CN104611312A - Culture medium for producing agarase by virtue of fermentation of vibrio natriegen and fermentation method for producing agarase by virtue of fermentation of vibrio natriegen - Google Patents
Culture medium for producing agarase by virtue of fermentation of vibrio natriegen and fermentation method for producing agarase by virtue of fermentation of vibrio natriegen Download PDFInfo
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- C12N9/2468—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
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- C12Y302/01081—Beta-agarase (3.2.1.81)
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Abstract
The invention discloses a culture medium for producing agarase by virtue of fermentation of vibrio natriegen. The culture medium comprises a strain activation culture medium, a seed culture medium and a fermentation culture medium. Meanwhile, the invention also discloses a fermentation method for producing the agarase by virtue of fermentation of the vibrio natriegen. According to the fermentation method disclosed by the invention, on the basis of shaking flask fermentation, fermentation culture is carried out on the screened vibrio natriegen by virtue of a fermentor, the fermentation conditions are optimized, a material feeding process is explored, and the enzyme producing regulation of the vibrio natriegen in the fermentor is studied, so that the yield of the agarase is effectively increased, pilot test amplification fermentation is finally carried out on the agarase produced by virtue of fermentation of the vibrio natriegen, and important technical parameter guidance is provided for mass production in future.
Description
Technical field
The present invention relates to the technical field of fermentation, particularly relate to a kind of substratum and fermentation process thereof of Vibrio natriegen fermentative production agarase.
Background technology
The Sargassum polysaccharides that agar-agar is made up of agarose and agaropectin, is widely used in fields such as biochemical, clinical, medical, food.Agaropectin oligose is degraded by agar-agar and obtains, have anti-oxidant, slow down Starch Hydrolysis, be easy to the pharmacological actions such as absorption of human body and biological activity, compare agar-agar and there is using value widely.
At present, agaropectin oligose is generally obtained by the method for traditional chemistry and mechanical degradation, but traditional method is prepared agaropectin oligose and be there is the problems such as difficult control of reaction conditions, degraded product heterogeneity, loss are many, contaminate environment, and enzyme process have efficient, special, reaction conditions is gentle, degradation process is easy to control, and the advantage such as environmental pollution is few.Therefore, degradation method such as replacing traditional chemistry is gradually become the best approach preparing agaropectin oligose by agarase enzymolysis agar-agar.
China has vast marine site, and marine algae resource abundant in ocean is that PRODUCTION TRAITS provides sufficient Sargassum polysaccharides.Main source due to agarase is marine microorganism, and the singularity of ocean environment result in that the stability of agarase is bad or vigor is not high, limits its application aborning.In view of this, the present inventor studies and devises a kind of substratum and fermentation process thereof of Vibrio natriegen fermentative production agarase, and this case produces thus.
Summary of the invention
The object of the present invention is to provide a kind of substratum of Vibrio natriegen fermentative production agarase, can the degrade Vibrio natriegen (Vibrio sp. natriegens) of agar-agar of a strain is utilized (to derive from Chinese industrial Culture Collection, deposit number: CICC 23820), after employing substratum of the present invention, effectively can improve the unit output of Vibrio natriegen fermentative production agarase.
Another object of the present invention is to provide the method utilizing above-mentioned substratum to carry out Vibrio natriegen fermentation agarase, by the step of amplification culture step by step that seed activation, shake flask fermentation, tank top fermentation and enlarged experiment ferment, determine best fermentation condition, and then improve the unit output of agarase in final fermented liquid.
For achieving the above object, the present invention solves the technical scheme of its technical problem and is:
A substratum for Vibrio natriegen fermentative production agarase, comprises strain activation and culture base, seed culture medium and fermention medium;
Described strain activation and culture base is: agar 20g/L, peptone 5g/L, yeast extract paste 5g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
Described seed culture medium is: peptone 5 g/L, yeast extract paste 1 g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
Described fermention medium is: agar 3.4 g/L, yeast extract paste 6.5 g/L, NaCl 20 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min.
The invention still further relates to described Vibrio natriegen utilizes substratum described above to produce the fermentation process of agarase, comprises the following steps:
The activation of seed and preparation: above-mentioned Vibrio natriegen bacterial classification is connected in strain activation and culture base and activates, the strain inoculation of activation is cultivated to the shaking flask that seed culture medium is housed, and obtains shake-flask seed liquid;
Shake flask fermentation: shake-flask seed liquid is seeded to be equipped with and is applicable to Vibrio natriegen and produces among the shaking flask of agarase fermention medium, cultivates 24 h for 25 DEG C, obtains the fermented liquid containing agarase;
Tank top fermentation: shake-flask seed liquid is seeded to and is equipped with in 7 L fermentor tanks of fermention medium, cultivates 36h for 27 DEG C, obtains the fermented liquid containing agarase;
Enlarged experiment ferments: shake-flask seed liquid is seeded in 20 L and 200 L fermentor tanks that fermention medium is housed, and cultivates 36h for 27 DEG C, obtains the fermented liquid containing agarase.
As the optimal way of embodiment, the activation of described seed and preparation process, the test tube slant that bacterium culture medium is housed is connected to after being thawed by the glycerine pipe bacterial classification of-20 DEG C of preservations, cultivate 24 h for 25 DEG C, obtain the Vibrio natriegen bacterial classification of activation, then from the cultured bacterial classification of picking test tube slant in the 250 mL shaking flasks that 30 mL seed culture mediums are housed, 25 DEG C, 180 r/min ferment 24 h, access is equipped with in 250 mL shaking flasks of 50 mL seed culture mediums again, 25 DEG C, 180 r/min ferment 12 h, obtain shake-flask seed liquid.
As the optimal way of embodiment, described shake flask fermentation, by cultured shake-flask seed liquid with 2% inoculum size access be equipped with in 250 mL shaking flasks of 50 mL fermention mediums, 25 DEG C, 180 r/min ferment 24 h.
As the optimal way of embodiment, described tank top fermentation, by cultured shake-flask seed liquid according to 2% inoculum size access be equipped with in 7 L fermentor tanks of 5 L fermention mediums, 27 DEG C, 500 r/min ferment 36 h.
As the optimal way of embodiment, the fermentation of described enlarged experiment, by cultured seed according to 2% inoculum size access be equipped with in 20 L tanks of 10 L fermention mediums, 27 DEG C, 150 r/min ferment 36 h.
As the optimal way of embodiment, described enlarged experiment fermentation, by cultured seed according to 2% inoculum size access be equipped with in 7 L tanks of 5 L seed culture mediums, 25 DEG C of fermentation 12 h, cultured seed liquor is equipped with in 200 L tanks of 100 L fermention mediums according to the inoculum size access of 2%, 27 DEG C, 100 r/min ferment 36 h.
After the present invention adopts technique scheme, on the basis of shake flask fermentation, the fermentation culture of fermentor tank has been carried out to the Vibrio natriegen that the present inventor screens, carry out probing into of fermentation condition optimization and supplying technics respectively, study its enzyme production law in fermentor tank, effectively improve agarase output.Finally agarase is produced to Vibrio natriegen fermentation and carried out enlarged experiment test, instruct for scale operation from now on provides important technical parameter.
Accompanying drawing explanation
Fig. 1 Vibrio natriegen culture plate;
Fig. 2 Vibrio natriegen produces the tank top fermentation curve of agarase;
Fig. 3 Vibrio natriegen produces the 20L tank top fermentation curve of agarase;
Fig. 4 Vibrio natriegen produces the 200L tank top fermentation curve of agarase.
Embodiment
the detection method adopted in the following example:
The mensuration of biomass: evenly draw 1.0 ml fermented liquids, centrifugal 10 min of 12000 r/min, uses distilled water gradient dilution to 6 times after removing supernatant liquor, and do blank with distilled water, 600 nm wavelength survey light absorption value.
The mensuration of agarase vigor: evenly get 1.0mL fermented liquid, the centrifugal 10min of 12000 r/min, accurately draws 0.3mL supernatant liquor, with 50 mmol/L NaH
2pO
4-Na
2hPO
4damping fluid (pH 7.0) dilution 4 times, gets 50 μ L diluents, adds 350 μ L and be dissolved in 50 mmol/L NaH
2pO
4-Na
2hPO
4damping fluid (pH 7.0) 0.5% agar (being incubated after high-temperature digestion in 40 DEG C of water-baths), 40 DEG C of temperature bath 20 min, add 0.6 mL DNS termination reaction, cool after boiling water bath 5 min, be diluted to 5 mL, survey the light absorption value at 540 nm places, then establishing criteria curve calculates the growing amount of reducing sugar in reaction solution, to detect the vigor of agarase in fermented liquid.With the enzyme liquid of boiling water bath deactivation 5 min in contrast.Be an enzyme activity unit (U) with the 1 min catalysis enzyme amount produced needed for 1 μm of ol reducing sugar under above-mentioned condition.
PH value measures: utilize pH meter to detect the pH value of fermenting process.
embodiment 1: the screening of Vibrio natriegen
The present invention first in mangrove forest earth sample be separated obtain a strain can degrade agar-agar Vibrio natriegen (Vibrio sp.) (Chinese industrial Microbiological Culture Collection number: CICC 23820), after adopting culture medium culturing of the present invention again, effectively can improve the unit output of Vibrio natriegen fermentative production agarase.
(1) sample thief mangrove forest earth 25g carries out gradient dilution in 225mL sterile artificial seawater, gets diluent and is coated with in the plate culture medium taking agar as single carbon source, and flat board cultivates 24 h ~ 36h in 25 DEG C of incubators.
(2) select on plate culture medium and form the darker bacterium colony of depression, as shown in Figure 1, transfer in slant medium and cultivate 24 ~ 36h, after being inoculated in artificial seawater culture medium culturing 24 ~ 36h, measure the agarase vigor of fermented liquid, final screening obtains enzyme the highest bacterial strain Vibrio natriegen alive.
embodiment 2: the shake flask fermentation of Vibrio natriegen produces agarase
(1) be connected to the test tube slant that strain activation and culture base is housed after being thawed by the glycerine pipe bacterial classification of-20 DEG C of preservations, 24 h are cultivated in inclined-plane in 25 DEG C of incubators, obtain the Vibrio natriegen bacterial classification of activation.Described strain activation and culture base is: agar 20g/L, peptone 5g/L, yeast extract paste 5g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(2) from the cultured bacterial classification of picking test tube slant in the 250 mL shaking flasks that 30 mL seed culture mediums are housed, 25 DEG C, 180 r/min ferment 24 h, access is equipped with in 250 mL shaking flasks of 50 mL seed culture mediums again, 25 DEG C, 180 r/min ferment 12 h, obtain shake-flask seed liquid.Described seed culture medium is: peptone 5 g/L, yeast extract paste 1 g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(3) by cultured shake-flask seed liquid with 2% inoculum size access be equipped with in 250 mL shaking flasks of 50 mL fermention mediums, 25 DEG C, 180 r/min ferment 24 h, obtain the fermented liquid containing agarase.Described fermention medium is: agar 3.4 g/L, yeast extract paste 6.5 g/L, NaCl 20 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(4) evenly get 1.0 mL containing agarase fermented liquid, the centrifugal 10min of 12000 r/min, the agarase vigor measuring gained clear liquid is 2.51 U/mL.
embodiment 3: agarase is produced in the tank top fermentation of Vibrio natriegen
(1) be connected to the test tube slant that strain activation and culture base is housed after being thawed by the glycerine pipe bacterial classification of-20 DEG C of preservations, 24 h are cultivated in inclined-plane in 25 DEG C of incubators, obtain the Vibrio natriegen bacterial classification of activation.Described strain activation and culture base is: agar 20g/L, peptone 5g/L, yeast extract paste 5g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(2) from the cultured bacterial classification of picking test tube slant in the 250 mL shaking flasks that 30 mL seed culture mediums are housed, 25 DEG C, 180 r/min ferment 24 h, access is equipped with in 250 mL shaking flasks of 50 mL seed culture mediums again, 25 DEG C, 180 r/min ferment 12 h, obtain shake-flask seed liquid.Described seed culture medium is: peptone 5 g/L, yeast extract paste 1 g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(3) by cultured shake-flask seed liquid according to 2% inoculum size access be equipped with in 7 L fermentor tanks of 5 L fermention mediums, 27 DEG C, 500 r/min ferment 36 h, obtain the fermented liquid containing agarase.Described fermention medium is: agar 3.4 g/L, yeast extract paste 6.5 g/L, NaCl 20 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(4) sample biomass, agarase vigor and the pH value change detecting fermenting process, result as shown in Figure 2.Fermentation to enzyme activity during 26 h reaches maximum value 2.69 U/mL.
embodiment 4: the enlarged experiment fermentation of Vibrio natriegen fermentative production agarase
(1) be connected to the test tube slant that strain activation and culture base is housed after being thawed by the glycerine pipe bacterial classification of-20 DEG C of preservations, 24 h are cultivated in inclined-plane in 25 DEG C of incubators, obtain the Vibrio natriegen bacterial classification of activation.Described strain activation and culture base is: agar 20g/L, peptone 5g/L, yeast extract paste 5g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(2) from the cultured bacterial classification of picking test tube slant in the 250 mL shaking flasks that 30 mL seed culture mediums are housed, 25 DEG C, 180 r/min ferment 24 h, access is equipped with in 250 mL shaking flasks of 50 mL seed culture mediums again, 25 DEG C, 180 r/min ferment 12 h, obtain shake-flask seed liquid.Described seed culture medium is: peptone 5 g/L, yeast extract paste 1 g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(3) by cultured shake-flask seed liquid according to 2% inoculum size access be equipped with in 20 L tanks of 10 L fermention mediums, 27 DEG C, 150 r/min ferment 36 h, obtain the fermented liquid containing agarase.Described fermention medium is: agar 3.4 g/L, yeast extract paste 6.5 g/L, NaCl 20 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(4) sampling detects the agarase vigour changes of fermenting process, and result as shown in Figure 3.Carry out the fermentation of three batches altogether, 20 L tank top fermentation enzyme production law and 7 L tanks basically identical, and its agarase output is further enhanced, and 20 L tank agarase vigor are up to 3.66 U/mL, improve 8.5% than 7 L tanks.
embodiment 5: the enlarged experiment fermentation of Vibrio natriegen fermentative production agarase
(1) be connected to the test tube slant that bacterium culture medium is housed after being thawed by the glycerine pipe bacterial classification of-20 DEG C of preservations, 24 h are cultivated in inclined-plane in 25 DEG C of incubators, obtain the Vibrio natriegen bacterial classification of activation.Described bacterium culture medium is: agar 20g/L, peptone 5g/L, yeast extract paste 5g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(2) from the cultured bacterial classification of picking test tube slant in the 250 mL shaking flasks that 30 mL seed culture mediums are housed, 25 DEG C, 180 r/min ferment 24 h, access is equipped with in 250 mL shaking flasks of 50 mL seed culture mediums again, 25 DEG C, 180 r/min ferment 12 h, obtain shake-flask seed liquid.Described seed culture medium is: peptone 5 g/L, yeast extract paste 1 g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(3) by cultured shake-flask seed liquid according to 2% inoculum size access be equipped with in 7 L tanks of 5 L seed culture mediums, 25 DEG C of fermentation 12 h, cultured seed liquor is equipped with in 200 L tanks of 100 L fermention mediums according to the inoculum size access of 2%, and 27 DEG C, 100 r/min ferment 36 h.Described fermention medium is: agar 3.4 g/L, yeast extract paste 6.5 g/L, NaCl 20 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
(4) sampling detects the agarase vigour changes of fermenting process, and result as shown in Figure 4.Carry out the fermentation of three batches altogether, 200 L tank top fermentation enzyme production law and 7 L tanks and 20 L tanks basically identical, 200 L tank agarase vigor are up to 3.73 U/mL, improve 10.5 % than 7 L tanks.
embodiment 6: utilize agarase to prepare fine jade oligosaccharides
Take agar powder and be dissolved in the NaH prepared
2pO
4-Na
2hPO
4damping fluid (pH 7.0,50 mmol/L) in, be mixed with the agar-agar solution of 5 g/L, therefrom take out 19 mL agar-agar solution in 50 mL tool plug triangular flasks, add agarase crude enzyme liquid in 1 mL embodiment 2 again, be hydrolyzed reaction under 40 DEG C of water bath with thermostatic control conditions, react 40 min after products and be mainly new fine jade disaccharides and Xin Qiong tetrose, prolongation in time afterwards, new fine jade tetrose in product is degraded into new fine jade disaccharides gradually, reacts product when proceeding to 24 h and substantially only has new fine jade disaccharides.
The present invention is on the basis of shake flask fermentation, the fermentation culture of fermentor tank has been carried out to the Vibrio natriegen that the present inventor screens, carry out probing into of fermentation condition optimization and supplying technics respectively, studied its enzyme production law in fermentor tank, effectively improve agarase output.Finally agarase is produced to Vibrio natriegen fermentation and carried out enlarged experiment test, instruct for scale operation from now on provides important technical parameter.
All distortion that those of ordinary skill in the art can directly derive from the disclosure of invention or associate, all should think protection scope of the present invention.
Claims (7)
1. a substratum for Vibrio natriegen fermentative production agarase, is characterized in that: comprise strain activation and culture base, seed culture medium and fermention medium;
Described strain activation and culture base is: agar 20g/L, peptone 5g/L, yeast extract paste 5g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
Described seed culture medium is: peptone 5 g/L, yeast extract paste 1 g/L, NaCl 30 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min;
Described fermention medium is: agar 3.4 g/L, yeast extract paste 6.5 g/L, NaCl 20 g/L, MgSO
47H
2o 5 g/L, KCl 1 g/L, CaCl
20.2 g/L, K
2hPO
40.1 g/L, FeSO
47H
2o 0.02 g/L, pH 7.5,121 DEG C of sterilizing 20 min.
2. Vibrio natriegen as claimed in claim 1 produces a fermentation process for agarase, it is characterized in that: comprise the steps:
The activation of seed and preparation: above-mentioned Vibrio natriegen bacterial classification is connected in strain activation and culture base and activates, the strain inoculation of activation is cultivated to the shaking flask that seed culture medium is housed, and obtains shake-flask seed liquid;
Shake flask fermentation: shake-flask seed liquid is seeded to be equipped with and is applicable to Vibrio natriegen and produces among the shaking flask of agarase fermention medium, cultivates 24 h for 25 DEG C, obtains the fermented liquid containing agarase;
Tank top fermentation: shake-flask seed liquid is seeded to and is equipped with in 7 L fermentor tanks of fermention medium, cultivates 36h for 27 DEG C, obtains the fermented liquid containing agarase;
Enlarged experiment ferments: shake-flask seed liquid is seeded in 20 L and 200 L fermentor tanks that fermention medium is housed, and cultivates 36h for 27 DEG C, obtains the fermented liquid containing agarase.
3. Vibrio natriegen as claimed in claim 2 produces the fermentation process of agarase, it is characterized in that: activation and the preparation process of described seed are as follows:
Be connected to the test tube slant that strain activation and culture base is housed after being thawed by the glycerine pipe bacterial classification of-20 DEG C of preservations, cultivate 24 h for 25 DEG C, obtain the Vibrio natriegen bacterial classification of activation; Then from the cultured bacterial classification of picking test tube slant in the 250 mL shaking flasks that 30 mL seed culture mediums are housed, 25 DEG C, 180 r/min ferment 24 h, access is equipped with in 250 mL shaking flasks of 50 mL seed culture mediums again, 25 DEG C, 180 r/min ferment 12 h, obtain shake-flask seed liquid.
4. Vibrio natriegen as claimed in claim 2 produces the fermentation process of agarase, it is characterized in that: described shake flask fermentation, by cultured shake-flask seed liquid with 2% inoculum size access be equipped with in 250 mL shaking flasks of 50 mL fermention mediums, 25 DEG C, 180 r/min ferment 24 h.
5. fermentation process as claimed in claim 2, is characterized in that: described tank top fermentation, by cultured shake-flask seed liquid according to 2% inoculum size access be equipped with in 7 L fermentor tanks of 5 L fermention mediums, 27 DEG C, 500 r/min ferment 36 h.
6. Vibrio natriegen as claimed in claim 2 produces the fermentation process of agarase, it is characterized in that: described enlarged experiment fermentation, by cultured seed according to 2% inoculum size access be equipped with in 20 L tanks of 10 L fermention mediums, 27 DEG C, 150 r/min ferment 36 h.
7. the fermentation process of a kind of Vibrio natriegen fermentative production agarase as claimed in claim 2, it is characterized in that: described enlarged experiment fermentation, by cultured seed according to 2% inoculum size access be equipped with in 7 L tanks of 5 L seed culture mediums, 25 DEG C of fermentation 12 h, cultured seed liquor is equipped with in 200 L tanks of 100 L fermention mediums according to the inoculum size access of 2%, 27 DEG C, 100 r/min ferment 36 h.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087427A (en) * | 2015-07-06 | 2015-11-25 | 华侨大学 | Vibrio natriegens for producing agarase and application of vibrio natriegens |
| CN110373442A (en) * | 2019-08-10 | 2019-10-25 | 青岛博智汇力生物科技有限公司 | A kind of preparation method and application with high anti-oxidation activity fine jade disaccharides |
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| CN101451113B (en) * | 2008-11-06 | 2010-10-06 | 国家***第二海洋研究所 | Vibrio natriegens and method for producing agarase by using the same |
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| CN101451113B (en) * | 2008-11-06 | 2010-10-06 | 国家***第二海洋研究所 | Vibrio natriegens and method for producing agarase by using the same |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087427A (en) * | 2015-07-06 | 2015-11-25 | 华侨大学 | Vibrio natriegens for producing agarase and application of vibrio natriegens |
| CN105087427B (en) * | 2015-07-06 | 2018-08-24 | 华侨大学 | Produce Vibrio natriegen and its application of agarase |
| CN110373442A (en) * | 2019-08-10 | 2019-10-25 | 青岛博智汇力生物科技有限公司 | A kind of preparation method and application with high anti-oxidation activity fine jade disaccharides |
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