CN1900277A - Alpha-galactosidase solid fermenting producing method - Google Patents
Alpha-galactosidase solid fermenting producing method Download PDFInfo
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- CN1900277A CN1900277A CN 200610052576 CN200610052576A CN1900277A CN 1900277 A CN1900277 A CN 1900277A CN 200610052576 CN200610052576 CN 200610052576 CN 200610052576 A CN200610052576 A CN 200610052576A CN 1900277 A CN1900277 A CN 1900277A
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- galactosidase
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- 102000005840 alpha-Galactosidase Human genes 0.000 title claims abstract description 40
- 108010030291 alpha-Galactosidase Proteins 0.000 title claims abstract description 40
- 239000007787 solid Substances 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 27
- 238000000855 fermentation Methods 0.000 claims abstract description 33
- 230000004151 fermentation Effects 0.000 claims abstract description 33
- 239000000725 suspension Substances 0.000 claims abstract description 21
- 238000002360 preparation method Methods 0.000 claims abstract description 19
- 230000008569 process Effects 0.000 claims abstract description 8
- 239000002609 medium Substances 0.000 claims description 45
- 239000007788 liquid Substances 0.000 claims description 41
- 238000011218 seed culture Methods 0.000 claims description 31
- 239000001963 growth medium Substances 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000002054 inoculum Substances 0.000 claims description 15
- 230000001954 sterilising effect Effects 0.000 claims description 15
- 238000004659 sterilization and disinfection Methods 0.000 claims description 15
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- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 12
- 238000001816 cooling Methods 0.000 claims description 10
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 claims description 8
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 8
- 235000015099 wheat brans Nutrition 0.000 claims description 8
- 241000228212 Aspergillus Species 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 7
- 150000008195 galaktosides Chemical class 0.000 claims description 7
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- 235000015097 nutrients Nutrition 0.000 claims description 6
- 238000010298 pulverizing process Methods 0.000 claims description 6
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- 102000004190 Enzymes Human genes 0.000 abstract description 17
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- 230000000694 effects Effects 0.000 abstract description 7
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
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- DLRVVLDZNNYCBX-ZZFZYMBESA-N beta-melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 DLRVVLDZNNYCBX-ZZFZYMBESA-N 0.000 description 2
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- RYVMUASDIZQXAA-UHFFFAOYSA-N pyranoside Natural products O1C2(OCC(C)C(OC3C(C(O)C(O)C(CO)O3)O)C2)C(C)C(C2(CCC3C4(C)CC5O)C)C1CC2C3CC=C4CC5OC(C(C1O)O)OC(CO)C1OC(C1OC2C(C(OC3C(C(O)C(O)C(CO)O3)O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OCC(O)C(O)C1O RYVMUASDIZQXAA-UHFFFAOYSA-N 0.000 description 1
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- AURFVNDXGLQSNN-UHFFFAOYSA-K trisodium 2-hydroxypropane-1,2,3-tricarboxylic acid phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O.OC(=O)CC(O)(C(O)=O)CC(O)=O AURFVNDXGLQSNN-UHFFFAOYSA-K 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The solid fermentation process of producing alpha-galactosidase includes the following steps: 1. preparing spore suspension; 2. preparing the first order seed; and 3. fermentation of alpha-galactosidase. Or, the solid fermentation process of producing alpha-galactosidase includes the following steps: 1. preparing spore suspension; 2. preparing the first order seed; 3. preparing the second order seed; and 4. fermentation of alpha-galactosidase. The alpha-galactosidase produced through the said process has high enzyme generating activity, simple preparation process and easy production application.
Description
Technical field
The present invention relates to biological technical field, particularly a kind of solid state fermentation is produced the method for feeding alpha-galactosidase.
Background technology
Alpha-galactosidase is a kind of non reducing end α-1 with specificity hydrolysis gala carbohydrate oligosaccharides and poly gala-(Portugal) mannosans, and 6-semi-lactosi glycosidic bond is the oligosaccharides lytic enzyme of feature, has great economic worth and using value.Since alpha-galactosidase can hydrolysis the polysaccharide of oligose such as melibiose, raffinose, stachyose and verbascose and polygalactomannan in the beans, can play effects such as improving carbohydrate metabolism, therefore be mainly used among the industry such as feed and food.
At present, fodder industry develops on a large scale along with the raising of livestock industry localization of production, mass-producing, feed proportion, and the contradiction that China's feed resource is in short supply becomes increasingly conspicuous; According to scholarly forecast, 2010-the year two thousand twenty, the difference of China's protein feed was 2,400 ten thousand ton-4,800 ten thousand tons.Dregs of beans is because of containing 43%～48% crude protein and by forming than equilibrated amino acid, and becomes the most important vegetable protein of livestock and poultry source; Yet blemish in an otherwise perfect thing is that the contained oligose of beans such as melibiose, raffinose, stachyose and a spot of verbascose can not be absorbed by the simple stomach animal digestion, but fermented by enteric microorganism, produce gases such as carbonic acid gas, methane and hydrogen, cause flatulence, cause power loss, increase the passage rate of chyme, reduce nutrition absorption, greatly influence the feeding value of dregs of beans, and caused the increase and the environmental pollution of aquaculture cost.
Data shows: raffinose and stachyose content are about 1.4% and 5.2% in the soybean meal, in order to eliminate the anti-oxidant action of these insoluble oligose, adopt method squeezing soyabeans such as high temperature, high pressure, high humidity and time expand for a long time always, perhaps adopt organic solvent extractionprocess to eliminate these antinutritional factor, handle soybean even take optimum complete processing, still there is a certain amount of alpha galactosides in the dregs of beans, and increased the production cost of dregs of beans.
After deliberation with facts have proved that adopting the achievement-zymin of modern biotechnology to improve usage quantity and the utilization ratio of the assorted dregs of rice in feed is to reduce feed cost at present and alleviate one of effective ways of environmental pollution, the requirement of also complying with the ecological agriculture.The application of China's alpha-galactosidase in animal-feed also is in the preliminary study stage; especially the compatibility research with other zymins does not appear in the newspapers; this shows, utilize microbe fermentation method industrialization, mass-producing ground production alpha-galactosidase that wide application development prospect is arranged.
Also utilize at present the report of microorganisms producing alpha-galactosidase, to produce enzyme level on the low side but existing alpha-galactosidase produces bacterium, and fermentation character, nutritive property and the fermentation technique of zymogenic bacteria do not carried out systematic research; Therefore it is alive high to explore enzyme, and the alpha-galactosidase production method of the also available solid seed inoculation of both available liquid seeds seems necessary.
Summary of the invention
At the deficiencies in the prior art part, the invention provides a kind of alpha-galactosidase solid fermenting producing method that enzyme activity height, technology are simple, be easy to production application that produces.
The present invention is to realize by such technical scheme: a kind of alpha-galactosidase solid fermenting producing method is provided, may further comprise the steps for reaching above purpose:
1), the preparation of spore suspension: with preserving number was that the smelly aspergillus of CGMCC No.1628 is inoculated on the slant medium, in 28 ℃ of constant temperature culture 7 days; Making concentration with sterilized water again is 10
6～10
7The spore suspension of individual/ml, stand-by; Described slant medium is the PDA slant medium;
2), first order seed preparation: at first with liquid seed culture medium through 121 ℃, 20min sterilization, cooling again; Being that 5% inoculum size inserts spore suspension in the above-mentioned cooled liquid seed culture medium according to volume ratio then, is to cultivate 18h on the shaking table of 200rpm in 28 ℃, rotating speed, liquid seeds; Described liquid seed culture medium is the PDA liquid nutrient medium;
3), the fermentation of alpha-galactosidase: at first with fermention medium through 121 ℃, 20min sterilization, cooling again; Inoculum size according to 1 milliliter/10 grams inserts the aforesaid liquid seed in the cooled fermention medium then, fully behind the stirring and evenly mixing, in 28 ℃ of cultivations 4～7 days, in culturing process, carries out the step of mixing oscillation and fermentation cultivation base when 20h; Described fermention medium is 2: 3 butt and the mixed back of distilled water, is transferred pH to 5.5～6.0 to make by weight ratio, and described every 100g butt is made up of 82.137g wheat bran, 17.843g dregs of beans, 0.01g manganous sulfate and 0.01g cupric sulfate pentahydrate.
Improvement as above-mentioned alpha-galactosidase solid fermenting producing method, carrying out post-processing step behind the described fermentation step: the product of fermentation step gained is dried to the moisture weight content under 40 ℃～45 ℃ low-temperature airflow be 10%, promptly gets the alpha galactosides zymin after the pulverizing.
The present invention also provides the another kind of alpha-galactosidase solid fermenting producing method that belongs to same inventive concept, may further comprise the steps:
1), the preparation of spore suspension: with preserving number was that the smelly aspergillus of CGMCC No.1628 is inoculated on the slant medium, in 28 ℃ of constant temperature culture 7 days; Making concentration with sterilized water again is 10
6～10
7The spore suspension of individual/ml, stand-by; Described slant medium is the PDA slant medium;
2), first order seed preparation: at first with liquid seed culture medium through 121 ℃, 20min sterilization, cooling again; Being that 5% inoculum size inserts spore suspension in the above-mentioned cooled liquid seed culture medium according to volume ratio then, is to cultivate 18h on the shaking table of 200rpm in 28 ℃, rotating speed, liquid seeds; Described liquid seed culture medium is the PDA liquid nutrient medium;
3) secondary seed preparation: the solid seed culture medium through 121 ℃, 20min sterilization, is cooled off again; Inoculum size according to 1 milliliter/10 grams inserts liquid seeds in the above-mentioned cooled solid seed culture medium then, cultivates 4～6 days in 28 ℃ of incubators, gets the solid seed; Described solid seed culture medium is 2: 3 butt and the mixed back of distilled water, is transferred pH to 5.5～6.0 to make by weight ratio, and described every 100g butt is made up of 82.137g wheat bran, 17.843g dregs of beans, 0.01g manganous sulfate and 0.01g cupric sulfate pentahydrate;
4), the fermentation of alpha-galactosidase: at first with fermention medium through 121 ℃, 20min sterilization, cooling again; Be that 1% inoculum size inserts above-mentioned solid seed in the cooled fermention medium according to weight ratio then, fully behind the stirring and evenly mixing, cultivated 4～7 days in 28 ℃; In culturing process, when 20h, carry out the mixing vibration; Described fermention medium is with the solid seed culture medium.
Improvement as above-mentioned alpha-galactosidase solid fermenting producing method, carrying out post-processing step behind the described fermentation step: the product of fermentation step gained is dried to the moisture weight content under 40 ℃～45 ℃ low-temperature airflow be 10%, promptly gets the alpha galactosides zymin after the pulverizing.
The selected preserving number of the present invention is the smelly aspergillus of CGMCC No.1628, and the applicant has applied for patent of invention " fetid aspergillic strain and uses thereof ", application number 200610050135.4 in 2006-3-31; This summary of the invention is to adopt the statistics optimization method that above-mentioned smelly aspergillar solid fermentation is produced the enzyme culture condition to have carried out system optimization, thereby obtains to be fit to the optimal culture condition and the technical parameter of suitability for industrialized production.
Adopt production method of the present invention, the level that bacterial strain produces alpha-galactosidase is 8～15 times that report under the existing solid fermentation condition.The optimum temperature of this enzyme is 60 ℃, and the same fermentoid of more domestic report is heat-resisting; Optimal pH is 5.0, and stability is very high under sour environment, is easy to use.The present invention adopts the coarse fodder substratum as fermentation raw material, has that technology is simple, enzymic activity is high, the unit enzyme is lived advantages such as low production cost, for the cost degradation scale operation of alpha-galactosidase provides foundation.
Embodiment
Embodiment 1, a kind of alpha-galactosidase production method, carry out following steps successively:
1), the preparation of spore suspension:
With preserving number be CGMCC No.1628 smelly aspergillus to the layout line method be inoculated on the slant medium, in 28 ℃ of constant temperature culture 7 days, this moment, bacterial strain was ripe; Wash the spore vibration with sterilized water and break up spore, making concentration is 10
6～10
7The spore suspension of individual/ml, stand-by; Described slant medium is the PDA slant medium, that is: fresh potato 200 restrains, glucose 20 grams, agar 20 grams, 1000 milliliters of distilled water, pH nature.
2), first order seed preparation:
At first with liquid seed culture medium through 121 ℃, 20min sterilization, cooling again; Being that 5% inoculum size inserts spore suspension in the above-mentioned cooled liquid seed culture medium according to volume ratio (V/V) then, is to cultivate 18h on the shaking table of 200rpm in 28 ℃, rotating speed, liquid seeds; Described liquid seed culture medium is the PDA liquid nutrient medium, that is: fresh potato 200 restrains, glucose 20 grams, 1000 milliliters of distilled water, pH nature.
3), the fermentation of alpha-galactosidase:
Packing in the triangular flask of 250ml is the butt of 10g, and this 10g butt is made up of 8.2137g wheat bran, 1.7843g dregs of beans, 0.001g manganous sulfate and 0.001g cupric sulfate pentahydrate.In bottle, add 15g water then, transfer pH to 5.5～6.0 with the Sodium phosphate dibasic of 0.5M again, make fermention medium.
The bottleneck of triangular flask covers with 6 layers of gauze, and fermention medium behind 121 ℃ of sterilization 20min, is cooled off again; Inoculum size according to 1 milliliter/10 grams inserts the aforesaid liquid seed in the cooled fermention medium then, fully behind the stirring and evenly mixing, cultivates 4 days in 28 ℃.In culturing process, when being cultured to 20h, the vibration triangular flask makes fermention medium and liquid seeds evenly mixed, leaves standstill until fermentation ends then.
4) aftertreatment:
The product of gained after the fermentation ends is dried to the moisture weight content under 40 ℃～45 ℃ low-temperature airflow be 10%, after the pulverizing, is packaged into the alpha galactosides zymin.
After measured, alpha-galactosidase is that enzyme work is the 2207.19U/g dry medium.
Embodiment 2: a kind of alpha-galactosidase production method, carry out following steps successively:
1), the preparation of spore suspension:
With preserving number be CGMCC No.1628 smelly aspergillus to the layout line method be inoculated on the slant medium, in 28 ℃ of constant temperature culture 7 days, this moment, bacterial strain was ripe; Wash the spore vibration with sterilized water and break up spore, making concentration is 10
6～10
7The spore suspension of individual/ml, stand-by.Described slant medium is the PDA slant medium that fresh potato-glucose-agar is made, with embodiment 1.
2), first order seed preparation:
At first with liquid seed culture medium through 121 ℃, 20min sterilization, cooling again; Being that 5% inoculum size inserts spore suspension in the above-mentioned cooled liquid seed culture medium according to volume ratio then, is to cultivate 18h on the shaking table of 200rpm in 28 ℃, rotating speed, liquid seeds.Described liquid seed culture medium is the PDA liquid nutrient medium that fresh potato-glucose is made; With embodiment 1.
3) secondary seed preparation:
The 10g butt of packing in the triangular flask of 250ml, this butt is made up of 8.2137g wheat bran, 1.7843g dregs of beans, 0.001g manganous sulfate and 0.001g copper sulfate.In bottle, add 15g water, transfer pH to 5.5～6.0 with the Sodium phosphate dibasic of 0.5M again, get the solid seed culture medium.
The bottleneck of triangular flask covers with 6 layers of gauze, and above-mentioned solid seed culture based on 121 ℃ of sterilization 20min, is cooled off again; Inoculum size according to 1 milliliter/10 grams inserts liquid seeds in the above-mentioned cooled solid seed culture medium then, cultivates 4 days in 28 ℃ of incubators behind vibration, the mixing, gets the solid seed.In culturing process, when being cultured to 20h, the vibration triangular flask makes liquid seeds and solid seed culture medium evenly mixed, leaves standstill then until cultivating and finishes.
4), the fermentation of alpha-galactosidase:
Packing in the triangular flask of 500ml is the butt of 40g, and this butt is made up of 32.8548g wheat bran, 7.1372g dregs of beans, 0.004g manganous sulfate and 0.004g cupric sulfate pentahydrate.In bottle, add 60g water again, transfer pH to 5.5～6.0 with the Sodium phosphate dibasic of 0.5M again, make fermention medium.
Bottleneck covers with 6 layers of gauze, and fermentation culture based on 121 ℃ of sterilization 20min, is cooled off again; Be that 1% inoculum size inserts above-mentioned solid seed in the cooled fermention medium according to weight ratio then, fully behind the stirring and evenly mixing, cultivated 7 days in 28 ℃ that in culturing process, with the vibration of substratum mixing, standing for fermentation is to finishing then when 20h.
5), aftertreatment
The product of fermentation step gained is dried to the moisture weight content under 40 ℃～45 ℃ low-temperature airflow be 10%, promptly gets the alpha galactosides zymin after the pulverizing.
After measured, alpha-galactosidase is that enzyme work is the 2352.74U/g dry medium.
Embodiment 3:
A kind of alpha-galactosidase production method, carry out following steps successively:
1), the preparation of spore suspension:
With embodiment 2.
2), first order seed preparation:
With embodiment 2.
3) secondary seed preparation:
The 40g butt of packing in the triangular flask of 500ml, this butt is made up of 32.855g wheat bran, 7.137g dregs of beans, 0.004g manganous sulfate and 0.004g copper sulfate.In bottle, add 60g water, transfer pH to 5.5～6.0 with the Sodium phosphate dibasic of 0.5M again, get the solid seed culture medium.
To cultivate fate and make into 6 days, all the other are with embodiment 2.
4), the fermentation of alpha-galactosidase:
With embodiment 2.
5), aftertreatment
With embodiment 2.
After measured, alpha-galactosidase is that enzyme work is the 1928.24U/g dry medium.
In the foregoing description, the extraction and the activity determination method of enzyme are as follows:
The solids of gained behind the fermentation step is stirred with stirrer, take by weighing about 1g, add 0.2M, Sodium phosphate dibasic-citrate buffer solution of pH5.0, the 30min that on 45 ℃ shaking bath, vibrates, filter paper filtering gets crude enzyme liquid.Live to measure enzyme with damping fluid dilution certain multiple.
The mensuration of alpha-galactosidase is as follows:
With the enzyme liquid 0.1ml that suitably dilutes, add citric acid-Sodium phosphate dibasic damping fluid 0.05ml of pH 5.0,10mmol/L p-nitrophenyl α-D-gala pyranoside 0.05ml under 50 ℃ of water bath condition, reacts 10min, adds the Na of the 0.25mol/L of 3.0ml then
2CO
3The solution termination reaction, OD value under 400nm.
Enzyme activity unit definition: at pH5.0, temperature is under 50 ℃ the condition, per minute generate 1 μ mol right-the required enzyme amount of nitrophenols is an enzyme activity unit.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (4)
1, alpha-galactosidase solid fermenting producing method, its feature may further comprise the steps:
1), the preparation of spore suspension: with preserving number was that the smelly aspergillus of CGMCC No.1628 is inoculated on the slant medium, in 28 ℃ of constant temperature culture 7 days; Making concentration with sterilized water again is 10
6～10
7The spore suspension of individual/ml, stand-by; Described slant medium is the PDA slant medium;
2), first order seed preparation: at first with liquid seed culture medium through 121 ℃, 20min sterilization, cooling again; Being that 5% inoculum size inserts spore suspension in the above-mentioned cooled liquid seed culture medium according to volume ratio then, is to cultivate 18h on the shaking table of 200rpm in 28 ℃, rotating speed, liquid seeds; Described liquid seed culture medium is the PDA liquid nutrient medium;
3), the fermentation of alpha-galactosidase: at first with fermention medium through 121 ℃, 20min sterilization, cooling again; Inoculum size according to 1 milliliter/10 grams inserts the aforesaid liquid seed in the cooled fermention medium then, fully behind the stirring and evenly mixing, in 28 ℃ of cultivations 4～7 days, in culturing process, carries out the step of mixing oscillation and fermentation cultivation base when 20h; Described fermention medium is 2: 3 butt and the mixed back of distilled water, is transferred pH to 5.5～6.0 to make by weight ratio, and described every 100g butt is made up of 82.137g wheat bran, 17.843g dregs of beans, 0.01g manganous sulfate and 0.01g cupric sulfate pentahydrate.
2, alpha-galactosidase solid fermenting producing method according to claim 1, it is characterized in that: carrying out post-processing step behind the described fermentation step: the product of fermentation step gained is dried to the moisture weight content under 40 ℃～45 ℃ low-temperature airflow be 10%, promptly gets the alpha galactosides zymin after the pulverizing.
3, alpha-galactosidase solid fermenting producing method, its feature may further comprise the steps:
1), the preparation of spore suspension: with preserving number was that the smelly aspergillus of CGMCC No.1628 is inoculated on the slant medium, in 28 ℃ of constant temperature culture 7 days; Making concentration with sterilized water again is 10
6～10
7The spore suspension of individual/ml, stand-by; Described slant medium is the PDA slant medium;
2), first order seed preparation: at first with liquid seed culture medium through 121 ℃, 20min sterilization, cooling again; Being that 5% inoculum size inserts spore suspension in the above-mentioned cooled liquid seed culture medium according to volume ratio then, is to cultivate 18h on the shaking table of 200rpm in 28 ℃, rotating speed, liquid seeds; Described liquid seed culture medium is the PDA liquid nutrient medium;
3) secondary seed preparation: the solid seed culture medium through 121 ℃, 20min sterilization, is cooled off again; Inoculum size according to 1 milliliter/10 grams inserts liquid seeds in the above-mentioned cooled solid seed culture medium then, cultivates 4～6 days in 28 ℃ of incubators, gets the solid seed; Described solid seed culture medium is 2: 3 butt and the mixed back of distilled water, is transferred pH to 5.5～6.0 to make by weight ratio, and described every 100g butt is made up of 82.137g wheat bran, 17.843g dregs of beans, 0.01g manganous sulfate and 0.01g cupric sulfate pentahydrate;
4), the fermentation of alpha-galactosidase: at first with fermention medium through 121 ℃, 20min sterilization, cooling again; Be that 1% inoculum size inserts above-mentioned solid seed in the cooled fermention medium according to weight ratio then, fully behind the stirring and evenly mixing, cultivated 4～7 days in 28 ℃; In culturing process, when 20h, carry out the mixing vibration; Described fermention medium is with the solid seed culture medium.
4, alpha-galactosidase solid fermenting producing method according to claim 3, it is characterized in that: carrying out post-processing step behind the described fermentation step: the product of fermentation step gained is dried to the moisture weight content under 40 ℃～45 ℃ low-temperature airflow be 10%, promptly gets the alpha galactosides zymin after the pulverizing.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286445A (en) * | 2011-08-01 | 2011-12-21 | 浙江大学 | Method for producing alpha-galactosidase by three-stage solid fermentation |
| CN102628056A (en) * | 2012-04-24 | 2012-08-08 | 中南林业科技大学 | Acid-resistant and high temperature-resistant beta-mannase gene and application thereof |
| CN104388405A (en) * | 2014-11-17 | 2015-03-04 | 中国农业大学 | Method for preparing alpha-galactosidase from rainbow conk |
| CN110066780A (en) * | 2019-03-29 | 2019-07-30 | 上海国龙生物技术集团有限公司 | A kind of feeding alpha-galactosidase solid fermenting production technology |
| CN110564712A (en) * | 2019-08-08 | 2019-12-13 | 河南省科学院生物研究所有限责任公司 | culture medium for producing alpha-galactosidase by liquid fermentation of filamentous fungi and application of culture medium |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK38893D0 (en) * | 1993-03-31 | 1993-03-31 | Novo Nordisk As | DNA |
| CN1693445A (en) * | 2004-08-25 | 2005-11-09 | 浙江省农业科学院 | Black aspergillus mutation of high active alpha-galactosldase and its ferment tech. on solid substrate |
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2006
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286445A (en) * | 2011-08-01 | 2011-12-21 | 浙江大学 | Method for producing alpha-galactosidase by three-stage solid fermentation |
| CN102286445B (en) * | 2011-08-01 | 2012-11-21 | 浙江大学 | Method for producing alpha-galactosidase by three-stage solid fermentation |
| CN102628056A (en) * | 2012-04-24 | 2012-08-08 | 中南林业科技大学 | Acid-resistant and high temperature-resistant beta-mannase gene and application thereof |
| CN104388405A (en) * | 2014-11-17 | 2015-03-04 | 中国农业大学 | Method for preparing alpha-galactosidase from rainbow conk |
| CN104388405B (en) * | 2014-11-17 | 2017-11-17 | 中国农业大学 | A kind of method that α galactosidases are prepared from rainbow conk |
| CN110066780A (en) * | 2019-03-29 | 2019-07-30 | 上海国龙生物技术集团有限公司 | A kind of feeding alpha-galactosidase solid fermenting production technology |
| CN110564712A (en) * | 2019-08-08 | 2019-12-13 | 河南省科学院生物研究所有限责任公司 | culture medium for producing alpha-galactosidase by liquid fermentation of filamentous fungi and application of culture medium |
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