CN105838697A - Fermentation method for producing salt-tolerant cellulase from marine aspergillus niger - Google Patents
Fermentation method for producing salt-tolerant cellulase from marine aspergillus niger Download PDFInfo
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- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
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- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
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- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
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- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01021—Beta-glucosidase (3.2.1.21)
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- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01091—Cellulose 1,4-beta-cellobiosidase (3.2.1.91)
Abstract
The invention provides a fermentation method for producing salt-tolerant cellulase from marine aspergillus niger .The salt-tolerant cellulase is obtained by means of preparation and preservation of slant pores, two-level expanded culture in a shake flask, scale-up culture in a seed tank and fermentation in a fermentation tank .The preservation number of marine aspergillus niger in the preservation center is CCTCC NO: M2010132, and the strain name is Aspergillus sp .ZJUBE 2010 .The salt-tolerant cellulase obtained through the method can be widely used in a high-salt industrial environment, and provides a new utilization way for salt-rich cellulose resources .According to the method, the adopted culture medium is cheap and low in cost, the culture process is simple, control and improvement are easy, and the activity of the cellulase is higher by regulating different temperatures at the growing stage and the cellulase production stage .The method provides a basis and foundation for factory production and application of the salt-tolerant cellulase.
Description
Technical field
The invention belongs to marine organisms field, relate to a kind of fermentation process utilizing ocean aspergillus niger to produce salt tolerant cellulase, completed by temperature section formula pilot scale liquid fermentation.
Background technology
The compound enzyme that cellulase is made up of three kinds of enzymes such as excision enzyme, restriction endonuclease and cellobiases, they make cellulose be degraded into cellobiose and glucose successively by a kind of synergism.Therefore cellulase purposes is very big, can remove textile surfaces fine hair in textile industry；The aspects such as alcoholic, soy sauce brewing and food processing in food service industry have important effect.And in terms of new forms of energy and environmental protection, also have the bright prospect of comparison.
But being as industrial expansion, general fibre element enzyme increasingly cannot meet some the resistance demands in commercial production, and such as, black liquor waste not only content of cellulose in paper mill is high but also salinity is high, under the conditions of general fibre element enzyme cannot be applicable to high salinity.And due to the environment of ocean uniqueness, there is multiple tolerance extreme condition high salt the most high temperature resistant, resistance to, the strain of resistance to extreme pH, this just for solve industrialized production also exists high salinity, high temperature, extreme pH environments and make cellulase activity reduce or even the problem of inactivation provides a new thinking and approach.Produced by the marine bacteria that Wang Fen etc. filter out from Huanghai Sea mud sample, the optimal reactive temperature of enzyme is 35 DEG C, hence it is evident that less than the optimal reactive temperature of general fibre element enzyme, and it also contains higher activity at about 10 DEG C.Trivedi etc. are cellulase produced by isolated bacillus from Sargassum, can alkaline environment also can at high salinity environment under keep high activity.In sum, the cellulase producing bacteria studying resistance to extreme condition has research and using value greatly.
Summary of the invention
It is an object of the invention to provide a kind of ocean aspergillus niger and produce the fermentation process of salt tolerant cellulase, realized by following steps:
(1) preparation of slant pore and preservation: ocean aspergillus niger is inoculated into PDA slant medium, cultivates 96 hours at 28 DEG C, obtains slant pore, preserves at 4 DEG C；Strain used: by screening the ocean aspergillus niger obtained in East China Sea Coastal Waters earth, be preserved in China typical culture collection center, address is China, and Wuhan, Wuhan University, deposit number is CCTCC
NO:M2010132, preservation date is on June 2nd, 2010, and strain name is Aspergillus sp.ZJUBE 2010；
(2) two grades of amplification culture of shaking flask: preparation 300 ~ 400ml shake-flask seed culture medium is dispensed in 1 L conical flask, 121 DEG C of sterilizing 20min, the distilled water eluting inclined-plane utilizing sterilizing obtains Aspergillus niger spores liquid, it is inoculated in shake-flask seed culture medium, inoculum concentration is 5 ~ 7%, and in 35 ~ 38 DEG C, cultivate under the conditions of 180 ~ 200r/min 24 ~ 36 hours and obtain two grades of expansion seed liquor；
(3) seed tank amplification culture: prepare 35 L seed tank culture bases, 121 DEG C of sterilizing 20min in 50 L seed fermentation tanks, the seed liquor taking shaking flask expansion is inoculated in seed tank, inoculum concentration is 3 ~ 5%, fermentation jar temperature is 35 ~ 38 DEG C, and ventilation ratio is 1 V/V min, and tank pressure is 0.02 ~ 0.1
Mpa, rotating speed 180r/min, obtain seed tank and amplify seed liquor after fermenting 48 hours；
(4) 500 L ferment tanks: prepare 350 L fermentation medium, 121 DEG C of sterilizing 20min in 500 L fermentation tanks, the seed liquor of seed tank is inoculated in seed tank, inoculum concentration is 10%, defoamer 0.03%, ventilation ratio is 0.5 ~ 2.5 V/V min, and tank pressure is 0.02 ~ 0.1
Mpa, rotating speed 180r/min, after 36 ~ 40 DEG C of conditions of temperature are fermented 36 ~ 60 hours, regulation temperature is 26 ~ 30 DEG C, obtains cellulase crude enzyme liquid, be salt tolerant cellulase after continuing fermentation 36 ~ 60 hours.
Described PDA culture medium consists of: 200 Rhizoma Solani tuber osis 1 L filtrate after 6 layers of filtered through gauze after 1 L water boil 20min, 20g glucose, and pH is natural.
Described shake-flask seed culture medium consists of: glucose 1%, Semen Maydis pulp 0.25%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80
0.2%。
Described seed tank culture is basis set to be become: wheat bran 5%, Semen Maydis pulp 1%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80
0.2%。
Described fermentation medium consists of: wheat bran 5%, Semen Maydis pulp 1%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80
0.2%。
In above culture medium, % is mass volume ratio.
The present invention used be bacterial strain be a kind of ocean aspergillus niger, it can produce the cellulase of salt tolerant, be widely used in the industrial production environment of high salinity.But, to the industrialized production of the cellulase of resistance to extreme condition and application correlational study and data few, therefore, how large-scale production and application salt tolerant cellulase are a urgent and valuable job.Ocean aspergillus niger used by the present invention can produce salt tolerant cellulase, can be widely used in industry hypersaline environment, also provide a kind of new utilization ways for the cellulose resource rich in salt；Method used medium is inexpensive, and cost is the highest；The inventive method incubation is simple, it is easy to controls and improves；Regulation and control growth stage and the different temperatures producing the enzyme stage so that cellulase activity is higher.Factorial praluction and application that the inventive method is salt tolerant cellulase provide foundation and basis.
Accompanying drawing explanation
Fig. 1 is embodiment 1 aspergillus niger biomass dry weight curve.
Fig. 2 is embodiment 2 aspergillus niger biomass dry weight curve.
Fig. 3 is embodiment 3 aspergillus niger biomass dry weight curve.
The embodiment 4 aspergillus niger biomass dry weight curve of Fig. 4.
Fig. 5 is embodiment 5 aspergillus niger biomass dry weight curve.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described further.
The culture medium that following example are used is as follows with method, and in all culture medium, % is mass volume ratio.
Described PDA culture medium consists of: 200 Rhizoma Solani tuber osis 1 L filtrate after 6 layers of filtered through gauze after 1 L water boil 20min, 20g glucose, and pH is natural.
Described shake-flask seed culture medium consists of: glucose 1%, Semen Maydis pulp 0.25%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80
0.2%。
Described seed tank culture is basis set to be become: wheat bran 5%, Semen Maydis pulp 1%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80
0.2%。
Described fermentation medium consists of: wheat bran 5%, Semen Maydis pulp 1%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80
0.2%。
Fermentation tank parametric measurement: 500 L full-automatic airlift fermentor liquid amount is 350 L, the optimal liquid culture medium prescription drawn using shake flask test is as fermentation medium, after a series of sterilizings, seed liquor in seed tank is linked in fermentation tank, is passed through filtrated air after regulating the parameters such as fermentation jar temperature, ventilation ratio, speed of agitator, pH and starts to control fermentation culture.
The mensuration of filter paper enzyme activity: take the fermentation liquid (control sample first to boil inactivation) that 0.5 mL suitably dilutes, join in 2.0 mL pH 4.8 sodium citrate-citric acid buffer, adds about 50 mg
Whatman filter paper (60 mm × 10 mm), 50 DEG C of insulation 30 min, add 2.5 mLDNS reagent termination reactions afterwards and boiling water bath processes 5 min, in 540 nm wavelength colorimetrics after cooling.Finally according to standard curve (y= 0.5839x+ 0.1687,R=0.9999, in formulaxFor optical density (A540nm),yFor concentration of glucose) calculate the glucose generation amount in sample, and utilize formula (U=(0.5839x+ 0.1687) * 1000/180/30) cellulase activity is obtained.Cellulase activity unit definition be per minute hydrolysis generate 1 μm ol glucose enzyme amount be 1 enzyme activity unit.
Embodiment
1
The preparation of slant pore and preservation: ocean aspergillus niger is inoculated into PDA slant medium, cultivate 96 hours at 28 DEG C, obtain slant pore, preserves at 4 DEG C.
Two grades of amplification culture of shaking flask: preparation 300ml shake-flask seed culture medium is dispensed in 1 L conical flask, 121 DEG C of sterilizing 20min, the distilled water eluting inclined-plane utilizing sterilizing obtains Aspergillus niger spores liquid, it is inoculated in shake-flask seed culture medium, inoculum concentration is 5%, and in 35 DEG C, cultivate under the conditions of 180r/min 24 hours and obtain two grades of expansion seed liquor.
Seed tank amplification culture: prepare 35 L seed tank culture bases in 50 L seed fermentation tanks, 121 DEG C of sterilizing 20min, the seed liquor taking shaking flask expansion is inoculated in seed tank, inoculum concentration is 3%, fermentation jar temperature is 35 DEG C, and ventilation ratio is 1 V/V min, and tank pressure is 0.02Mpa, rotating speed 180r/min, obtains seed tank and amplifies seed liquor after fermenting 48 hours.
500 L ferment tanks: prepare 350 L fermentation medium in 500 L fermentation tanks, 121 DEG C of sterilizing 20min, are inoculated into the seed liquor of seed tank in seed tank, and inoculum concentration is 10%, defoamer 0.03%, ventilation ratio is 1.5 V/V min, and tank pressure is 0.02Mpa, rotating speed 180r/min, after the conditions such as temperature 36 DEG C are fermented 96 hours, regulation temperature is 28 DEG C, obtains cellulase crude enzyme liquid after continuing fermentation 48 hours, and measuring its filter paper enzyme activity is 0.76
U/ml, dry weight Dependence Results is shown in Fig. 1, reached peak at 96 hours.
Embodiment
2
The preparation of slant pore and preservation: ocean aspergillus niger is inoculated into PDA slant medium, cultivate 96 hours at 28 DEG C, obtain slant pore, preserves at 4 DEG C.
Two grades of amplification culture of shaking flask: preparation 300ml shake-flask seed culture medium is dispensed in 1 L conical flask, 121 DEG C of sterilizing 20min, the distilled water eluting inclined-plane utilizing sterilizing obtains Aspergillus niger spores liquid, it is inoculated in shake-flask seed culture medium, inoculum concentration is 7%, and in 38 DEG C, cultivate under the conditions of 200r/min 36 hours and obtain two grades of expansion seed liquor.
Seed tank amplification culture: prepare 35 L seed tank culture bases in 50 L seed fermentation tanks, 121 DEG C of sterilizing 20min, the seed liquor taking shaking flask expansion is inoculated in seed tank, inoculum concentration is 5%, fermentation jar temperature is 38 DEG C, and ventilation ratio is 1 V/V min, and tank pressure is 0.1 Mpa, rotating speed 180r/min, obtains seed tank and amplifies seed liquor after fermenting 48 hours.
500 L ferment tanks: prepare 350 L fermentation medium in 500 L fermentation tanks, 121 DEG C of sterilizing 20min, are inoculated into the seed liquor of seed tank in seed tank, and inoculum concentration is 10%, defoamer 0.03%, ventilation ratio is 1.5 V/V min, and tank pressure is 0.1 Mpa, rotating speed 180r/min, after 38 DEG C of conditions of temperature are fermented 84 hours, regulation temperature is 30 DEG C, obtains cellulase crude enzyme liquid after continuing fermentation 60 hours, and measuring its filter paper enzyme activity is 0.85
U/ml, dry weight Dependence Results is shown in Fig. 2, reached peak at 96 hours.
Embodiment
3
The preparation of slant pore and preservation: ocean aspergillus niger is inoculated into PDA slant medium, cultivate 96 hours at 28 DEG C, obtain slant pore, preserves at 4 DEG C.
Two grades of amplification culture of shaking flask: preparation 300ml shake-flask seed culture medium is dispensed in 1 L conical flask, 121 DEG C of sterilizing 20min, the distilled water eluting inclined-plane utilizing sterilizing obtains Aspergillus niger spores liquid, it is inoculated in shake-flask seed culture medium, inoculum concentration is 6%, and in 35 DEG C, cultivate under the conditions of 180r/min 36 hours and obtain two grades of expansion seed liquor.
Seed tank amplification culture: prepare 35 L seed tank culture bases in 50 L seed fermentation tanks, 121 DEG C of sterilizing 20min, the seed liquor taking shaking flask expansion is inoculated in seed tank, inoculum concentration is 5%, fermentation jar temperature is 36 DEG C, and ventilation ratio is 1 V/V min, and tank pressure is 0.06 Mpa, rotating speed 180r/min, obtains seed tank and amplifies seed liquor after fermenting 48 hours.
500 L ferment tanks: prepare 350 L fermentation medium in 500 L fermentation tanks, 121 DEG C of sterilizing 20min, are inoculated into the seed liquor of seed tank in seed tank, and inoculum concentration is 10%, defoamer 0.03%, ventilation ratio is 1.5 V/V min, and tank pressure is 0.07Mpa, rotating speed 180r/min, after 38 DEG C of conditions of temperature are fermented 72 hours, regulation temperature is 28 DEG C, obtains cellulase crude enzyme liquid after continuing fermentation 72 hours, and measuring its filter paper enzyme activity is 0.89
U/ml, dry weight Dependence Results is shown in Fig. 3, reached peak at 84 hours.
Embodiment
4
The preparation of slant pore and preservation: ocean aspergillus niger is inoculated into PDA slant medium, cultivate 96 hours at 28 DEG C, obtain slant pore, preserves at 4 DEG C.
Two grades of amplification culture of shaking flask: preparation 400ml shake-flask seed culture medium is dispensed in 1 L conical flask, 121 DEG C of sterilizing 20min, the distilled water eluting inclined-plane utilizing sterilizing obtains Aspergillus niger spores liquid, it is inoculated in shake-flask seed culture medium, inoculum concentration is 5%, and in 35 ~ 38 DEG C, cultivate under the conditions of 180 ~ 200r/min 24 ~ 36 hours and obtain two grades of expansion seed liquor.
Seed tank amplification culture: prepare 35 L seed tank culture bases in 50 L seed fermentation tanks, 121 DEG C of sterilizing 20min, the seed liquor taking shaking flask expansion is inoculated in seed tank, inoculum concentration is 5%, fermentation jar temperature is 38 DEG C, and ventilation ratio is 1 V/V min, and tank pressure is 0.07 Mpa, rotating speed 180r/min, obtains seed tank and amplifies seed liquor after fermenting 48 hours.
500 L ferment tanks: prepare 350 L fermentation medium in 500 L fermentation tanks, 121 DEG C of sterilizing 20min, are inoculated into the seed liquor of seed tank in seed tank, and inoculum concentration is 10%, defoamer 0.03%, ventilation ratio is 1.5 V/V min, and tank pressure is 0.07Mpa, rotating speed 180r/min, after 38 DEG C of conditions of temperature are fermented 60 hours, regulation temperature is 28 DEG C, obtains cellulase crude enzyme liquid after continuing fermentation 84 hours, and measuring its filter paper enzyme activity is 0.88
U/ml, dry weight Dependence Results is shown in Fig. 4, reached peak at 96 hours.
Embodiment
5
The preparation of slant pore and preservation: ocean aspergillus niger is inoculated into PDA slant medium, cultivate 96 hours at 28 DEG C, obtain slant pore, preserves at 4 DEG C.
Two grades of amplification culture of shaking flask: preparation 400ml shake-flask seed culture medium is dispensed in 1 L conical flask, 121 DEG C of sterilizing 20min, the distilled water eluting inclined-plane utilizing sterilizing obtains Aspergillus niger spores liquid, it is inoculated in shake-flask seed culture medium, inoculum concentration is 5%, and in 35 DEG C, cultivate under the conditions of 180 ~ 200r/min 24 ~ 36 hours and obtain two grades of expansion seed liquor.
Seed tank amplification culture: prepare 35 L seed tank culture bases, 121 DEG C of sterilizing 20min in 50 L seed fermentation tanks, the seed liquor taking shaking flask expansion is inoculated in seed tank, inoculum concentration is 3%, fermentation jar temperature is 37 DEG C, and ventilation ratio is 1 V/V min, and tank pressure is 0.02 ~ 0.1
Mpa, rotating speed 180r/min, obtain seed tank and amplify seed liquor after fermenting 48 hours.
500 L ferment tanks: prepare 350 L fermentation medium in 500 L fermentation tanks, 121 DEG C of sterilizing 20min, the seed liquor of seed tank is inoculated in seed tank, inoculum concentration is 10%, defoamer 0.03%, ventilation ratio is 2.5 V/V min, tank pressure is 0.09Mpa, rotating speed 180r/min, after 38 DEG C of conditions of temperature are fermented 48 hours respectively, regulation temperature is 28 DEG C, cellulase crude enzyme liquid is obtained after continuing fermentation 96 hours, measuring its filter paper enzyme activity is 0.90 U/ml, and dry weight Dependence Results is shown in Fig. 5, reaches peak at 108 hours.
Claims (6)
1. the fermentation process of an ocean aspergillus niger production salt tolerant cellulase, it is characterised in that realized by following steps:
(1) preparation of slant pore and preservation: the ocean aspergillus niger of outsourcing is inoculated into PDA slant medium, cultivates 96 hours at 28 DEG C, obtains slant pore, preserves at 4 DEG C；
(2) two grades of amplification culture of shaking flask: preparation 300 ~ 400ml shake-flask seed culture medium is dispensed in conical flask, 121 DEG C of sterilizings, the distilled water eluting inclined-plane utilizing sterilizing obtains Aspergillus niger spores liquid, it is inoculated in shake-flask seed culture medium, and in 35 ~ 38 DEG C, cultivate under the conditions of 180 ~ 200r/min 24 ~ 36 hours and obtain two grades of expansion seed liquor；
(3) seed tank amplification culture: prepare 35 L seed tank culture bases in seed fermentation tank, 121 DEG C of sterilizings, the seed liquor taking shaking flask expansion is inoculated in seed tank, fermentation jar temperature is 35 ~ 38 DEG C, ventilation ratio is 1 V/V min, tank pressure is 0.02 ~ 0.1 Mpa, rotating speed 180r/min, obtains seed tank and amplify seed liquor after fermenting 48 hours；
(4) 500 L ferment tanks: prepare 350 L fermentation medium in fermentation tank, 121 DEG C of sterilizings, the seed liquor of seed tank being inoculated in seed tank, defoamer 0.03%, ventilation ratio is 0.5 ~ 2.5 V/V min, tank pressure is 0.02 ~ 0.1 Mpa, rotating speed 180r/min, after 36 ~ 40 DEG C of conditions of temperature are fermented 36 ~ 60 hours, regulation temperature is 26 ~ 30 DEG C, obtain cellulase crude enzyme liquid after continuing fermentation 36 ~ 60 hours, be described salt tolerant cellulase.
A kind of ocean the most according to claim 1 aspergillus niger produces the fermentation process of salt tolerant cellulase, it is characterized in that, PDA culture medium described in step (1) consists of: 200 Rhizoma Solani tuber osis 1 L filtrate after 6 layers of filtered through gauze after 1 L water boil 20min, 20g glucose, and pH is natural.
A kind of ocean the most according to claim 1 aspergillus niger produces the fermentation process of salt tolerant cellulase, it is characterised in that the shake-flask seed culture medium described in step (2) consists of: glucose 1%, Semen Maydis pulp 0.25%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80 0.2%.
A kind of ocean the most according to claim 1 aspergillus niger produces the fermentation process of salt tolerant cellulase, it is characterised in that seed tank culture described in step (3) is basis set to be become: wheat bran 5%, Semen Maydis pulp 1%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80 0.2%.
A kind of ocean the most according to claim 1 aspergillus niger produces the fermentation process of salt tolerant cellulase, it is characterised in that the fermentation medium described in step (4) consists of: wheat bran 5%, Semen Maydis pulp 1%, sodium chloride 2%, potassium dihydrogen phosphate 0.3%, calcium chloride 0.03%, magnesium sulfate 0.08%, ferrous sulfate 0.005%, manganese sulfate 0.0016%, zinc sulfate 0.0014%, cobaltous chloride 0.0037%, Tween 80 0.2%.
A kind of ocean the most according to claim 1 aspergillus niger produces the fermentation process of salt tolerant cellulase, it is characterized in that, ocean aspergillus niger used by step (1), being preserved in China typical culture collection center, address is China, Wuhan, Wuhan University, deposit number is CCTCC NO:M2010132, and preservation date is on June 2nd, 2010, and strain name is Aspergillus sp.ZJUBE 2010.
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| CN106801045A (en) * | 2017-02-15 | 2017-06-06 | 浙江大学舟山海洋研究中心 | A kind of method of recycled fiber element enzyme in fermentation of Aspergillus niger liquid from ocean |
| CN107488595A (en) * | 2017-08-14 | 2017-12-19 | 浙江大学舟山海洋研究中心 | A kind of ocean aspergillus niger ZJUBE2010 and its application |
| CN110564625A (en) * | 2019-08-13 | 2019-12-13 | 内蒙古世洪农业科技有限公司 | Saline-alkali resistant aspergillus flavus and separation method and application thereof |
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| CN110564625A (en) * | 2019-08-13 | 2019-12-13 | 内蒙古世洪农业科技有限公司 | Saline-alkali resistant aspergillus flavus and separation method and application thereof |
| CN110564625B (en) * | 2019-08-13 | 2023-01-24 | 内蒙古世洪农业科技有限公司 | Saline-alkali resistant aspergillus flavus and separation method and application thereof |
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