CN108018226A - A kind of high-yield extracellular polysaccharide strains and its polysaccharide fermentation production method - Google Patents
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Abstract
A kind of high-yield extracellular polysaccharide strains and its polysaccharide fermentation production method, strain was named Leuconostoc mesenteroides (Leuconstoc mesenteroides) DRP105.The present invention separates the lactic acid bacteria of one plant of high-yield extracellular polysaccharide from the nature sauerkraut zymotic fluid of northeast, is identified as Leuconostoc mesenteroides (Leuconstoc mesenteroides).Leuconostoc mesenteroides (Leuconstoc mesenteroides) DRP105 is inoculated in the production sugar fermentation medium containing 5% sucrose, it is 5~8 in initial medium pH value, temperature is 20~35 DEG C, under the conditions of 0~170rpm of shaking speed, cultivate 36~72h, zymotic fluid is obtained, zymotic fluid is extracted, obtains Thick many candies；Thick many candies are purified, up to exocellular polysaccharide.Exocellular polysaccharide content is 20.12 ± 1.87g/L.This method fermentation period is short, cost is low, output of sugar is high, easy to operate.
Description
Technical field
The present invention relates to a kind of high-yield extracellular polysaccharide strains and its polysaccharide fermentation production method.
Background technology
Exocellular polysaccharide (exopolysaccharide, EPS) is microorganism (such as bacterium, yeast, fungi) in growth course
It is secreted into extracellular mucus or capsular polysaccharide.Microbial exopolysaccharides are because with mild condition needed for deriving from a wealth of sources, react, easy
It is always the main source of exocellular polysaccharide theory and practice research in isolate and purify the advantages that.In the micro- of numerous extracellular polysaccharide
In biology, lactic acid bacteria (lactic acid bacteria, LAB) has the superiority such as fermentation period is short, nutritional requirement is simple, into
For the ideal source of Microbial exopolysaccharides.Successfully developed by leuconostoc mesenteroide (Leuconostoc from the forties
Mesenteroides) since fermenting and producing dextran (dextran), in world wide the research of Microbial exopolysaccharides with
Exploitation achieves the achievement that attracts people's attention, the exploitations of new Microbial exopolysaccharides become industrial microorganism research hot spot it
One.Lactic acid bacteria is the common name of a kind of bacterium that fermentable sugars can be utilized to produce a large amount of lactic acid, as the probiotics of generally recognized as safe,
Lactic acid bacteria can promote the growth of intestinal beneficial microbes, equal to diarrhea, intestines easily swash disease, allergy, lactose intolerance, stimulate the reaction etc.
There is the effect of certain, have high application value in the key areas closely related with human lives such as industry, agricultural, medicine.
Exopolysaccharides Produced by Lactic Acid Bacteria is used in the production of the fermented dairy products such as Yoghourt, cheese as stabilizer, thickener.It is in addition, newborn
Sour bacterium exopolysaccharide also have reduce cholesterol, it is anti-oxidant, antitumor, to it is gastral adjust etc. effect.Compared with other polysaccharide,
The research of Exopolysaccharides Produced by Lactic Acid Bacteria has more real value, and lactobacter growth is extremely rapid, its exocellular polysaccharide extraction process is more
Simply, security is higher, cheap, and the mankind are more suitable for than other microbial polysaccharides.Therefore, excavate what is enriched in nature
Resource of lactic bacteria database, finds the lactobacillus bacteria bacterial strain of high-yield extracellular polysaccharide, is obtained with short cycle, low cost more extracellular more
Sugar, is always the research hotspot of microbiological art.
The content of the invention
The purpose of the invention is to expand the microorganism fungus kind source of extracellular polysaccharide, there is provided a kind of lactic acid bacteria is extracellular
The new method of polysaccharide.
The extracellular polysaccharide strains are leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105, institute
State leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 and be preserved in China on 04 18th, 2017
(No. 3 Chinese Academy of Sciences of institute of Chaoyang District, Beijing City Beichen Lu 1 are micro- for Microbiological Culture Collection administration committee common micro-organisms center
Biological study institute), deposit number is CGMCC NO:14046.The bacterial strain through 16S rRNA gene sequencing the result is shown in sequence table
SEQ ID NO:1。
The extracellular polysaccharide strains are gram-positive bacterium, do not produce gemma and flagellum, amphimicrobian can be with a variety of sugar
Class carries out lactic fermentation for carbon source.Bacterium colony is circular, opaque, canescence, neat in edge, slightly projection, positive and negative solid colour,
Center is consistent with edge color.The sour aerogenesis of glucose fermentation production, catalase is negative, does not hydrolyze arginine, does not produce indoles.Microorganism
Physiological and biochemical test illustrates the characteristic with bright string strain Pseudomonas (Leuconostoc).
Exopolysaccharide Production From The Fermentation production method of the present invention, carries out according to the following steps：
1) leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 is inoculated in containing 5% sucrose
It is 5~8 in initial medium pH value, temperature is 20~35 DEG C, shaking speed 0~170rpm conditions in MRS basal mediums
Under, 36~72h is cultivated, obtains zymotic fluid.
2) zymotic fluid is subjected to low-temperature and high-speed centrifugation, removes thalline, take supernatant.
3) the supernatant ethanol precipitation after removing thalline will be centrifuged, stood, centrifugation, takes precipitation, i.e. Thick many candies.
4) Thick many candies are purified, obtains pure exocellular polysaccharide.
In step 1), leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 is extracellular
Polysaccharide, optimal fermentation culture conditions are that initial medium pH value is 6.5, and temperature is 30 DEG C, shaking speed 100rpm, cultivates 48h.
Exocellular polysaccharide content is 27.67 ± 2.54g/L, and fermentation period is short, and output of sugar is high.
In step 2), the centrifugal rotational speed is preferably 12000rpm, and centrifugation time is preferably 40min.At this time thalline and
Exocellular polysaccharide can be separated well.
In step 3), the ethanol percentage concentration is preferably 95%~100%, by volume, supernatant:Ethanol is most
It is 1 well:3, the time of repose is preferably 24h.12000rpm centrifuges 30~40min.
In step 4), the purifying can purify polysaccharide using gel permeation chromatography, can apply phend-sulphuric acid
Exocellular polysaccharide content is measured, exocellular polysaccharide content is 20.12 ± 1.87g/L.
The present invention obtains strain fermentation product exocellular polysaccharide by range of biochemical technology, using modern extraction technique pair
Broth extraction purifying obtains high-purity polysaccharide.Verified by serial experiment, illustrate that obtained bacterial strain has Leuconostoc
Characteristic, under the induction of substrate sucrose, produces a large amount of exocellular polysaccharides.Therefore, leuconostoc mesenteroide (Leuconostoc
Mesenteroides) DRP105, containing exocellular polysaccharide, has production in the sugared fermentation medium of production in zymotic fluid obtained by fermentation
The advantages of sugared cycle is short, output of sugar is high.
Brief description of the drawings
Fig. 1 leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 colonial morphologies；
Fig. 2 is that initial pH value produces sugar to leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105
Influence；
Fig. 3 is that cultivation temperature produces sugar to leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105
Influence；
Fig. 4 is that shaking speed produces sugar to leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105
Influence；
Fig. 5 is leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 growths and production sugar after optimization
Process；
Fig. 6 is leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 exocellular polysaccharide gel filtrations
Chromatograph collection of illustrative plates；
Fig. 7 is the pure exocellular polysaccharide ultraviolet lights of leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105
Spectrogram.
Embodiment
Embodiment 1：A kind of separation screening of high-yield extracellular polysaccharide strains
1) culture medium
MRS basal mediums：Glucose 20g, tryptone 10g, beef extract 10g, yeast extract 5g, ammonium citrate
2g, K2HPO42g, CH3COONa 5g, MgSO4·7H2O 0.58g, MnSO4·H2O 0.25g, Tween 80 1mL, distilled water
1000mL, 6.5,115 DEG C of sterilizing 20min of pH value.
The sugared fermentation medium of MRS productions：Sucrose 50g, tryptone 10g, beef extract 10g, yeast extract 5g, K2HPO4
2g, CH3COONa 5g, ammonium citrate 2g, MgSO4·7H2O 0.58g, MnSO4·H2O 0.25g, Tween 80 1mL, distilled water
1000mL, 6.5,115 DEG C of sterilizing 20min of pH value.
2) sample pretreatment
Take northeast sauerkraut zymotic fluid 1mL to add in the sterile saline equipped with 9mL, be made 10-1The sample liquid of concentration, shake
Suitable concentration, stand for standby use are diluted to step by step after swinging mixing.
3) production Acarasiales falls primary dcreening operation
The 50 μ L of bacterium solution of above-mentioned each dilution factor are taken to be coated in production sugar culture-medium agar plate, in 37 DEG C of constant incubators
Cultivate 24-48h.Viscous bacterium colony region is produced on picking tablet, in three ride repeatedly on production sugar culture-medium agar plate, is obtained
The tablet of pure culture is placed in 4 DEG C of refrigerators and preserves.
4) bacterial strain secondary screening
The pure culture obtained from tablet is inoculated into MRS fluid nutrient mediums and cultivates 18h, then connecing by 2% (v/v)
Kind amount is inoculated into production sugar liquors culture medium, and 48h are cultivated in 30 DEG C.Take zymotic fluid to heat 10min in 90 DEG C of water-baths, remove bacterium
The enzyme of possible degradation of polysaccharide, is cooled to room temperature in liquid.80% trichloroacetic acid (TCA) solution is added into zymotic fluid to final concentration of
5% (m/v), stirs 2h, 12000 × g, 4 DEG C of centrifugation 40min, remove cell and albumen precipitation at room temperature.Supernatant loads dialysis
In bag (molecular cut off 14000Da), ultra-pure water dialysis 2d in 4 DEG C of refrigerators, a water is changed per 8h.After constant volume, measure bacterial strain production
The yield of exocellular polysaccharide.The mucus concentration fallen with reference to production Acarasiales, chooses the higher bacterial strain of polysaccharide yield.
Embodiment 2：A kind of identification of high-yield extracellular polysaccharide strains
Classical microbial biochemical examines explanation, and the high-yield extracellular polysaccharide strains obtained have Leuconostoc
(Leuconostoc) characteristic, gram-positive bacterium, does not produce gemma and flagellum, amphimicrobian.Bacterium colony is circle, impermeable
Bright, canescence, neat in edge omits projection, and positive and negative solid colour is central consistent with edge color.
This bacterial strain azymic galactolipin, lactose and raffinose.Glucose can be utilized to produce sour aerogenesis, catalase is negative, no
Arginine is hydrolyzed, does not produce indoles.Arginine production ammonia test, H2S is tested, and Starch Hydrolysis experiment, gelatin liquefaction test is feminine gender.
According to its physio-biochemical characteristics and 16S rRNA sequence analyses as a result, through China General Microbiological culture presevation pipe
Reason center (CGMCC) is accredited as (Leuconostoc mesenteroides) DRP105 bacterial strains, and carries out preservation, deposit number
For CGMCC NO:14046.
Embodiment 3：A kind of zymotechnique of the extracellular polysaccharide of high-yield extracellular polysaccharide strains fermentation
A kind of zymotechnique of the extracellular polysaccharide of high-yield extracellular polysaccharide strains fermentation of the present embodiment, carries out according to the following steps：
1) seed liquor is prepared
Leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 is inoculated in basic MRS culture mediums
In, it is 1.0 × 10 to make initial cell density in culture medium8A/mL, is seed liquor.
2) sugared fermentation condition optimization is produced
By leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 with the inoculation of 2.0% (V/V)
Amount, is inoculated in the 100/250mL culture mediums that initial pH value is 5.5,6.0,6.5,7.0,7.5 and 8.5,80r/min, 25 respectively
DEG C constant-temperature shaking culture 30h, measures fermented supernatant fluid polyoses content using Phenol sulfuric acid procedure, determines optimal initial pH value.
By leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 with the inoculum concentration of 2.0% (V/V)
In 100/250mL culture mediums after access optimization, the 80r/min shaken cultivations 30h at a temperature of 20,25,30 and 35, sharp respectively
Fermented supernatant fluid polyoses content is measured with Phenol sulfuric acid procedure, determines optimal cultivation temperature.
By leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 with the inoculum concentration of 2.0% (V/V)
In 100/250mL culture mediums after access optimization, respectively at 0,50,80,100,120,150 and the shaking table of 170r/min rotating speeds
In, 30 DEG C of constant-temperature shaking culture 30h, fermented supernatant fluid polyoses content is measured using Phenol sulfuric acid procedure, determines that most suitable shaking table turns
Speed.
Using the optimal culture condition fermentation leuconostoc mesenteroide (Leuconostoc mesenteroides) after optimization
DRP105, is placed in 100r/min, 30 DEG C of constant-temperature shaking culture 72h, and fermented supernatant fluid polyoses content is measured using Phenol sulfuric acid procedure,
Determine optimal production sugar fermentation time.All test process are respectively provided with three repetitions.
Initial pH value be 6.5, temperature is 30 DEG C, under the conditions of shaking speed 120rpm, cultivate 48h, obtain zymotic fluid, it is right
Zymotic fluid is extracted, and obtains exocellular polysaccharide.The content that exocellular polysaccharide is obtained in the zymotic fluid that the present embodiment obtains is 27.67
±2.54g/L。
Embodiment 4：A kind of separation purifying technique of the extracellular polysaccharide of high-yield extracellular polysaccharide strains fermentation
1) prepared by Thick many candies
By the leuconostoc mesenteroide activated (Leuconostoc mesenteroides) DRP105 bacterium solutions by 2% (v/
V) inoculum concentration is inoculated into the sugared fermentation medium of production, 30 DEG C, and 120rpm cultures 48h obtains Exopolysaccharide Production From The Fermentation liquid.By 500mL
4 DEG C of zymotic fluid, 12000rpm centrifugation 40min, removes thalline.Supernatant adds 95% ethanol of precooling of 3 times of volumes, 4 DEG C of hatchings
Overnight with precipitate polysaccharides.4 DEG C, 12000rpm centrifugations 50min collects polysaccharide precipitation, more with 30~40 DEG C of dissolvings of 250mL ultra-pure waters
Sugar precipitation, adds 250mL (isometric with the ultra-pure water used in dissolving polysaccharide) 10% solution of trichloroacetic acid and removes deproteinized, 4 DEG C quiet
10h is put, 4 DEG C, 12000rpm centrifugations 40min is collected supernatant.Add 3 times of volumes 95% ethanol of precooling, 4 DEG C of incubated overnights with
Precipitate polysaccharides.4 DEG C, 12000rpm centrifugations 40min collects polysaccharide precipitation, is dissolved in again in ultra-pure water (about 200 mL), loads saturating
Analyse in bag (molecular cut off 14000Da), 4 DEG C of distilled water dialysis 2d, a water is changed per 8h.Finally obtain the water-soluble of Thick many candies
Liquid, 4 DEG C of refrigerators preserve.
2) polysaccharide purification
Take the aqueous solution of above-mentioned Thick many candies to carry out gel permeation chromatography directly or after first appropriateness dilution, merge collecting pipe polysaccharide
Solution, is freeze-dried 24h, obtains holosaccharide, and purifying exocellular polysaccharide content is measured using Phenol sulfuric acid procedure.
3) purity of polysaccharide is identified
The accurate polysaccharide 5mg weighed after purification, is dissolved in 5mL ultra-pure waters, the polysaccharide solution of 1mg/mL is made, with it is ultraviolet can
See that spectrometer carries out UV scanning in 190-350nm wave bands, detect purity of polysaccharide.
The content that exocellular polysaccharide is obtained in the zymotic fluid that the present embodiment obtains is 20.12 ± 1.87g/L, no albumen and nucleic acid
Pollution, purity are high.
Sequence table
<110>University Of Tianjin
<120>A kind of high-yield extracellular polysaccharide strains and its polysaccharide fermentation production method
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1224
<212> DNA
<213>Artificial sequence
<400> 1
agactgatcg gccacattgg gactgagaca cggcccaaac tcctacggca ggctgcagta 60
gggaatcttc cacaatgggc gaaagcctga tggagcaacg ccgcgtgtgt gatgaaggct 120
ttcgggtcgt aaagcactgt tgtatgggaa gaacagctag aataggaaat gattttagtt 180
tgacggtacc ataccagaaa gggacggcta aatacgtgcc agcagccgcg gtaatacgta 240
tgtcccgagc gttatccgga tttattgggc gtaaagcgag cgcagacggt ttattaagtc 360
tgatgtgaaa gcccggagct caactccgga attgcattgg aaactggtta acttgagtgc 420
agtagaggta agtggaactc catgtgtagc ggtggaatgc gtagatatat ggaagaacac 480
cagtggcgaa ggcggcttac tggactgcaa ctgacgttga ggctcgaaag tgtgggtagc 540
aaacaggatt agataccctg gtagtccaca ccgtaaacga tgaacactag gtgttaggag 600
gtttccgcct cttagtgccg aagctaacgc attaagtgtt ccgcctgggg agtacgaccg 660
caaggttgaa actcaaagga attgacgggg acccgcacaa gcggtggagc atgtggttta 720
attcgaagca acgcgaagaa ccttaccagg tcttgacatc ctttgaagct tttagagata 780
gaagtgttct cttcggagac aaagtgacag gtggtgcatg gtcgtcgtca gctcgtgtcg 840
tgagatgttg ggttaagtcc cgcaacgagc gcaaccctta ttgttagttg ccagcattca 900
gatgggcact ctagcgagac tgccggtgac aaaccggagg aaggcgggga cgacgtcaga 960
tcatcatgcc ccttatgacc tgggctacac acgtgctaca atggcgtata caacgagttg 1020
ccagcccgcg agggtgagct aatctcttaa agtacgtctc agttcggatt gtagtctgca 1080
actcgactac atgaagtcgg aatcgctagt aatcgcggat cagcacgccg cggtgaatac 1140
gttcccgggt cttftacaca ccgcccgtca caccatggga gtttgtaatg cccaaagccg 1200
gtggcctaac cttcgggaag gagc 1224
Claims (9)
1. a kind of high-yield extracellular polysaccharide strains, it is leuconostoc mesenteroide (Leuconostoc mesenteroides)
DRP105, in preservation on the 18th in 04 month in 2017 to China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation
Numbering is CGMCC NO:14046.
2. the polysaccharide fermentation production method of a kind of high-yield extracellular polysaccharide strains, it is characterised in that take as claimed in claim 1
A kind of extracellular polysaccharide strains, comprise the following steps：
1) leuconostoc mesenteroide (Leuconostoc mesenteroides) DRP105 is inoculated in the MRS bases containing 5% sucrose
It is 5~8 in initial medium pH value in basal culture medium, temperature is 20~35 DEG C, under the conditions of 0~170rpm of shaking speed, culture
36~72h, obtains zymotic fluid；
2) zymotic fluid is subjected to low-temperature and high-speed centrifugation, removes thalline, take supernatant；
3) the supernatant ethanol precipitation after removing thalline will be centrifuged, stood, centrifugation, takes precipitation, i.e. Thick many candies；
4) Thick many candies are purified, obtains pure exocellular polysaccharide.
A kind of 3. polysaccharide fermentation production method of high-yield extracellular polysaccharide strains according to claim 2, it is characterised in that step
It is rapid 1) described in initial medium pH value be 6.5, temperature is 20~35 DEG C, under the conditions of 0~170rpm of shaking speed, culture 36~
72h。
A kind of 4. polysaccharide fermentation production method of high-yield extracellular polysaccharide strains according to claim 2, it is characterised in that step
It is rapid 1) described in initial medium pH value be 6.5, temperature is 30 DEG C, under the conditions of 0~170rpm of shaking speed, cultivates 36~72h.
A kind of 5. polysaccharide fermentation production method of high-yield extracellular polysaccharide strains according to claim 2, it is characterised in that step
It is rapid 1) described in initial medium pH value be 6.5, temperature is 30 DEG C, under the conditions of shaking speed 100rpm, cultivates 36~72h.
A kind of 6. polysaccharide fermentation production method of high-yield extracellular polysaccharide strains according to claim 2, it is characterised in that step
It is rapid 1) described in initial medium pH value be 6.5, temperature is 30 DEG C, under the conditions of shaking speed 100rpm, cultivates 48h.
A kind of 7. polysaccharide fermentation production method of high-yield extracellular polysaccharide strains according to claim 2, it is characterised in that step
Rapid 2) described centrifugal rotational speed is 12000rpm, and centrifugation time is 30~40min.
A kind of 8. polysaccharide fermentation production method of high-yield extracellular polysaccharide strains according to claim 2, it is characterised in that step
Rapid 3) described ethanol percentage concentration is 95%~100%, by volume, supernatant:Ethanol is 1:3 add ethanol, stand
24h, 12000rpm centrifuge 30~40min.
A kind of 9. polysaccharide fermentation production method of high-yield extracellular polysaccharide strains according to claim 2, it is characterised in that step
Rapid 4) described purifying purifies polysaccharide using gel permeation chromatography.
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| CN114196564A (en) * | 2021-10-08 | 2022-03-18 | 沈阳农业大学 | Tetragenococcus halophilus and application thereof in production of anti-cancer extracellular polysaccharide |
| CN114437985A (en) * | 2022-02-18 | 2022-05-06 | 南京工业大学 | Enterobacter aerogenes and application thereof in synthesizing microbial polysaccharide |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1692158A (en) * | 2002-08-06 | 2005-11-02 | 丹尼斯科公司 | Use of lactobacillus to produce exopolysaccharides in food and pharmaceutical compositions |
| CN1695469A (en) * | 2005-05-17 | 2005-11-16 | 田星 | Flavor rancid milk beneficial for health and preparation method |
| CN101654664A (en) * | 2009-09-25 | 2010-02-24 | 厦门百拓生物工程有限公司 | Fermentation production method and application of high-yield exopolysaccharide strain and amylase thereof |
| CN103013891A (en) * | 2012-12-27 | 2013-04-03 | 光明乳业股份有限公司 | Leuconostoc mesenteroides, as well as exopolysaccharide and application thereof |
| CN105349477A (en) * | 2015-12-21 | 2016-02-24 | 光明乳业股份有限公司 | Leuconostoc mesenteroides as well as preparation method and application thereof |
| CN105440153A (en) * | 2015-12-21 | 2016-03-30 | 光明乳业股份有限公司 | Water-insoluble exopolysaccharide of leuconostoc mesenteroides and preparation method thereof |
-
2017
- 2017-06-24 CN CN201710489399.8A patent/CN108018226A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1692158A (en) * | 2002-08-06 | 2005-11-02 | 丹尼斯科公司 | Use of lactobacillus to produce exopolysaccharides in food and pharmaceutical compositions |
| CN1695469A (en) * | 2005-05-17 | 2005-11-16 | 田星 | Flavor rancid milk beneficial for health and preparation method |
| CN101654664A (en) * | 2009-09-25 | 2010-02-24 | 厦门百拓生物工程有限公司 | Fermentation production method and application of high-yield exopolysaccharide strain and amylase thereof |
| CN103013891A (en) * | 2012-12-27 | 2013-04-03 | 光明乳业股份有限公司 | Leuconostoc mesenteroides, as well as exopolysaccharide and application thereof |
| CN105349477A (en) * | 2015-12-21 | 2016-02-24 | 光明乳业股份有限公司 | Leuconostoc mesenteroides as well as preparation method and application thereof |
| CN105440153A (en) * | 2015-12-21 | 2016-03-30 | 光明乳业股份有限公司 | Water-insoluble exopolysaccharide of leuconostoc mesenteroides and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 王英等: "产葡聚糖明串珠菌的分离鉴定及发酵条件研究", 《食品与生物技术学报》 * |
| 范娟等: "一株肠膜明串珠菌的分离筛选及其合成葡聚糖培养条件的优化", 《食品科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114196564A (en) * | 2021-10-08 | 2022-03-18 | 沈阳农业大学 | Tetragenococcus halophilus and application thereof in production of anti-cancer extracellular polysaccharide |
| CN114437985A (en) * | 2022-02-18 | 2022-05-06 | 南京工业大学 | Enterobacter aerogenes and application thereof in synthesizing microbial polysaccharide |
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