CN108220258A - A kind of preparation method of the Stropharia rugoso-annulata activated protein with laccase activity - Google Patents
A kind of preparation method of the Stropharia rugoso-annulata activated protein with laccase activity Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of the Stropharia rugoso-annulata activated protein with laccase activity.The activated protein of the present invention is prepared using Stropharia rugoso-annulata fructification as raw material by techniques such as crushing, Ultrasonic Wave-Assisted Extraction, ammonium sulfate precipitation, ultrafiltration concentration, ion-exchange chromatography, gel permeation chromatography, vacuum freeze dryings.The relative molecular mass of the Stropharia rugoso-annulata activated protein of the present invention is 40KD, has proliferation inhibition activity to breast cancer cell MCF7 in vitro.
Description
Technical field
The invention belongs to biotechnologies, and in particular to a kind of system of the Stropharia rugoso-annulata activated protein with laccase activity
Preparation Method.
Background technology
Stropharia rugoso-annulata（Stropharia rugoso-annulata）Also known as Stropharia rugoso annulata, wrinkle Stropharia rugoso-annulata, claret ball
The false black umbrella of lid mushroom, Fei Shi Stropharia rugoso-annulatas, Fei Shi, belongs to Basidiomycota, Hymenomycetes, Agaricales, Strophariaceae, Stropharia.1922
Year, Stropharia rugoso-annulata is found and names for the first time in the U.S., also finds its distribution, wild big spherical cap in succession in Europe, Asia thereafter
Mushroom is distributed mainly on the ground such as Yunnan, Jilin, Tibet in China.In the 1960s, Germany carries out artificial domesticating cultivation earliest, I
State starts introducing and planting the nineties.
Stropharia rugoso-annulata is a kind of sharp and clear edible and medicinal fungi beautiful in colour, soft, and fructification, which contains, health
The substances such as protein, carbohydrate, minerals, the vitamin of benefit, for amino acid content up to 17 kinds, essential amino acid is complete, is one
The free of contamination safety food of kind.Compared with other culturing edible fungus, Stropharia rugoso-annulata is with planting material is wide, cultivation technique is easy, suitable
The advantages such as kind season length are that one of ten big mushroom classes of international mushroom class trade market and FAO (Food and Agriculture Organization of the United Nation) are Chinese to development
One of new mushroom species of cultivation are recommended by family.
Laccase（Laccase, EC 1.10.3.2）It is the polyphenol oxidase of a kind of cupric, in 1883 by Japanese scholars
Yoshida has found for the first time in japanese lacquer tree paint solution, belongs to covellite oxidation enzyme family.Over more than 100 years, people carry out laccase
Numerous studies, it is found that it is widely existed in a variety of fungies and plant.Ma Qianqian etc. is from rainbow conk（Coriolus
versicolor）A kind of laccase, molecular weight 67kDa are separated in zymotic fluid, N-terminal amino acid sequence is
GIGPVADLTITNAAV, the enzyme have inhibiting effect, IC to breast cancer MCF7 cells and hepatoma Hep G 2 cells50Respectively 2.3 μM
With 4.4 μM.Hu etc. is from agrocybe（Agrocybe cylindracea）A kind of laccase is separated in fructification, molecular weight is
58kDa, N-terminal amino acid sequence are SDAQKPFVNL, which has breast cancer MCF7 cells and liver cancer HePG2 cells inhibition to make
With IC50Respectively 6.5 μM and 5.6 μM.Zhao etc. is from coprinus comatus（Coprinus comatus）It is separated in mycelium fermentation broth
A kind of laccase, molecular weight 64kDa, N-terminal amino acid sequence are AIGPVADLKV, and the enzyme is to breast cancer MCF7 cells and liver cancer
HePG2 cells have inhibiting effect, IC50Respectively 4.95 μM and 3.46 μM.
Stropharia rugoso-annulata has wide potentiality to be exploited and market prospects as a kind of emerging rare high-quality edible mushroom.But
It is that since China's introducing and planting is later, domestic related research is less, and existing research is concentrated mainly on breeding, cultivation etc., right
The report of its active constituent is less.Therefore, in Stropharia rugoso-annulata activated protein and its function discussion and research, for its protein
The development and utilization of resource is of great significance.
Invention content
The purpose of the present invention is to provide a kind of preparation methods of the Stropharia rugoso-annulata activated protein with laccase activity.
The activated protein of the present invention assists flooding, sulphur using Stropharia rugoso-annulata fructification as raw material, by crushing, ultrasonic wave
The techniques such as acid ammonium salt analysis, ultrafiltration concentration, ion-exchange chromatography, gel permeation chromatography, vacuum freeze drying are prepared.The present invention
Stropharia rugoso-annulata activated protein relative molecular mass be 40KD.
Stropharia rugoso-annulata activated protein of the present invention is prepared as follows：
（1）Dry Stropharia rugoso-annulata fructification is crushed, crosses 20 ~ 100 mesh sieve, i.e. fructification coarse powder；
（2）Fructification coarse powder is pressed 1:20~1:40（W/V）Ratio, be dissolved in 0.9% sodium chloride solution, be placed in ultrasonic wave and carry
It takes in device, ultrasonic frequency control centrifuges 10 ~ 30min, receive in 200W-400W, 2 ~ 10 DEG C of processing 4 ~ 8h, 6000 ~ 10000rpm
Collect supernatant, i.e. fructification leaching liquor；
（3）Into fructification leaching liquor add in ammonium sulfate to saturation degree for 60 ~ 80%, 2 ~ 10 DEG C processing 4 ~ 8h, 6000 ~
10000rpm centrifuges 10 ~ 30min, collects precipitation, i.e. protein crude extract administration；
（4）Protein crude extract administration is pressed 1:5~1:15（W/V）Ratio be dissolved in deionized water, 2 ~ 10 DEG C, 3000 ~ 4000rpm
20 ~ 40min of centrifugal concentrating, repeatedly 2 ~ 4 times, i.e. protein concentrated solution；
（5）Protein concentrated solution is pressed 1:10~1:20（W/V）Ratio, be dissolved in 10 ~ 100mM Tris, pH 7.0 ~ 9.0
In Tris A buffer solutions, carry out anion exchange chromatography, with 10 ~ 100mM Tris, 1M NaCl, pH 7.0 ~ 9.0 Tris
B buffer solution for gradient elution collects the eluting peak with laccase activity；
（6）Eluting peak is subjected to gel permeation chromatography, with 10 ~ 100mM Tris, 100 ~ 500mM NaCl, pH7.0 ~ 9.0
Tris C buffer solutions elute, and collect the eluting peak with laccase activity；
（7）Eluting peak under -20 DEG C ~ -40 DEG C, the vacuum of 8 ~ 12 pas is lyophilized, obtains the big spherical cap with laccase activity
Mushroom activated protein.
The Stropharia rugoso-annulata activated protein with laccase activity of the present invention can inhibit breast cancer cell MCF7, lung in vitro
Cancer cell A549, hepatocellular carcinoma H22 proliferation, wherein the antiproliferative activity to breast cancer cell MCF7 is stronger, antitumor work
Property is related to inducing cell apoptosis.
The present invention has the advantage that and advantageous effect：
For the present invention using Stropharia rugoso-annulata fructification as material, the albumen with laccase activity that therefrom separation obtains is natural extraction
Object, Product Safety are good；Activated protein has antitumor activity, can be applied to health food and drug.
Description of the drawings
Fig. 1 is the ion-exchange chromatography figure of the Stropharia rugoso-annulata activated protein of the present invention.
Fig. 2 is the gel permeation chromatography figure of the Stropharia rugoso-annulata activated protein of the present invention.
Fig. 3 is the SDS-PAGE electrophoresis of the Stropharia rugoso-annulata activated protein of the present invention.
Specific embodiment
With reference to specific embodiment, the present invention is further illustrated.
The preparation of 1 Stropharia rugoso-annulata activated protein of embodiment
（1）Dry Stropharia rugoso-annulata fructification is crushed, crosses 20 mesh sieve, i.e. fructification coarse powder；
（2）Fructification coarse powder is pressed 1:20（W/V）Ratio, be dissolved in 0.9% sodium chloride solution, be placed in ultrasonic extractor
In, ultrasonic frequency control collects supernatant, i.e. fructification leaching liquor in 200W, 2 DEG C of processing 8h, 6000rpm centrifugation 30min；
（3）It is 60% that ammonium sulfate to saturation degree is added in into fructification leaching liquor, and 2 DEG C of processing 8h, 6000rpm centrifugation 30min are received
Collection precipitation, i.e. protein crude extract administration；
（4）Protein crude extract administration is pressed 1:5（W/V）Ratio be dissolved in deionized water, 2 DEG C, 3000rpm centrifugal concentrating 40min,
4 times, i.e. protein concentrated solution repeatedly；
（5）Protein concentrated solution is pressed 1:10（W/V）Ratio, be dissolved in 10mM Tris, pH 7.0 Tris A buffer solutions in,
Carry out anion exchange chromatography, with 10mM Tris, 1M NaCl, pH 7.0 Tris B buffer solution for gradient elution, collect tool
There is the eluting peak of laccase activity；
（6）Eluting peak is subjected to gel permeation chromatography, is washed with the Tris C buffer solutions of 10mM Tris, 100mM NaCl, pH7.0
It is de-, collect the eluting peak with laccase activity；
（7）Eluting peak under -20 DEG C, the vacuum of 12 pas is lyophilized, obtains the Stropharia rugoso-annulata activity egg with laccase activity
In vain.
The preparation of 2 Stropharia rugoso-annulata activated protein of embodiment
（1）Dry Stropharia rugoso-annulata fructification is crushed, crosses 60 mesh sieve, i.e. fructification coarse powder；
（2）Fructification coarse powder is pressed 1:30（W/V）Ratio, be dissolved in 0.9% sodium chloride solution, be placed in ultrasonic extractor
In, ultrasonic frequency control collects supernatant, i.e. fructification leaching liquor in 300W, 6 DEG C of processing 6h, 8000rpm centrifugation 20min；
（3）It is 70% that ammonium sulfate to saturation degree is added in into fructification leaching liquor, and 6 DEG C of processing 6h, 8000rpm centrifugation 20min are received
Collection precipitation, i.e. protein crude extract administration；
（4）Protein crude extract administration is pressed 1:10（W/V）Ratio be dissolved in deionized water, 6 DEG C, 3500rpm centrifugal concentratings
30min, repeatedly 3 times, i.e. protein concentrated solution；
（5）Protein concentrated solution is pressed 1:15（W/V）Ratio, be dissolved in 50mM Tris, pH 8.0 Tris A buffer solutions in,
Carry out anion exchange chromatography, with 50mM Tris, 1M NaCl, pH 8.0 Tris B buffer solution for gradient elution, collect tool
There is the eluting peak of laccase activity；
（6）Eluting peak is subjected to gel permeation chromatography, is washed with the Tris C buffer solutions of 50mM Tris, 300mM NaCl, pH8.0
It is de-, collect the eluting peak with laccase activity；
（7）Eluting peak under -30 DEG C, the vacuum of 10 pas is lyophilized, obtains the Stropharia rugoso-annulata activity egg with laccase activity
In vain.
The preparation of 3 Stropharia rugoso-annulata activated protein of embodiment
（1）Dry Stropharia rugoso-annulata fructification is crushed, is sieved with 100 mesh sieve, i.e. fructification coarse powder；
（2）Fructification coarse powder is pressed 1:40（W/V）Ratio, be dissolved in 0.9% sodium chloride solution, be placed in ultrasonic extractor
In, ultrasonic frequency control collects supernatant, i.e. fructification extracts in 400W, 10 DEG C of processing 4h, 10000rpm centrifugation 10min
Liquid；
（3）It is 80% that ammonium sulfate to saturation degree is added in into fructification leaching liquor, 10 DEG C of processing 4h, 10000rpm centrifugation 10min,
Collect precipitation, i.e. protein crude extract administration；
（4）Protein crude extract administration is pressed 1:15（W/V）Ratio be dissolved in deionized water, 10 DEG C, 4000rpm centrifugal concentratings
20min, repeatedly 2 times, i.e. protein concentrated solution；
（5）Protein concentrated solution is pressed 1:20（W/V）Ratio, be dissolved in 100mM Tris, pH 9.0 Tris A buffer solutions in,
Carry out anion exchange chromatography, with 100mM Tris, 1M NaCl, pH 9.0 Tris B buffer solution for gradient elution, collect tool
There is the eluting peak of laccase activity；
（6）Eluting peak is subjected to gel permeation chromatography, is washed with the Tris C buffer solutions of 100mM Tris, 500mM NaCl, pH9.0
It is de-, collect the eluting peak with laccase activity；
（7）Eluting peak under -40 DEG C, the vacuum of 8 pas is lyophilized, obtains the Stropharia rugoso-annulata activity egg with laccase activity
In vain.
The laccase activity of 4 Stropharia rugoso-annulata activated protein of embodiment
3mL reaction systems contain one sodium acetate of 0.2mL 3mmol/L ABTS, 1.8mL 0.1mol/L acetic acid（pH 4.5）Buffering
Liquid and 1mL samples to be tested, mixing, 25 DEG C of reaction 5min measure light absorption value at 420nm wavelength.Enzyme activity is defined as：1 enzyme activity
Unit（U）Refer under certain condition, the required enzyme amount of 1 μm of ol ABTS oxidation of catalysis per minute in reaction system.
Laccase can be catalyzed ABTS and aoxidize, and the oxidized form ABTS of generation has reduced form ABTS not had at 420 nm
Strong absworption peak, pass through whereby the light absorption value that detects 420 nm places increase measure the vigor of laccase.As shown in Table 1, have
The Stropharia rugoso-annulata activated protein of laccase activity is isolated and purified through ammonium sulfate precipitation, ion-exchange chromatography, gel permeation chromatography, purifying
Multiple is 15.83, the rate of recovery 25.90%.
1 Stropharia rugoso-annulata activated protein of table isolates and purifies
The antitumor activity of 5 Stropharia rugoso-annulata activated protein of embodiment
（1）Breast cancer cell MCF7, lung cell A549, hepatocellular carcinoma H22 are pressed 1 × 105The concentration of cfu/mL is inoculated in
96 orifice plates, per 100 μ L of hole, 37 DEG C are cultivated 24 hours；
（2）10 μ L various concentrations are added in per hole（1.0μM、2.0μM、4.0μM、8.0μM、16.0μM）Stropharia rugoso-annulata activity egg
In vain, it cultivates 48 hours for 37 DEG C；
（3）25 μ L trichloroacetic acids are added in per hole（500mg/mL）, 4 DEG C are placed 1 hour；
（4）It distills water washing 5 times, adds in 4mg/mL SRB per hole, dye 30min；
（5）Acetic acid washs 5 times, and 100 μ L Tris buffer solutions are added in per hole（10mmol/L）, measure light absorption value at 490nm.
503nhibiting concentration（IC50）Be evaluate tumor cell proliferation inhibition activity common counter, Stropharia rugoso-annulata activated protein
To breast cancer cell MCF7, lung cell A549, hepatocellular carcinoma H22 IC50Respectively 4.65 μM, 11.28 μM, 8.74 μM,
Show the Stropharia rugoso-annulata activated protein of the present invention has stronger antiproliferative activity to breast cancer cell MCF7.
6 Stropharia rugoso-annulata activated protein inducing cell apoptosis of embodiment
（1）Breast cancer cell MCF7 is pressed 1 × 105The concentration of cfu/mL is inoculated in 12 orifice plates, and per hole 1mL, 37 DEG C of cultures 24 are small
When；
（2）10 μ L various concentrations are added in per hole（1.0μM、2.0μM、4.0μM、8.0μM）Stropharia rugoso-annulata activated protein, 37 DEG C training
It supports 24 hours；
（3）Collect 1 × 105Cfu cells add in 75% ethyl alcohol of 0.5mL precoolings, and 4 DEG C are fixed 24 hours；
（4）1000rpm centrifuges 5min, collects cell, adds in 0.5mL PI dyeing liquors, 37 DEG C of incubation 30min, flow cytometer inspection
It surveys.
Fluorescent dye PI（Propidium iodide）It is a kind of nuclei dyeing color reagent, it is red glimmering that release can be combined with double-stranded DNA
Light utilizesFlow cytometerThe fluorescence intensity of PI and DNA compounds are detected, calculates the different cell cycles（The G1/G0 phases, the S phases and
The G2/M phases）DNA content, wherein Sub-G1 represents the cell of apoptosis, so as to detect Apoptosis.From table 2 it can be seen that with
The increase of Stropharia rugoso-annulata activated protein concentration, the ratio of apoptotic cell gradually increase, and show that the processing of Stropharia rugoso-annulata activated protein is drawn
MCF7 Apoptosis is played, and in dose-effect relationship.
2 Stropharia rugoso-annulata activated protein inducing cell apoptosis of table
| Cell proportion（%） | The G1/G0 phases | The S phases | The G2/M phases | Sub-G1 |
| 1.0μM | 51.03 | 28.61 | 20.36 | 1.85 |
| 2.0μM | 55.32 | 28.57 | 16.11 | 6.37 |
| 4.0μM | 60.58 | 29.14 | 10.28 | 14.71 |
| 8.0μM | 67.15 | 27.96 | 4.89 | 23.96 |
It these are only the preferred embodiment of the present invention, be not intended to restrict the invention, those skilled in the art is come
It says, any modification, equivalent substitution, improvement and etc. done all within the spirits and principles of the present invention should be included in the present invention
Protection domain within.
Claims (5)
1. a kind of preparation method of the Stropharia rugoso-annulata activated protein with laccase activity, it is characterised in that the Stropharia rugoso-annulata is lived
Property albumen relative molecular mass for 40KD, preparation method includes the following steps：
（1）Dry Stropharia rugoso-annulata fructification is crushed, crosses 20 ~ 100 mesh sieve, i.e. fructification coarse powder；
（2）Fructification coarse powder is pressed 1:20~1:40（W/V）Ratio, be dissolved in 0.9% sodium chloride solution, be placed in ultrasonic wave and carry
It takes in device, ultrasonic frequency control centrifuges 10 ~ 30min, receive in 200W-400W, 2 ~ 10 DEG C of processing 4 ~ 8h, 6000 ~ 10000rpm
Collect supernatant, i.e. fructification leaching liquor；
（3）Into fructification leaching liquor add in ammonium sulfate to saturation degree for 60 ~ 80%, 2 ~ 10 DEG C processing 4 ~ 8h, 6000 ~
10000rpm centrifuges 10 ~ 30min, collects precipitation, i.e. protein crude extract administration；
（4）Protein crude extract administration is pressed 1:5~1:15（W/V）Ratio be dissolved in deionized water, 2 ~ 10 DEG C, 3000 ~ 4000rpm
20 ~ 40min of centrifugal concentrating, repeatedly 2 ~ 4 times, i.e. protein concentrated solution；
（5）Protein concentrated solution is pressed 1:10~1:20（W/V）Ratio, be dissolved in 10 ~ 100mM Tris, pH 7.0 ~ 9.0
In Tris A buffer solutions, carry out anion exchange chromatography, with 10 ~ 100mM Tris, 1M NaCl, pH 7.0 ~ 9.0 Tris
B buffer solution for gradient elution collects the eluting peak with laccase activity；
（6）Eluting peak is subjected to gel permeation chromatography, with 10 ~ 100mM Tris, 100 ~ 500mM NaCl, pH7.0 ~ 9.0
Tris C buffer solutions elute, and collect the eluting peak with laccase activity；
（7）Eluting peak under -20 DEG C ~ -40 DEG C, the vacuum of 8 ~ 12 pas is lyophilized, obtains the big spherical cap with laccase activity
Mushroom activated protein.
2. a kind of Stropharia rugoso-annulata activated protein, it is characterised in that method is prepared according to claim 1.
3. purposes of the Stropharia rugoso-annulata activated protein in the drug for antitumor activity is prepared described in claim 2.
4. according to claim 3 purposes, it is characterised in that the tumour is breast cancer.
5. purposes according to claim 4, it is characterised in that breast cancer is breast cancer cell MCF7.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110790814A (en) * | 2019-11-25 | 2020-02-14 | 天津市林业果树研究所 | Extraction method and application of pholiota nameko protein |
| CN117987279A (en) * | 2024-02-19 | 2024-05-07 | 中国农业科学院农业资源与农业区划研究所 | Composition and culture medium for stropharia rugoso-annulata mother culture and preparation method thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH11315096A (en) * | 1997-08-08 | 1999-11-16 | New Food Creation Gijutsu Kenkyu Kumiai | Antitumoral protein derived from pleurotus cornucopiae and its gene |
| CN101790938A (en) * | 2010-03-02 | 2010-08-04 | 福建省农业科学院农业工程技术研究所 | Method for improving content and extraction rate of flavonoids in fruit bodies of Stropharia rugoso-annulata |
| CN102225995A (en) * | 2011-04-20 | 2011-10-26 | 南京大学 | Single-type melanins and their preparation methods and applications |
| CN102268414A (en) * | 2011-06-16 | 2011-12-07 | 北京市农林科学院 | Coprinus comatus laccase with activity of inhibiting tumor cell proliferation and preparation method thereof |
| CN103102399A (en) * | 2013-03-01 | 2013-05-15 | 南开大学 | Anti-tumor protein extracted from pholiota nameko and preparation method of antineoplastic protein |
| CN104855142A (en) * | 2015-06-12 | 2015-08-26 | 河北大学 | Method for using cattail crumbs to cultivate stropharia rugoso-annulata |
| CN104974991A (en) * | 2015-07-09 | 2015-10-14 | 福建农林大学 | Method for preparing solid laccase through pleurotus eryngii waste fungus chaff |
| CN105062983A (en) * | 2015-08-05 | 2015-11-18 | 北京农学院 | Cerrena unicolor laccase with dye degradation activity and preparation method thereof |
| CN105062984A (en) * | 2015-09-16 | 2015-11-18 | 福建师范大学 | Method for preparing laccase by fungus fermentation |
| US20160000754A1 (en) * | 2004-01-06 | 2016-01-07 | Paul Edward Stamets | Antiviral Activity from Medicinal Mushrooms and their Active Constituents |
-
2016
- 2016-12-15 CN CN201611157373.5A patent/CN108220258B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH11315096A (en) * | 1997-08-08 | 1999-11-16 | New Food Creation Gijutsu Kenkyu Kumiai | Antitumoral protein derived from pleurotus cornucopiae and its gene |
| US20160000754A1 (en) * | 2004-01-06 | 2016-01-07 | Paul Edward Stamets | Antiviral Activity from Medicinal Mushrooms and their Active Constituents |
| CN101790938A (en) * | 2010-03-02 | 2010-08-04 | 福建省农业科学院农业工程技术研究所 | Method for improving content and extraction rate of flavonoids in fruit bodies of Stropharia rugoso-annulata |
| CN102225995A (en) * | 2011-04-20 | 2011-10-26 | 南京大学 | Single-type melanins and their preparation methods and applications |
| CN102268414A (en) * | 2011-06-16 | 2011-12-07 | 北京市农林科学院 | Coprinus comatus laccase with activity of inhibiting tumor cell proliferation and preparation method thereof |
| CN103102399A (en) * | 2013-03-01 | 2013-05-15 | 南开大学 | Anti-tumor protein extracted from pholiota nameko and preparation method of antineoplastic protein |
| CN104855142A (en) * | 2015-06-12 | 2015-08-26 | 河北大学 | Method for using cattail crumbs to cultivate stropharia rugoso-annulata |
| CN104974991A (en) * | 2015-07-09 | 2015-10-14 | 福建农林大学 | Method for preparing solid laccase through pleurotus eryngii waste fungus chaff |
| CN105062983A (en) * | 2015-08-05 | 2015-11-18 | 北京农学院 | Cerrena unicolor laccase with dye degradation activity and preparation method thereof |
| CN105062984A (en) * | 2015-09-16 | 2015-11-18 | 福建师范大学 | Method for preparing laccase by fungus fermentation |
Non-Patent Citations (9)
| Title |
|---|
| DIETMAR等: "Laccase-catalyzed oxidation of Mn(2+) in the presence of natural Mn(3+) chelators as a novel source of extracellular H(2)O(2) production and its impact on manganese peroxidase.", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
| 于萍，等: ""大球盖菇栽培期间胞外酶活性变化研究"", 《中国食用菌》 * |
| 刘淑珍等: "担子菌漆酶的分离纯化及其性质研究", 《微生物学报》 * |
| 叶汉玲等: "新鲜香菇子实体漆酶的纯化与性质", 《南京林业大学学报》 * |
| 张琪林等: "大球盖菇菌丝几种酶的性质研究", 《江苏农业科学》 * |
| 杨娟等: "白灵侧耳漆酶分离纯化及其酶学性质研究", 《菌物学报》 * |
| 王方忠等: "香菇漆酶的纯化及部分性质研究", 《北方园艺》 * |
| 王红等: "大球盖菇液体培养胞外酶初步测定", 《食用菌》 * |
| 马茜茜等: "云芝漆酶的分离纯化和理化性质及抗增殖活性", 《食用菌学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110790814A (en) * | 2019-11-25 | 2020-02-14 | 天津市林业果树研究所 | Extraction method and application of pholiota nameko protein |
| CN117987279A (en) * | 2024-02-19 | 2024-05-07 | 中国农业科学院农业资源与农业区划研究所 | Composition and culture medium for stropharia rugoso-annulata mother culture and preparation method thereof |
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