CN104673766A - Method for performing simulated solid-state fermentation on tannase - Google Patents
Method for performing simulated solid-state fermentation on tannase Download PDFInfo
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- CN104673766A CN104673766A CN201510077373.3A CN201510077373A CN104673766A CN 104673766 A CN104673766 A CN 104673766A CN 201510077373 A CN201510077373 A CN 201510077373A CN 104673766 A CN104673766 A CN 104673766A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- C12N9/18—Carboxylic ester hydrolases (3.1.1)
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/01—Carboxylic ester hydrolases (3.1.1)
- C12Y301/0102—Tannase (3.1.1.20)
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Abstract
The invention discloses a method for performing simulated solid-state fermentation on tannase. The method comprises the following steps: step 1, preparation of a simulated solid-state fermentation culture medium, namely selecting, drying and sieving tealeaf stem powder, then uniformly mixing the tealeaf stem powder with deionized water, performing stirring extraction, filtering, collecting a filter liquid, so as to prepare tea stem extracting liquid, selecting and cutting a polymeric sponge into particles, drying the particles to prepare sponge particles, preparing a nutritive solution and uniformly mixing the nutritive solution with the sponge particles to prepare an initial fermentation culture medium; step 2, inoculation and fermentation culture, namely inoculating aspergillus niger-spores in each culture bottle, uniformly stirring and culturing at a constant temperature; and step 3, crude enzyme extraction, namely adding citric acid-sodium citrate buffer liquid into each culture bottle to perform oscillating extraction, and centrifuging or filtering to prepare the crude enzyme. According to the method, the polymeric sponge is used as an inertia matrix to culture the aspergillus niger-spores; after the solid-state fermentation is finished, the biomass of aspergillus niger can be accurately measured, and the separation and purification of tannase become easier.
Description
Technical field
The present invention relates to the technical field of fermentable, particularly relate to a kind of method simulating solid state fermentation tannase.
Background technology
Utilize fermentable to produce tannase and can carry out large-scale industrial production, become the Main Means of current acquisition tannase.Because having, solid state fermentation obtains that extracellular enzyme vigor is high, equipment is simple, with low cost, saving water resource and pollute the advantage such as little and become one of focus of current tannase research.
Utilize agriculture and forestry by-product cheap and easy to get to have the advantage of significant cost as solid-state fermentation substrate, but also bring a series of problem that cannot avoid simultaneously.First be that the existence of a large amount of insoluble solid substance in substratum causes microbial bacteria body burden wherein directly not measure, and the biomass method in all indirect measurement solid mediums all costs dearly and process is loaded down with trivial details, and be difficult to the growing state that reflects that microorganism is real accurately; Its two, along with fermenting process continue carry out, the matrix in substratum, constantly by microbial consumption, causes the oxygen transfer of substratum and hot mass transfer to change, is unfavorable for process control; They are three years old, the existence of the agriculture and forestry by-product solid substance of a large amount of nutritive ingredient complexity also makes the metabolism of microorganism become more complicated, while output object meta-bolites, also generate other materials many, bring certain difficulty to the separation and purification of object metabolite.
In view of this, the present inventor studies and devises a kind of method simulating solid state fermentation tannase, and this case produces thus.
Summary of the invention
The object of the present invention is to provide a kind of method simulating solid state fermentation tannase, aspergillus niger spore is cultivated as solid-state carrier by inert base, after solid state fermentation terminates, the biomass of aspergillus niger can be measured accurately, make the separation and purification of tannase become simpler simultaneously.
For achieving the above object, the present invention solves the technical scheme of its technical problem and is:
Simulate a method for solid state fermentation tannase, comprise the following steps:
Step one, prepare Preliminary fermentation substratum:
Get tea stalks powder, cross 80 mesh sieves after drying, mix with deionized water and stir evenly, after stirring and leaching, collecting by filtration filtrate, obtained tea stalk vat liquor;
Get polyurethane sponge, cut into particle, dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, dries to constant weight;
The tea stalk vat liquor of every gram of dry sponge preparation 9-19 gram, take glucose and the ammonium chloride of 0.06 gram respectively, be added in described tea stalk vat liquor, adjust initial pH to 4.0-5.0, after high pressure steam sterilization, obtained nutritive medium, aseptically mixes described nutritive medium with the sponge particles in described Erlenmeyer flask, obtained Preliminary fermentation substratum;
Step 2, inoculation and fermentation culture:
Every Erlenmeyer flask inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h;
Step 3, crude enzyme liquid extract:
Add the citric acid-sodium citrate damping fluid that 50 mL concentration are 0.05 mol/L, pH5.0 in every Erlenmeyer flask, 180 rpm vibration lixiviate 1h under 25oC, qualitative filter paper filters, obtained crude enzyme liquid.
As the optimal way of embodiment, in described step one, get tea stalks powder, 80 mesh sieves are crossed, with deionized water according to 1:20(w/v after drying) ratio mixes and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtained tea stalk vat liquor.
As the optimal way of embodiment, in described step one, get polyurethane sponge, cut into particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight.
As the optimal way of embodiment, in described step one, in described nutritive medium, be also added with Weibull composition.
As the optimal way of embodiment, in described step one, in described nutritive medium, be also added with ammonium sulfate and yeast extract composition.
As the optimal way of embodiment, select polyurethane sponge as the inert support of absorption nutritive medium, described polyurethane sponge is vesicular sponge material, and density is 40 kg/m
3, open pore size is 300-500 μm, specific surface area is 380.6 m
2/ g.
As the optimal way of embodiment, the diameter of described sponge particles is 0.2-1.0cm.
As the optimal way of embodiment, described polyurethane sponge, through the recycling of 5 circulations, still has good enzymatic productivity.
Simulation solid state fermentation (Modified solid state fermentation, MSSF) be a kind of novel process for solid state fermentation, it is that matrix is adsorbed specific nutritive medium and prepared fermention medium culturing micro-organisms fermentative production object meta-bolites usually with inert support.It had both remained all advantages of solid state fermentation, effectively can solve again the several difficult problems be present in solid state substrate fermentation, further can further investigate the mechanism of solid state fermentation.
Polyurethane sponge (Polyurethane foam sponge, PUFS) be a kind of flexible PU foam (Flexible polyurethane foam, FPUF), there is fabulous morphological structure stability, high specific surface area, good ventilation property, excellent absorption and water retention capacity.It can not bring disadvantageous effect also can not to microorganism growth metabolism by microbial consumption, keeps very stable state, ensure that the stable of substratum oxygen transfer and hot mass transfer in whole fermenting process.Take PUFS as inert base, simply, directly can measure the content of microorganisms in substratum.
After the present invention adopts technique scheme, adopt polyurethane sponge to cultivate aspergillus niger spore as inert base, after solid state fermentation terminates, the biomass of aspergillus niger can be measured accurately, make the separation and purification of tannase become simpler simultaneously.
Accompanying drawing explanation
Fig. 1 is high density polyurethane sponge of the present invention and aspergillus niger morphological specificity figure thereof;
Fig. 2 be polyurethane sponge particle of the present invention recycling on tannase synthesis affect diagram.
Embodiment
the detection method adopted in the following example:
Tannase vigour-testing method: 0.01 mol/L Tenox PG 0.25 mL, crude enzyme liquid 0.25 mL, 6.67 g/L methyl alcohol rhodanine solution 0.3 mL, 0.7 mol/L KOH 0.2 mL is added after 30 DEG C of water-bath 5 min, 30 DEG C of water-bath 5 min, add distilled water 4 mL again, 30 DEG C of water-bath 10 min, survey light absorption value at 520 nm places.The tannase vigor Y(U/gds that every gram of butt matter obtains)=12.5 × (Δ A-0.0524)
Biomass estimation: record the sponge dry weight m in each fermentation flask
1, the filter weight m for lixiviate that record is corresponding
2.After fermentation ends, damping fluid lixiviate crude enzyme liquid, the non-permeate of vacuum decompression collecting by filtration 80 DEG C is dried to constant weight m
3.Biomass (dry cell weight) M(mg)=m
3-m
2-m
1
High density polyurethane sponge characterizes: SEM observes sponge particles internal morphology feature, pore size; Analyze determinator and measure sponge specific surface area; The sponge of getting certain volume carries out quality determination and bulk density on analytical balance.
The inner specific surface area thalline of polyurethane sponge characterizes: SEM observes the inner thalline shape characteristic of substratum.
embodiment 1
Test the high density polyurethane sponge density adopted and be about 40 kg/m
3, open pore size is about 300-500 μm, specific surface area is 380.6 m
2/ g.Shown in the inner and inner thalline mode of appearance result Fig. 1 of sem observation sponge.
The external morphology of high density polyurethane sponge can be observed clearly by Fig. 1 a and 1b.Sponge interior smooth, smooth, opening is good and aperture is more homogeneous.The growth metabolism activity that this is aspergillus niger provides the huge support interface of area, also makes it have excellent absorption property simultaneously.In fermenting process, the mechanical property Biostatic performance of polyurethane sponge excellence is enough to ensure that in fermenting process, oxygen transfer and hot mass transfer remain stable.
Fig. 1 c and 1d shows, and aspergillus niger is attached to uniformly on sponge inwall and carries out growth and breeding, and this shows that aspergillus niger can use the nutritive substance in matrix adequately and uniformly, can also touch sufficient oxygen simultaneously.Sponge internal voids is obvious, and not because the growth of aspergillus niger causes the decline of permeability, this also shows that highdensity polyurethane sponge can provide the growth metabolism environment of high-quality for aspergillus niger, is a kind of inert support of excellent performance.
embodiment 2
Get tea stalks powder, cross 80 mesh sieves after drying, with deionized water according to 1:20(w/v) ratio mix and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtains tea stalk vat liquor; Get polyurethane sponge, cut into 0.2 cm diameter particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight; Every gram of dry sponge prepares the tea stalk vat liquor of 9 grams, takes glucose and the ammonium chloride of 0.06 gram respectively, is added in described tea stalk vat liquor, adjusts initial pH to 4.0, after high pressure steam sterilization, and obtained nutritive medium; Described nutritive medium is mixed with described sponge particles, obtained simulation solid-state fermentation culture medium; Every culturing bottle inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h; Add citric acid-sodium citrate damping fluid in every culturing bottle, vibration lixiviate, qualitative filter paper filters, and obtained crude enzyme liquid, measuring biomass is 46.4 mg/gds, and tannase vigor is 3.02 U/gds.
embodiment 3
Get tea stalks powder, cross 80 mesh sieves after drying, with deionized water according to 1:20(w/v) ratio mix and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtains tea stalk vat liquor; Get polyurethane sponge, cut into 1.0 cm diameter particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight; Every gram of dry sponge prepares the tea stalk vat liquor of 9 grams, takes glucose and the ammonium chloride of 0.06 gram respectively, is added in described tea stalk vat liquor, adjusts initial pH to 4.0, after high pressure steam sterilization, and obtained nutritive medium; Described nutritive medium is mixed with described sponge particles, obtained simulation solid-state fermentation culture medium; Every culturing bottle inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h; Add citric acid-sodium citrate damping fluid in every culturing bottle, vibration lixiviate, qualitative filter paper filters, obtained crude enzyme liquid.Measuring biomass is 61 mg/gds, and tannase vigor is 1.53 U/gds.
embodiment 4
Last Batch fermentation is cultivated the carrier having surveyed biomass after terminating and again prepare fermention medium, inoculated aspergillus niger 96 h that ferment measure tannase vigor and biomasss, repeat 5 times continuously.As shown in Figure 2, through the recycling of 5 circulations, polyurethane sponge still has good enzymatic productivity.To 5th batch, biomass is 56 mg/gds, and tannase vigor is 3 U/gds.
embodiment 5
Get tea stalks powder, cross 80 mesh sieves after drying, with deionized water according to 1:20(w/v) ratio mix and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtains tea stalk vat liquor; Get polyurethane sponge, cut into 0.2 cm diameter particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight; Every gram of dry sponge prepares the tea stalk vat liquor of 19 grams, takes glucose and the ammonium chloride of 0.06 gram respectively, is added in described tea stalk vat liquor, adjusts initial pH to 4.0, after high pressure steam sterilization, and obtained nutritive medium; Described nutritive medium is mixed with described sponge particles, obtained simulation solid-state fermentation culture medium; Every culturing bottle inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h; Add citric acid-sodium citrate damping fluid in every culturing bottle, vibration lixiviate, 10000 rpm are centrifugal, obtained crude enzyme liquid.Measuring biomass is 117 mg/gds, and tannase vigor is 5.1 U/gds.
embodiment 6
Get tea stalks powder, cross 80 mesh sieves after drying, with deionized water according to 1:20(w/v) ratio mix and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtains tea stalk vat liquor; Get polyurethane sponge, cut into 0.2 cm diameter particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight; Every gram of dry sponge prepares the tea stalk vat liquor of 9 grams, takes glucose and the ammonium chloride of 0.06 gram respectively, is added in described tea stalk vat liquor, adjusts initial pH to 5.0, after high pressure steam sterilization, and obtained nutritive medium; Described nutritive medium is mixed with described sponge particles, obtained simulation solid-state fermentation culture medium; Every culturing bottle inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h; Add citric acid-sodium citrate damping fluid in every culturing bottle, vibration lixiviate, 10000 rpm are centrifugal, obtained crude enzyme liquid.Measuring biomass is 58 mg/gds, and tannase vigor is 3 U/gds.
embodiment 7
Get tea stalks powder, cross 80 mesh sieves after drying, with deionized water according to 1:20(w/v) ratio mix and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtains tea stalk vat liquor; Get polyurethane sponge, cut into 0.2 cm diameter particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight; Every gram of dry sponge prepares the tea stalk vat liquor of 9 grams, takes Weibull and the ammonium chloride of 0.06 gram respectively, is added in described tea stalk vat liquor, adjusts initial pH to 4.0, after high pressure steam sterilization, and obtained nutritive medium; Described nutritive medium is mixed with described sponge particles, obtained simulation solid-state fermentation culture medium; Every culturing bottle inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h; Add citric acid-sodium citrate damping fluid in every culturing bottle, vibration lixiviate, qualitative filter paper filters, and obtained crude enzyme liquid, measuring biomass is 72 mg/gds, and tannase vigor is 6.9 U/gds.
embodiment 8
Get tea stalks powder, cross 80 mesh sieves after drying, with deionized water according to 1:20(w/v) ratio mix and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtains tea stalk vat liquor; Get polyurethane sponge, cut into 0.2 cm diameter particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight; Every gram of dry sponge prepares the tea stalk vat liquor of 9 grams, takes glucose and the ammonium sulfate of 0.06 gram respectively, is added in described tea stalk vat liquor, adjusts initial pH to 4.0, after high pressure steam sterilization, and obtained nutritive medium; Described nutritive medium is mixed with described sponge particles, obtained simulation solid-state fermentation culture medium; Every culturing bottle inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h; Add citric acid-sodium citrate damping fluid in every culturing bottle, vibration lixiviate, qualitative filter paper filters, and obtained crude enzyme liquid, measuring biomass is 82 mg/gds, and tannase vigor is 5 U/gds.
After the present invention adopts technique scheme, adopt polyurethane sponge to cultivate aspergillus niger spore as inert base, after solid state fermentation terminates, the biomass of aspergillus niger can be measured accurately, make the separation and purification of tannase become simpler simultaneously.
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 (10)
1. simulate a method for solid state fermentation tannase, it is characterized in that: comprise the following steps:
Step one, preparation simulation solid-state fermentation culture medium:
Get tea stalks powder, cross 80 mesh sieves after drying, mix with deionized water and stir evenly, after stirring and leaching, collecting by filtration filtrate, obtained tea stalk vat liquor;
Get polyurethane sponge, cut into particle, dry, after constant weight, be cooled to room temperature, obtained sponge particles; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, dries to constant weight;
Interpolation glucose and ammonium chloride are dissolved in tea stalk vat liquor, and after high pressure steam sterilization, obtained nutritive medium, aseptically mixes described nutritive medium with the sponge particles in described Erlenmeyer flask, obtained simulation solid-state fermentation culture medium;
Step 2, inoculation and fermentation culture:
Every Erlenmeyer flask inoculation 6.4 × l0
7individual aspergillus niger spore, stirs, under 30oC condition, and constant temperature culture 96h;
Step 3, acquisition tannase enzyme liquid:
Add the citric acid-sodium citrate damping fluid that 50 mL concentration are 0.05 mol/L, pH 5.0 in every Erlenmeyer flask, under 25oC, 180 rpm vibrate lixiviate 1 h, and the filtrate obtained through the centrifugal supernatant liquor that obtains or filtration is tannase crude enzyme liquid.
2. a kind of method simulating solid state fermentation tannase as claimed in claim 1, it is characterized in that: adsorb specific nutritive medium using inert support polyurethane sponge for matrix and cultivate fermentation of Aspergillus niger production tannase as fermention medium, exist without insoluble solid substance except inert support polyurethane sponge in simulation solid-state fermentation culture medium.
3. a kind of method simulating solid state fermentation tannase as claimed in claim 1, it is characterized in that: in described step one, get tea stalks powder, 80 mesh sieves are crossed after drying, with deionized water according to 1:20(w/v) ratio mixes and stirs evenly, be placed on magnetic stirring apparatus and be heated to 80 DEG C of stirring and leaching 1 h, with vacuum pump pressure collecting by filtration filtrate, obtained tea stalk vat liquor.
4. a kind of method simulating solid state fermentation tannase as claimed in claim 1, it is characterized in that: in described step one, get polyurethane sponge, cut into particle, 60 DEG C of warm water wash 3 times, then after using deionized water wash 3 times, are placed in 80 DEG C of baking ovens and dry, room temperature is cooled to, obtained sponge particles after constant weight; Every only sponge particles described in 250 mL Erlenmeyer flask packing 1g, after gauze sealing sterilizing, is placed in 80 DEG C of baking ovens and dries to constant weight.
5. a kind of method simulating solid state fermentation tannase as claimed in claim 1, it is characterized in that: in described step one, the tea stalk vat liquor of every gram of dry sponge preparation 9-19 gram, take glucose and the ammonium chloride of 0.06 gram respectively, be added in described tea stalk vat liquor, adjust initial pH to 4.0-5.0, after high pressure steam sterilization, nutritive medium and sponge particles are aseptically mixed to obtain and simulate solid-state fermentation culture medium.
6. a kind of method simulating solid state fermentation tannase as claimed in claim 1, is characterized in that: in described step one, in described nutritive medium, is also added with Weibull composition.
7. a kind of method simulating solid state fermentation tannase as claimed in claim 1, is characterized in that: in described step one, in described nutritive medium, is also added with ammonium sulfate composition.
8. a kind of method simulating solid state fermentation tannase as claimed in claim 1, is characterized in that: the length of described sponge particles is 0.2-1.0cm.
9. a kind of method simulating solid state fermentation tannase as claimed in claim 1, it is characterized in that: described polyurethane sponge is vesicular sponge material, density is 40 kg/m
3, open pore size is 300-500 μm, specific surface area is 380.6 m
2/ g.
10. a kind of method simulating solid state fermentation tannase as claimed in claim 1, is characterized in that: described polyurethane sponge, through the recycling of 5 circulations, still has good enzymatic productivity.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107904218A (en) * | 2017-12-29 | 2018-04-13 | 集美大学 | A kind of tannase solid-state fermentation culture medium preparation method and applications |
| CN107904219A (en) * | 2017-12-29 | 2018-04-13 | 集美大学 | A kind of tannase solid-state fermentation culture medium preparation method and applications |
| CN110226385A (en) * | 2019-06-11 | 2019-09-13 | 深圳市铁汉生态环境股份有限公司 | A kind of type of seeding of pteridophyte |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101705260A (en) * | 2009-11-16 | 2010-05-12 | 天津北洋百川生物技术有限公司 | Method for producing gamma-polyglutamic acid by inert carrier solid state fermentation method |
| CN102796716A (en) * | 2012-08-10 | 2012-11-28 | 集美大学 | Method for preparing tannase |
-
2015
- 2015-02-13 CN CN201510077373.3A patent/CN104673766A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101705260A (en) * | 2009-11-16 | 2010-05-12 | 天津北洋百川生物技术有限公司 | Method for producing gamma-polyglutamic acid by inert carrier solid state fermentation method |
| CN102796716A (en) * | 2012-08-10 | 2012-11-28 | 集美大学 | Method for preparing tannase |
Non-Patent Citations (2)
| Title |
|---|
| J. VAN DE LAGEMAAT等: ""Solid-state fermentation and bioremediation: development of a continuous process for the production of fungal tannase"", 《CHEMICAL ENGINEERING JOURNAL》 * |
| 刘建国 等: ""以聚氨酯泡沫为载体固态发酵纤维素酶的工艺条件研究"", 《现代化工》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107904218A (en) * | 2017-12-29 | 2018-04-13 | 集美大学 | A kind of tannase solid-state fermentation culture medium preparation method and applications |
| CN107904219A (en) * | 2017-12-29 | 2018-04-13 | 集美大学 | A kind of tannase solid-state fermentation culture medium preparation method and applications |
| CN107904218B (en) * | 2017-12-29 | 2020-07-28 | 集美大学 | Preparation method and application of tannase solid-state fermentation medium |
| CN110226385A (en) * | 2019-06-11 | 2019-09-13 | 深圳市铁汉生态环境股份有限公司 | A kind of type of seeding of pteridophyte |
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