国)有限公司 | Method for fermenting ginseng by using lactobacillus |
| CN113621673A (en) * | 2021-09-03 | 2021-11-09 | 北京思维晶格科技有限公司 | Method for fermenting ginseng by using compound strain, fermentation product and application |
| CN113621673B (en) * | 2021-09-03 | 2023-07-25 | 北京思维晶格科技有限公司 | Method for fermenting ginseng by composite strain, fermentation product and application |
| CN114525322A (en) * | 2021-12-29 | 2022-05-24 | 杭州娃哈哈科技有限公司 | Application of lactobacillus plantarum in preparation of ginseng fermentation liquor |
| CN115029385A (en) * | 2022-07-25 | 2022-09-09 | 华南理工大学 | Method for extracting tea saponin from tea seed meal by microbial fermentation |
| CN115029385B (en) * | 2022-07-25 | 2024-05-24 | 华南理工大学 | Method for extracting tea saponin from tea seed cake by microbial fermentation |
| CN115444792A (en) * | 2022-08-10 | 2022-12-09 | 山东朵拉朵尚生物科技有限公司 | Ginseng fermentation liquor with anti-wrinkle and anti-firmness effects and preparation method and application thereof |
| CN115444792B (en) * | 2022-08-10 | 2023-08-18 | 山东朵拉朵尚生物科技有限公司 | Ginseng fermentation liquor with anti-wrinkle tightening effect and preparation method and application thereof |
| CN117547491A (en) * | 2024-01-10 | 2024-02-13 | 拉芳家化股份有限公司 | Microbial conversion ginseng fermentation liquor for promoting scalp health and preparation method and application thereof |
| CN117547491B (en) * | 2024-01-10 | 2024-05-03 | 拉芳家化股份有限公司 | Microbial conversion ginseng fermentation liquor for promoting scalp health and preparation method and application thereof |
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