CN109182141B - A method of improving white-rot fungi producing enzyme enzyme activity - Google Patents
A method of improving white-rot fungi producing enzyme enzyme activity Download PDFInfo
- Publication number
- CN109182141B CN109182141B CN201811119511.XA CN201811119511A CN109182141B CN 109182141 B CN109182141 B CN 109182141B CN 201811119511 A CN201811119511 A CN 201811119511A CN 109182141 B CN109182141 B CN 109182141B
- Authority
- CN
- China
- Prior art keywords
- blue light
- enzyme
- white
- enzyme activity
- fermentation medium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
- 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
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- 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
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
-
- C—CHEMISTRY; METALLURGY
- 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)
- C12N9/24—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)
- C12N9/2405—Glucanases
- C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
- C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
Abstract
The present invention relates to microorganisms technical fields, disclose a kind of method for improving white-rot fungi producing enzyme enzyme activity.White-rot fungi is inoculated into fermentation medium by the method for the invention, is fermented using blue light illumination.The present invention replaces traditional dark surrounds with blue light illumination and carries out producing enzyme fermentation to white-rot fungi, can significantly improve the enzyme activity of institute's cellulase-producing and hemicellulase；This fit on adds copper ion and/or manganese ion also in the fermentation medium can improve the enzyme activity of produced lignin-degrading enzymes simultaneously.
Description
Technical field
The present invention relates to microorganisms technical fields, and in particular to a method of improve white-rot fungi producing enzyme enzyme activity.
Background technique
2017, all kinds of stalk annual outputs were about 1,000,000,000 tons in China's agricultural production, were contained in these agricultural wastes
Renewable resource abundant.It is current for making full use of these processing feedstuff with stalks, fertilizer and bioenergy (ethyl alcohol, biogas etc.)
The research hotspot that straw lignocellulose orientation bioconversion utilizes.
Lignocellulosic main component is cellulose, hemicellulose and lignin.Restrict the high-valued comprehensive utilization of stalk
Main bottleneck is effective degradation of lignocellulosic.Up to the present, lignin degradation method mainly includes physical method, chemistry
Method, physical-chemical process and biological degradation method.Wherein, biodegrade becomes and grinds as environmentally protective lignin processing method
Study carefully hot spot.Biodegrade mainly has microbial degradation method and enzyme edman degradation Edman.But microbial degradation method is long (general there are the time
Need 21-60 days) and growth conditions require the problem of harsh (sterile, to exclude the interference of extraneous miscellaneous bacteria), therefore limit it
Commercial applications in actual production.Therefore, emphasis is transferred in enzyme degradation by researcher.Therefore, it is fine to improve microorganism
Dimension element, hemicellulose and lignin degradation enzymatic activity become research emphasis.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of methods for improving white-rot fungi producing enzyme enzyme activity, so that described
Method can improve the cellulase of white-rot fungi production and the enzyme activity of hemicellulase simultaneously；
Another object of the present invention is that a kind of method for improving white-rot fungi producing enzyme enzyme activity is provided, so that the side
Method can improve the enzyme activity of the cellulase of white-rot fungi production, hemicellulase and lignin-degrading enzymes simultaneously；
To achieve the goals above, the invention provides the following technical scheme:
A method of white-rot fungi producing enzyme enzyme activity is improved, white-rot fungi is inoculated into fermentation medium, using blue light
Irradiation is fermented.
White-rot fungi normal fermentation producing enzyme is in a dark environment, but there is a problem of that enzyme activity is lower.For this problem,
The present invention induces fermented by white rot fungus producing enzyme using the method for blue light illumination, improves institute's cellulase-producing and hemicellulase
Enzyme activity.
Preferably, the white-rot fungi is one in Pleurotus eryngii, oyster mushroom, Phanerochaete chrysosporium and Irpex lacteus
Kind is two or more.Wherein, Pleurotus eryngii, oyster mushroom, Phanerochaete chrysosporium, Irpex lacteus can be common by CICC or China
It is obtained at Microbiological Culture Collection administrative center, preferably, the blue light is 440-470nm blue light, intensity is 50-100 μ
mol m-2s-1, 455nm blue light is selected in the specific embodiment of the invention, intensity is 80 μm of ol m-2s-1。
Preferably, the temperature of the fermentation is 25-30 DEG C；Can choose in the specific implementation process for 25 DEG C, 26 DEG C,
It 27 DEG C, 28 DEG C, 29 DEG C or 30 DEG C, can also be fluctuated within the scope of 25-30 DEG C.
In the specific embodiment of the invention, respectively with Pleurotus eryngii, oyster mushroom, Phanerochaete chrysosporium and Irpex lacteus
Four kinds of representative different white-rot fungis ferment under dark and blue light environment, the results show that the white-rot fungi under blue light illumination
The enzyme activity of institute's cellulase-producing and hemicellulase is obviously improved；Meanwhile present invention employs cellulase-producings and half fiber
The Trichoderma harzianum for tieing up plain enzyme verifies the specificity of blue light illumination effect again, as the result is shown Trichoderma harzianum institute's cellulase-producing and half
The enzyme activity of cellulase is not significantly improved, and dark surrounds and the enzyme activity of blue light illumination are almost consistent, this shows that blue light shines
The measure penetrated is not appropriate for all microbial bacterias.
In addition, the present invention provides preferred embodiment with regard to fermentation medium used in fermentation process, i.e., the described fermented and cultured
Base is the fermentation medium containing ramie stalk；It is further preferred that the fermentation medium include glucose, ramie stalk,
(NH4)2SO4, urea, peptone, KH2PO4、CaCl2、MgSO4、FeSO4、MnSO4、ZnSO4、CoCl2, Tween and water；More preferably
Ground, the fermentation medium are as follows:
Glucose 0.5-2g/L, ramie stalk 15-60g/L, (NH4)2SO41.1-4.4g/L, urea 0.25-1g/L, egg
White peptone 0.5-2g/L, KH2PO4 1-4g/L、CaCl2 0.15-0.6g/L、MgSO4 0.04-0.16g/L、FeSO4 0.0025-
0.01g/L、MnSO4 0.0008-0.0032g/L、ZnSO4 0.0007-0.0028g/L、CoCl2 0.00185-0.0074g/L、
Tween 1-4 drop/L, excess water.
In the specific implementation process, following fermentation medium can arbitrarily be selected:
(1) glucose 0.5g/L, ramie stalk 60g/L, (NH4)2SO41.1g/L, urea 1g/L, peptone 0.5g/L,
KH2PO4 4g/L、CaCl2 0.15g/L、MgSO4 0.16g/L、FeSO4 0.0025g/L、MnSO4 0.0032g/L、ZnSO4
0.0007g/L、CoCl21 drop of 0.0074g/L, Tween/L, excess water；
(2) glucose 1g/L, ramie stalk 30g/L, (NH4)2SO42.2g/L, urea 0.5g/L, peptone 1g/L,
KH2PO42g/L、CaCl2 0.3g/L、MgSO4 0.08g/L、FeSO4 0.005g/L、MnSO4 0.0016g/L、ZnSO4
0.0014g/L、CoCl22 drops of 0.0037g/L, Tween/L, excess water；
(3) glucose 2g/L, ramie stalk 15g/L, (NH4)2SO44.4g/L, urea 0.25g/L, peptone 2g/L,
KH2PO4 1g/L、CaCl2 0.6g/L、MgSO4 0.04g/L、FeSO4 0.01g/L、MnSO4 0.0008g/L、ZnSO4
0.0028g/L、CoCl24 drops of 0.00185g/L, Tween/L, excess water；
(4) glucose 1.5g/L, ramie stalk 45g/L, (NH4)2SO43g/L, urea 0.5g/L, peptone 1.5g/L,
KH2PO4 3g/L、CaCl2 0.2g/L、MgSO4 0.1g/L、FeSO4 0.006g/L、MnSO4 0.0025g/L、ZnSO4
0.001g/L、CoCl23 drops of 0.0025g/L, Tween/L, excess water.
On preferred fermentation medium provided by the invention, the enzymatic production under conventional black environment, each enzyme obtained
Enzyme activity is substantially higher the normal fermentation culture medium in existing patent, then cooperates blue light illumination, can obtain and more preferably improve effect
Fruit.
In addition, the method for the invention further includes that the group of copper ion or manganese ion or both is added in the fermentation medium
It closes, the enzyme activity of lignin-degrading enzymes thus can also be significantly improved on the basis of aforementioned techniques effect；Wherein, the copper ion
Final concentration of 0.1mM-0.5mM, the final concentration of 0.1mM-0.5mM of the manganese ion；In the specific embodiment of the invention, institute
State the final concentration of 0.2mM of copper ion, the final concentration of 0.2mM of the manganese ion.Wherein, copper ion can pass through addition
CuSO4It realizes, manganese ion passes through addition MnSO4To realize.
According to the technical effect of above-mentioned various aspects, the invention also provides blue light illuminations or blue light illumination joint, and copper is added
Application of the combined fermentation medium of ion or manganese ion or both in fermented by white rot fungus producing enzyme.
From the above technical scheme, the present invention replaces traditional dark surrounds with blue light illumination and produces to white-rot fungi
Enzyme fermentation can significantly improve the enzyme activity of institute's cellulase-producing and hemicellulase；This fit on is added in the fermentation medium
Copper ion and/or manganese ion can also improve the enzyme activity of produced lignin-degrading enzymes simultaneously.
Specific embodiment
The invention discloses a kind of method for improving white-rot fungi producing enzyme enzyme activity, those skilled in the art can be used for reference herein
Content is suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications are to those skilled in the art
It is it will be apparent that they are considered as being included in the present invention for member.The method of the invention has passed through preferred embodiment
It is described, related personnel can obviously not depart from the content of present invention, carried out in spirit and scope to method described herein
Change or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
Before white-rot fungi inoculation fermentation culture medium, it generally can be first seeded to seed culture medium activation, seed liquor is made
Inoculation, such as glucose can be added as seed culture medium using PDA culture medium, it is as follows:
Configuration PDA culture medium (takes peeled potatoes 200g, is cut into small pieces, 1000mL boiling is added to boil 20min.Filter off Ma Ling
Potato wedge, and filtrate is complemented into 1000mL.Glucose 20g is added, is dispensed after dissolving, 115 DEG C, sterilize 30min), it is inoculated with white rot
Fungi slant strains (20-30mm2Every piece, it is inoculated with 2-3 block, 150-170r/min is cultivated 1-2 days on 25-30 DEG C of shaking table.
The present invention also provides a kind of seed culture medium, i.e. glucose 0.5-2g/L, corn flour 15-60g/L, (NH4)2SO4
1.1-4.4g/L, urea 0.25-1g/L, peptone 0.5-2g/L, KH2PO4 1-4g/L、CaCl2 0.15-0.6g/L、MgSO4
0.04-0.16g/L、FeSO4 0.0025-0.01g/L、MnSO4 0.0008-0.0032g/L、ZnSO4 0.0007-0.0028g/
L、CoCl20.00185-0.0074g/L, Tween 1-4 drop/L, excess water；Then White-Rot Fungi slant strains (20-
30mm2Every piece, it is inoculated with 2-3 block, 150-170r/min is cultivated 1-2 days on 25-30 DEG C of shaking table.
Prepared seed liquor is seeded to fermentation medium enzymatic production with the inoculum concentration of 10-20%.
In every comparative test of the invention, each group is removed outside due difference, remaining experimental enviroment and material keep one
It causes.
Enzyme activity determination method of the present invention is using following three kinds of methods detection cellulase, hemicellulase and lignin drop
Solve the enzyme activity of enzyme:
(1) CMC cellulase activity measuring method
By CMC as in 5.0 citric acid sodium citrate buffer solution of pH, it is configured to 0.5% (w/v) reaction substrate solution.It takes
0.5mL crude enzyme liquid is added after 1.5mL substrate mixes in 50 DEG C of reaction 1h.Addition 3mL DNS solution boils after mixing after reaction
Water-bath 15min surveys its light absorption value after chromogenic reaction.
It is an enzyme-activity unit that CMC enzyme activity, which is defined as every 1min enzyme amount needed for releasing 1 μm of ol reduced sugar in substrate,.
(2) xylanase activity amylograph
The diluted enzyme solution of 1mL is taken, 5mL buffer solution is added, boiling boils 10min (inactivation zytase) and substrate wood is added afterwards
Glycan solution.Processing after a certain period of time, is added DNS colour developing 8min, obtains reference liquid.Xylan solution 1mL is separately taken to try in scale
Guan Zhong is added the buffer solution 5mL of different pH value, balances 5min in the water-bath of certain temperature.Xylan after dilution is added
Enzyme solutions 1mL shakes up and starts timing.Enzymatic treatment for a period of time after, be rapidly added DNS reagent 5mL, and scale test tube is put into
Develop the color 8min in boiling water bath, cooling, is settled to 25mL.Using reference liquid as blank, measured at 540nm using spectrophotometer molten
Liquid absorbance.If absorbance is excessive, tested after suitably dilution can be carried out.1mL enzyme is under certain temperature and pH value condition, 1h
It decomposes xylan and generates 1mgD (+)-xylose, be considered as an enzyme activity unit U.
C=(A-n)/m；
U=(C × 0.025L)/(t × 1mL)；
In formula: the mass concentration (mg/L) of C- generation xylose；
A- solution absorbance；
U- xylanase activity；
Constant volume after 0.025L- colour developing；
The t- enzyme reaction time (h)；
1mL- zytase volume；
(3) laccase activity measuring method:
ABTS is aoxidized using Lac, measures 0-3min internal absorbance value at 420nm with visible-ultraviolet specrophotometer, at any time
Between variation, to learn enzyme reaction speed, and then calculate enzyme activity.With isometric-two (3- ethyl of 0.5mmol/L2,2- azine
Benzothiazole -6- sulfonic acid) (ABTs) solution and enzyme solution reaction, measurement react before in 3min at 420nm light absorption value increase, every point
1 unit of activity (U) of enzyme amount needed for clock makes 1umolABTS conversion.
Reagent needed for enzyme activity determination: 0.5mmol/LABTS；0.2mol/L NaAc_HAc buffer solution (pH=5.0) with
And a certain amount of crude enzyme liquid；
Enzyme activity determination step:
Blank sample (4mL): 2mL0.2mol/L buffer -0.4mLABTS-1.6mL deionized water；
Sample (4mL): 2mL0.2mol/L buffer -0.4mLABTS-1.2mL deionized water -0.4mL enzyme solution is with ultraviolet point
Light photometer detects absorption value variation at 420nm, and 30 seconds interval readings record absorption value variation in 210 seconds.
Enzyme activity calculation formula is
Just a kind of method for improving white-rot fungi producing enzyme enzyme activity provided by the present invention is described further below.
Embodiment 1: the method for the invention (while improving cellulase and hemicellulase enzyme activity)
White-rot fungi (Pleurotus eryngii, oyster mushroom, Phanerochaete chrysosporium or Irpex lacteus) seed liquor is prepared, according to 10%,
15% or 20% inoculum concentration is seeded in fermentation medium, and cultivation temperature is 25-30 DEG C, is placed in 455nm, 80 μm of ol m-2s-1
Enzymatic production under blue light.
Fermentation medium: glucose 1g/L, ramie stalk 30g/L, (NH4)2SO42.2g/L, urea 0.5g/L, peptone
1g/L、KH2PO4 2g/L、CaCl2 0.3g/L、MgSO4 0.08g/L、FeSO4 0.005g/L、MnSO4 0.0016g/L、
ZnSO4 0.0014g/L、CoCl22 drops of 0.0037g/L, Tween/L, excess water；
Embodiment 2: the enzyme activity comparative test of blue light and dark surrounds
According to the method for embodiment 1, it is inoculated with Pleurotus eryngii, oyster mushroom, Phanerochaete chrysosporium and Irpex lacteus kind respectively
Sub- liquid is placed in blue light or lower culture 7-28 days dark, periodically to cellulase, hemicellulase and lignin degradation enzyme activity
Property is detected, and preparation method, inoculum concentration, the fermented and cultured temperature of seed liquor are consistent, and the results are shown in Table 1 and table 2；
Table 1
Table 2
By Tables 1 and 2, it is apparent that under blue light environment, the cellulase and hemicellulose of each white-rot fungi
Enzyme enzyme activity is improved significantly.In addition, the present invention also changes blue light wavelength and intensity carries out above-mentioned experiment, the results show that respectively
The cellulase and hemicellulase enzyme activity of white-rot fungi are improved significantly, relative to dark surrounds raising degree with
Tables 1 and 2 result is consistent, and each group blue light wavelength and intensity are as follows:
440nm、50μmol m-2s-1Blue light；
460nm、70μmol m-2s-1Blue light；
470nm、100μmol m-2s-1Blue light.
Embodiment 3: the enzyme activity comparative test under Trichoderma harzianum blue light and dark surrounds
Entirely by reference to the test method and experimental enviroment of embodiment 2, difference is only that strain is changed to Trichoderma harzianum, ferments
Incubation time is 7 days, counts cellulose enzyme activity, the results are shown in Table 3；
Table 3
| Cellulose enzyme activity (CMC cellulase activity measuring method) (IU/mL) | |
| It is dark | 1.023 |
| Blue light | 0.963 |
As can be seen from Table 3, no matter in dark surrounds or blue light environment, the cellulose enzyme activity of Trichoderma harzianum is not bright
Aobvious to improve, the two is almost the same, shows and not all microbes producing cellulase can improve cellulose enzyme activity under blue light environment.
Embodiment 4: the method for the invention (while improving cellulase, hemicellulase and lignin-degrading enzymes enzyme activity)
And compliance test result
1, method
CuSO is added into fermentation medium on the basis of 1 method of embodiment4To the final concentration of 0.1mM-0.5mM of copper ion
Or addition MnSO4The purpose of addition copper ion or manganese ion is realized to the final concentration of 0.1mM-0.5mM of manganese ion.
2, comparative test
In dark, blue light, dark+manganese ion (final concentration 0.2mM), blue light+manganese ion (final concentration 0.2mM), dark+copper
It is wooden that white-rot fungi production is carried out under ion (final concentration 0.2mM) and blue light+various combination situations of manganese ion (final concentration 0.2mM)
The enzyme activity determination of plain degrading enzyme, the results are shown in Table 4.
Table 4
As can be seen from Table 4, no matter under dark surrounds or under blue light environment, increase manganese ion in the fermentation medium
Or copper ion can significantly improve the enzyme activity of lignin-degrading enzymes, wherein optimal with the enzyme activity under blue light environment.In addition, this
Invention replacement copper chloride and manganese chloride simultaneously adjust separately the above-mentioned experiment of final concentration of 0.1mM and 0.5mM repetition, as a result with above-mentioned table
4 results are consistent.
Embodiment 5: enzyme activity comparative test of the different fermentations culture medium under conventional dark surrounds
Subjects:
(1) glucose 0.5g/L, ramie stalk 60g/L, (NH4)2SO41.1g/L, urea 1g/L, peptone 0.5g/L,
KH2PO4 4g/L、CaCl2 0.15g/L、MgSO4 0.16g/L、FeSO4 0.0025g/L、MnSO4 0.0032g/L、ZnSO4
0.0007g/L、CoCl21 drop of 0.0074g/L, Tween/L, excess water；
(2) glucose 1g/L, ramie stalk 30g/L, (NH4)2SO42.2g/L, urea 0.5g/L, peptone 1g/L,
KH2PO4 2g/L、CaCl2 0.3g/L、MgSO4 0.08g/L、FeSO4 0.005g/L、MnSO4 0.0016g/L、ZnSO4
0.0014g/L、CoCl22 drops of 0.0037g/L, Tween/L, excess water；
(3) glucose 2g/L, ramie stalk 15g/L, (NH4)2SO44.4g/L, urea 0.25g/L, peptone 2g/L,
KH2PO4 1g/L、CaCl2 0.6g/L、MgSO4 0.04g/L、FeSO4 0.01g/L、MnSO4 0.0008g/L、ZnSO4
0.0028g/L、CoCl24 drops of 0.00185g/L, Tween/L, excess water；
(4) glucose 1.5g/L, ramie stalk 45g/L, (NH4)2SO43g/L, urea 0.5g/L, peptone 1.5g/L,
KH2PO4 3g/L、CaCl2 0.2g/L、MgSO4 0.1g/L、FeSO4 0.006g/L、MnSO4 0.0025g/L、ZnSO4
0.001g/L、CoCl23 drops of 0.0025g/L, Tween/L, excess water.
(5) culture medium in existing patent CN104419692A: glucose 4.5g/L, microcrystalline cellulose 30g/L, corn pulp
25g/L, KH2PO42g/L, (NH4)2SO41.4g/L, MgSO4·7H2O0.3g/L, CaCl20.45g/L, FeSO4·7H2O5mg/
L, MnSO4·H2O1.6mg/L, ZnSO4·7H2O1.4mg/L, CoCl2·6H2O 0.37mg/L, adjusting pH value is 4.5.
Above-mentioned 5 kinds of fermentation mediums use identical white-rot fungi seed liquor, in identical inoculum concentration and fermented and cultured temperature
Under degree, it is placed in dark surrounds fermented and cultured, detects the enzyme activity of cellulase, hemicellulase and lignin-degrading enzymes, as a result
It is shown in the following table；Wherein table 5- table 8 be followed successively by fermentation medium (1), (3), (4), (5) as a result, the knot of fermentation medium (2)
Result is corresponded in fruit reference table 1-2 and table 4；
Table 5
Table 6
Table 7
Table 8
Compare each table result, it is apparent that use fermentation medium of the present invention, can be bright under conventional dark surrounds
The aobvious enzyme activity for improving cellulase, hemicellulase and lignin-degrading enzymes, and the effect of the culture medium of existing patent is poor.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of method for improving white-rot fungi producing enzyme enzyme activity, which is characterized in that white-rot fungi is inoculated into fermentation medium,
It is fermented using blue light illumination, the blue light is 440-470nm blue light, and intensity is 50-100 μm of ol m-2s-1。
2. method according to claim 1, which is characterized in that the white-rot fungi is Pleurotus eryngii, oyster mushroom, yellow archespore Mao Pingge
One or more of bacterium and Irpex lacteus.
3. method according to claim 1, which is characterized in that the temperature of the fermentation is 25-30 DEG C.
4. method according to claim 1, which is characterized in that the fermentation medium is the fermented and cultured containing ramie stalk
Base.
5. method according to claim 4, which is characterized in that the fermentation medium include glucose, ramie stalk,
(NH4)2SO4, urea, peptone, KH2PO4、CaCl2、MgSO4、FeSO4、MnSO4、ZnSO4、CoCl2, Tween and water.
6. method according to claim 5, which is characterized in that the fermentation medium are as follows:
Glucose 0.5-2g/L, ramie stalk 15-60g/L, (NH4)2SO41.1-4.4g/L, urea 0.25-1g/L, peptone
0.5-2g/L、KH2PO4 1-4g/L、CaCl2 0.15-0.6g/L、MgSO40.04-0.16g/L、FeSO4 0.0025-0.01g/
L、MnSO4 0.0008-0.0032g/L、ZnSO40.0007-0.0028g/L、CoCl2 0.00185-0.0074g/L、Tween
1-4 drop/L, excess water.
7. -5 any one the method according to claim 1, which is characterized in that further include in the fermentation medium be added copper from
The combination of son or manganese ion or both, the final concentration of 0.1mM-0.5mM of the copper ion, the manganese ion it is final concentration of
0.1mM-0.5mM。
8. the combined fermentation medium of copper ion or manganese ion or both is added in white rot in blue light illumination or blue light illumination joint
Application in fungi fermentation producing enzyme；Wherein, the blue light is 440-470nm blue light, and intensity is 50-100 μm of ol m-2s-1；It is described
The final concentration of 0.1mM-0.5mM of copper ion, the final concentration of 0.1mM-0.5mM of the manganese ion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811119511.XA CN109182141B (en) | 2018-09-25 | 2018-09-25 | A method of improving white-rot fungi producing enzyme enzyme activity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811119511.XA CN109182141B (en) | 2018-09-25 | 2018-09-25 | A method of improving white-rot fungi producing enzyme enzyme activity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109182141A CN109182141A (en) | 2019-01-11 |
| CN109182141B true CN109182141B (en) | 2019-09-10 |
Family
ID=64909877
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811119511.XA Active CN109182141B (en) | 2018-09-25 | 2018-09-25 | A method of improving white-rot fungi producing enzyme enzyme activity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109182141B (en) |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101857879A (en) * | 2010-03-16 | 2010-10-13 | 唐东升 | Method for promoting and inducing Pleurotus eryngii mycelium to produce and accumulate triterpenoid |
-
2018
- 2018-09-25 CN CN201811119511.XA patent/CN109182141B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN109182141A (en) | 2019-01-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Elisashvili et al. | Lignocellulose-degrading enzyme production by white-rot Basidiomycetes isolated from the forests of Georgia | |
| Camassola et al. | Biological pretreatment of sugar cane bagasse for the production of cellulases and xylanases by Penicillium echinulatum | |
| Thygesen et al. | Production of cellulose and hemicellulose-degrading enzymes by filamentous fungi cultivated on wet-oxidised wheat straw | |
| Díaz et al. | Enhance hydrolytic enzymes production by Aspergillus awamori on supplemented grape pomace | |
| CN110066746B (en) | High-temperature-resistant bacillus bacterium NJAU-ND8 for accelerating compost maturity and application thereof | |
| CN114381378B (en) | Penicillium chrysogenum for degrading lignin and application thereof | |
| CN101855973B (en) | Fungus strain irpex iacteus for producing laccase, and culturing method and application thereof | |
| CN101210228A (en) | Bacillus licheniformis for degrading cellulose and application thereof in straw fermentation | |
| CN104988077A (en) | Eupenicillium parvum producing high temperature cellulase and xylanase and application thereof | |
| Bilal et al. | Biodegradation of agrowastes by lignocellulolytic activity of an oyster mushroom, Pleurotus sapidus | |
| CN104789492A (en) | Bacillus megaterium strain and application thereof | |
| Ogbonna et al. | Isolation of amylase and cellulase producing fungi from decaying tubers and optimization of their enzyme production in solid and submerged cultures | |
| CN104789491B (en) | Lichem bacillus strain and its application | |
| Oyewole et al. | Extracellular enzyme activities during cassava fermentation for ‘fufu’production | |
| Ezeonu et al. | Enhanced availability of biofuel and biomass components in Aspergillus niger and Aspergillus fumigatus treated rice husk | |
| CN109182141B (en) | A method of improving white-rot fungi producing enzyme enzyme activity | |
| CN101857859A (en) | Preparation method of xylanase or lignin peroxidase and application thereof to industrial bleaching field | |
| CN103131639B (en) | Trichoderma longibrachiatum strain and application thereof | |
| CN101638645A (en) | Method for producing xylanase by solid mechanical fermentation | |
| CN109055335A (en) | It is a kind of improve white-rot fungi producing enzyme enzyme activity fermentation medium and its application | |
| CN103060206B (en) | Fermentation bacteria agent and preparation method and application thereof | |
| CN110527634A (en) | One plant of Tibet source produces trichoderma harzianum strain and its application of cellulase | |
| CN109161535A (en) | A kind of fermentation medium and its application for improving white-rot fungi and producing lignin-degrading enzymes enzyme activity | |
| CN102559511B (en) | Hypocrea for producing mesophile ethanol-tolerant beta-glucosidase highly and application of hypocrea | |
| CN104893988A (en) | Trichoderma atroviride strain capable of producing high-temperature-resistant feruloyl esterase and high-temperature-resistant cellulase and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |