CN104087514A - Aspergillus clavatus for producing feruloyl esterase and application thereof - Google Patents
Aspergillus clavatus for producing feruloyl esterase and application thereof Download PDFInfo
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- CN104087514A CN104087514A CN201410299853.XA CN201410299853A CN104087514A CN 104087514 A CN104087514 A CN 104087514A CN 201410299853 A CN201410299853 A CN 201410299853A CN 104087514 A CN104087514 A CN 104087514A
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- aspergillus
- feruloyl esterase
- wheat bran
- fermentation
- excellent aspergillus
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- 101001065065 Aspergillus awamori Feruloyl esterase A Proteins 0.000 title claims abstract description 22
- 241000228193 Aspergillus clavatus Species 0.000 title claims abstract description 22
- 235000015099 wheat brans Nutrition 0.000 claims abstract description 29
- 238000000855 fermentation Methods 0.000 claims abstract description 23
- 230000004151 fermentation Effects 0.000 claims abstract description 23
- 241000228212 Aspergillus Species 0.000 claims description 49
- 239000002253 acid Substances 0.000 claims description 33
- 235000019504 cigarettes Nutrition 0.000 claims description 21
- 244000025254 Cannabis sativa Species 0.000 claims description 20
- 241000894006 Bacteria Species 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 10
- 239000001963 growth medium Substances 0.000 claims description 7
- 238000011218 seed culture Methods 0.000 claims description 7
- 238000004321 preservation Methods 0.000 claims description 5
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- 239000007787 solid Substances 0.000 claims description 2
- 238000004611 spectroscopical analysis Methods 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 12
- 241000196324 Embryophyta Species 0.000 abstract description 7
- 235000001785 ferulic acid Nutrition 0.000 abstract description 7
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 abstract description 6
- 229940114124 ferulic acid Drugs 0.000 abstract description 6
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 abstract description 6
- 150000007965 phenolic acids Chemical class 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 6
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 abstract description 6
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- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 abstract description 3
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- -1 ferulic acid ester Chemical class 0.000 description 3
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention provides a strain of Aspergillus clavatus for producing feruloyl esterase and application thereof. The Aspergillus clavatus has the category name of Aspergillus clavatus fumigatus, is collected by China center for type culture collection, called CCTCC for short, at December 8, 2013, has the collection number of CCTCCNO: M2013641 and is applied to the production of feruloyl esterase. According to the strain provided by the invention, a method that feruloyl esterase is produced through fermenting by taking wheat bran as a fermentation substrate and bound-type phenolic and ferulic acids are obtained through rupturing ester bonds between the polysaccharide and lignin of plant cell walls and ferulic acid by the feruloyl esterase, is adopted, and the condition that microorganisms are short in growth cycle, are not restricted to climate and are easily subjected to large-scale production is utilized, so that a novel biological method for extracting bound-type phenolic acid substances, such as ferulic acid is provided.
Description
Technical field
The invention belongs to biological technical field, be specifically related to excellent aspergillus and the application thereof of a strain product feruloyl esterase.
Background technology
Wheat is cultivated area one of crop the most widely in the world, account for 26% of plant of grain crops area, China's wheat annual production is high, the annual production of Testa Tritici can reach more than 2,000 ten thousand tons, but all the time less for the exploitation of wheat bran, substantially only to be used as feedstuff raw material.In Testa Tritici, mainly containing the Multiple components such as non-starch polysaccharide, starch, fat, xylogen, protein, VITAMIN and aldehydes matter,, taking araboxylan as main non-starch polysaccharide is containing 46%, is wherein the main component of Wheat cell wall.In wheat bran, also contain the phenolic acid of some amount, as compositions such as forulic acid (ferulic acid), P-coumaric acid (p-coumaricacid), coffic acid (caffeicacid), sinapinic acid (sinapicacid) and syringic acids (syringicacid), wherein, taking forulic acid as main component, be about 0.4%～0.7% of wheat bran quality.Forulic acid, mainly by ester bond and cell wall polysaccharides and xylogen necklace, generally can be used highly basic ester bond is disconnected and discharge.
Phenolic acid group will be present in wheat bran cortex, and it has stronger anti-oxidant activity and antitumor action, and the toxic substance in environment is had to anti-mutagenic activity as polycyclic aromatic hydrocarbons and inferior ammonium nitrate and mycotoxins.Studies have reported that discovery, in the isolated fermentation test of wheat-bran dietary fiber, faecal flora can discharge in connection with the forulic acid on wheat-bran dietary fiber by fermentative action.And forulic acid can significantly reduce viability, survival and the antioxidant levels of human cervical carcinoma cell, improve the reactive oxygen species of cancer cells in protecting simultaneously and increase lipid peroxidation and the DNA damage effect of cancer cells.Separately there is report to think that forulic acid can be by the formation that is used for regulating blood vessel to vascular endothelial growth factor (VEGF), platelet derived growth factor (PDGF) and hypoxia inducible factor (HIF).
At present, research shows that aldehydes matter is mainly present in plant body with several forms such as sequestered, mating type and bound forms, and in general, free property and mating type aldehydes matter are solubility phenols; Bound form is insoluble aldehydes matter, combines with other materials (comprising protein, carbohydrate, organic acid etc.) with ester bond, glycosidic link and ether glycosidic bond form.
For the extraction of aldehydes matter, generally adopting organic solvent is medium, is aided with ultrasonic, microwave, the method such as subcritical is carried out.Because bound form aldehydes matter is generally connected with different of bondings from other materials, traditional extracting method is not enough to ester bond, glycosidic link or ether bond rupture, therefore the extraction of bound form aldehydes matter is mainly adopted to strong acid and strong base or biological method (enzyme process and fermentation method), its main purpose is all to destroy the structure of cell walls fibre composition densification, so that constraint row aldehydes matter discharges.
Although chemical process is simple to operate, efficiency is high, the follow-up processing extraction waste liquid that needs, is unfavorable for sustainable development.Therefore compare, adopt biological method more to meet the demand for development of green, environmental protection.Carry out focusing mostly in fields such as xylo-oligosaccharide production and inulinase-producing activities of fermentation aspect for wheat bran, in addition existing lot of documents report wheat bran after fermentation by the time food fibre functional property have significantly before than fermentation and improve, as retentiveness, hold oiliness and the binding ability to objectionable impurities etc.
Forulic acid is as the main phenolic acid in wheat bran, at present the mode of its acquisition is mainly to enzyme process, less with the report of fermentation mode, special needs to be pointed out is, the form of the substantially suitable bound form of the forulic acid in wheat bran exists, its sequestered: mating type: the ratio of bound form is 0.1:1:100.In recent years, the research of aldehydes matter existence form is become to new study hotspot gradually.The report of the aspects such as current extraction, mensuration and anti-oxidant activity to some sequestereds, esterification type, glucoside type, bound form aldehydes matter.The exist shape and its content and anti-oxidant activity of aldehydes matter have important relationship, bound form aldehydes matter not only content far away higher than sequestered and mating type, and antioxidant activity and suppress DNA damage aspect will be higher than other two types.
The present invention is intended to utilize Microbial resources to adopt fermentation engineering agricultural resource to be transformed to the material that forms human society demand, solves the problem of antioxidant shortage of resources.
Summary of the invention
Technical problem to be solved by this invention is: for the new raw material of providing of existing anti-oxidant supply and method, and large plants such as wheat are recycled, provide a strain to produce excellent aspergillus and the application thereof of feruloyl esterase, it can utilize Testa Tritici fermentative production forulic acid, because the microorganism growth cycle is short, not climate restriction, be easy to realize scale operation, thereby solve the problems such as forulic acid deficiency.
The excellent aspergillus of feruloyl esterase is produced in a strain provided by the invention, derives from South China Botanical Garden soil, and through screening, separation and purification obtains.This its Classification And Nomenclature of rod aspergillus be excellent aspergillus (
aspergillus clavatus) cigarette green grass or young crops, by the center preservation of Chinese Typical Representative culture collection, be called for short CCTCC, deposit number is CCTCC NO:M2013641, and its ITS sequence table is as shown in sequence table 1, and preservation date is on December 8th, 2013, preservation address is Wuhan, China Wuhan University, Chinese Typical Representative culture collection center.
Gained rod aspergillus of the present invention (
aspergillus clavatus) flat board of cigarette green grass or young crops observe feature and microscopical character as follows: excellent aspergillus (Aspergillus clavatus) cigarette green grass or young crops colony growth rate on czapek's solution is slower, cultivates 7d diameter 30mm; Quality velvet shape is to granular, white mycelium, and conidium structure is dark blue green, without transudate.On wort agar, colony growth rate is very fast, cultivates 7d diameter 45mm, smooth, quality granular, and conidium structure is many, greyish-green, bacterium colony reverse side sorrel.Cultivate 7d diameter 40mm at CheckShi yeast extract paste agar colony, quality velvet shape, bottle is slightly thick; Conidium structure is many, is dusty blue, and edge is rarer, is white in color, without transudate.Can observe conidial head children under microscope time, be clavate, conidiophore betides matrix, and falx stem is different in size; Top capsule is expanded gradually and is become club shape by falx top, and diameter 50～60mm produces spore structure individual layer, and intensive this of bottle stalk is born in all surfaces of top capsule, and a bottle stalk size is 8～12 μ m × 2～3 μ m; Conidium is oval, and conidium size is 4～5 μ m × 3 μ m, and wall is smooth.
Described excellent aspergillus (
aspergillus clavatus) ITS of cigarette green grass or young crops and excellent aspergillus (
aspergillus clavatus) homology reaches 100%.
A kind of preparation method of the excellent aspergillus that produces feruloyl esterase, excellent aspergillus of the present invention derives from South China Botanical Garden soil, adopts different substratum to screen, and screens multiple bacterial strains, carry out one by one ferulaic acid content and Oxford cup transparent circle determination experiment, obtain high yield feruloyl esterase excellent aspergillus (
aspergillus clavatus) cigarette green grass or young crops, comprising the steps: to take pedotheque 10g puts into the 300mL triangular flask that the aseptic physiological saline containing granulated glass sphere of 90 ~ 100 mL is housed and sways 18 ~ 20min, make 10 times of even liquid 1:10 of ascending series dilute sample, 1:100,1:1000 and 1:10000, get the even liquid 1:100 of dilute sample, 1:1000 and the each 1.0mL of 1:10000 and join respectively aseptic plate; Adding 10～15mL PDA substratum mixes again, leave standstill 1 ~ 2 min, be inverted and cultivate 70 ~ 72h in 28 DEG C ± 1 DEG C, bacterium colony single on PDA substratum is transferred to 28 DEG C ± 1 DEG C cultivation 70 ~ 72h on inclined-plane PDA substratum, proceed to again primary dcreening operation substratum, in 28 DEG C ± 1 DEG C cultivation 58 ~ 60h, with the experiment of Oxford cup transparent circle, excellent aspergillus (Aspergillus clavatus) the cigarette green grass or young crops that screening obtains producing feruloyl esterase; Described pedotheque gathers from South China Botanical Garden.
In aforesaid method, the excellent aspergillus that obtains product feruloyl esterase from primary dcreening operation substratum is inoculated in triangular flask seed culture medium, in 28 DEG C ± 1 DEG C cultivation 58 ~ 60h, is excellent aspergillus seed; Again excellent aspergillus seed is inoculated in fermention medium and is cultivated, for producing the extract of excellent aspergillus of forulic acid.
Primary dcreening operation substratum: FeSO
47H
2o 0.01g, NaNO
32.0 g, KCl 0.5 g, MgSO
47H
2o 0.5 g, K
2hPO
41.0g, sucrose 30 g, agar 20 g, distilled water 1000m L, pH value nature; 121 DEG C of sterilizing 20 min.After sterilizing, substratum is as cold as about the 60 DEG C dimethyl formamide solution 100mL that add aseptic 10% Ferulic acid ethylester, shakes up to uniform oyster white and is down flat plate.
Seed culture medium: FeSO
47H
2o 0.01g, NaNO
32.0 g, KCl 0.5 g, MgSO
47H
2o 0.5 g, K
2hPO
41.0g, wheat bran 1000 g, distilled water 1000m L, pH value nature.121 DEG C of sterilizing 20 min.
PDA substratum: PDA(potato agar) 40g, be settled to 1L, pH nature.121 DEG C of insulated sterilizing 20min.
Fermention medium: wheat bran: water is weight ratio 1:1, pH value nature.
Another object of the present invention is to provide the fermentation application of the production forulic acid of described excellent aspergillus cigarette green grass or young crops.Described excellent aspergillus cigarette green grass or young crops is applied to product feruloyl esterase.
Above-mentioned application, comprise the steps: to utilize the direct loop-carrier aspergillus of wheat bran (Aspergillus clavatus) cigarette green grass or young crops, in fermention medium, carry out koji plate fermentation, obtain fermented product extract, utilize the forulic acid burst size of wheat bran in high-efficient liquid phase color spectrometry fermention medium.
In above-mentioned application, the condition of described fermentation is: on koji tray, carry out solid fermentation with wheat bran, wheat bran gauge control is in 2 ~ 3cm, and excellent aspergillus seed inoculum size is that 0.5% (bacteria containing amount is 1～2 × 10
9individual/g), culture temperature is 25～28 DEG C, carries out aerlbic culture, fermentation time is 48～55h; PH nature.Described wheat bran is natural wheat bran.Described excellent aspergillus seed is inoculated in triangular flask seed culture medium by excellent aspergillus, cultivates 58 ~ 72h obtain in 28 DEG C ± 1 DEG C.
Compared with prior art, the invention has the beneficial effects as follows:
The invention provides a kind of new excellent aspergillus (
aspergillus clavatus) cigarette green grass or young crops, described bacterial strain has the ability of producing feruloyl esterase, has supplemented greatly the database of extracting bound form forulic acid bacterium; Described excellent aspergillus (
aspergillus clavatus) cigarette green grass or young crops from organic sphere separation and purification out, and be easy to cultivate and preserve, rapidly, energy for growth is strong in breeding.
The enzyme that feruloyl esterase is produced in the fermentation of the present invention's rod aspergillus is alive very high, can be used as the biological bacteria of extracting bound form phenolic acid from plant.
The invention provides excellent aspergillus (
aspergillus clavatus) application of cigarette green grass or young crops bound form forulic acid biological process in plant, for release and the follow-up anti-oxidant engineering of bound form phenolic acid, provide new biological method and strong technical support.
Brief description of the drawings
Fig. 1 is the transparent loop graph of the ferulic acid ester enzyme activity of bacterial strain of the present invention and other bacterial strain fermentation liquors.
Fig. 2 is the excellent aspergillus cigarette green grass or young crops podocyte aspect graph of 400 times of observations under the microscope.
Fig. 3 is the excellent aspergillus cigarette green grass or young crops conidial head aspect graph of 400 times of observations under the microscope.
Fig. 4 is the bacterium colony figure of excellent aspergillus cigarette green grass or young crops in PDA cultivation.
Fig. 5 is the color atlas of the forulic acid standard substance of HPLC mensuration.
Fig. 6 is the typical curve of the forulic acid standard substance of HPLC mensuration.
Fig. 7 is excellent aspergillus fermentation color atlas.
Embodiment
Below in conjunction with embodiment, the present invention is further described.
The culture medium prescription of using in following examples:
Primary dcreening operation substratum: FeSO
47H
2o 0.01g, NaNO
32.0 g, KCl 0.5 g, MgSO
47H
2o 0.5 g, K
2hPO
41.0g, sucrose 30 g, agar 20 g, distilled water 1000m L, pH value nature; 121 DEG C of sterilizing 20 min.After sterilizing, substratum is as cold as about the 60 DEG C dimethyl formamide solution 100mL that add aseptic 10% Ferulic acid ethylester, shakes up to uniform oyster white and is down flat plate.
Seed culture medium: FeSO
47H
20 0.01g, NaNO
32.0 g, KCl 0.5 g, MgSO
47H
20 0.5 g, K
2hPO
41.0g, wheat bran 1000 g, distilled water 1000m L, pH value nature.121 DEG C of insulated sterilizing 20min.
PDA substratum: PDA(potato agar) 40g, be settled to 1L, pH nature.121 DEG C of insulated sterilizing 20min.
Fermention medium: wheat bran: water weight ratio is 1:1, pH nature.121 DEG C of insulated sterilizing 20min.
Embodiment 1
The primary dcreening operation of rod aspergillus tubigensis, Oxford cup transparent circle sieve bacterium experiment screening superior strain.
Gather pedotheque from South China Botanical Garden, taking sample 10g puts into and the aseptic 300mL triangular flask containing the physiological saline of granulated glass sphere of 90mL is housed sways 20 min, make 10 times of even liquid 1:10 of ascending series dilute sample, 1:100,1:1000 and 1:10000, get the even liquid 1:100 of dilute sample, 1:1000 and the each 1.0mL of 1:10000 and add respectively aseptic plate; Add again 15mL PDA substratum and mix, leave standstill 1min, be inverted and cultivate 72h in 28 DEG C, bacterium colony single on substratum is transferred to 28 DEG C of cultivation 72h on inclined-plane PDA substratum, proceed to the 28 DEG C of cultivation 60h of primary dcreening operation substratum containing Ferulic acid ethylester, the bacterial strain of ferulic acid ester is produced in screening, for subsequent use again;
Do and again sieve bacterium with the experiment of plate Oxford cup transparent circle, by separate obtain product ferulic acid ester bacterial strain adopt liquid fermentation process in fermention medium 28 DEG C cultivate 2 days, obtain fermented liquid, get 200uL supernatant liquor by after centrifugal fermented liquid 6000r/min, be put on primary dcreening operation substratum in the cup of Oxford, be placed in 37 DEG C of incubators and cultivate 5-27h observation transparent circle, the supernatant liquor of fermented liquid is cleared up substrate and is produced transparent circle, the height that produces ferulaic acid esterase activity can be described, in the bacterial strain that produces feruloyl esterase, contrast screening transparent circle with this method, the bacterial strain of what transparent circle was large be high yield feruloyl esterase.See shown in accompanying drawing 1, wherein a bacterial classification is gained rod aspergillus (Aspergillus clavatus) cigarette green grass or young crops of the present invention, and its transparent circle is large than other bacterial strains.
Embodiment 2
The Microbiological Characteristics of bacterial strain and qualification
Rod aspergillus colony growth rate on czapek's solution is slower, cultivates 7d diameter 30mm; Quality velvet shape is to granular, white mycelium, and conidium structure is dark blue green, without transudate.On wort agar, colony growth rate is very fast, cultivates 7d diameter 45mm, smooth, quality granular, and conidium structure is many, greyish-green, bacterium colony reverse side sorrel.Cultivate 7d diameter 40mm at CheckShi yeast extract paste agar colony, quality velvet shape, bottle is slightly thick; Conidium structure is many, is dusty blue, and edge is rarer, is white in color, without transudate.
Can observe conidial head children under microscope time, be clavate, conidiophore betides matrix, and falx stem is different in size; Top capsule is expanded gradually and is become club shape by falx top, and diameter 50～60mm produces spore structure individual layer, and intensive this of bottle stalk is born in all surfaces of top capsule, bottle stalk size 8～12 μ m × 2～3 μ m; Conidium is oval, spore size 4～5 μ m × 3 μ m, wall smooth (seeing shown in accompanying drawing 2-4).
Described excellent aspergillus (
aspergillus clavatus) ITS of cigarette green grass or young crops and excellent aspergillus (
aspergillus clavatus) homology reaches 100%.
Embodiment 3
Solid state fermentation is measured bound form forulic acid burst size
By seed culture based formulas, claim the 10g wheat bran 10g that adds water to be mixed into 500mL triangular flask, loop-carrier aspergillus after seed culture medium sterilizing (
aspergillus clavatus) the blue or green inclined-plane of cigarette bacterial strain, inoculum size is 0.5%, 28 DEG C cultivates after 3d, obtains excellent aspergillus seed;
Press fermentative medium formula, claim 200g wheat bran add water 200g mix after sterilizing, put porcelain dish cooling, wheat bran gauge control is in 2.5cm, excellent aspergillus seed inoculum size is that 0.5% (bacteria containing amount is 1～2 × 10
9individual/g), culture temperature is 28 DEG C, carries out aerlbic culture, fermentation time is 48h.By solid state fermentation wheat bran, add 90mL water ratio in 40 DEG C of water-baths after lixiviate 60min in 15g, in 4 DEG C, centrifugal 20 min of 12 000 r/min, get supernatant liquor enzyme liquid and are placed in refrigerator and preserve.
Get supernatant liquor 3mL and enter 10mL centrifuge tube and add 3mL methanol mixed, 10000 r/min centrifuging and taking supernatant liquors enter sample injection bottle Refrigerator store with 0.45 μ m membrane filtration, measure and use as liquid phase.Sample does blank, and to be that bran mass is azymous according to said method process and get final product.HPLC adopts Venusil XBP C
18(L) (4.6mm × 250mm, m), guard column is C to 5 μ
18(4.6 × 7.5 mm, 5 μ m) condition are: 10 μ L sample sizes, moving phase is 1% glacial acetic acid: acetonitrile (4:1), wavelength 320nm, 20 DEG C of column temperatures, flow velocity is 0.9mLmin
-1.Get 1.0mg forulic acid standard substance, through methanol constant volume to 10mL, be mixed with respectively the solution that concentration is 40,60,80,100,120,140 μ g/mL, measure (seeing that Fig. 5 is certain concentration forulic acid standard substance color atlas) through above-mentioned chromatographic condition, and be X-coordinate (x) by forulic acid concentration, peak area is ordinate zou (Y), finally obtains regression equation to be: Y=311134x+202717, R
2=0.9969 (seeing that Fig. 6 is the typical curve of forulic acid standard substance), then the Y value (see figure 7) of the forulic acid of measuring per sample, by Y value substitution regression equation Y=311134x+202717, obtain x value, is sample forulic acid concentration.Utilize the bound form forulic acid burst size of liquid chromatogram measuring to reach 0.3-0.5%, and in pertinent literature, the content of bound form forulic acid is 0.3%-0.7%.Prove to have obtained great release by bound form forulic acid after fermenting.This bacterial strain is high yield feruloyl esterase and related enzyme systems the bacterial strain that can obtain by the method for fermentation bound form forulic acid from plant.
Sequence table 1
SEQUENCE LISTING
<110> South China Science & Engineering University
Excellent aspergillus and application thereof that feruloyl esterase is produced in <120> mono-strain
<130>
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 594
<212> DNA
<213> rod aspergillus (Aspergillus clavatus) cigarette green grass or young crops
<400> 1
tttccgtagg tgaacctgcg gaaggatcat taccgagtgc gggccctctg ggtccaacct 60
cccacccgtg tttatcgtac cttgttgctt cggcgggccc gccgtcttcg gacggccgcc 120
ggggaggcct ccgcgccccc gggcccgcgc ccgccgaaga ccacaacatg aactctgttc 180
tgaagttttg cagtctgagt tgattatcat aatcagttaa aactttcaac aacggatctc 240
ttggttccgg catcgatgaa gaacgcagcg aaatgcgata actaatgtga attgcagaat 300
tcagtgaatc atcgagtctt tgaacgcaca ttgcgccccc tggtattccg gggggcatgc 360
ctgtccgagc gtcattgctg ccctcaagca cggcttgtgt gttgggcccc cgtccccggt 420
ttcccgggga cgggcccgaa aggcagcggc ggcaccgcgt ccggtcctcg agcgtatggg 480
gctttgtcac ccgctcttgt agggccggcc ggcgcctgtc gacaccaacc caatttttct 540
aaggttgacc tcggatcagg tagggatacc cgctgaactt aagcatatca ataa 594
Claims (5)
1. the excellent aspergillus of feruloyl esterase is produced in a strain, it is characterized in that, the Classification And Nomenclature of described excellent aspergillus be excellent aspergillus (
aspergillus clavatus) cigarette green grass or young crops, by the center preservation of Chinese Typical Representative culture collection, deposit number is CCTCC NO:M2013641, and preservation date is on December 8th, 2013, and its ITS sequence table is as shown in sequence table 1.
2. described in claim 1, excellent aspergillus is applied in product feruloyl esterase.
3. application according to claim 2, it is characterized in that, comprise the steps: to utilize the direct loop-carrier aspergillus of wheat bran (Aspergillus clavatus) cigarette green grass or young crops, in fermention medium, carry out koji plate fermentation, obtain fermented product extract, utilize the forulic acid burst size of wheat bran in high-efficient liquid phase color spectrometry fermention medium.
4. application according to claim 3, is characterized in that, the condition of described koji plate fermentation is: on koji tray, carry out solid fermentation with wheat bran, wheat bran gauge control is in 2 ~ 3cm, and excellent aspergillus seed inoculum size is 0.5% to be that bacteria containing amount is 1～2 × 10
9individual/g, culture temperature is 25～28 DEG C, carries out aerlbic culture, fermentation time is 48～55h; PH nature.
5. application according to claim 4, is characterized in that, described excellent aspergillus seed is inoculated in triangular flask seed culture medium by excellent aspergillus, cultivates 58 ~ 60h obtain in 28 DEG C ± 1 DEG C.
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| CN103641791A (en) * | 2013-11-15 | 2014-03-19 | 浙江大学 | Cyclopeptide compound clavatustide B, and preparation method and application thereof |
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| CN116396868A (en) * | 2021-11-23 | 2023-07-07 | 北京工商大学 | Microbial agent for preparing high-quality dietary fibers by fermenting peanut shells and method thereof |
| CN116396868B (en) * | 2021-11-23 | 2023-09-22 | 北京工商大学 | Microbial agent for preparing high-quality dietary fibers by fermenting peanut shells and method thereof |
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