CN105670945A - High-hesperidinase-yield aspergillus niger KH005 and application thereof - Google Patents
High-hesperidinase-yield aspergillus niger KH005 and application thereof Download PDFInfo
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- C12R2001/685—Aspergillus niger
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- 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)
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- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
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Abstract
The invention discloses a high-hesperidinase-yield aspergillus niger KH005 and application thereof. The aspergillus niger KH005 with a preservation name being KH005 and a preservation number being CGMCC12101 is preserved in the China General Microbiological Culture Collection Center. The aspergillus niger is used for production of hesperidinase, degradation of hesperidin and production of hesperetin. The aspergillus niger KH005 is high in hesperidinase production efficiency, the activity is up to 2548U, valuable strain resources are provided for hesperidinase production, the produced hesperidinase can be used for degradation of hesperidin and production of hesperetin, and high conversion efficiency, mild and environment-friendly conditions and suitableness for industrial production are realized. In addition, the hesperidinase produced by the aspergillus niger KH005 has the advantages of high heat resistance, high enzyme activity and easiness in extraction and storage, thereby providing a new choice for production of high-purity commercial enzyme preparations and having high application and popularization values.
Description
Technical field
The present invention relates to microorganism field, be specifically related to aspergillus niger KH005 and the application thereof of a kind of high yield hesperidinase.
Background technology
Hesperidinase is hydrolysis Hesperidin key enzyme, has significantly high using value. Utilize hesperidinase hydrolysis Hesperidin can obtain hesperetin monoglycosides, hesperetin, rhamnose and glucose. In addition hesperidinase is added in Fructus Citri tangerinae lobe canned food, be hydrolyzed Hesperidin therein, it is possible to prevent mandarin orange can from producing white precipitate.
Hesperidinase can be produced by multiple-microorganism, such as penicillium, aspergillus oryzae, aspergillus niger and Bacteroides etc. Domestic also fewer about the research producing hesperidinase bacterial strain at present, the bacterial strain producing hesperidinase having been reported mostly is mycete.
At present, the research of hesperidinase substantially all rests on laboratory stage at home, can't carry out commercial production; Commercial enzyme preparation relies primarily on import, and price is (1200 yuan/more than kg) also costly. Adopt existing bacterial strain to produce hesperidinase cost higher, be extremely difficult to the requirement of industrialized production. The strain enzyme-producing efficiency having been reported is all not high, and incubation time is generally all at 5-7 days; And the heat stability of enzyme is poor, when processing about 30min in the water-bath more than 45 DEG C, enzyme is dexterous occurs in that obvious decline. Therefore, enzyme efficiency height is produced in screening, and the thermally-stabilised good bacterial strain of enzyme is that hesperidinase can industrial applications key.
Summary of the invention
Present invention aim at providing the aspergillus niger KH005 of a kind of high yield hesperidinase, solve existing bacterial strain and produce the problem of poor heat stability of the hesperidinase that the efficiency of hesperidinase is low, produce, reduce the production cost of hesperidinase, to realize industrialized production.
Additionally, the present invention also provides for the application of aspergillus niger KH005.
The present invention is achieved through the following technical solutions:
The aspergillus niger KH005 of a kind of high yield hesperidinase, described aspergillus niger KH005 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation name is called KH005, and its preserving number is: CGMCC12101.
The preservation information of the aspergillus niger KH005 relating to preservation in the present invention is as follows: depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center; Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica; Preservation date: on January 25th, 2016; Deposit number: CGMCC12101, preservation name is called KH005; Classification And Nomenclature: Aspergillusniger.
The qualification of KH005 bacterial strain:
1), DNA extraction:
Collect thalline, be dissolved in 5mL Extraction buffer (100mMTris Cl, 100mMNa2EDTA, 200mMNaCl, 2%CTAB, pH8.0) in, 37 DEG C of vibration 45min. Add 0.75mL20%SDS, 65 DEG C of water-bath 1h. 12,000rpm, centrifugal 10min, collects supernatant. The isopyknic phenol of supernatant: chloroform: isoamyl alcohol (25:24:1) extracting 2 times, adds the NaAC (pH5.2) of final concentration 0.3M and the dehydrated alcohol of 2 times of volumes, precipitation at room temperature 1h. 4 DEG C, 12,000rpm, centrifugal 20min, collects precipitation, with 70% ethanol rinse 2 times, is dissolved in 50 μ LTE (10mmTris-Hcl, 1mmNa after drying2EDTA), in, STb gene is obtained.
2), amplification ITS sequence:
With above-mentioned STb gene for template, fungus universal primer ITS1 (SEQIDNo.2:TCCGTAGGTGAACCTGCGG) and ITS4 (SEQIDNo.3:TCCTCCGCTTATTGATATGC) respectively forward primer and reverse primer expand ITS sequence. Amplification reaction system is 50 μ L:10 × Buffer5 μ L, dNTP1 μ L, Taq enzyme 0.5 μ L (2U), each 1 μ L of forward and reverse primer, template DNA 1 μ L, ddH2O40.5 μ L. Amplification reaction condition: 94 DEG C of 4min denaturations; 94 DEG C of 0.5min, 55 DEG C of 1min, 72 DEG C of 0.5min, 30 circulations; 72 DEG C extend 7min.
3), ITS sequence analysis:
The ITS fragment of amplification is served the raw work order-checking in sea, shown in amplified fragments sequencing result SEQIDNo.1; Acquisition homology analysis result is compared as shown in table 1 by the blast search program at American National Biotechnology Information center:
The sequence analysis analysis of table 1 amplified fragments
| Similar strain | Homology |
| Aspergillus sp. (aspergillosis) | 99% |
| Aspergillus niger (aspergillus niger) | 99% |
Further, aspergillus niger KH005 is with Fructus Aurantii Immaturus for raw material, picks out through just screening in the medium and produces the bacterial strain that hesperidinase ability is strong, is obtained through screening of repeatedly transferring in the medium by the bacterial strain just filtered out.
Described immature bitter orange raw material is bought by market and is obtained.
Further, the composition of culture medium is: 2.0g sodium nitrate, 0.05g potassium chloride, 1.0g potassium dihydrogen phosphate, 18.0g Hesperidin, 20.g agar and 1L distilled water.
Further, the original ph of culture medium is 6.0-6.5.
The screening process of aspergillus niger KH005:
1), the preparation of culture medium: 2.0g sodium nitrate, 0.05g potassium chloride, 1.0g potassium dihydrogen phosphate, 18.0g Hesperidin, 20.g agar are dissolved and in 1L distilled water, and adjust pH to be 6.0-6.5 with the hydrochloric acid of 1M, subpackage triangular flask, 121 DEG C of sterilizing 15min, when culture medium is cooled to 50-55 DEG C, it is down flat plate, standby after cooling.
2), the primary dcreening operation of bacterial strain: take 20 grams of Fructus Aurantii Immaturuss under aseptic condition in equipped with, in 200mL sterilized water triangular flask, putting shaken at room temperature 30 minutes on bed. Take bacterium solution by the gradient dilution of 10 times to 10-6Take each dilution factor bacterium solution 1mL and put in sterilized petri dishes, the culture medium of 50 DEG C it is cooled to after pouring sterilizing into, mixing, put into 30 DEG C of incubators after to be solidified to cultivate 5 days, the size observing periphery of bacterial colonies transparent circle carries out primary dcreening operation, and picking has the bacterial strain guarantor of transparent circle to be saved in slant medium after being further purified;Then cultivating 5 days in the inoculation after purification to plating medium, measure the diameter of transparent circle, result is as shown in table 2. Produce to form transparent circle in periphery of bacterial colonies because hesperidinase decomposes Hesperidin, so can tentatively judge that bacterial strain produces the size of hesperidinase ability by the size of transparent circle. The single bacterium colony having transparent circle around choosing colony preserves. The bacterial strain that primary dcreening operation obtains finds to be mycete through morphologic observation.
Table 2 primary dcreening operation obtains colonial morphology and the transparent circle diameter of bacterial strain
3), the multiple sieve of bacterial strain:
Being found after repeatedly switching by the bacterial strain of primary dcreening operation, big portion number bacterial strain colony growth becomes slow gradually, and the transparent circle of formation is also more and more less. Through too much generation screening, it is finally obtained the bacterial strain KH005 that a strain grows rapidly, pigment is few and product hesperidinase ability is stable, is initially identified as mycete according to form.
Further, the colony characteristics of aspergillus niger KH005 is as follows: the bacterium colony of described aspergillus niger is radiation growth in PDA culture medium, and 30 DEG C are cultivated 3 days diameters is 30-40mm; Be just white, after become black heavy fleece shape, around have white haloing, the bacterium colony back side is shallow brown (colonial morphology figure is as shown in Figure 1); Its conidial head is spherical in shape, and microgonidium head is in loose cylindrical arrangement; Conidiophore is directly born in substrate or is born in aerial hyphae, and conidiophore is spherical or subsphaeroidal, brown, and spore surface smooths.
The 4 DEG C of bacterial strain preserved PDA slant mediums are activated 3 days, then with the colony inoculation on Inoculating needle picking inclined-plane on PDA plate culture medium, cultivate 3 days in 30 DEG C, observe colonial morphology, in the form (as shown in Figure 2) of basis of microscopic observation conidium and sporophore of attending to anything else. Use scanning electron microscope QuantaTM450FEG observes shown in the ultrastructure (such as Fig. 3 and Fig. 4) of thalline.
The application of the aspergillus niger KH005 of a kind of high yield hesperidinase, aspergillus niger KH005 is used for producing hesperidinase.
By in KH005 inoculation to fermentation medium, the component of fermentation medium is as shown in table 3, under 30～35 DEG C of conditions, and 150～200rpm, fermentation culture 6～8 days; Then filtering, remove thalline residue, collect and obtain crude enzyme liquid, 4 DEG C of stored refrigerated are standby.
Table 3 fermentation medium component
| Title | NaNO3 | KCl | K2HPO4 | MgSO4.7H2O | FeSO4.7H2O | Hesperidin | Wheat bran |
| Content (g) | 2 | 0.05 | 1 | 0.5 | 0.01 | 10 | 12 |
It has been experienced that, KH005 bacterial strain of the present invention produces the efficiency of hesperidinase compared with existing bacterial strain, and high (application number such as announced is the patent of invention of 200610051963.x, when it adopts liquid to cultivate, the enzyme of hesperidinase is lived as 14000U/g, when adopting solid state rheology, the enzyme of hesperidinase is lived as 6000U/g; And the input amount of raw material is relatively big, it is unfavorable for industrialized production), and the Heat stability is good of the hesperidinase produced, the input amount of raw material is few, being easier to realize industrialized production, the efficiency producing hesperidinase is learnt by measuring hesperidinase activity in above-mentioned crude enzyme liquid, and concrete mensuration process is as follows:
Test tube adds the Acetic acid-sodium acetate buffer of 5mlpH4.5,4.5ml concentration be 0.2% Hesperidin substrate solution, 5min it is incubated under 45 DEG C of water-baths, it is subsequently adding the above-mentioned crude enzyme liquid of 0.5ml, fully shake up, accurate timing, 45 DEG C of reaction 20min, move into rapidly inactivation 10min in boiling water bath again, adopting high performance liquid chromatography to record residue amount of substrate, draw substrate minimizing amount, living thus measuring enzyme.
Computing formula is as follows:
Enzyme activity (U/ml)=(b-a)/(20 × 0.5) of crude enzyme liquid;Wherein, a (μ g) is for remaining substrate content after enzymolysis; B (μ g) is the substrate content before reaction.
Measurement result shows that this crude enzyme liquid hesperidinase activity is 2218U-2548U.
The calculation lived for enzyme mainly includes two kinds: one is to calculate U/ml according to liquid volume; Another kind is the Mass Calculation U/g according to thrown raw material, enzyme work of the present invention enzyme work in 3-4 days after optimizing (according to liquid volume calculating U/ml) just can reach more than 2000U, if according to the Mass Calculation of thrown raw material, our bacterial strain enzyme work can reach more than 80000U/g.
The culture medium that the present invention adopts is different from the culture medium of issued patents 200610051963.x, and the mass ratio putting into raw material is less, through conversion reality we put into that the quality of raw material feeds intake less than issued patents 1/7. We find in actual production, feed intake too many, and when liquid fermentation, culture medium can very thickness, it is easy to causes that sterilizing is thorough, and produces the ventilation of substantial amounts of aeration, is unfavorable for the enzymatic production of microorganism on the contrary. Our bacterial strain is when inventory is fewer, it is possible to obtain reasonable product enzyme effect, it is easier to realize commercial production.
The enzyme of hesperidinase is lived definition: 45 DEG C, pH4.5 when, the enzyme amount needed for conversion 1 microgram Hesperidin substrate per minute is an enzyme activity unit (U).
The different article of definition that enzyme is lived or patent have certain difference, but the definition lived of our enzyme adopts is a kind of definition relatively generally acknowledged at present. May refer to document (Shen Lijing, Wang Zhao. the preparation of hesperidinase and catalytic Quality Research [D] thereof. Zhejiang Polytechnical University, 2006:63; Meng Na, Wei Shenghua, Zheng Changlong. fermentation of Aspergillus niger produces the process optimization [J] of hesperidinase. biotechnology, 2011,21 (2): 77-80).
Further, the heat stability of the hesperidinase in the crude enzyme liquid of preparation is measured:
Cultivate by hesperidinase being placed under uniform temperature, the enzyme measuring hesperidinase at set intervals is lived, live with the enzyme of initial hesperidinase and be compared to judge the heat stability of hesperidinase, applicant is experimentally confirmed: by the Heat stability is good of the aspergillus niger KH005 of the present invention hesperidinase produced: after reacting 72h under 50-55 DEG C of condition, enzyme work can preserve about 50%; Under 60-65 DEG C of condition, reaction 24h enzyme is lived and still can be preserved about 50%, namely under 50-55 DEG C of condition, the half-life of enzyme is 72h, it is 24h 60-65 DEG C of condition half-life, and the heat stability of existing the produced enzyme of bacterial strain is poor, when processing about 30min in the water-bath more than 45 DEG C, enzyme is dexterous occurs in that obvious decline, substantially inactivates more than 2h enzyme.
The application of the aspergillus niger KH005 of a kind of high yield hesperidinase, aspergillus niger KH005 is used for Hesperidin of degrading.
The application of the aspergillus niger KH005 of a kind of high yield hesperidinase, aspergillus niger KH005 is used for producing hesperetin.
By in KH005 inoculation to fermentation medium, the component of fermentation medium as shown in Table 3 above, under 30～35 DEG C of conditions, 150～200rpm, fermentation culture 6～8 days. Then filtering, remove thalline residue, it is standby that collection obtains crude enzyme liquid.
Crude enzyme liquid adds the Hesperidin of 1.5%; Under 45～50 DEG C of conditions, 150～200rpm, after reaction 24h, adopt the amount that the remaining content of hesperidin of high effective liquid chromatography for measuring and hesperetin generate. Result shows that the resolution ratio of Hesperidin reaches more than 82%, the hesperetin content generated reaches more than 5.83g/L, it is fully able to and meets industrial needs, and further demonstrate the activity height of the hesperidinase prepared by KH005 bacterial strain of the present invention, Heat stability is good.
The present invention compared with prior art, has such advantages as and beneficial effect:
1, the aspergillus niger KH005 of a kind of high yield hesperidinase of the present invention; This aspergillus niger KH005 produces the efficiency height of hesperidinase, enzyme lives height, production for hesperidinase provides valuable strain resource, and the hesperidinase of production can be used in degraded Hesperidin, produces hesperetin, and transformation efficiency is high, mild condition environmental protection, is suitable to industrialized production.
2, the aspergillus niger KH005 of a kind of high yield hesperidinase of the present invention and application thereof, the hesperidinase that this aspergillus niger KH005 produces is heat-resist, enzyme lives height, being prone to extract and store, the commercial enzyme preparation of production of high purity of making a living provides new selection, has application and promotional value greatly.
Accompanying drawing explanation
Accompanying drawing described herein is used for providing being further appreciated by the embodiment of the present invention, constitutes the part of the application, is not intended that the restriction to the embodiment of the present invention. In the accompanying drawings:
Fig. 1 is the colonial morphology figure of bacterial strain of the present invention;
Fig. 2 is the aspect graph of the conidium of bacterial strain of the present invention and conidiophore;
Fig. 3 is the ultrastructure figure of bacterial strain conidial head of the present invention;
Fig. 4 is conidial ultrastructure figure of bacterial strain of the present invention.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearly understand, below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, and exemplary embodiment and the explanation thereof of the present invention are only used for explaining the present invention, not as a limitation of the invention.
Embodiment 1:
Aspergillus niger KH005 is used for producing hesperidinase:
Aspergillus niger KH005 is inoculated in fermentation medium, under 30 DEG C of conditions, 150rpm, to ferment 6 days, then filter, remove thalline residue, collect and obtain crude enzyme liquid, 4 DEG C of stored refrigerated are standby; Consisting of of described culture medium: 2.0gNaNO3、0.05gKCl、1.0gK2HPO4、0.5gMgSO4.7H2O、0.01gFeSO4.7H2O, 10.0g Hesperidin, 12.0g wheat bran, 1000mL distilled water.
In the crude enzyme liquid surveyed, the enzyme work of hesperidinase is: 2349U.
The detection of the heat stability of hesperidinase in crude enzyme liquid: cultivated 72 hours at 50 DEG C by the hesperidinase produced, measures enzyme and lives as 1358U; The hesperidinase produced is cultivated 72 hours at 55 DEG C, measures enzyme and live as 1267U; The hesperidinase produced is cultivated 24 hours at 60 DEG C, measures enzyme and live as 1189U; The hesperidinase produced is cultivated 24 hours at 65 DEG C, measures enzyme and live as 1176U.
Embodiment 2:
Aspergillus niger KH005 is used for producing hesperidinase:
Aspergillus niger KH005 is inoculated in fermentation medium, under 35 DEG C of conditions, 200rpm, to ferment 8 days, then filter, remove thalline residue, collect and obtain crude enzyme liquid, 4 DEG C of stored refrigerated are standby; Consisting of of described culture medium: 2.0gNaNO3、0.05gKCl、1.0gK2HPO4、0.5gMgSO4.7H2O、0.01gFeSO4.7H2O, 10.0g Hesperidin, 12.0g wheat bran, 1000mL distilled water.
In the crude enzyme liquid surveyed, the enzyme work of hesperidinase is: 2548U.
The detection of the heat stability of hesperidinase in crude enzyme liquid: cultivated 72 hours at 50 DEG C by the hesperidinase produced, measures enzyme and lives as 1459U; The hesperidinase produced is cultivated 72 hours at 55 DEG C, measures enzyme and live as 1320U; The hesperidinase produced is cultivated 24 hours at 60 DEG C, measures enzyme and live as 1278U; The hesperidinase produced is cultivated 24 hours at 65 DEG C, measures enzyme and live as 1288U.
Embodiment 3:
Aspergillus niger KH005 is used for producing hesperidinase:
Aspergillus niger KH005 is inoculated in fermentation medium, under 33 DEG C of conditions, 180rpm, to ferment 7 days, then filter, remove thalline residue, collect and obtain crude enzyme liquid, 4 DEG C of stored refrigerated are standby;Consisting of of described culture medium: 2.0gNaNO3、0.05gKCl、1.0gK2HPO4、0.5gMgSO4.7H2O、0.01gFeSO4.7H2O, 10.0g Hesperidin, 12.0g wheat bran, 1000mL distilled water.
In the crude enzyme liquid surveyed, the enzyme work of hesperidinase is: 2218U.
The detection of the heat stability of hesperidinase in crude enzyme liquid: cultivated 72 hours at 50 DEG C by the hesperidinase produced, measures enzyme and lives as 1105U; The hesperidinase produced is cultivated 72 hours at 55 DEG C, measures enzyme and live as 985U; The hesperidinase produced is cultivated 24 hours at 60 DEG C, measures enzyme and live as 1150U; The hesperidinase produced is cultivated 24 hours at 65 DEG C, measures enzyme and live as 1089U.
Embodiment 4:
Aspergillus niger KH005 is used for degrade Hesperidin, production hesperetin:
1000mL embodiment 1 adds in the crude enzyme liquid of preparation 15g Hesperidin, under 45 DEG C of conditions, 150rpm, stand 24 hours, adopt the amount that the remaining content of hesperidin of high effective liquid chromatography for measuring and hesperetin generate.
High-performance liquid chromatogram determination result: Hesperidin residue 2.69g, the resolution ratio of Hesperidin is 82.1%, and hesperetin content reaches 5.83g/L.
Embodiment 5:
Aspergillus niger KH005 is used for degrade Hesperidin, production hesperetin:
1000mL embodiment 2 adds in the crude enzyme liquid of preparation 15g Hesperidin, under 50 DEG C of conditions, 200rpm, stand 24 hours, adopt the amount that the remaining content of hesperidin of high effective liquid chromatography for measuring and hesperetin generate.
High-performance liquid chromatogram determination result: Hesperidin residue 2.48g, the resolution ratio of Hesperidin is 83.5%, and hesperetin content reaches 6.13g/L.
Embodiment 6:
Aspergillus niger KH005 is used for degrade Hesperidin, production hesperetin:
1000mL embodiment 3 adds in the crude enzyme liquid of preparation 15g Hesperidin, under 48 DEG C of conditions, 180rpm, stand 24 hours, adopt the amount that the remaining content of hesperidin of high effective liquid chromatography for measuring and hesperetin generate.
High-performance liquid chromatogram determination result: Hesperidin residue 2.39g, the resolution ratio of Hesperidin is 84.1%, and hesperetin content reaches 5.98g/L.
By embodiment 1 to embodiment 3: the efficiency that the enzyme of the hesperidinase that aspergillus niger KH005 of the present invention produces height alive and aspergillus niger KH005 produce hesperidinase is high; And the Heat stability is good of the hesperidinase of aspergillus niger KH005 production.
By embodiment 4-embodiment 6: the efficiency ability high, production hesperetin of the hesperidinase degraded Hesperidin that aspergillus niger KH005 of the present invention produces is strong, is suitable to commercial production.
Above-described detailed description of the invention; the purpose of the present invention, technical scheme and beneficial effect have been further described; it is it should be understood that; the foregoing is only the specific embodiment of the present invention; the protection domain being not intended to limit the present invention; all within the spirit and principles in the present invention, any amendment of making, equivalent replacement, improvement etc., should be included within protection scope of the present invention.
Claims (8)
1. the aspergillus niger KH005 of a high yield hesperidinase, it is characterised in that described aspergillus niger KH005 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation name is called KH005, its preserving number is: CGMCC12101.
2. the aspergillus niger KH005 of a kind of high yield hesperidinase according to claim 1, it is characterized in that, described aspergillus niger KH005 is with Fructus Aurantii Immaturus for raw material, pick out through just screening in the medium and produce the bacterial strain that hesperidinase ability is strong, the bacterial strain just filtered out is obtained through screening of repeatedly transferring in the medium.
3. the aspergillus niger KH005 of a kind of high yield hesperidinase according to claim 2, it is characterised in that the composition of described culture medium is: 2.0g sodium nitrate, 0.05g potassium chloride, 1.0g potassium dihydrogen phosphate, 18.0g Hesperidin, 20.g agar and 1L distilled water.
4. the aspergillus niger KH005 of a kind of high yield hesperidinase according to claim 2, it is characterised in that the original ph of described culture medium is 6.0-6.5.
5. the aspergillus niger KH005 of a kind of high yield hesperidinase according to claim 1, it is characterized in that, the colony characteristics of described aspergillus niger KH005 is as follows: the bacterium colony of described aspergillus niger KH005 radiation growth in PDA culture medium, and 30 DEG C are cultivated 3 days diameters is 30-40mm; Be just white, after become black heavy fleece shape, around have white haloing, the bacterium colony back side is shallow brown; Its conidial head is spherical in shape, and microgonidium head is in loose cylindrical arrangement; Conidiophore is directly born in substrate or is born in aerial hyphae, and conidiophore is spherical or subsphaeroidal, brown, and spore surface smooths.
6. the application of the aspergillus niger KH005 of a kind of high yield hesperidinase as described in as arbitrary in claim 1 to 5, it is characterised in that described aspergillus niger KH005 is used for producing hesperidinase.
7. the application of the aspergillus niger KH005 of a kind of high yield hesperidinase as described in as arbitrary in claim 1 to 5, it is characterised in that described aspergillus niger KH005 is used for Hesperidin of degrading.
8. the application of the aspergillus niger KH005 of a kind of high yield hesperidinase as described in as arbitrary in claim 1 to 5, it is characterised in that described aspergillus niger KH005 is used for producing hesperetin.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110438015A (en) * | 2019-09-04 | 2019-11-12 | 桂林理工大学 | The method that the dried immature fruit of citron orange endogenetic fungus and its fermentation for producing hesperidinase produce hesperidinase |
| CN110438015B (en) * | 2019-09-04 | 2022-09-16 | 桂林理工大学 | Immature bitter orange endophytic fungus for producing hesperidinase and method for producing hesperidinase by fermenting immature bitter orange endophytic fungus |
| CN113980821A (en) * | 2021-11-15 | 2022-01-28 | 广东省农业科学院蚕业与农产品加工研究所 | Aspergillus niger capable of transforming hesperidin and application thereof |
| CN113980821B (en) * | 2021-11-15 | 2023-11-10 | 广东省农业科学院蚕业与农产品加工研究所 | Aspergillus niger capable of converting hesperidin and application thereof |
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| CN105670945B (en) | 2019-04-23 |
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