CN104263705A - Method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium - Google Patents

Method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium Download PDF

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CN104263705A
CN104263705A CN201410471747.5A CN201410471747A CN104263705A CN 104263705 A CN104263705 A CN 104263705A CN 201410471747 A CN201410471747 A CN 201410471747A CN 104263705 A CN104263705 A CN 104263705A
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final concentration
lignin
substratum
degrading enzymes
phanerochaete chrysosporium
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文湘华
喻国策
钱易
王建龙
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Tsinghua University
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Tsinghua University
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    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01013—Manganese peroxidase (1.11.1.13)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01014—Lignin peroxidase (1.11.1.14)

Abstract

The invention relates to a method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium. The invention provides a method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium, which comprises the following steps: fermenting phanerochaete chrysosporium in a culture medium containing an immobilized culture carrier in a non-immersed state, and collecting a fermentation product, namely, a lignin-degrading enzyme. Experiments show that when the method is adopted for carrying out fermentation on a lignin-degrading enzyme, the lignin-degrading enzyme can be selectively obtained under air conditions without feeding pure oxygen or air containing high-concentration oxygen in the process of culturing but just by controlling the concentration of a carbon and nitrogen source in the culture medium and adding a carrier in a non-immersed state. The method disclosed by the invention is simple in condition control, and the obtained lignin-degrading enzyme is strong in selectivity.

Description

A kind of method utilizing Phanerochaete chrysosporium selectivity to produce lignin-degrading enzymes
Technical field
The present invention relates to biological technical field, particularly relate to a kind of method utilizing Phanerochaete chrysosporium selectivity to produce lignin-degrading enzymes.
Background technology
White-rot fungi is a kind of high filamentous fungus, and in classification, great majority belong to Basidiomycetes, grow on trees or timber, gains the name because causing wooden white to rot.White rot fungus degrading xylogen, is because it can produce lignin-degrading enzymes, and is secreted into extracellular.The degraded of lignin-degrading enzymes to xylogen is the chain reaction process based on free radical, there is the non-specific and oxidisability of extremely strong substrate, this degradation mechanism makes lignin-degrading enzymes can not only lignin degrading, can also to degrade many xenobionticses in environment and persistence toxic organic pollutant (Barr & Aust, 1994; Cameron et al., 2000; Gao et al., 2010).Therefore, white-rot fungi and lignin-degrading enzymes thereof have important using value in environment pollution control and biological restoration etc. thereof.
Phanerochaete chrysosporium (Phanerochaete chrysosporium) is type culture (Wesenberg et al., 2003 that white-rot fungi produces lignin-degrading enzymes and the research of degraded environmental pollutant; Singh & Chen, 2008), belong to Aphyllophorales (Phyllophorales), Fu Geke (Corticiaceae) and aobvious lead fungi and belong to (Phanerochaete).P.chrysosporium Ligninolytic Enzymes system mainly comprises lignin peroxidase (LiP, EC1.11.1.14), manganese peroxidase (MnP, EC1.11.1.13) and produces H 2o 2oxydase.Lignin peroxidase and manganese peroxidase are the multiple isozyme of glycosylated iron content, lignin peroxidase can catalysis to the oxidation of non-phenolic lignin pattern compound and aromatic pollution, manganese peroxidase can catalysis to the oxidation of the xylogen pattern compound of xylogen, lignin derivative and a lot of phenols.The synthesis of Phanerochaete chrysosporium lignin-degrading enzymes is subject to the complexity adjustment of multiple nutrients and environmental factors, lignin-degrading enzymes can be subject to nitrogen, carbon or sulphur nutrition restriction time excite generation, performance active (Faison & Kirk, 1985 under hyperoxic conditions; Dosoretz et al., 1990).Hyperoxic conditions is considered to improve the mass transfer of oxygen in mycelia, particularly when exocellular polysaccharide accumulates, thus the synthesis of inducible enzyme.It is generally acknowledged that Phanerochaete chrysosporium does not produce laccase.
The application of lignin-degrading enzymes depends on the high level fermentation of enzyme.The research of Phanerochaete chrysosporium lignin degradation enzymic fermentation, mainly comprises following aspect (Ikehata et al., 2004; Singh & Chen, 2008): the nutritional condition (comprising suitable carbon source, nitrogenous source, trace element, dissolved oxygen and inductor etc.) that (1) ferments; (2) envrionment conditions (as temperature, pH, stirring and fixing condition etc.) of fermenting; (3) research of reactor fermentation scale-up.Lignin peroxidase and manganese peroxidase are the enzymes be closely related, and usually produce in white-rot fungi simultaneously.But, the catalyst mechanism of the two is different, has different degradation capability (Cameron et al., 2000 to the substrate of different structure; Martinez, 2002).The degree that generation due to two kinds of peroxidase is subject to nutrition and such environmental effects there are differences, and the synthesis of two kinds of enzymes may exist respective feature outside common accommodation property.Therefore, the difference utilizing nutrition and envrionment conditions to regulate enzymic synthesis, the selectivity likely realizing two kinds of peroxidase produces.The selectivity of peroxidase produces the research contributing to lignin degradation enzymic synthesis and regulation mechanism, has important application value for meeting the degraded needs with different structure characteristic organic pollutants simultaneously.So far, utilize the report of Phanerochaete chrysosporium selectivity generation lignin-degrading enzymes also very rare.Bonnarme and Jeffries (1990a, b) finds mn ion (Mn 2+) to the enzymic synthesis of Phanerochaete chrysosporium lignin degradation, there is different mediating effect+6, by controlling Mn in substratum 2+the selectivity that concentration achieves Phanerochaete chrysosporium and manganese peroxidase in airlift reactor produces, and but, requires pure oxygen to pass into be a restriction of its fermenting process.Li Huazhong etc. (2002) have studied Phanerochaete chrysosporium equally at Mn 2+under concentration adjustment, the selectivity of lignin-degrading enzymes produces, but effect is still not ideal enough, and still by the restriction of supplied with pure oxygen.
Except Mn 2+control, at present, also do not see the report utilizing other regulating measures to realize the generation of Phanerochaete chrysosporium lignin degradation enzyme selectivity, especially in air ambient.The effective synthesis of lignin-degrading enzymes under air conditions has cost benefit and larger feasibility for the large scale fermentation of enzyme.
Summary of the invention
An object of the present invention is to provide a kind of method utilizing Phanerochaete chrysosporium to produce lignin-degrading enzymes.
Method provided by the invention, comprise the steps: containing be in non-submerged state Immobilized culture carrier substratum in ferment Phanerochaete chrysosporium, collect tunning, namely obtain lignin-degrading enzymes;
The substratum of the described Immobilized culture carrier containing being in non-submerged state is add for cultivating in the liquid nutrient medium of white-rot fungi by described Immobilized culture carrier, and make its piling height higher than described liquid nutrient medium liquid level, be in non-submerged state, the substratum obtained.
In aforesaid method, described containing be in non-submerged state Immobilized culture carrier substratum in the content of Immobilized culture carrier be more than or equal to 1.8g/100mL.
In aforesaid method, described containing be in non-submerged state Immobilized culture carrier substratum in the content of Immobilized culture carrier be 1.8g/100mL-3.0g/100mL, be specially 1.8g/100mL, 2.2g/100mL, 2.6g/100mL or 3.0g/100mL.
In aforesaid method, described liquid nutrient medium is carbon confined liquid substratum, is lignin peroxidase with the lignin-degrading enzymes that its fermentation produces;
The mol ratio that described carbon confined liquid substratum is specially C atom and atom N is less than or equal to the carbon confined liquid substratum of 3.8.
In aforesaid method, the C atom source in described carbon confined liquid substratum is in glucose, and atom N derives from ammonium tartrate;
Carbon restriction substratum described in every 1L by final concentration be 5.04g/L glucose, final concentration is 4.05g/L ammonium tartrate, final concentration is 2.0g/L KH 2pO 4, final concentration is 0.5g/L MgSO 4, final concentration is 0.1g/L CaCl 2, final concentration is 1mg/L vitamins B 1, final concentration is 1.5mM veratryl alcohol, pH 4.4 and final concentration is 20mM acetate buffer and 70mL/L trace element solution composition;
Described trace element solution is 3g/L MgSO by final concentration 4, final concentration is 0.5g/L MnSO 4, final concentration is 1.0g/L NaCl, final concentration is 0.1g/L FeSO 47H 2o, final concentration are 0.1g/L CoCl 2, final concentration is 0.1g/L ZnSO 47H 2o, final concentration are 0.1g/L CuSO 4, final concentration is 10mg/L AlK (SO 4) 212H 2o, final concentration are 10mg/L H 3bO 3, final concentration is 10mg/L Na 2moO 42H 2o, final concentration are 1.5g/L nitrilotriacetic acid(NTA) salt and water composition.
In aforesaid method, described liquid nutrient medium is nitrogen confined liquid substratum, is manganese peroxidase with the lignin-degrading enzymes that its fermentation produces;
The mol ratio that described nitrogen confined liquid substratum is specially C atom and atom N is more than or equal to the nitrogen confined liquid substratum of 152.7.
In aforesaid method, the C atom source in described nitrogen confined liquid substratum is in glucose, and atom N derives from ammonium tartrate;
Nitrogen confined liquid substratum described in every 1L be with final concentration be 10.08g/L glucose, final concentration is that 0.203g/L ammonium tartrate replaces glucose in described carbon confined liquid substratum and ammonium tartrate, all the other components are identical with described carbon confined liquid substratum.
In aforesaid method, the pH value of described liquid nutrient medium is 4.4;
Described Immobilized culture carrier is inert material, is specially polyurethane foamed blocks; The length of side of described polyurethane foamed blocks is especially specially 0.5cm.
Another object of the present invention is to provide a kind of test kit utilizing Phanerochaete chrysosporium to produce lignin-degrading enzymes.
Test kit provided by the invention, comprises the above-mentioned substratum of Immobilized culture carrier containing being in non-submerged state.
In aforesaid method, described lignin-degrading enzymes is lignin peroxidase or manganese peroxidase.
In aforesaid method or test kit, described Phanerochaete chrysosporium is BKM-F-1767, and the pH of described substratum is 4.0-5.0, and described fermentation culture conditions is: temperature is 37oC, and rotating speed is 160rpm fermentation culture.
Experiment of the present invention proves, the present invention adopts Phanerochaete chrysosporium as lignin-degrading enzymes fermentation strain, and fermention medium adopts carbon confined liquid substratum, and (C/N ratio is not more than 3.8, i.e. glucose/NH 4 +be not more than 28/44mM), or nitrogen confined liquid substratum (C/N ratio is not less than 152.7, i.e. glucose/NH 4 +be not less than 56/2.2mM), adopt inert material as Immobilized culture carrier, carrier is made to be in non-submerged state in the medium, shaking culture under air ambient, in carbon restriction nutrient solution, mainly obtain lignin peroxidase, in nitrogen restriction nutrient solution, mainly obtain manganese peroxidase.Present method is adopted to carry out lignin degradation enzymic fermentation, pure oxygen or the air containing high-concentration oxygen need not be passed in culturing process, only need control the concentration of carbon and nitrogen sources in substratum and the carrier adding non-submerged amount, lignin-degrading enzymes can be obtained in selectivity under air conditions.The inventive method condition controls simple, and the lignin degradation enzyme selectivity of acquisition is strong.
Accompanying drawing explanation
Fig. 1 is Phanerochaete chrysosporium generation of lignin peroxidase under carbon restriction during Immobilized culture under air ambient.
Fig. 2 is Phanerochaete chrysosporium generation of lignin-degrading enzymes under nitrogen restriction during Immobilized culture under air ambient. (a) lignin peroxidase; (b) manganese peroxidase.
The stereoscan photograph (FEI QUANTA 200) that Fig. 3 is Phanerochaete chrysosporium under carbon restriction on polyurethane foamed blocks during Immobilized culture. (a) 30 ×; (b) 100 ×.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
In following embodiment, the pH of substratum is 4.0-5.0.
Embodiment 1, Phanerochaete chrysosporium selectivity under carbon restriction produces lignin peroxidase
1, the preparation of carbon confined liquid substratum
Adopt carbon confined liquid substratum, substratum revises a little based on typical Tien and Kirk substratum (1988) to form, and culture medium C/N ratio (mol ratio) is 3.8, specifically composed as follows:
Carbon confined liquid substratum by final concentration be 5.04g/L (28mM) glucose, final concentration is 4.05g/L (NH 4 +44mM) ammonium tartrate, final concentration are 2.0g/L KH 2pO 4, final concentration is 0.5g/L MgSO 4, final concentration is 0.1g/L CaCl 2, final concentration is 1mg/L vitamins B 1, final concentration is 1.5mM veratryl alcohol, final concentration is 20mM (pH 4.4) acetate buffer, 70mL/L trace element solution and deionized water composition; PH is 4.4.
Trace element solution is 3g/L MgSO by final concentration 4, final concentration is 0.5g/L MnSO 4, final concentration is 1.0g/L NaCl, final concentration is 0.1g/L FeSO 47H 2o, final concentration are 0.1g/L CoCl 2, final concentration is 0.1g/L ZnSO 47H 2o, final concentration are 0.1g/L CuSO 4, final concentration is 10mg/L AlK (SO 4) 212H 2o, final concentration are 10mg/L H 3bO 3, final concentration is 10mg/L Na 2moO 42H 2o, final concentration are 1.5g/L nitrilotriacetic acid(NTA) salt and water composition.
2, Phanerochaete chrysosporium selectivity under carbon restriction produces lignin peroxidase
Adopt the polyurethane foamed blocks of length of side 0.5cm as Immobilized culture carrier, the carrier amount adopted is respectively 1.0,1.4,1.8,2.2,2.6 and 3.0g, carrier is placed in the 250mL Erlenmeyer flask that 100mL carbon confined liquid substratum is housed, the piling height of carrier correspond to different submergences and non-submerged state; Be divided into following two groups of training methods:
Different carriers amount submerged culture: add 1.0 and 1.4g Immobilized culture carrier respectively in 100mL carbon confined liquid substratum, and make its piling height lower than liquid level, be in submerged state, obtain the submerged culture system of different carriers amount;
The non-submerged of different carriers amount is cultivated: in 100mL carbon confined liquid substratum, add 1.8,2.2,2.6 and 3.0g Immobilized culture carrier respectively, and make its piling height higher than liquid level, be in non-submerged state, obtain the non-submerged culture system of different carriers amount;
The non-submerged culture system of the submerged culture system of above-mentioned different carriers amount and different carriers amount is distinguished sterilizing (115 DEG C, 30min), to grow at 30 DEG C of PDA flat boards (200g potato leach liquor/L, 20g glucose/L, 20g agar/L) on Phanerochaete chrysosporium BKM-F-1767 spore (Tien, M., Kirk, T.K.Lignin peroxidase of Phanerochaete chrysosporium.Methods in Enzymology.1988, 161:238-249, the public can obtain from Tsing-Hua University) aseptic access substratum, after inoculation, spore concentration is about 1.0 × 10 5spore/mL.
By the non-submerged culture system of the submerged culture system of the different carriers amount of above-mentioned access spore with the different carriers amount of access spore, air ambient (not being filled with pure oxygen in addition), temperature is 37 DEG C, rotating speed is that 160rpm condition bottom fermentation is cultivated, and obtains nutrient solution and is tunning lignin degradation enzyme solution.Nutrient solution sample obtains supernatant liquor, for lignin-degrading enzymes Enzyme activity assay through centrifugal (9000r/min, 10min, 4 DEG C).
Supernatant liquor is carried out lignin peroxidase activity analysis, and lignin peroxidase activity employing is that the method for substrate measures with veratryl alcohol, and every 1min is oxidized 1 μm of ol veratryl alcohol and becomes the enzyme amount needed for veratryl aldehyde to be defined as 1 enzyme activity unit.
Bearing results as shown in Figure 1 of lignin peroxidase.
Carrier amount be 1.0 and the submerged culture system of 1.4g cultivate the fermented supernatant fluid of BKM-F-1767, do not detect that enzyme is lived;
Carrier amount is not less than the fermented supernatant fluid of the non-submerged culture system cultivation BKM-F-1767 of 1.8g, and maximum enzyme work increases with carrier amount and increases, and when 3.0g carrier, obtains the highest enzyme and lives as 87.5U/L.
Different culture condition bottom fermentation product supernatant liquor is carried out manganese peroxidase Enzyme assay simultaneously, and manganese peroxidase enzymic activity adopts with Mn 2+for the methods analyst of substrate, every 1min is oxidized 1 μm of ol Mn 2+required enzyme amount is defined as 1 enzyme activity unit.No matter adopt which kind of carrier amount, culturing process does not all have manganese peroxidase to produce.
Therefore, Phanerochaete chrysosporium during non-submerged Immobilized culture, selectivity can produce lignin peroxidase under air ambient under carbon restriction.
The stereoscan photograph of Phanerochaete chrysosporium under carbon restriction on polyurethane foamed blocks during Immobilized culture, as shown in Figure 3.
Embodiment 2, Phanerochaete chrysosporium selectivity under nitrogen restriction produces manganese peroxidase
1, the preparation of nitrogen confined liquid substratum
Adopt nitrogen confined liquid substratum, culture medium C/N ratio (mol ratio) is 152.7, specifically composed as follows:
Nitrogen confined liquid culture medium prescription is for being 10.08g/L (56mM) glucose, 0.203g/L (NH by final concentration 4 +2.2mM) ammonium tartrate replaces glucose in carbon confined liquid substratum and ammonium tartrate, and all the other components are identical with the carbon confined liquid substratum in embodiment 1; PH is 4.4.
2, Phanerochaete chrysosporium selectivity under nitrogen restriction produces manganese peroxidase
Culture condition is identical with embodiment 1 with analytical procedure, is only that substratum replaces with nitrogen confined liquid substratum.
Lignin-degrading enzymes bears results as shown in Figure 2, detects that lignin peroxidase enzyme is lived (during 2.2g carrier, enzyme is lived maximum) when carrier amount is not less than 1.8g; When carrier amount is less than 1.8g, do not detect that lignin peroxidase enzyme is lived, but the highest enzyme is lived and is all no more than 20U/L (Fig. 2 a) under all carrier conditions;
In contrast, when carrier amount is not less than 1.8g, manganese peroxidase significantly generates, and when 2.2g carrier, the highest enzyme is lived and reached 338.7U/L (Fig. 2 b); When carrier amount is less than 1.8g, do not detect that manganese peroxidase enzyme is lived.
Therefore, Phanerochaete chrysosporium during non-submerged Immobilized culture, selectivity can produce manganese peroxidase under air ambient under nitrogen restriction.

Claims (10)

1. utilize Phanerochaete chrysosporium to produce the method for lignin-degrading enzymes, comprise the steps: containing be in non-submerged state Immobilized culture carrier substratum in ferment Phanerochaete chrysosporium, collect tunning, namely obtain lignin-degrading enzymes;
The substratum of the described Immobilized culture carrier containing being in non-submerged state is add for cultivating in the liquid nutrient medium of white-rot fungi by described Immobilized culture carrier, and make its piling height higher than described liquid nutrient medium liquid level, be in non-submerged state, the substratum obtained.
2. method according to claim 1, is characterized in that: described containing be in non-submerged state Immobilized culture carrier substratum in the content of Immobilized culture carrier be more than or equal to 1.8g/100mL.
3. method according to claim 2, is characterized in that:
Described containing be in non-submerged state Immobilized culture carrier substratum in the content of Immobilized culture carrier be 1.8g/100mL, 2.2g/100mL, 2.6g/100mL or 3.0g/100mL.
4., according to described method arbitrary in claim 1-3, it is characterized in that:
Described liquid nutrient medium is carbon confined liquid substratum, is lignin peroxidase with the lignin-degrading enzymes that its fermentation produces;
The mol ratio that described carbon confined liquid substratum is specially C atom and atom N is less than or equal to the carbon confined liquid substratum of 3.8.
5. method according to claim 4, is characterized in that:
C atom source in described carbon confined liquid substratum is in glucose, and atom N derives from ammonium tartrate;
Carbon restriction substratum described in every 1L by final concentration be 5.04g/L glucose, final concentration is 4.05g/L ammonium tartrate, final concentration is 2.0g/L KH 2pO 4, final concentration is 0.5g/L MgSO 4, final concentration is 0.1g/L CaCl 2, final concentration is 1mg/L vitamins B 1, final concentration is 1.5mM veratryl alcohol, pH 4.4 and final concentration is 20mM acetate buffer and 70mL/L trace element solution composition;
Described trace element solution is 3g/L MgSO by final concentration 4, final concentration is 0.5g/L MnSO 4, final concentration is 1.0g/L NaCl, final concentration is 0.1g/L FeSO 47H 2o, final concentration are 0.1g/L CoCl 2, final concentration is 0.1g/L ZnSO 47H 2o, final concentration are 0.1g/L CuSO 4, final concentration is 10mg/L AlK (SO 4) 212H 2o, final concentration are 10mg/L H 3bO 3, final concentration is 10mg/L Na 2moO 42H 2o, final concentration are 1.5g/L nitrilotriacetic acid(NTA) salt and water composition.
6., according to described method arbitrary in claim 1-5, it is characterized in that:
Described liquid nutrient medium is nitrogen confined liquid substratum, is manganese peroxidase with the lignin-degrading enzymes that its fermentation produces;
The mol ratio that described nitrogen confined liquid substratum is specially C atom and atom N is more than or equal to the nitrogen confined liquid substratum of 152.7.
7. method according to claim 6, is characterized in that:
C atom source in described nitrogen confined liquid substratum is in glucose, and atom N derives from ammonium tartrate;
Nitrogen confined liquid substratum described in every 1L be with final concentration be 10.08g/L glucose, final concentration is that 0.203g/L ammonium tartrate replaces glucose in described carbon confined liquid substratum and ammonium tartrate, all the other components are identical with described carbon confined liquid substratum.
8., according to described method arbitrary in claim 1-7, it is characterized in that:
The pH value of described liquid nutrient medium is 4.4;
Described Immobilized culture carrier is inert material, is specially polyurethane foamed blocks; The length of side of described polyurethane foamed blocks is especially specially 0.5cm.
9. utilizing Phanerochaete chrysosporium to produce the test kit of lignin-degrading enzymes, comprising the described substratum of Immobilized culture carrier containing being in non-submerged state in claim 1-8.
10. test kit according to claim 9, is characterized in that: described lignin-degrading enzymes is lignin peroxidase or manganese peroxidase.
CN201410471747.5A 2014-09-16 2014-09-16 Method for selectively producing lignin-degrading enzymes by using phanerochaete chrysosporium Pending CN104263705A (en)

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CN104830741A (en) * 2015-05-28 2015-08-12 陕西科技大学 Preparing method of compound microorganism agent for papermaking wastewater
CN105524891A (en) * 2016-02-02 2016-04-27 清华大学 Method for improving activity of lignin-degrading enzyme generated by phanerochaete chrysosporium after enzyme activity decay
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104830741A (en) * 2015-05-28 2015-08-12 陕西科技大学 Preparing method of compound microorganism agent for papermaking wastewater
CN105754889A (en) * 2015-12-11 2016-07-13 清华大学 Enterococcus faecium and application of enterococcus faecium to producing hydrogen by means of anaerobic fermentation
CN105754889B (en) * 2015-12-11 2019-07-02 清华大学 A kind of enterococcus faecium and its application in hydrogen production through anaerobic fermentation
CN105524891A (en) * 2016-02-02 2016-04-27 清华大学 Method for improving activity of lignin-degrading enzyme generated by phanerochaete chrysosporium after enzyme activity decay
CN106036007A (en) * 2016-06-08 2016-10-26 莫竹承 Biodegradation method for spartina alterniflora lignin
CN109161535A (en) * 2018-09-25 2019-01-08 中国农业科学院麻类研究所 A kind of fermentation medium and its application for improving white-rot fungi and producing lignin-degrading enzymes enzyme activity
CN114606211A (en) * 2022-04-28 2022-06-10 西安交通大学 Vitamin-induced lignin degrading enzyme, method for improving enzyme activity and application
CN114606211B (en) * 2022-04-28 2023-08-29 西安交通大学 Vitamin-induced lignin degrading enzyme, and method and application for improving enzyme activity
CN114908024A (en) * 2022-06-23 2022-08-16 合肥工业大学 Method for promoting white rot fungi to directionally produce enzyme and degrade plastic pollutants
CN114908024B (en) * 2022-06-23 2023-05-19 合肥工业大学 Method for promoting white rot fungi to directionally produce enzyme to degrade plastic pollutants

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