CN104371933A - Strain of sclerotium rolfsii and application thereof in fermentation production of scleroglucan - Google Patents
Strain of sclerotium rolfsii and application thereof in fermentation production of scleroglucan Download PDFInfo
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Abstract
The invention discloses a strain of sclerotium rolfsii suitable for industrial production of scleroglucan. The strain is called sclerotium rolfsii SCL2010; the strain has been collected in China Center for Type Culture Collection on July 6, 2014 with the collection number of CCTCC NO.M2014325. The invention also discloses the application of the sclerotium rolfsii in fermentation production of scleroglucan. Perfect combination of rapid growth of the sclerotium rolfsii and efficient generation of the scleroglucan is realized by controlling the fermentation conditions and the fermentation process stage by stage; the stable operation of the fermentation process is guaranteed, the fermentation yield and the production strength are enabled to reach 35g/L and 0.44g/L/h, respectively, and the yield is remarkably higher than that of the existing process; the prepared product scleroglucan is high in viscosity, excellent in stability and low in viscosity change, and has excellent application performance.
Description
Technical field
The present invention relates to a kind of microbial polysaccharide Sclerotium gum and produce bacterial strain, particularly relate to a strain Sclerotium rolfsii and the application in fermentative production Sclerotium gum thereof.
Background technology
Sclerotium gum (also referred to as scleroglucan or Scleroglucan) is the non-ionic water-soluble homopolysaccharide produced by filamentous fungus.Its molecule is made up of the linear β-D-(1,3) containing β-D-(1,6)-glucosyl residue side chain-glucosyl residue chain.The remarkable characteristic of Sclerotium gum is good rheological and stability, and it all has satisfactory stability in pH (1 ~ 12), salinity (0 ~ 200,000ppm) and temperature (130 DEG C) widely.
Sclerotium gum all has good using value in industry such as food, medicine, makeup, oil.At present in world market, Sclerotium gum is generally applied in makeup as moisturizing, anti-inflammatory, thickening composition, and the international well-known makeup brand of Chang Bei adopted.Sclerotium gum can also improve oil recovery factor, current generally enter three times, four times recover the oil trend under, it has broad application prospects.
Fermentation production rate due to Sclerotium gum is not high causes it to hold at high price, and the exploitation that this severely limits its market (particularly petroleum industry field) is expanded.Therefore, how research improves the output of Sclerotium gum is principal concern.The research about Sclerotium gum fermentative production of current report focuses mostly in the optimization of fermention medium, but it still exists fermentation time long (more than 96h), productive rate low (about 20g/l), the defect that production cost is higher.Although the published domestic and international patent CN201110006769.0 about raising Sclerotium gum fermentative production, CN201310283916.8 and EP 1417325A2 productive rate reach as high as 32g/L, its technology controlling and process is complicated, and stability is not enough.
Summary of the invention
For the deficiencies in the prior art, the present invention to deal with problems be to provide a strain have unique fermentating law be applicable to industrialized Sclerotium gum high-efficiency fermenting produce bacterial classification---Sclerotium rolfsii and the application in fermentative production Sclerotium gum thereof.
The Sclerotium rolfsii being applicable to industrial production Sclerotium gum of the present invention, it is characterized in that: this bacterial strain is called Sclerotium rolfsii (Sclerotium rolfsii) SCL2010, bacterial strain is deposited in China typical culture collection center (China Center for Type Culture Collection on July 6th, 2014, be called for short CCTCC, address: China. Wuhan. Wuhan University), its deposit number is CCTCC NO.M2014325.
Above-mentioned Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 colonial morphology and microscopic features as follows:
Described bacterium grows soon on PDA substratum, and cultivate 3 days under 28 DEG C of conditions, mycelia covers with culture dish, and mycelia is luxuriant, flocculence, and gas is raw, white.
Mycelia wall is thin, and diameter 2.5 ~ 6.5 μm, branch, has clamp connexion.
Colony edge produce sclerotium, be just oyster white, after gradually become light yellow to brown or black, spherical or oval, size 0.5 ~ 2mm, smooth surface tool gloss.
Sclerotium rolfsii of the present invention (Sclerotium rolfsii) SCL2010 is measured to the result display of the gene order (PCR universal primer is NS1F and NS8R) of 18S rDNA, its gene order length is 1694bp, and concrete nucleotide sequence is as shown in SEQ ID NO.1.
Wherein: described PCR universal primer NS1F and NS8R sequence as follows:
NS1F：5’-GTAGTCATATGCTTGTCTC-3’
NS8R：5’-TCCGCAGGTTCACCTACGGA-3’
The basic skills of Sclerotium rolfsii of the present invention (Sclerotium rolfsii) SCL2010 strain improvement is: from Shouguang City of Weifang City of Shandong Province, Gaomi City, Linqu County infects the potato plant of southern blight and stem tuber collects southern blight sclerotium, take sclerotium as parting material, sclerotium is inoculated on PDA solid medium under aseptic condition and cultivates, after sclerotial germination mycelia grows, carry out isolation and purification culture, transfer and to preserve on PDA slant medium.
The mycelia of separation and purification being transferred on PDA substratum is transferred respectively on new PDA flat board, cultivate Continuous Observation for 28 DEG C, treat that mycelial growth is paved with flat board and bears sclerotium, till sclerotium variable color maturation, record mycelial growth situation, tuberculosis and sclerotium changing conditions, determine Sclerotium rolfsii bacterial strain wherein in conjunction with microscopic examination and carry out strain number; To be defined as the sclerotium switching of Sclerotium rolfsii with on PDA flat board, observed and recorded sclerotial germination situation, germination rate, mycelial growth situation, knot sclerotium time and sclerotium amount.Choose that sclerotial germination is fast, germination rate >=75%, knot sclerotium time fast (about 72h), bacterial strain that sclerotium amount is many carry out the experiment of Sclerotium gum shake flask fermentation, pass through great many of experiments, the Sclerotium rolfsii of final acquisition high yield Sclerotium gum, called after Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 bacterial strain, this bacterial strain sclerotial germination rate is up to 80%, fermentation final pH≤2.5, productive rate can reach 35g/L, and passage assays shows that this bacterial strain genetic stability is good.
The application of Sclerotium rolfsii of the present invention (Sclerotium rolfsii) SCL2010 in fermentative production Sclerotium gum.
Realize smoothly for ease of fermenting process, reduce production cost, the preferred technical scheme of application method is: by Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 after solid slope and seed liquor enlarged culturing, be inoculated in fermentation culture according to the amount of fermentation culture mass percent 4 ~ 10%, grading-controlling fermentation condition, fermentation obtains Sclerotium gum.
Wherein, described application method relate to concrete steps order as follows:
(1) Sclerotium gum produces bacterial classification Sclerotium rolfsii eggplant-shape bottle inclined-plane solid culture:
Be inoculated on eggplant-shape bottle inclined-plane solid medium by the ripe sclerotium of Sclerotium rolfsii (Sclerotium rolfsii) SCL2010,30 ~ 34 DEG C of quiescent culture 5 ~ 7 days, obtain a large amount of ripe Sclerotium rolfsii, prepare for fermentative production seed liquor.
Above-mentioned inclined-plane solid PDA forms (unit: g/L): potato 200 (filtering after poach is rotten), glucose 20, agar 20.
(2) Sclerotium gum fermentative production seed liquor preparation:
Wash by cultivating ripe Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 stroke-physiological saline solution, amount is inoculated in seed culture fluid routinely, pH to 4 ~ 5 of adjustment seed culture fluid, 30 ~ 34 DEG C are carried out aeration-agitation and cultivate 30 ~ 36h, obtain containing a large amount of mycelial Sclerotium gum fermentative production seed liquor.
Wherein: described seed culture fluid is made up of (unit: g/L) the component of following massfraction:
Glucose: 10 ~ 20, yeast extract paste 3 ~ 6, SODIUMNITRATE 2 ~ 3, dipotassium hydrogen phosphate 2 ~ 5, magnesium sulfate 1 ~ 3, polyether antifoam agent 0.2 ~ 1, other are softening water, pH 4 ~ 5.
(3) Sclerotium gum fermentative production and grading-controlling fermentation condition technique:
Measure in the fermentation culture be inoculated into containing carbon source routinely by cultivating ripe Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 fermentative production seed liquor, the pH of adjustment nutrient solution, carry out aeration-agitation fermentation culture, by grading-controlling fermentation condition, produce and obtain Sclerotium gum.
Wherein, described fermentation culture is made up of (unit: g/L) the component of following mass percent:
Carbon source: 45 ~ 60, bean cake powder 1 ~ 3, corn steep liquor 0.5 ~ 2, SODIUMNITRATE 1 ~ 3, dipotassium hydrogen phosphate 1 ~ 2, magnesium sulfate 0.5 ~ 1, polyether antifoam agent 0.2 ~ 1, other are softening water, pH 4 ~ 5.Carbon source in nutrient solution is selected from the mixture of a kind of or its any part by weight in starch, sucrose and glucose.
Be divided into the thalli growth phase due to fermenting process and produce the sugar phase, and carry out along with fermentation, fermentation broth viscosity rises fast, therefore preferred technical scheme carries out the stage control of culture temperature, pH, ventilating ratio, dissolved oxygen (DO) at different fermentation stages, effectively to improve fermentation production rate.Preferred fermentating controling condition is:
0 ~ 24h: temperature 30 ~ 34 DEG C, pH 4.0 ~ 5.0, ventilation 0.1 ~ 0.4v.v.m., pressure are 0.04 ~ 0.05MPa, dissolved oxygen 30% ~ 50%.
24h ~ put tank (60 ~ 72h): temperature 26 ~ 28 DEG C, pH 2.5, ventilation 0.5 ~ 2.0v.v.m., pressure are 0.04 ~ 0.05MPa, dissolved oxygen 5% ~ 15%.
Sclerotium gum performance index detected result prepared by application aforesaid method is as follows:
(1) productive rate measures
As follows according to the method adopted general in document:
A. instrument: thermostat container, analytical balance (0.001g).
B. measuring method: the fermented liquid accurately measuring certain volume, regulates pH to 7.0, adopts the alcohol settling polysaccharide of 90% ~ 95%, uses the washing with alcohol one time of 90% ~ 95% after filtration again, is filtered dry to be placed in 105 DEG C of thermostat containers to be dried to constant weight, weighs after cooling.
Productive rate=sample dry weight/fermentating liquid volume × 100%
C. detected result: the productive rate adopting Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 and above fermentation process gained Sclerotium gum is 32 ~ 35g/L, higher than now studies have reported that.
D. document: Liu Rulin, weighs refined, Zhao great Jian, Zhou Yuliang. by research [J] Nankai University journal (natural science) of Starch Production scleroglycan, and 1990,4:15-22.
Y.Wang，B.McNeil.Effect of temperature on scleroglucan synthesis and organic acidproduction by Sclerotium glucanicum[J].Enzyme and Microbial Technology，1995，17:893-899.
Shrikant A.Survase，Parag S.Saudagar，Rekha S.Singhal.Enhanced production ofscleroglucan by Sclerotium rolfsii MTCC2156by use of metabolic precursors[J].BioresourceTechnology，2007，98:410-415.
(2) product viscosity measures
(1) instrument: NDJ-1 type rotational viscosimeter, 3# rotor, 60r/min, 20 DEG C of mensuration.
(2) measuring method: accurately take the Sclerotium gum product 2.0g adopting the inventive method to prepare and be dissolved in 200.0mL distilled water, after abundant swelling 60min, adopt high-shear homogenizing machine homogeneous 3 ~ 5min, standing 30min carries out viscosimetric analysis.
(3) measurement result: adopt the viscosity of Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 and above fermentation process gained Sclerotium gum product 1% (w/v) at more than 800mPa.s.
Substantive distinguishing features acquired by the present invention and significant technique effect are:
1, the invention discloses a strain can the Sclerotium rolfsii of fermentative production Sclerotium gum, i.e. Sclerotium rolfsii (Sclerotiumrolfsii) SCL2010CCTCC No.M2014325.
2, adopt Sclerotium rolfsii of the present invention and application method fermentative production Sclerotium gum thereof, yield is significantly higher than existing technique, and prepared product viscosity is high, has good stability, and viscosity B coefficent is little, has good application performance.
3, fermentation culture fluid component of the present invention adopts carbon source based on starch, sucrose and glucose, low price; Nitrogenous source then adopts the common organic nitrogen source such as bean cake powder, corn steep liquor, and simple to operate, safety is cheap.
4, the present invention adopts grading-controlling fermentation condition (especially the control of temperature, pH and dissolved oxygen) technique according to the functional characteristics of different steps in fermenting process, achieve thalline to grow fast and the perfect adaptation of efficiently producing sugar, the steady running of the fermenting process of guarantee.
Accompanying drawing explanation
Fig. 1: Sclerotium rolfsii of the present invention (Sclerotium rolfsii) SCL2010 plated growth figure.
Embodiment
Carry out complete describing in detail to the technical scheme in the embodiment of the present invention below, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
Embodiment 1: the seed selection of Sclerotium rolfsii (Sclerotium rolfsii) SCL2010
From Shouguang City of Weifang City of Shandong Province, Gaomi City, Linqu County infects the potato plant of southern blight and stem tuber collects southern blight sclerotium, take sclerotium as parting material, cultivate on sclerotium inoculation and PDA solid medium under aseptic condition, after sclerotial germination mycelia grows, carry out isolation and purification culture, transfer and to preserve on PDA slant medium.
The mycelia of separation and purification being transferred on PDA substratum is transferred respectively on new PDA flat board, cultivate Continuous Observation for 28 DEG C, treat that mycelial growth is paved with flat board and bears sclerotium, till sclerotium variable color maturation, record mycelial growth situation, tuberculosis and sclerotium changing conditions, determine Sclerotium rolfsii bacterial strain wherein in conjunction with microscopic examination and carry out strain number; To be defined as the sclerotium switching of Sclerotium rolfsii with on PDA flat board, observed and recorded sclerotial germination situation, germination rate, mycelial growth situation, knot sclerotium time and sclerotium amount.Choose that sclerotial germination is fast, germination rate >=75%, knot sclerotium time fast (about 72h), bacterial strain that sclerotium amount is many carry out the experiment of Sclerotium gum shake flask fermentation, pass through great many of experiments, through further separation screening, seed selection obtains a strain stable hereditary property, sclerotial germination rate up to 80%, Sclerotium gum productive rate is up to the Sclerotium rolfsii of 37g/L, and this Strain Designation is Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 bacterial strain.
Above-mentioned bacterial strains Sclerotium rolfsii (Sclerotium rolfsii) SCL2010, be deposited in China typical culture collection center (China Center for Type Culture Collection on July 6th, 2014, be called for short CCTCC, address: China. Wuhan. Wuhan University), its deposit number is CCTCC NO.M2014325.
Above-mentioned slant medium consists of (unit: g/L): potato 200 (filtering after poach is rotten), glucose 20, agar 20.
Above-mentioned seed culture medium composition (unit: g/L): glucose 10, yeast extract paste 3, dipotassium hydrogen phosphate 2.5, magnesium sulfate 1.5, polyether antifoam agent 0.2, other are softening water, pH 4 ~ 5.
The composition (unit: g/L) of above-mentioned fermention medium: glucose 55, bean cake powder 1, corn steep liquor 2, dipotassium hydrogen phosphate 2, magnesium sulfate 1, polyether antifoam agent 0.8, other are softening water, pH 4 ~ 5.
Embodiment 2: the gene order order-checking of Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 bacterial strain 18S rDNA
Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 bacterial strain embodiment 1 seed selection obtained and CCTCC NO.M2014325 bacterial strain entrust Hua Da gene sequencing.
Experimental technique:
(1) a certain amount of new knot sclerotium physiological saline is washed in seed culture medium, cultivate 36h for 28 DEG C, after sclerotial germination produces a large amount of mycelia, the centrifugal acquisition mycelium of 8000r/min;
(2) adopting Takara9765 fungal DNA extraction kits from the mycelium obtained, extract postgenome send order-checking company to check order.
Sequencing result: the gene order length of Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 bacterial strain 18S rDNA is 1694bp, and concrete nucleotide sequence is as shown in SEQ ID NO.1.
By using U.S.'s Biotechnology Information center (National Center for Biotechnology Information, NCBI) BLASTN program comparison, find that many strains Sclerotium rolfsii (Sclerotium rolfsii) the 18S rDNA sequence that CCTCC NO.M2014325 bacterial strain 18S rDNA sequence and NCBI register has high homology, illustrate that CCTCCNO.M2014325 bacterial strain is a strain Sclerotium rolfsii (Sclerotium rolfsii).Further, the sequence in its 18S rDNA sequence between base is not again identical, and product application is also different, illustrates that bacterial strain of the present invention is not same bacterial strain with disclosing bacterial strain.
Above-mentioned seed culture medium composition (unit: g/L): glucose 10, yeast extract paste 3, dipotassium hydrogen phosphate 2.5, magnesium sulfate 1.5, polyether antifoam agent 0.5, other are softening water, pH 4 ~ 5.
Embodiment 3: Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 strain fermentation produces the method for Sclerotium gum
(1) be inoculated on eggplant-shape bottle inclined-plane solid medium by the ripe sclerotium of Sclerotium rolfsii CCTCC No.M2014325,30 ~ 34 DEG C of quiescent culture 5 ~ 7 days, to obtain a large amount of ripe sclerotium for seed culture.
(2) washing down cultivating ripe Sclerotium rolfsii CCTCC No.M2014325 sclerotium stroke-physiological saline solution, being inoculated in seed culture medium in right amount, 30h is cultivated in 34 DEG C of aeration-agitations.
(3) Sclerotium gum fermentation manufacturing technique:
Be inoculated in right amount in fermention medium as fermented bacterium by Sclerotium rolfsii CCTCC No.M2014325 by seed culture maturation, aeration-agitation controls fermentation to produce Sclerotium gum.
Preferred fermentating controling condition:
0 ~ 24h: temperature 32 ~ 34 DEG C, pH 4.0 ~ 5.0, ventilation 0.1v.v.m., pressure are 0.04 ~ 0.05MPa, DO 40%.
24 ~ put tank (60 ~ 72h): temperature 26 ~ 28 DEG C, pH 2.5, ventilation 1.5v.v.m., pressure are 0.04 ~ 0.05MPa, DO 10%.
(4) the technique Sclerotium gum yield of the present embodiment is 35g/L, and ferment strength is 0.49g/L/h, and 1% (w/v) viscosity is 1100mPa.s.
Above-mentioned eggplant-shape bottle slant medium consists of (unit: g/L): potato 200 (filtering after poach is rotten), glucose 20, agar 20.
Above-mentioned seed culture medium composition (unit: g/L): glucose 20, yeast extract paste 3, SODIUMNITRATE 3, dipotassium hydrogen phosphate 2; Magnesium sulfate 3, polyether antifoam agent 0.5; Other are softening water, pH 4 ~ 5.
Above-mentioned fermention medium consists of (unit: g/L): sucrose 35, glucose 25, bean cake powder 3, corn steep liquor 0.5, dipotassium hydrogen phosphate 1, magnesium sulfate 0.5, polyether antifoam agent 0.5, and other are softening water, pH 4 ~ 5.
Embodiment 4: Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 strain fermentation produces the method for Sclerotium gum
(1) be inoculated on eggplant-shape bottle inclined-plane solid medium by the ripe sclerotium of Sclerotium rolfsii CCTCC No.M2014325,30 ~ 34 DEG C of quiescent culture 5 ~ 7 days, to obtain a large amount of ripe sclerotium for seed culture.
(2) washing down cultivating ripe Sclerotium rolfsii CCTCC No.M2014325 sclerotium stroke-physiological saline solution, being inoculated in seed culture medium in right amount, 30h is cultivated in 34 DEG C of aeration-agitations.
(3) Sclerotium gum fermentation manufacturing technique:
Be inoculated in right amount in fermention medium as fermented bacterium by Sclerotium rolfsii CCTCC No.M2014325 by seed culture maturation, aeration-agitation controls fermentation to produce Sclerotium gum.
Preferred fermentating controling condition:
0 ~ 24h: temperature 32 ~ 34 DEG C, pH 4.0 ~ 5.0, ventilation 0.2v.v.m., pressure are 0.04 ~ 0.05MPa, DO 50%.
24 ~ put tank (60 ~ 72h): temperature 26 ~ 28 DEG C, pH 2.5, ventilation 1.5v.v.m., pressure are 0.04 ~ 0.05MPa, DO 12%.
(4) the technique Sclerotium gum yield of the present embodiment is 32g/L, and ferment strength is 0.44g/L/h, and 1% (w/v) viscosity is 1220mPa.s.
Above-mentioned eggplant-shape bottle slant medium consists of (unit: g/L): potato 200 (filtering after poach is rotten), glucose 20, agar 20.
Above-mentioned seed culture medium composition (unit: g/L): glucose 15, yeast extract paste 4, SODIUMNITRATE 2.5, dipotassium hydrogen phosphate 2; Magnesium sulfate 2, polyether antifoam agent 0.5; Other are softening water, pH 4 ~ 5.
Above-mentioned fermention medium consists of (unit: g/L): starch 25, glucose 25, bean cake powder 1, corn steep liquor 2, SODIUMNITRATE 1.5, dipotassium hydrogen phosphate 2, magnesium sulfate 1, polyether antifoam agent 0.8, and other are softening water, pH 4 ~ 5.
Embodiment 5: Sclerotium rolfsii (Sclerotium rolfsii) SCL2010 strain fermentation produces the method for Sclerotium gum
(1) be inoculated on eggplant-shape bottle inclined-plane solid medium by the ripe sclerotium of Sclerotium rolfsii CCTCC No.M2014325,30 ~ 34 DEG C of quiescent culture 5 ~ 7 days, to obtain a large amount of ripe sclerotium for seed culture.
(2) washing down cultivating ripe Sclerotium rolfsii CCTCC No.M2014325 sclerotium stroke-physiological saline solution, being inoculated in seed culture medium in right amount, 33h is cultivated in 32 ~ 34 DEG C of aeration-agitations.
(3) Sclerotium gum fermentation manufacturing technique:
Be inoculated in right amount in fermention medium as fermented bacterium by Sclerotium rolfsii CCTCC No.M2014325 by seed culture maturation, aeration-agitation controls fermentation to produce Sclerotium gum.
Preferred fermentating controling condition:
0 ~ 24h: temperature 32 ~ 34 DEG C, pH 4.0 ~ 5.0, ventilation 0.3v.v.m., pressure are 0.04 ~ 0.05MPa, DO 40%.
24 ~ put tank (60 ~ 72h): temperature 26 ~ 28 DEG C, pH 2.5, ventilation 1.2v.v.m., pressure are 0.04 ~ 0.05MPa, DO 7%.
(4) the technique Sclerotium gum yield of the present embodiment is 34g/L, and ferment strength is 0.47g/L/h, and 1% (w/v) viscosity is 1016mPa.s.
Above-mentioned eggplant-shape bottle slant medium consists of (g/L): potato 200 (filtering after poach is rotten), glucose 20, agar 20.
Above-mentioned seed culture medium composition (g/L): glucose 10, yeast extract paste 2, SODIUMNITRATE 2, dipotassium hydrogen phosphate 2; Magnesium sulfate 3, polyether antifoam agent 1; Other are softening water, pH 4 ~ 5.
Above-mentioned fermention medium consists of (g/L): starch 10, sucrose 45, bean cake powder 15, corn steep liquor 15, SODIUMNITRATE 1, dipotassium hydrogen phosphate 1.5, magnesium sulfate 1, polyether antifoam agent 1, and other are softening water, pH 4 ~ 5.
Claims (3)
1. a strain is applicable to the Sclerotium rolfsii of industrial production Sclerotium gum, it is characterized in that: this bacterial strain is called Sclerotium rolfsii (Sclerotium rolfsii) SCL2010, bacterial strain is deposited in China typical culture collection center on July 6th, 2014, and its deposit number is CCTCC NO.M2014325.
2. the application of Sclerotium rolfsii described in claim 1 in fermentative production Sclerotium gum.
3. apply as claimed in claim 2, it is characterized in that: in fermentative production Sclerotium gum process, adopt grading-controlling fermentation condition technique, that is:
During fermentation 0 to 24h, fermentation condition is: temperature 30 ~ 34 DEG C, pH 4.0 ~ 5.0, ventilation 0.1 ~ 0.4v.v.m., pressure are 0.04 ~ 0.05MPa, dissolved oxygen 30% ~ 50%;
Fermentation 24h to put tank fermentation 60 ~ 72h time fermentation condition be: temperature 26 ~ 28 DEG C, pH 2.5, ventilation 0.5 ~ 2.0v.v.m., pressure are 0.04 ~ 0.05MPa, dissolved oxygen 5% ~ 15%;
In above-mentioned fermentative production Sclerotium gum process, fermention medium used consists of:
Carbon source: 45 ~ 60g/L; Bean cake powder 1 ~ 3g/L; Corn steep liquor 0.5 ~ 2g/L; SODIUMNITRATE 1 ~ 3g/L; Dipotassium hydrogen phosphate 1 ~ 2g/L; Magnesium sulfate 0.5 ~ 1g/L; Polyether antifoam agent 0.2 ~ 1g/L; Surplus is softening water; PH 4 ~ 5; Carbon source in nutrient solution be selected from starch, sucrose, glucose, in a kind of mixture of or its any part by weight.
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| CN201410572695.0A Active CN104371933B (en) | 2014-10-23 | 2014-10-23 | Strain of sclerotium rolfsii and application thereof in fermentation production of scleroglucan |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087402A (en) * | 2015-10-09 | 2015-11-25 | 上海市农业科学院 | Liquid submerged fermentation production method of ganoderma lucidum mycelia |
| CN105754872A (en) * | 2016-03-30 | 2016-07-13 | 广东省农业科学院作物研究所 | Method for fast separating, identifying and storing sweet potato sclerotium rolfsii |
| CN106337071A (en) * | 2016-01-28 | 2017-01-18 | 通辽市黄河龙生物工程有限公司 | Sclerotium rolfssii scleroglucan industrial batch production fermentation and extraction technology |
| CN108441429A (en) * | 2018-03-22 | 2018-08-24 | 江南大学 | A kind of method of pyrenomycetes and its fermenting and producing scleroglucan |
| CN112266907A (en) * | 2020-10-28 | 2021-01-26 | 江南大学 | Sclerotium rolfsii endogenous sclerotium rolfsii hydrolase and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103361278B (en) * | 2013-07-06 | 2015-08-05 | 河北恒标生物科技有限公司 | A kind of Roche Ah too bacterium and adopt this bacterial classification to produce the method for Sclerotium gum fermented liquid |
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- 2014-10-23 CN CN201410572695.0A patent/CN104371933B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| SHRIKANT A. SURVASE ET AL: "Use of complex media for the production of scleroglucan by Sclerotium rolfsii MTCC 2156", 《BIORESOURCE TECHNOLOGY》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087402A (en) * | 2015-10-09 | 2015-11-25 | 上海市农业科学院 | Liquid submerged fermentation production method of ganoderma lucidum mycelia |
| CN105087402B (en) * | 2015-10-09 | 2018-05-08 | 上海市农业科学院 | A kind of method of liquid submerged fermentation production ganoderma lucidum mycelium |
| CN106337071A (en) * | 2016-01-28 | 2017-01-18 | 通辽市黄河龙生物工程有限公司 | Sclerotium rolfssii scleroglucan industrial batch production fermentation and extraction technology |
| CN105754872A (en) * | 2016-03-30 | 2016-07-13 | 广东省农业科学院作物研究所 | Method for fast separating, identifying and storing sweet potato sclerotium rolfsii |
| CN108441429A (en) * | 2018-03-22 | 2018-08-24 | 江南大学 | A kind of method of pyrenomycetes and its fermenting and producing scleroglucan |
| CN112266907A (en) * | 2020-10-28 | 2021-01-26 | 江南大学 | Sclerotium rolfsii endogenous sclerotium rolfsii hydrolase and application thereof |
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| CN104371933B (en) | 2017-05-03 |
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