CN109810905A - The interior raw Xylaria sp. fungus UT-X bacterial strain and application thereof of one plant of production polysaccharide - Google Patents
The interior raw Xylaria sp. fungus UT-X bacterial strain and application thereof of one plant of production polysaccharide Download PDFInfo
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- CN109810905A CN109810905A CN201910068660.6A CN201910068660A CN109810905A CN 109810905 A CN109810905 A CN 109810905A CN 201910068660 A CN201910068660 A CN 201910068660A CN 109810905 A CN109810905 A CN 109810905A
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- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 70
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 70
- 150000004676 glycans Chemical class 0.000 title claims abstract description 69
- 230000001580 bacterial effect Effects 0.000 title claims abstract description 23
- 241001557892 Xylaria sp. Species 0.000 title claims abstract description 18
- 238000004519 manufacturing process Methods 0.000 title claims description 6
- 238000000855 fermentation Methods 0.000 claims abstract description 14
- 230000004151 fermentation Effects 0.000 claims abstract description 14
- 241000468405 Xylariaceae sp. Species 0.000 claims abstract description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 claims abstract description 3
- 239000006228 supernatant Substances 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 9
- 230000001376 precipitating effect Effects 0.000 claims description 7
- 235000019441 ethanol Nutrition 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 4
- OAABHEHWRQAHEJ-UHFFFAOYSA-N butan-1-ol;chloroform Chemical compound ClC(Cl)Cl.CCCCO OAABHEHWRQAHEJ-UHFFFAOYSA-N 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000005374 membrane filtration Methods 0.000 claims description 3
- 244000267222 Brasenia schreberi Species 0.000 abstract description 54
- 235000006506 Brasenia schreberi Nutrition 0.000 abstract description 53
- 230000008901 benefit Effects 0.000 abstract description 3
- 238000013459 approach Methods 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 230000002950 deficient Effects 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 19
- 241000894006 Bacteria Species 0.000 description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- 238000010521 absorption reaction Methods 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 8
- 239000002609 medium Substances 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 238000002329 infrared spectrum Methods 0.000 description 4
- 241000233866 Fungi Species 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 3
- 241001523965 Xylaria Species 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 239000008239 natural water Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000003595 spectral effect Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 description 2
- 241000186046 Actinomyces Species 0.000 description 2
- 229920002498 Beta-glucan Polymers 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 108091023242 Internal transcribed spacer Proteins 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
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- JFTUSFFYSRNFBA-UHFFFAOYSA-N 3-amino-5-nitrosalicylic acid Chemical compound NC1=CC([N+]([O-])=O)=CC(C(O)=O)=C1O JFTUSFFYSRNFBA-UHFFFAOYSA-N 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000019155 Radiation injury Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
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- 229960000935 dehydrated alcohol Drugs 0.000 description 1
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- -1 hydroxy radical Polysaccharide Chemical class 0.000 description 1
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- 238000003760 magnetic stirring Methods 0.000 description 1
- 229960002523 mercuric chloride Drugs 0.000 description 1
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
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- 229920001277 pectin Polymers 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
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- 238000002360 preparation method Methods 0.000 description 1
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- 238000007480 sanger sequencing Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
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- 238000010183 spectrum analysis Methods 0.000 description 1
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- 238000012916 structural analysis Methods 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
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- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The present invention provides a kind of interior raw Xylaria sp. fungus UT-X bacterial strains for producing polysaccharide, belong to Xylaria sp. fungus (Xylariaceae sp.strain) raw in one kind, which is preserved in China typical culture collection center, address: China, Wuhan, Wuhan University；Postcode 430072, the deposit date is on January 17th, 2019, deposit numbers are as follows: CCTCC M 2019055；The 16S rDNA sequence of the bacterial strain is as shown in SEQ ID NO.1.Above-mentioned interior raw Xylaria sp. fungus UT-X bacterial strain can be used in fermentation and prepare polysaccharide.Bacterial strain provided by the invention can carry out large scale fermentation culture in a short time, and fermentation costs are lower, it is not limited again by conditions such as region, seasons, therefore the increasingly deficient status of water shield natural resources can be solved by the approach of microbial fermentation, to realize higher economic benefit.
Description
Technical field
The present invention relates to Xylaria strain raw in one kind, which can produce polysaccharide, belong to microorganisms technical field.
Background technique
The research of endophyte of plant is gradually taken seriously at present, and the various endophytes of different plants are found in succession.Interior life
The diversity of strain class also gives the diversity of its metabolite, this, which is also implied, excavates biology using microbial metabolism, changes
The new resources such as, medicine have huge potentiality.It is a large amount of studies have shown that endophyte of plant can produce it is identical as host plant or
Similar active material and precursor, this discovery have pushed directly on the especially economical endophyte of plant research of endophyte of plant
Expansion extensively.Gradually research with people to economical endophyte of plant generates corresponding active constituent using that can be metabolized
Endophyte produces novel, efficient, inexpensive active medicine, solves rare economical plant resources status in short supply, will
Emphasis as economical endophyte of plant research in the future.
Although the concern increasingly by numerous scholars in recent years of the especially economical endophyte of plant of endophyte of plant,
But the endophyte research of several hundred kinds of plants is still only carried out at present.Water shield (Brasenia schreberi) also known as horseshoe
Dish, lake dish, water certain herbaceous plants with big flowers, dew certain herbaceous plants with big flowers, water lotus leaf, flower case dish etc. are that perennial root floats leaf water plant, belong to Nymphaeceae water shield category.Water shield
The stickiness pectin main component on dish surface is stickiness polysaccharide, has high nutritive value.Water shield is a kind of important aquatic warp
Ji plant, the requirement to water quality are very high.It is worsening instantly in the wild border environment of water shield, find a kind of Polysaccharide of Brasenia Schreberi production
Alternative route be particularly important.
Water shield endophyte just starts to be concerned by people in recent years, on the whole, especially about water shield endophyte
Can be metabolized generate corresponding Polysaccharide of Brasenia Schreberi ingredient research it is still relatively fewer, rarely have about produce Polysaccharide of Brasenia Schreberi water shield endophyte
Report.
Summary of the invention
The present invention solves the problems in background technique, provides a kind of interior raw Xylaria sp. fungus UT-X bacterial strain for producing polysaccharide, should
Bacterial strain can produce Polysaccharide of Brasenia Schreberi.
The present inventor screens one plant of novel strain, which is named as UT-X, belongs to raw Xylaria sp. fungus in one kind
(Xylariaceae sp.strain), which is preserved in China typical culture collection center, address: China, and Wuhan is military
Chinese university；Postcode 430072, the deposit date is on January 17th, 2019, deposit numbers are as follows: CCTCC M 2019055；The bacterial strain
16S rDNA sequence is as shown in SEQ ID NO.1.
Above-mentioned interior raw Xylaria sp. fungus UT-X bacterial strain can be used in fermentation and prepare polysaccharide.
The method of polysaccharide is prepared using above-mentioned interior raw Xylaria sp. fungus UT-X bacterial strain the following steps are included: first by interior raw Xylaria sp. fungus
UT-X strain inoculated is in LB liquid medium, the overnight shaking culture 2 days of 37 DEG C of 200r/min revolving speed, by zymocyte liquid in
12000r/min centrifugation, takes supernatant；Supernatant and chloroform-butanol solution are mixed in centrifuge tube, 20-30min is vibrated,
Supernatant is taken after being centrifuged again, by supernatant with twice of membrane filtration of 0.45 μm, heating is concentrated into original volume at 70 DEG C after filtering
1/2-2/3,95% ethyl alcohol of 4 times of volumes is added, be put into refrigerator freezing stay overnight, finally with 12000r/min revolving speed be centrifuged
10min, precipitating generated are polysaccharide.
Compared with prior art, the invention has the following advantages that interior raw Xylaria sp. fungus UT-X bacterial strain energy provided by the present invention
Enough metabolism generate polysaccharide.It is metabolized the polysaccharide generated and the polysaccharide HPLC wave having the same isolated and purified from natural water shield
Spectral peak and similar polysaccharide infrared spectrum characteristic peak and similar or identical chemical functional group.And prepared by the strain fermentation
Polysaccharide biological activity compared with natural Polysaccharide of Brasenia Schreberi, natural Polysaccharide of Brasenia Schreberi is significantly higher than to the removing of hydroxy radical effect,
With better antioxidant activity.
Bacterial strain provided by the invention can carry out large scale fermentation culture in a short time, and fermentation costs are lower, but not by
The limitation of the conditions such as region, season, therefore showing for water shield natural resources increasingly scarcity can be solved by the approach of microbial fermentation
Shape, to realize higher economic benefit.
Detailed description of the invention
Fig. 1 is the colonial morphology figure of bacterial strain provided by the invention in the medium；
Fig. 2 is efficient liquid phase (HPLC) analysis of Polysaccharide of Brasenia Schreberi extract and strain fermentation product polysaccharide provided by the invention
Figure；
Fig. 3 is the infrared spectrum analysis figure of strain fermentation product polysaccharide provided by the invention.
Specific embodiment
Detailed specific description done to the present invention combined with specific embodiments below, but protection scope of the present invention not office
It is limited to following embodiment.
The separation of water shield endophyte
Stem, the Ye Hegen for taking fresh water shield are first rinsed with distilled water with mercuric chloride disinfection several again under sterile environment
Time, it is inoculated on LB solid medium, is protected from light culture.Go out well-grown bacterium colony from culture medium picking and moves to new LB culture
In base plate, is purified, numbered respectively.The Strain Designation provided in the present embodiment is UT-X bacterial strain, by the bacterium of the bacterial strain
It falls and moves to well-grown in new LB culture medium flat plate, colonial morphology figure is as shown in Figure 1, Fig. 1 medium scale=2mm.From Fig. 1
In it can be seen that colony diameter is about 4-6mm, bacterium colony protuberance, edge is rough, it is seen that mycelia, color are opaque white.
The identification of water shield endophyte
The endophyte isolated and purified (is extracted DNA or boil processing) after processing, amplification fungi ITS sequence (is used
ITS1 and ITS4 primer), amplified production carry out sanger sequencing, sequencing result the website NCBI (https: //
Blast.ncbi.nlm.nih.gov/Blast.cgi analysis) is compared, with homology 97% or more and the highest object of homology
Kind is judged as the kind of endophyte.
ITS sequence amplimer sequence:
1 ITS1:5 '-TCCGTAGGTGAACCTGCGG-3 ' of primer
Primer 2 ITS4:5 '-TCCTCCGCTTATTGATATGC-3 '
Amplified fragments size about 400-700bp
According to the standard sequencing methods of fungi, the sequence of above-mentioned water shield endosymbiosis bacterium is measured as shown in SEQ ID NO.1, is surveyed
Sequence result is compared on the website NCBI, the results showed that, the water shield endosymbiosis bacterium provided in the present embodiment is fungi, interior
The Latin generic name of raw bacterium is interior raw Xylaria sp. fungus (Xylariaceae sp.strain).
The bacterial strain is preserved in China typical culture collection center by applicant, address: China, Wuhan, Wuhan University；Postal
430072 are compiled, the deposit date is on January 17th, 2019, patent culture presevation numbers are as follows: CCTCC M 2019055.
The preparation of water shield endophyte polysaccharide
The strain inoculated for being partially transferred to purifying is picked them separately in LB liquid medium from the bacterium in LB culture medium flat plate,
Zymocyte liquid is centrifuged 5min in 12000r/min, takes supernatant by the overnight shaking culture 2 days of 37 DEG C of 200r/min revolving speed.Configuration
Supernatant is mixed by 3:1 with centrifuge tube with organic solution, is vibrated by chloroform-butanol solution (chloroform: n-butanol=4:1)
Supernatant is taken after 20-30min, 8000r/min centrifugation 10min, pays attention to the protein precipitation that cannot be drawn between two-phase.Continue by upper
It states operation and carries out 5 removing proteins.By supernatant with twice of membrane filtration of 0.45 μm, heating is concentrated into original at 70 DEG C after filtering
95% ethyl alcohol of 4 times of volumes is added in the 1/2-2/3 of volume, be put into refrigerator freezing stay overnight, second day with 12000r/min revolving speed from
Heart 10min, precipitating are endophyte polysaccharide.
Whether the fermentation liquid for water shield endophyte more provided by the invention contains polysaccharide, internally raw fermented liquid extract
Measurement of the polysaccharide content is carried out.
Thick many candies are extracted and isolated first from natural water shield as object of reference, the method is as follows: are taken water shield tender shoots, be put into
Extraction in NaOH (0.1mol/L), solid-liquid ratio 1:2 (w/v), 25 DEG C of lye adjust pH to 7.0 after extracting 1.5h.Choose water shield bud,
It is centrifugated residue with centrifuge, with magnetic stirring apparatus by supernatant in 60 DEG C of concentration 1h.Then it is handled repeatedly with sevage method
The crude extract of concentration removes free protein, is again concentrated to 2/3 of original volume or so.Four times of volume dehydrated alcohol precipitatings are added
Overnight, mixture is then centrifuged 10min with 4000r/min to obtain sediment, is concentrated again after being precipitated with the dissolution of a small amount of water
Four times of volume ethanol precipitations are added to stay overnight, repetitive operation is three times, finally dry to obtain Polysaccharide of Brasenia Schreberi (BSP).
Measuring method is as follows: by polysaccharide after sulphuric acid hydrolysis, measuring bacterium using DNS method (3- amino -5-NITROSALICYLIC ACID)
Secretion sugared content in liquid is tried by the glucose standard of various concentration and DNS first using D-Glucose aldehydic acid as standard
Absorbance after agent, sulfuric acid reaction makes standard curve.After measured, standard curve is y=0.0368x-0.0436 (R2=
0.9807).According to standard curve, the content of polysaccharide is measured.The experimental results showed that in BSP sample polysaccharide content be 0.30 ±
0.01mg/mL, the polyoses content in interior raw Xylaria actinomyces UT-X strain fermentation extract are 0.07 ± 0.001mg/mL.
The Performance Testing of polysaccharide component
Whether to compare water shield endophyte polysaccharide and Polysaccharide of Brasenia Schreberi there are also identical polysaccharide component, the present embodiment is carried out
HPLC analysis.As a result as shown in Fig. 2, wherein upper figure is endophyte polysaccharide in the present embodiment, the following figure is the natural water shield extracted
Polysaccharide, from figure 2 it can be seen that interior raw Xylaria actinomyces polyoses extract has a wave spectrum peak in 19min, in 21min
There is a subwave spectral peak, it is notable that the HPLC appearance time class at secondary pop peak and Polysaccharide of Brasenia Schreberi that 21min occurs
Seemingly, show water shield endosymbiosis bacterium in addition to generating specific polysaccharide peak value, also have the generation polysaccharide similar with Polysaccharide of Brasenia Schreberi at
The ability of part.
The structural analysis of Polysaccharide of Brasenia Schreberi is compared with the carbohydrate of endophyte
To further determine that water shield endophyte polysaccharide and Polysaccharide of Brasenia Schreberi whether also containing identical chemical functional group, this implementation
Further progress infrared spectrum analysis in example.
The strain inoculated for being partially transferred to purifying is picked them separately in LB liquid medium from the bacterium in LB culture medium flat plate,
The overnight shaking culture 2 days of 37 DEG C of 200r/min revolving speed.Bacterium solution is centrifuged, supernatant removes deproteinized with sevage method, then with 4 times of bodies
Product alcohol chromatography, is separated by filtration precipitating, whether has glycosidic bond in infrared spectrum analysis precipitating or alcohol analysis liquid.Use infrared spectroscopy
The polysaccharide extracted from water shield is analyzed, is made comparisons with the map of the precipitating containing glycosidic bond or alcohol analysis liquid.
Infrared spectrogram is as shown in figure 3, analysis shows, water shield endophyte polysaccharide provided in this embodiment has 9 in Fig. 3
Characteristic absorption peak.It can be seen that from infrared spectrogram, 3360.00cm-1The absorption peak at place is the stretching vibration absworption peak of-OH,
2924.09cm-1It is the stretching vibration absworption peak of C-H, this is the typical wave spectrum peak of two polysaccharide, the typical wave of this and Polysaccharide of Brasenia Schreberi
Spectral peak is similar；2854.65cm-1、1556.55cm-1And 1456.26cm-The absorption peak at place is that the stretching vibration of the C-H on phenyl ring is inhaled
Peak is received, is water shield endosymbiosis bacterium distinctive polysaccharide wave spectrum peak；1651.07cm-1The absorption peak at place is that C=O bond (C=O) inhales
Receive peak；1404.18cm-1, 551.64cm-1The absorption peak at place is the stretching vibration absworption peak of C-H.It is specifically intended that
1083.99cm-1The absorption peak at place is absorption peak associated with glycosidic inkage C-O-H, is the characteristic peak of beta glucan.
Compared with Polysaccharide of Brasenia Schreberi, water shield endosymbiosis bacterium produces polysaccharide in 3360.00cm-1And 2924.09cm-1It is having the same
Polysaccharide absorption peak, while having unique characteristic peak (in 2854.65cm again-1、1556.55cm-1And 1456.26cm-1).Water shield
Endosymbiosis bacterium and Polysaccharide of Brasenia Schreberi, in 1083.99cm-1There is the characteristic peak of identical beta glucan at place.Separate sources polysaccharide infrared light
Spectrum analysis and its characteristic peak parsing are as shown in the table:
The active measurement of polysaccharide anti-oxidative
The present embodiment detects Polysaccharide of Brasenia Schreberi extract using the hydroxy radical kit that biology offer is built up in Nanjing.
Polysaccharide sample is uniformly mixed with reagent, in 37 DEG C of accurate response 1min, color developing agent is added immediately and terminates reaction, is uniformly mixed
Afterwards, it is placed at room temperature for 20min.Spectrophotometer is returned to zero with distilled water, the absorbance value of each pipe is surveyed at 550nm.Pass through experiment
As the result is shown to the Scavenging activity of hydroxy radical, the Partial Antioxidation activity of Polysaccharide of Brasenia Schreberi is obtained.
The importance of polysaccharide is embodied on its physiological activity.Hydroxy radical (·It OH) is a kind of very strong freedom of oxidability
Base, property is very active, and the rate for aoxidizing various organic matters and inorganic matter is exceedingly fast, and is to cause tissue lipid peroxidating, nucleic acid disconnected
It splits, protein and how glycolytic principal element, it is related with body aging, tumour, radiation injury and cell phagocytic activity.For into
One step determines the biological activity of water shield endosymbiosis bacterium polysaccharide, generates system in the present embodiment with hydroxy radical and compares in water shield
The antioxidant activity of symbiosis granulose.Result of study shows that the supression of generation of the water shield endosymbiosis bacterium polysaccharide to hydroxy radical is made
Reach in the case of 1.0mg/mL most strong (as shown in the table), and the inhibition that Polysaccharide of Brasenia Schreberi generates hydroxy radical with
The raising of concentration and increase (as shown in the table).Experimental result shows that water shield endosymbiosis bacterium polysaccharide imitates the removing of hydroxy radical
Polysaccharide of Brasenia Schreberi should be significantly higher than.
Sequence table
<110>South-Center University For Nationalities
The interior raw Xylaria sp. fungus UT-X bacterial strain and application thereof of<120>one plants of production polysaccharide
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 599
<212> DNA
<213>in raw Xylaria sp. fungus (Xylariaceae sp. strain)
<400> 1
gtaggttgaa cctgcggagg gatcattact gagttctaca aaaaactccc aaccctttgt 60
gaaccttacc gtcgttgcct cggcgccgag cggcggctac cctggagaag ctacccggga 120
gccacctacc ctgtaggtgg ctaccctgga gctaccctgt agtagtttgc attctacgct 180
ccgccggcgg accttctaca ctctgttttg tatagtgtat ctctgaaacc tataacgtaa 240
tacgttaaaa ctttcaacaa cggatctctt ggttctggca tcgatgaaga acgcagcgaa 300
atgcgatacg taatgtgaat tgcagaattc agtgaatcat cgaatctttg aacgcatatt 360
gcgcccatta gtattctagt gggcatgcct attcgagcgt catttcaacc cttacgcccc 420
tgttgcgtag tgttgggaac ctacaggcct gtaaaaagga cctgtagctc cctaaaggta 480
gtggcggtgt taggtacact cgtagcgtag taacatcttt tctcgctcct gcagtgtacc 540
taaggcctgc cgtgaaaaac cccctataac ttctagtggt tgacttcgga ttaggtagg 599
Claims (3)
1. interior raw Xylaria sp. fungus (Xylariaceae sp.strain) the UT-X bacterial strain of one plant of production polysaccharide, the bacterial strain are preserved in China
Type Tissue Collection, deposit number are as follows: CCTCC M 2019055, the 16S rDNA sequence such as SEQ ID of the bacterial strain
Shown in NO.1.
2. the purposes of interior raw Xylaria sp. fungus UT-X bacterial strain described in claim 1, it is characterised in that: prepare polysaccharide for fermentation.
3. utilizing the method that interior raw Xylaria sp. fungus UT-X bacterial strain prepares polysaccharide described in claim 1, it is characterised in that including following step
It is rapid: first by interior raw Xylaria sp. fungus UT-X strain inoculated in LB liquid medium, the 37 DEG C of overnight shaking cultures 2 of 200r/min revolving speed
It, zymocyte liquid is centrifuged in 12000r/min, takes supernatant；Supernatant and chloroform-butanol solution are mixed in centrifuge tube
In, 20-30min is vibrated, then takes supernatant after being centrifuged, by supernatant with twice of membrane filtration of 0.45 μm, after filtering at 70 DEG C
Heating is concentrated into the 1/2~2/3 of original volume, and 95% ethyl alcohol of 4 times of volumes is added, and is put into refrigerator freezing and stays overnight, finally uses
12000r/min revolving speed is centrifuged 10min, and precipitating generated is polysaccharide.
Priority Applications (1)
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| CN118792170A (en) * | 2024-08-21 | 2024-10-18 | 贵州农业职业学院 | A strain of Xylaria officinalis UJ3-2 and its application |
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