CN108130277A - A kind of white-rot fungi culture medium and its preparation method based on lignin - Google Patents

A kind of white-rot fungi culture medium and its preparation method based on lignin Download PDF

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Publication number
CN108130277A
CN108130277A CN201711250061.3A CN201711250061A CN108130277A CN 108130277 A CN108130277 A CN 108130277A CN 201711250061 A CN201711250061 A CN 201711250061A CN 108130277 A CN108130277 A CN 108130277A
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China
Prior art keywords
lignin
culture medium
white
finished product
rot fungi
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CN201711250061.3A
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姬丹丹
臧立华
侯立鹏
李肖玲
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Qilu University of Technology
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Qilu University of Technology
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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
    • 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

Abstract

The present invention provides a kind of white-rot fungi culture mediums based on finished product lignin and preparation method thereof.White-rot fungi relates generally to Phanerochaete chrysosporium, rainbow conk, mushroom, oyster mushroom.The culture medium raw material and its concentration are as follows：0.05 0.20g/L of finished product lignin, glucose 3 6g/L, KH2PO41 3g/L, MgSO40.20 0.30g/L, CaCl20.05 0.15g/L, MnSO40.005g/L, VB10.005g/L, 0.1 0.3g/L of ammonium tartrate, 130 160mL/L of trace element.The present invention solves the problems, such as that existing white-rot fungi culture medium technique is complicated, the period is long, long potential difference.A kind of nutrition equilibrium is provided simultaneously, conducive to the culture medium of growth；The main body of culture medium has used finished product lignin, can use the lignin in different waste liquid waste materials as the standard medium of lignin processing.

Description

A kind of white-rot fungi culture medium and its preparation method based on lignin
Technical field
The invention belongs to Liginon Resourceization using technical field, more particularly to a kind of based on finished product lignin White-rot fungi culture medium and its preparation method.
Background technology
Photosynthesis of plant generates the biomass that total amount reaches hundred million tons of 1440-1800 every year on earth, wherein the overwhelming majority It is the lignocellulose-like biomass in plant cell wall component.Lignocellulosic is considered as to exist most widely on the earth Recyclability biomass resource, main component are cellulose, hemicellulose and lignin.In lignocellulosic, cellulose Microfibre is formed, and then forms the mesh skeleton of fiber finer cell wall, and lignin and hemicellulose knot in the form of covalent bond It closes, cellulose is embedded therein, a kind of natural cover for defense is formed, microorganism is made to be not easy to contact with cellulose.
It can mitigate environmental pollution with bioanalysis lignin degrading, save energy consumption, can also turn waste into wealth, realize discarded plant Object resource reutilization.The biodegradation of lignin research shows that, white-rot fungi (White rotfungi) be it is currently the only can will A quasi-microorganism of the lignin degradation for carbon dioxide and water, gone back while secreting lignin-degrading enzymes eccrine fiber element enzyme, The enzymes such as hemicellulase, pectase are more suitable for decomposing natural lignocellulosic materials than trichoderma, aspergillus etc., therefore utilize white Rotten fungi Selective lignin-degradation is the hot spot in current biotechnology research.
Invention content
The technical problem to be solved in the present invention is to provide a kind of white-rot fungi culture medium based on finished product lignin, and Preparation method is provided.
In order to achieve the above objectives, the technical scheme is that：
A kind of fluid nutrient medium of the present invention, culture medium include following components by concentration：
Preferably, the pH value of the culture medium is 4.5-7.0.
The fungi is white-rot fungi, and the lignin in waste is utilized for degrading.
The VB1For vitamin B1Abbreviation.
The culture medium raw material and its concentration are as follows：Finished product lignin (Vinsol or dealkalize lignin) 0.08-0.16g/ L, glucose 3-5g/L, KH2PO41-3g/L, MgSO40.22-0.28g/L, CaCl20.08-0.12g/L, MnSO40.005g/ L, VB10.005g/L, ammonium tartrate 0.1-0.3g/L, trace element solution 135-155mL/L.
Preferably, the trace element uses as a solution.Trace element solution component and a concentration of：NaCl 1.0g/L, FeSO4`7H2O 100mg/L, CoSO4`7H2O 100mg/L, ZnSO4`7H2O 100mg/L, CuSO4`5H2O 10mg/ L, KAl (SO4)2100mg/L, H3BO310mg/L, Na2MoO4 10mg/L。
The preparation method of the culture medium is：
(1) it pre-processes：Finished product lignin purity is measured with Klason methods；
(2) said components are sufficiently mixed according to the proportioning；
(3) it after adding in a certain amount of water according to concentration, adjusts pH value and can be used for 4.5-7.0.
The finished product lignin is common lignin：Alkali lignin, dealkalize lignin, Vinsol.It is it is furthermore preferred that described Lignin is Vinsol.
The finished product lignin can be the lignin in various sources, such as the lignin extracted in waste liquid waste material；Described Content is the content for being converted into Vinsol or dealkalize lignin.
The present invention also provides the purposes of the culture medium, for the culture medium of white-rot fungi.
Preferably, the purposes of the culture medium will make the culture medium that finishes, aseptically White-Rot Fungi, And it is put into constant temperature culture oscillator, 37 DEG C of set temperature, rotating speed 120rpm.
White-rot fungi applied in the present invention is：Phanerochaete chrysosporium (Phanerochaetechrysosporium), rainbow conk (Coriolus versicolor), mushroom (Lentinusedodes), Oyster mushroom (Pleurotusostreatus).
White-rot fungi culture medium according to the present invention can ensure the supply of nutrient needed for white-rot fungi growth course Optimization culture technique simultaneously shortens cultivation cycle, reduces production cost and is conducive to promote white-rot fungi Biological Pretreatment work The utilization of industry.
The present invention has the advantages that following：(1) it is equal to provide a kind of nutrition according to the growth demand of white-rot fungi by the present invention Weighing apparatus, fungi culture medium that is cheap and being conducive to strain producing enzyme；It is complicated, all to solve existing white-rot fungi culture medium technique The problem of phase length, long potential difference.(2) preparation of white-rot fungi culture medium according to the present invention can ensure white-rot fungi in life Optimization culture technique, shortening cultivation cycle reduce production cost, are conducive to promote while nutrient needed for supply in growth process The industrialized utilization of white-rot fungi Biological Pretreatment lignin waste.(3) culture medium provided by the present invention is wooden with finished product Based on element, using white-rot fungi to the selection degradability of lignin, lignin degradation efficiency is improved, is given up for different waste liquids Lignin biodegradation in material provides theoretical foundation.The present invention solves existing white-rot fungi culture medium technique complexity, period The problem of long, long potential difference.Simultaneously according to white-rot fungi nutritional need, a kind of nutrition equilibrium is provided, conducive to the training of growth Support base；The main body of culture medium has used finished product lignin, can use difference as the standard medium of lignin processing Lignin in waste liquid waste material, biodegradation provide theoretical foundation.
Specific embodiment
Embodiment 1：
A kind of Phanerochaete chrysosporium (Phanerochaetechrysosporium) of the 50mg/L of content containing Vinsol Culture medium
Include the component of following weight/volume：
Finished product Vinsol 0.05g, glucose 5g, KH2PO42g, trace element solution 150mL, MgSO40.25g, wine Stone acid ammonium 0.2g, CaCl20.1g, MnSO40.005g, VB1 0.005g。
Trace Elements match：NaCl 1.0g/L, FeSO4`7H2O 100mg/L, CoSO4`7H2O 100mg/L, ZnSO4`7H2O 100mg/L, CuSO4`5H2O 10mg/L, KAl (SO4)2100mg/L, H3BO310mg/L, Na2MoO4 10mg/L。
Preparation method is：Measure finished product lignin purity with Klason methods, with deionized water dissolving, add in above-mentioned quality or Each material of volume fully dissolves and water is added to be settled to 1L, and it is 6.0 to correct pH value, you can is used.In incubation, culture medium is put In 37 DEG C of constant temperature culture oscillators, rotating speed 120rpm.
Aseptically, the culture medium 20mL through cultivating after a period of time is taken out, can be measured according to potassium ferricyanide method method Content is compared and both can obtain Lignin degradation rate in incubation before content of lignin, with culture in matrix.
In the pure lignin culture medium of the present invention not only comprising contained by conventional medium carbon source, nitrogen source, inorganic salts (including Trace element) and nutrients such as vitamin, and also lignin is as uniquely can be degradable by white-rot fungi in nature Biomass, sufficient nutrient can be provided, white-rot fungi is promoted to generate a variety of degrading enzymes, so as to make white-rot fungi big Amount breeding.
By content of lignin in matrix before and after measure culture it is known that culture medium provided by the present invention can be trained efficiently Support Phanerochaete chrysosporium and lignin degrading, while much fuller than the growing way of conventional medium of mycelium growth vigor, and bacterium The speed of growth of silk shifts to an earlier date 8-10 days.
Embodiment 2：
A kind of Phanerochaete chrysosporium (Phanerochaetechrysosporium) of the 200mg/L of content containing Vinsol Culture medium
Include the component of following weight (volume) part：
Finished product Vinsol 0.20g, glucose 3g, KH2PO43g, trace element solution 135mL, ammonium tartrate 0.1g, MgSO40.25g, CaCl20.1g, MnSO40.005g, VB1 0.005g。
Trace Elements match：NaCl 1.0g/L, FeSO4`7H2O 100mg/L, CoSO4`7H2O 100mg/L, ZnSO4`7H2O 100mg/L, CuSO4`5H2O 10mg/L, KAl (SO4)2100mg/L, H3BO310mg/L, Na2MoO4 10mg/L。
Preparation method is：Finished product lignin purity is measured with Klason methods, with deionized water dissolving, it is each to add in above-mentioned quality Material fully dissolves and water is added to be settled to 1L, and it is 6.0 to correct pH value, you can is used.In incubation, culture medium is placed in 37 DEG C In constant temperature culture oscillator, rotating speed 120rpm.
Phanerochaete chrysosporium incubation Enzymatic characteristic is measured in incubation.Specific embodiment：
Phanerochaete chrysosporium can generate the lignin that extracellular oxidizing ferment comes in degrading straw, mainly include manganese peroxide Compound enzyme (MnP), lignin peroxidase (LiP), laccase (Lac).It is illustrated by taking manganese peroxidase as an example herein.
Take the Azure B solutions of sodium tartrate buffer solution (pH=3.0) 1mL, 0.160mmol/L of 125mmol/L 0.5mL, lignin liquid medium 0.5mL；The hydrogenperoxide steam generator 0.5mL that 2mmol/L is added at 30 DEG C starts reaction；With purple Outer spectrophotometric determination is reacted in initial 3min, absorbance change at wavelength 651nm.(1 enzyme activity unit is with per minute Lignin liquid medium increases 0.1OD to represent)
1 Phanerochaete chrysosporium of table LiP enzymatic activitys activity change (U/mL) on different culture media
200mg/L lignin culture medium in table 1, each ingredient as mass fraction, finished product lignin 3.2%, glucose 47.6%th, KH2PO431.7%th, trace element 8.6%, MgSO44.0%th, ammonium tartrate 3.2%, CaCl21.6%th, MnSO4 0.08%th, VB10.08%.
Common white rot fungi culture medium in table 1, each ingredient as mass fraction, glucose 61.7%, KH2PO425.2%th, Trace element 6.2%, MgSO43.1%th, ammonium tartrate 2.5%, CaCl21.3%th, MnSO40.06%th, VB10.06%.
It can analyze to obtain from table, 2d after incubation, 200mg/L lignin culture mediums start to show Lip work Property, and raised trend is presented in two kinds of culture mediums, peaks in the 7d of culture, 200mg/L lignin culture medium and Common white rot fungi culture medium enzyme activity is up to 0.05U/mL and 0.03U/mL respectively.
Liquid fungal culture medium provided by the present invention in incubation compared with conventional medium, identical incubation time The yield of interior lignin oxidation's object enzyme significantly improves, and illustrates that it is more preferable to lignin selection degradability in matrix, yellow archespore hair is put down Lead fungi growth is more vigorous, and biomass is more abundant.And mycelium growth vigor is equally denseer than the growing way of conventional solid culture medium It is close, sturdy, and the speed of growth of mycelia is slightly in advance.

Claims (10)

1. a kind of fluid nutrient medium, culture medium includes following components by concentration：
Finished product lignin 0.05-0.20g/L；
Glucose 3-6g/L；
KH2PO41-3g/L；
Trace element solution 130-160mL/L；
Ammonium tartrate 0.1-0.3g/L；
MgSO40.20-0.30g/L；
CaCl20.05-0.15g/L；
MnSO40.005g/L；
VB1 0.005g/L。
2. fluid nutrient medium as described in claim 1, which is characterized in that the pH value of the culture medium is 4.5-7.0.
3. fluid nutrient medium as claimed in claim 1 or 2, which is characterized in that the finished product lignin is：Alkali lignin takes off Alkali lignin or Vinsol.
4. the fluid nutrient medium as described in right wants 3, which is characterized in that the lignin is Vinsol.
5. such as claim 1-4 any one of them fluid nutrient mediums, which is characterized in that the culture medium raw material and its concentration are such as Under：Finished product lignin (Vinsol or dealkalize lignin) 0.08-0.16g/L, glucose 3-5g/L, KH2PO41-3g/L, MgSO40.22-0.28g/L, CaCl20.08-0.12g/L, MnSO40.005g/L, VB10.005g/L, ammonium tartrate 0.1- 0.3g/L, trace element solution 135-155mL/L.
6. such as claim 1-5 any one of them fluid nutrient mediums, which is characterized in that the trace element is as a solution It uses；Trace element solution component and a concentration of：NaCl 1.0g/L, FeSO4`7H2O 100mg/L, CoSO4`7H2O 100mg/ L, ZnSO4`7H2O 100mg/L, CuSO4`5H2O 10mg/L, KAl (SO4)2100mg/L, H3BO310mg/L, Na2MoO4 10mg/L。
7. such as the preparation method of claim 1-6 any one of them culture mediums, include the following steps：
(1) it pre-processes：Finished product lignin purity is measured with Klason methods；
(2) said components are sufficiently mixed according to the proportioning；
(3) it after adding in a certain amount of water according to concentration, adjusts pH value and can be used for 4.5-7.0.
8. such as the purposes of claim 1-6 any one of them culture mediums, for the culture medium of white-rot fungi.
9. it is true to be aseptically inoculated with white rot for the purposes of culture medium as claimed in claim 8, the culture medium that making is finished Bacterium, and be put into constant temperature culture oscillator, 37 DEG C of set temperature, rotating speed 120rpm.
10. the purposes of culture medium as claimed in claim 8 or 9, the white-rot fungi are：Phanerochaete chrysosporium (Phanerochaetechrysosporium), rainbow conk (Coriolus versicolor), mushroom (Lentinusedodes), Oyster mushroom (Pleurotusostreatus).
CN201711250061.3A 2017-12-01 2017-12-01 A kind of white-rot fungi culture medium and its preparation method based on lignin Pending CN108130277A (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109055335A (en) * 2018-09-25 2018-12-21 中国农业科学院麻类研究所 It is a kind of improve white-rot fungi producing enzyme enzyme activity fermentation medium and its application
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
CN109997689A (en) * 2019-04-19 2019-07-12 广西南岜仔科技有限公司 A method of cultivating polyploid succulent
CN113396777A (en) * 2021-06-17 2021-09-17 齐鲁工业大学 Method for treating agricultural organic solid waste by using ammonia fiber expansion in cooperation with white rot fungi
CN114105714A (en) * 2021-12-15 2022-03-01 桂润环境科技股份有限公司 Soil remediation agent, preparation method and application thereof, and soil remediation method

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104711236A (en) * 2015-03-11 2015-06-17 湖南大学 Lignin-degrading enzyme regulator and application thereof
CN106520569A (en) * 2016-10-28 2017-03-22 齐鲁工业大学 White rot fungi medium taking crop straws as main body and preparation method of white rot fungi medium
CN107435053A (en) * 2016-05-25 2017-12-05 东北林业大学 A kind of white-rot fungi pretreatment agricultural crop straw quickly produces the fermentation process of biogas

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104711236A (en) * 2015-03-11 2015-06-17 湖南大学 Lignin-degrading enzyme regulator and application thereof
CN107435053A (en) * 2016-05-25 2017-12-05 东北林业大学 A kind of white-rot fungi pretreatment agricultural crop straw quickly produces the fermentation process of biogas
CN106520569A (en) * 2016-10-28 2017-03-22 齐鲁工业大学 White rot fungi medium taking crop straws as main body and preparation method of white rot fungi medium

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
YULING HAN等: "Azo Dye Biodecolorization Enhanced by Echinodontium taxodii Cultured with Lignin", 《PLOS ONE》 *
张晶等: "天然浸出液对自分离白腐真菌产木质素降解酶的影响", 《清华大学学报(自然科学版)》 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109055335A (en) * 2018-09-25 2018-12-21 中国农业科学院麻类研究所 It is a kind of improve white-rot fungi producing enzyme enzyme activity fermentation medium and its application
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
CN109055335B (en) * 2018-09-25 2021-11-12 中国农业科学院麻类研究所 Fermentation medium for improving enzyme production activity of white rot fungi and application thereof
CN109997689A (en) * 2019-04-19 2019-07-12 广西南岜仔科技有限公司 A method of cultivating polyploid succulent
CN113396777A (en) * 2021-06-17 2021-09-17 齐鲁工业大学 Method for treating agricultural organic solid waste by using ammonia fiber expansion in cooperation with white rot fungi
CN114105714A (en) * 2021-12-15 2022-03-01 桂润环境科技股份有限公司 Soil remediation agent, preparation method and application thereof, and soil remediation method

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