CN115888652A Preparation method of hypha-based biochar

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A kind of preparation method of mycelia-based biochar

technical field

The invention belongs to the technical field, and in particular relates to a method for preparing mycelia-based
biochar.

Background technique

Biochar is a solid by-product produced by high-temperature pyrolysis of biomass materials in anoxic or low-
oxygen environments. It has the characteristics of high aromaticity, high carbon content, and strong resistance to
biological and abiotic degradation.

Biochar is very versatile and can be applied to soil to increase soil fertility and reduce greenhouse gas emissions.
In addition, biochar has a large specific surface area and high porosity, which can absorb and degrade toxic and
harmful substances in the environment, including various heavy metal ions and organic pollutants, and play an
important role in the control of environmental pollution and ecological restoration. role.

Fungal hyphae are rich in chitin, lignin compounds, aromatic compounds, etc., and the growth rate is fast, and
the source is relatively rich. Mycelia-based biochar prepared from fungal mycelium has the characteristics of
wide sources, low cost and high adsorption activity, and can be widely used in the removal of various pollutants
in the environment.

Pleurotus eryngii (Pleurotus eryngii) is a common edible fungus whose mycelium is easy to cultivate and can be
produced on a large scale. The invention uses Pleurotus eryngii mycelium as a raw material, uses sodium
hydroxide to activate the mycelium, and then prepares the mycelium-based biochar under high temperature
conditions. The mycelium-based biochar has high adsorption performance and can effectively adsorb and
remove heavy metal ions in the environment.

Contents of the invention

The purpose of the present invention is to propose a method for preparing mycelium-based biochar, which has
high adsorption performance and can effectively adsorb and remove heavy metal ions in the environment.

In order to achieve the above object, the technical scheme adopted in the present invention is as follows:

A method for preparing mycelia-based biochar, characterized in that: the method is as follows:

Step (1) Make a PDA slant medium, insert a 0.5 cm-diameter strain block into the PDA slant medium, and
incubate at 25 °C for 8 days until the hyphae basically cover the entire slant;

Step (2) Insert a slant strain into 100 mL of seed liquid, and culture it on a shaker at 25 °C for 7 days, and this is
the seed liquid;

Step (3) Use the fermentation medium to inoculate the seed solution into a pneumatic fermenter with an
inoculum size of 2%, culture with aeration and stirring, and cultivate at 20 °C for 7 days;

Step (4) After the fermentation and cultivation, the culture medium was taken out from the fermenter, and
centrifuged at 3000 rpm for 10 minutes; then washed three times with distilled water, and centrifuged at 3000
rpm for 10 minutes each time to obtain pure Pleurotus eryngii mycelium;
Step (5) Freeze-dry the mycelium by using a vacuum freeze-dryer, and store it at 4°C for later use;

Step (6) Weigh the mycelium of step (5), grind it and pass it through a 100-mesh sieve, then soak it in 200 mL
of 2% sodium hydroxide solution for 2 h, wash it with ultrapure water until neutral, and dry it at 80 °C 12 hours;

Step (7) Put the modified mycelium into a crucible and place it in a muffle furnace; carbonize for 2 h at 400°C-
600°C as the preparation temperature;

Step (8) Grinding and pulverizing the biochar obtained in step (7), passing it through a 100-mesh sieve, and
storing it in a desiccator for later use;

Step (9) Using a fully automatic specific surface area analyzer to analyze the specific surface area of the biochar
by the BET method to determine the optimal carbonization temperature and high-quality biochar.

Further, the PDA medium raw materials in step (1) include: 200 g of potatoes, 20 g of glucose, 20 g of agar, and
1000 mL of distilled water.

Further, the seed liquid raw materials in step (2) include: 30 g of glucose, 2 g of peptone, 5 g of yeast powder,
0.5 g of potassium dihydrogen phosphate, 0.5 g of magnesium sulfate, 0.01 g of vitamin B1, and 1000 mL of
distilled water.

Further, the fermentation medium raw materials in step (3) include: 15 g of glucose, 3 g of soybean meal, 3 g of
corn flour, 1 g of bran, 0.5 g of peptone, 0.5 g of potassium dihydrogen phosphate, 0.5 g of magnesium sulfate,
and 0.2 g of vegetable oil mL, distilled water 1000 mL, pH6.0~6.5.

Furthermore, 600 °C in step (7) is a more suitable carbonization temperature, and its specific surface area is
102.37 m2/g.

An application of mycelium-based biochar. Mycelium-based biochar has good adsorption performance and can
be used to remove heavy metal ions in the environment.

The above technical scheme can obtain the following beneficial effects:

The invention uses Pleurotus eryngii mycelium as a raw material, uses sodium hydroxide to activate the
mycelium, and then prepares the mycelium-based biochar under high temperature conditions. The mycelium-
based biochar has high adsorption performance and can effectively adsorb and remove heavy metal ions in the
environment.

Detailed ways

The present invention will be further described below in conjunction with embodiment:

1 Cultivation of mycelium:

1.1 culture medium

(1) PDA medium: 200 g potato, 20 g glucose, 20 g agar, 1000 mL distilled water.

(2) Seed medium: 30 g of glucose, 2 g of peptone, 5 g of yeast powder, 0.5 g of potassium dihydrogen
phosphate, 0.5 g of magnesium sulfate, 10.01 g of vitamin B, and 1000 mL of distilled water.

(3) Fermentation medium: glucose 15 g, soybean meal 3 g, corn flour 3 g, bran 1 g, peptone 0.5 g, potassium
dihydrogen phosphate 0.5 g, magnesium sulfate 0.5 g, vegetable oil 0.2 mL, distilled water 1000 mL, pH 6.0～
6.5.

1.2 training process

(1) Make a PDA slant medium, insert a 0.5 cm-diameter strain block into the PDA slant medium, and incubate
at 25°C for 8 days until the hyphae basically cover the entire slant.
(2) Introduce a slant strain into 100 mL seed liquid, and culture it on a shaker at 25 °C for 7 days, and this is the
seed liquid.

(3) Using the fermentation medium, inoculate the seed solution into a pneumatic fermenter with an inoculum
size of 2%, culture with aeration and stirring, and cultivate at 20 °C for 7 days.

(4) After the fermentation and cultivation, the culture medium was taken out from the fermenter and centrifuged
at 3000 rpm for 10 minutes. Then use distilled water to wash three times, and centrifuge at 3000 rotation speed
for 10 minutes each time to obtain pure Pleurotus eryngii mycelium.

(5) Freeze-dry the mycelium using a vacuum freeze-dryer, and store it at 4°C for later use.

2 Preparation of mycelium-based biochar

(1) Accurately weigh 15 g of mycelium, grind it and pass it through a 100-mesh sieve, then soak it in 200 mL of
2% sodium hydroxide solution for 2 h, wash it with ultrapure water until neutral, and dry it at 80 °C for 12 h .

(2) The modified mycelium was put into a crucible and placed in a muffle furnace; carbonization was carried
out at 400 °C, 500 °C, and 600 °C for 2 h.

(3) The obtained biochar was ground and pulverized, passed through a 100-mesh sieve, and stored in a
desiccator for later use.

(4) Using a fully automatic specific surface area analyzer, the specific surface area of biochar was analyzed by
BET method, and the results are shown in Table 1. In the process of biochar preparation, the carbonization
temperature has a great influence on the internal structure of biochar. It is found that 600 ℃ is a more suitable
carbonization temperature, and its specific surface area is 102.37 m2/g.

Table 1 Specific surface area of mycelium-based biochar at different temperatures

Preparation temperature (℃) ⟨![CDATA[specific surface area (m ⟨sup ⟩2 ⟨/sup ⟩/g)]] ⟩ 400 25.48 500 56.24 600
102.37

3 Application of mycelium-based biochar

(1) Prepare 20 mg/L Cu2+, Pb2+, and Cd2+ solutions respectively, adjust the pH to 6.0, and then take 50 mL
into a 250 mL Erlenmeyer flask.

(2) Add 0.05 g of mycelium-based biochar, then place the flask in a shaker and shake it at 180 r/min for 24 h.

(3) After the adsorption is completed, take the supernatant and pass it through a 0.22 micron filter membrane,
and use an atomic absorption spectrophotometer to measure the concentrations of Cu2+, Pb2+, and Cd2+
remaining in the filtrate.

(4) Mycelia-based biochar has a strong ability to adsorb heavy metal ions. When the dosage is 1 g/L, the
removal rates of mycelium-based biochar for Cu2+, Pb2+ and Cd2+ are 91.38%, 92.67% and 90.28%,
respectively. %(Table 2).

Table 2 The removal rate of heavy metal ions by mycelium-based biochar

Heavy metal category⟨![CDATA[Cu⟨sup⟩2+⟨/sup⟩]]⟩ ⟨![CDATA[Pb⟨sup⟩2+⟨/sup⟩]]⟩ ⟨![CDATA[Cd⟨sup⟩2+

⟨ /sup⟩]]⟩ Removal rate (%) 91.38% 92.67% 90.28%

The present invention uses Pleurotus eryngii mycelium as a raw material, uses sodium hydroxide to activate the
mycelium, and then prepares mycelium-based biochar under high temperature conditions, and 600° C. is a
relatively suitable carbonization temperature.

The mycelium-based biochar has good adsorption performance and can effectively remove heavy metal ions in
the environment.
The above description of the disclosed embodiments is provided to enable any person skilled in the art to make
or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the
art, and the general principles defined herein may be implemented in other embodiments without departing from
the spirit or scope of the invention. Therefore, the present invention will not be limited to the embodiments
shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed
herein.