CN109652479A - A method of improving dendrobium polysaccharide oxidation resistance - Google Patents
A method of improving dendrobium polysaccharide oxidation resistance Download PDFInfo
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- CN109652479A CN109652479A CN201910073553.2A CN201910073553A CN109652479A CN 109652479 A CN109652479 A CN 109652479A CN 201910073553 A CN201910073553 A CN 201910073553A CN 109652479 A CN109652479 A CN 109652479A
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- dendrobium
- oxidation resistance
- polysaccharide
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- 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
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
Abstract
The invention discloses a kind of microbe fermentation methods for improving dendrobium polysaccharide oxidation resistance, belong to active components of plants functional promotion field.The present invention passes through inoculation beneficial microbe, the big polysaccharide of molecular weight is degraded to the micromolecular polysaccharide absorbed conducive to organism using microbial fermentation, and confirms that this method effectively increases the inoxidizability of dendrobium polysaccharide by the total antioxidant capacity of measurement fermentation liquid and stoste.Method and process of the invention is simple, easy to operate, and reaction condition is mild, environmentally friendly, easy to implement and popularization；And it is small by the dendrobium polysaccharide molecular weight that this method obtains, it is conducive to organism and absorbs, oxidation resistance is strong, has broad application prospects in fields such as the research and development of functional medicines and food.
Description
Technical field
The present invention relates to active components of plants functional promotion field more particularly to a kind of raising dendrobium polysaccharide oxidation resistances
Method.
Background technique
Dendrobium nobile (Dendrobium) is one of the orchid recorded earliest in China's ancient documents, has reinforcing stomach reg fluid, grows
The effect of negative heat-clearing, main active are polysaccharide.Nearly ten years, relevant research work is concentrated mainly on dendrobium candidum
Extracting method, chemical component, pharmacological action and finger-print etc. research (morning, the progress of Dendrobium officinale polysaccharide,
China National folk medicine, 2015,24 (15): 6-7).
Polysaccharide refers to as 10 or more monosaccharide through polymer made of glucosides key connection, is widely present in animal, plant
In microorganism cultures, it is one of base substance for sustaining life and running well (Cheng Gaomin etc., anti-skin of Chinese medicinal plant polysaccharide
Skin aging).Research finds that polysaccharide and saccharide complex participate in and mediated adjusting (Liu Yao etc., the plant of the various biological phenomenas of cell
Polysaccharide bioactivity research progress, Jiangsu's agriculture science, 2013,41 (1): 1-4).
There are many free radicals in human body, have extremely strong oxidability, the unsaturated fatty acid of various biomembranes can be made to send out
Raw peroxidating, forms lipid peroxide, and nucleic acid is caused to be crosslinked mistake, breaks down proteins, the weight consequence such as DNA mutation, with body
Aging and a variety of diseases are related.Scientific research is related to many diseases, it was also found that oxidative stress increases.People are general at present
Free radical, anti-aging effects are removed all over thinking that plant polyose has.Its mechanism of action, it is possible explain there are several types of: (1) it is straight
It connects and acts on ROS itself.(2) polysaccharide molecule acts on antioxidase.(3) polysaccharide molecule complexing generates metal necessary to ROS
Ion.(4) SOD is promoted to discharge from cell surface.As people go deep into polysaccharide researches, it has been found that polysaccharide has many lifes
Object active function.It is antitumor, improve immune, antiviral, anti-inflammatory, hypoglycemic, reducing blood lipid, anti-aging and in terms of
All show gratifying effect.This may show related (Liu of stronger antioxidant activity with polysaccharide in various experimental systems
Yao etc., plant polyose bioactivity research progress, Jiangsu's agriculture science, 2013,41 (1): 1-4).
In the polysaccharide leaching liquor typically directly extracted, polysaccharide molecular weight is big, and water-soluble effect is poor, is unfavorable for organism suction
Income in vivo, plays its bioactivity, this limits the application of polysaccharide to a certain extent, and a degree of degradation is able to maintain
The biological activity of polysaccharide is even improved, thus is of great significance.In Degradation of Polysaccharides, microbial degradation method is not needed
A large amount of reaction reagents, reaction condition is mild, and environmentally friendly.Catabolite safety can be used as raw material for cosmetics, food
And the production of drug.
Summary of the invention
The object of the present invention is to provide a kind of methods for improving polysaccharide anti-oxidative ability, by way of microbial fermentation,
Polysaccharide anti-oxidative ability is improved, reduces polysaccharide molecular weight while not influencing polysaccharide main component, is conducive to organism and absorbs
With the raising of bioactivity, it is not required to a large amount of reaction reagent, reaction condition is mild, environmentally friendly.
The object of the invention is achieved by the following technical programs:
A method of improving dendrobium polysaccharide oxidation resistance, comprising the following steps:
(1) Hot water extraction extracts dendrobium polysaccharide；
(2) microorganism fungus kind is activated；
(3) seed liquor is prepared；
(4) inoculation fermentation.
Preferably, the microorganism is bacillus subtilis.It is protected by Agricultural University Of South China's plant germplasm and research on utilization
Center saves, the bacillus subtilis for being 1.821 purchased from China General Microbiological culture presevation administrative center number.
Preferably, detailed process is as follows for step (1): taking fresh dendrobium nobile, is sliced, drying to constant weight, sieving is ground, by 1:80
(g:mL) dendrobium nobile dry powder and distilled water is added in solid-liquid ratio, filters after 80~90 DEG C of 1~2h of heating water bath, and filter residue repeats to extract one
It is secondary, merge extracting solution twice, addition dehydrated alcohol to final concentration of the 80% of alcohol filters to obtain Thick many candies precipitating overnight, will be slightly more
Sugared precipitating is redissolved with distilled water, redissolve after solution concentration be adjusted to 10mg/mL, take 50mL redissolve after solution into reaction vessel,
It sterilizes spare.
Preferably, detailed process is as follows for step (2): the bacterium colony saved with transfer needle picking, in flat lining out, constant temperature training
Support 37 DEG C of cultures of case for 24 hours, picking single bacterium kind does inclined-plane and saves or continue to draw plate amplification.
Preferably, detailed process is as follows for step (3): scraping 3 ring strains in the nutrition of 100mL from the plate cultivated for 24 hours
Broth bouillon, 37 DEG C of constant-temperature table, 180r/min culture obtains seed liquor for 24 hours.
Preferably, detailed process is as follows for step (4): 1mL seed liquor is added in wait the polysaccharide solution that ferments, constant-temperature table training
Support 37 DEG C of case, after 180r/min, 16h, 121 DEG C of sterilizing 20min.
Preferably, the dendrobium nobile is dendrobium candidum, HERBA DENDROBII, Dendrobium Chrysotoxum Lindl, Dendrobium fimbriatum Hook, Dendrobium crepidatum lindl et paxt., big luxuriant sheath
Dendrobium nobile, swollen arthrolith dry measure used in former times.
The invention has the following advantages:
The present invention solves in the certain situation of application cost condition, reduces the dosage of dendrobium nobile and possesses same antioxygen originally
The property changed, this will greatly reduce polysaccharide dosage, provide basic data for the development and utilization of dendrobium polysaccharide, increase dendrobium polysaccharide processing enterprise
The economic benefit of industry.Meanwhile after microbial fermentation, macromolecular polysaccharide will be degraded to the small molecule for being conducive to organism and absorbing, will
Improve the medical value of dendrobium polysaccharide, and has great importance for the research and development of functional medicines and food.The method operation
Simply, low in cost, it is not required to a large amount of reaction reagent, reaction condition is mild, easy to operate.
Detailed description of the invention
Fig. 1 is polysaccharide of the embodiment 1 through biofermentation compared with polysaccharide stoste oxidation resistance.
Fig. 2 is polysaccharide of the embodiment 2 through biofermentation compared with polysaccharide stoste oxidation resistance.
Specific embodiment
It is clearly and completely described below in conjunction with the technical solution in the embodiment of the present invention, it is clear that described reality
Applying example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is general
Logical technical staff every other embodiment obtained without making creative work belongs to what the present invention protected
Range.
In order to make the foregoing objectives, features and advantages of the present invention clearer and more comprehensible, With reference to embodiment
The present invention is described in further detail.
Embodiment 1
Agar Plating: agar powder (dry powder) 20g/L, beef extract 3g/L, peptone 10g/L, NaCl 5g/L, pH
Value is 7.4 ± 0.2；Fluid nutrient medium: beef extract 3g/L, peptone 10g/L, NaCl 5g/L, pH value are 7.4 ± 0.2.It is used
Microorganism be bacillus subtilis, purchased from China General Microbiological culture presevation administrative center number be 1.821 withered grass bud
Spore bacillus, ferments in the steps below:
(1) Hot water extraction extracts Dendrobium officinale polysaccharide: taking fresh dendrobium nobile 100g, is sliced, is placed in 105 DEG C of dryer progress
Drying and processing, after 5min, 65 DEG C dry to constant weight (about 41.31g), milled 40 mesh, and stone is added by 1:80 (g:mL) solid-liquid ratio
Dry measure used in former times dry powder 20g and distilled water 1600mL and, filtered after 80 DEG C of heating water bath 1h, filter residue repeats to extract primary, and merging is extracted twice
Dehydrated alcohol is added to final concentration of the 80% of alcohol in liquid, about 3.00L, and 4 DEG C overnight, filters to obtain Thick many candies precipitating about 13.77g, adds
5L distilled water redissolves, and solution concentration after redissolving is adjusted to 10mg/mL, solution is placed in into 100mL conical flask after taking 50mL to redissolve
121 DEG C, sterilizing 20min is spare.
(2) microorganism fungus kind is activated: the bacterium colony saved with transfer needle picking, in flat lining out, 37 DEG C of constant incubator
For 24 hours, picking single bacterium kind does inclined-plane and saves or continue to draw plate amplification for culture.
(3) it prepares seed liquor: scraping 3 ring strains from the plate cultivated for 24 hours in the nutrient broth medium of 100mL, perseverance
37 DEG C of warm shaking table, 180r/min culture obtain seed liquor for 24 hours.
(4) inoculation fermentation: being added 1mL seed liquor in wait the polysaccharide solution that ferments, and 37 DEG C of constant-temperature table incubator, 180r/
After min, 16h, 121 DEG C of sterilizing 20min.
Measurement fermentation liquid total antioxidant capacity: surveying the height of its total economic value using ABTS free radical scavenging ability, with
Polysaccharide stoste without everfermentation is as control, the result is shown in Figure 1.Stable bluish-green color substance is generated after ABTS oxidation, in 734nm
There is maximum absorption band at place.And measured object oxidation resistance makes the bluish-green color substance that different degrees of colour fading occur, according to light absorption value
Variation, calculate measured matter total antioxidant capacity.
It will be seen from figure 1 that comparing with stoste, the polysaccharide total antioxidant capacity through biofermentation is enhanced.Work as fermentation
Time is 16h, and when polysaccharide concentration is 2.5mg/mL, the clearance rate to ABTS free radical is 14.48%, with higher anti-oxidant
Ability.Stoste is 6.21% to the clearance rate of ABTS free radical.Compared with stoste, the total antioxidant capacity for the 16h that ferments is improved
2.3 again.
Embodiment 2
Agar Plating: agar powder (dry powder) 20g/L, beef extract 3g/L, peptone 10g/L, NaCl 5g/L, pH
Value is 7.4 ± 0.2；Fluid nutrient medium: beef extract 3g/L, peptone 10g/L, NaCl 5g/L, pH value are 7.4 ± 0.2.It is used
Microorganism be bacillus subtilis, purchased from China General Microbiological culture presevation administrative center number be 1.821 withered grass bud
Spore bacillus, ferments in the steps below:
(1) Hot water extraction extracts Dendrobium officinale polysaccharide: fresh dendrobium nobile 200g is taken, is sliced, 105 DEG C, after 5min, and 65 DEG C of bakings
To constant weight (about 50.34g), dendrobium nobile dry powder 30g and distilled water is added by 1:80 (g:mL) solid-liquid ratio in milled 40 mesh
It is filtered after 2400mL, 90 DEG C of heating water bath 1h, filter residue repeats to extract once, merges extracting solution, about 4.60L twice, is added anhydrous
For ethyl alcohol to final concentration of the 80% of alcohol, 4 DEG C overnight, filters to obtain Thick many candies precipitating about 18.10g, 5L distilled water is added to redissolve precipitating
Afterwards, concentration is adjusted to 10mg/mL, in the conical flask for taking 50mL to 100mL, is placed in 121 DEG C, sterilizing 20min is spare.
Remaining step is the same as embodiment 1.
Figure it is seen that comparing with stoste, the polysaccharide total antioxidant capacity through biofermentation is enhanced.Work as fermentation
Time is 16h, and when polysaccharide concentration is 2.5mg/mL, the clearance rate to ABTS free radical is 15.13%, with higher anti-oxidant
Ability.Stoste is 6.81% to the clearance rate of ABTS free radical.Compared with stoste, the total antioxidant capacity for the 16h that ferments is improved
2.22 again.
Embodiment described above is only that preferred embodiment of the invention is described, and is not carried out to the scope of the present invention
It limits, without departing from the spirit of the design of the present invention, those of ordinary skill in the art make technical solution of the present invention
Various changes and improvements, should all fall into claims of the present invention determine protection scope in.
Claims (7)
1. a kind of method for improving dendrobium polysaccharide oxidation resistance, it is characterised in that: the following steps are included:
(1) Hot water extraction extracts dendrobium polysaccharide；
(2) microorganism fungus kind is activated；
(3) seed liquor is prepared；
(4) inoculation fermentation.
2. the method according to claim 1 for improving dendrobium polysaccharide oxidation resistance, it is characterised in that: the microorganism is
Bacillus subtilis.
3. the method according to claim 1 for improving dendrobium polysaccharide oxidation resistance, it is characterised in that:
Detailed process is as follows for step (1): taking fresh dendrobium nobile, is sliced, drying to constant weight, sieving is ground, by 1:80 (g:mL) feed liquid
Than dendrobium nobile dry powder and distilled water is added, filtered after 80~90 DEG C of 1~2h of heating water bath, filter residue repeats to extract once, merges twice
Extracting solution is added dehydrated alcohol to final concentration of the 80% of alcohol, overnight, filters to obtain Thick many candies precipitating, Thick many candies are precipitated to use and are steamed
Distilled water is redissolved, and solution concentration after redissolving is adjusted to 10mg/mL, solution sterilizes spare into reaction vessel after taking 50mL to redissolve.
4. the method according to claim 1 for improving dendrobium polysaccharide oxidation resistance, it is characterised in that:
Detailed process is as follows for step (2): the bacterium colony saved with transfer needle picking, in flat lining out, 37 DEG C of constant incubator trainings
It supports for 24 hours, picking single bacterium kind does inclined-plane and saves or continue to draw plate amplification.
5. a kind of method for improving dendrobium polysaccharide oxidation resistance according to claim 1, it is characterised in that:
Detailed process is as follows in step (3)；It is permanent in the nutrient broth medium of 100mL from scraping strain on the plate of culture for 24 hours
37 DEG C of warm shaking table is cultivated under the conditions of 180r/min and obtains seed liquor for 24 hours.
6. the method according to claim 1 for improving dendrobium polysaccharide oxidation resistance, it is characterised in that:
Detailed process is as follows in step (4): 2% seed liquor is added in conical flask, 37 DEG C of constant-temperature table incubator, and 180r/
Min, 37 DEG C of constant-temperature table incubator, after 180r/min, 16h, 121 DEG C of sterilizing 20min.
7. the method according to claim 1 for improving dendrobium polysaccharide oxidation resistance, it is characterised in that: the dendrobium nobile is iron
Skin dendrobium nobile, HERBA DENDROBII, Dendrobium Chrysotoxum Lindl, Dendrobium fimbriatum Hook, Dendrobium crepidatum lindl et paxt., big luxuriant sheath dendrobium nobile, swollen arthrolith dry measure used in former times.
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| CN111196864A (en) * | 2020-03-06 | 2020-05-26 | 遵义医科大学 | Dendrobium nobile polysaccharide with moisturizing effect and preparation method and application thereof |
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| CN111196864B (en) * | 2020-03-06 | 2021-09-14 | 遵义医科大学 | Dendrobium nobile polysaccharide with moisturizing effect and preparation method and application thereof |
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