Morchella ZJYDJ002 capable of absorbing lead ions and / or cadmium ions and its application

Abstract

The present application discloses a Morchella ZJYDJ002 capable of absorbing lead ions and / or cadmium ions and its application, which relates to the field of microbial technology and is classified as Morchella Morchella sp. , the preservation number is CGMCC No. 40977. The Morchella strain ZJYDJ002 can effectively absorb lead ions and cadmium ions in sewage.

Description

Technical Field

[0001] The present application relates to the field of microbial technology, and in particular to a Morchella ZJYDJ002 capable of absorbing lead ions and / or cadmium ions and its application. Background Art

[0002] Water is one of the important energy sources for human survival and plays a very important role in human life activities. Different types of heavy metals have different toxicity. Among them, heavy metals such as cadmium (Cd) and lead (Pb) have significant biological toxicity, are easy to migrate in the water environment, and can be enriched in organisms. Trace amounts can have toxic effects on the human body, and excessive amounts can cause mutations, carcinogenesis, and teratogenesis.

[0003] At present, the main methods for sewage treatment include chemical precipitation, membrane separation, dialysis, solvent extraction, reverse osmosis, distillation, resin ion exchange and activated carbon adsorption. Although the above methods have achieved good results to a certain extent, the wastewater still needs to be discharged during the treatment process, and there is a common problem of secondary pollution, and the treatment cost is high. The biological adsorption method is efficient and cheap, adapts to a wide range of pH values ​​and temperatures, has little pollution, no secondary pollution, and can fundamentally achieve effective treatment of heavy metals such as cadmium (Cd) and lead (Pb), and is widely used in sewage treatment.

[0004] The existing fungi used to treat heavy metal cadmium and lead include: agricultural Bacillus species (CN202210089530.2), arbuscular mycorrhizal fungi (CN201210071205.X), radioresistant Methylobacterium (CN202111001835.5), and Burkholderia (CN200710132243.0).

[0005] There has been no research on using Morchella liquid fermentation to absorb heavy metal ions in sewage, nor has there been any research on using edible fungi liquid fermentation to absorb heavy metal ions in sewage.

[0006] The information disclosed in the background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0007] In response to the above technical problems, the present application provides a Morchella strain ZJYDJ002 capable of absorbing lead ions and / or cadmium ions and its application. The Morchella strain ZJYDJ002 can effectively absorb lead ions and cadmium ions in sewage.

[0008] The present application provides a Morchella sp. ZJYDJ002 capable of adsorbing lead ions and / or cadmium ions, which is classified and named as Morchella sp. and has a preservation number of CGMCC No. 40977.

[0009] Preferably, the adsorption rate of lead ions by Morchella strain ZJYDJ002 is 29.40-82.35%; the adsorption rate of cadmium ions by Morchella strain ZJYDJ002 is 92.95-96.90%.

[0010] Preferably, the adsorption rate of lead ions in high-concentration sewage by Morchella strain ZJYDJ002 is 82.35%, and the adsorption rate of cadmium ions is 92.95%; the various indicators of high-concentration sewage are: cadmium ion content is 4 mg / L; lead ion content is 4 mg / L.

[0011] Another aspect of the present application provides a method for treating wastewater containing at least lead ions and / or cadmium ions, comprising the following steps: inoculating the above-mentioned Morchella ZJYDJ002 into a culture solution containing wastewater to obtain a first culture and then culturing it; or adding wastewater to a culture solution containing at least the mycelium of the above-mentioned Morchella ZJYDJ002 to obtain a second culture and then culturing it.

[0012] Preferably, the amount of sewage added is such that the concentration of lead ions in the mixed solution obtained after adding the sewage is less than 4 mg / L; the concentration of cadmium ions is less than 4 mg / L.

[0013] Preferably, the amount of sewage added is such that the concentration of lead ions in the mixed solution obtained after the sewage is added is 0.5-4 mg / L; the concentration of cadmium ions is 0.5-4 mg / L.

[0014] Preferably, the culture conditions of the first culture or the second culture are shaking culture at 20-25° C. for at least 7 days. Specifically, the liquid culture conditions are: at 20-25° C., at a shaking rate of 140-180 r / min, and the culture time is 5-7 days.

[0015] Preferably, the mycelium culture solution is obtained by inoculating Morchella ZJYDJ002 into a culture solution and shaking and culturing at 20-30° C. for at least 7 days.

[0016] The mycelium grows fast and has a good adsorption effect on lead ions and / or cadmium ions. It only takes 7 days of cultivation to achieve an adsorption effect of more than 80%.

[0017] Preferably, the culture solution is 5-6 parts by weight of corn flour, 6-7 parts by weight of soybean meal, 20-21 parts by weight of glucose, 1-2 parts by weight of peptone, 1 part by weight of potassium dihydrogen phosphate, 0.5 parts by weight of magnesium sulfate, and 3 parts by weight of yeast powder, and water is added to 1000 mL, and the pH is adjusted naturally.

[0018] Preferably, the inoculation amount of Morchella ZJYDJ002 in the culture solution is at least 4 pieces / 150 mL. Specifically, the inoculation amount of Morchella ZJYDJ002 in the culture solution is 4 to 6 pieces / 150 mL.

[0019] Preferably, the inoculated Morchella ZJYDJ002 is an activated test tube strain as described above.

[0020] Preferably, the test tube seed preparation method is as follows: strain ZJYDJ002 is inoculated into a comprehensive PDA medium slant and cultured at 16-20° C. for at least 3 days to obtain a test tube seed. Specifically, the test tube seed is cultured at 16-20° C. for at least 3-5 days to obtain a test tube seed.

[0021] Preferably, the comprehensive PDA medium is composed of 200 parts by weight of potato, 18 parts by weight of glucose, 18 parts by weight of agar, 1 part by weight of potassium dihydrogen phosphate, and 0.5 parts by weight of magnesium sulfate, and water is added to make the volume to 1000 mL, and the pH is natural.

[0022] The beneficial effects of this application include:

[0023] 1) The Morchella ZJYDJ002 capable of adsorbing lead ions and / or cadmium ions provided in this application and its application, the strain has a highly efficient adsorption effect and can quickly adsorb lead ions and cadmium ions in sewage. The liquid culture time of this strain is only 7 days, and the lead ions and cadmium ions in the fermentation broth can be efficiently adsorbed. It can be used for biological adsorption of lead ions and cadmium ions in sewage, providing a basis for the selection of efficient strains in urban comprehensive heavy metal sewage treatment systems. This provides a basis for the selection of efficient strains in urban comprehensive heavy metal sewage treatment systems, and also provides a reference for the research and development of microbial agents.

[0024] 2) The Morchella ZJYDJ002 capable of adsorbing lead ions and / or cadmium ions provided in this application and its application, when the strain is cultured on a plate, it can be seen that the mycelium of the strain is thick and strong, and there is no interference from foreign bacteria during the culture process, indicating that there is a certain resistance to foreign bacteria. After the slant test tube strain is inoculated, it only needs to be cultured for 3 to 5 days before it can be used for liquid culture inoculation, indicating that the mycelium grows fast, with a growth rate of 2 to 3 cm / d and fast fungus growth, which can effectively shorten the culture time and production cycle. The strain has fast fungus growth, less pollution, strong mycelium, and a short production cycle. During the fermentation culture process, the strain can effectively reduce the content of lead ions and cadmium ions added to the culture solution. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1A schematic diagram of the inclined test tube culture of Morchella ZJYDJ002 capable of adsorbing lead ions and / or cadmium ions in Example 1 provided in the present application, wherein the test tubes are numbered from left to right as test tube No. 1, test tube No. 2, test tube No. 3, test tube No. 4, test tube No. 5, test tube No. 6, test tube No. 7, and test tube No. 8;

[0026] Figure 2 A schematic diagram of the plate culture of Morchella ZJYDJ002 capable of adsorbing lead ions and / or cadmium ions in Example 2 provided in the present application;

[0027] Figure 3 A schematic diagram of the plate culture comparison of Morchella ZJYDJ002 capable of adsorbing lead ions and / or cadmium ions and other Morchella strains in Example 2 provided in this application; a) a plate inoculated with commercially available Morchella fruiting bodies; b) a plate inoculated with Morchella ZJYDJ002;

[0028] Figure 4 A schematic diagram of the liquid culture mycelium pellets of the treatment group of Morchella ZJYDJ002 capable of adsorbing lead ions and / or cadmium ions in Example 3 is provided for the present application; the conical flasks in the figure are numbered from left to right as conical flask No. 1, conical flask No. 2, conical flask No. 3, conical flask No. 4, conical flask No. 5, and conical flask No. 6. DETAILED DESCRIPTION

[0029] The present invention is further described in detail below in conjunction with the accompanying drawings and embodiments, but the present invention is not limited in any way. Any changes or improvements made based on the teachings of the present invention fall within the protection scope of the present invention. Example

[0030] Unless otherwise specified, the materials and instruments used in the following examples were obtained from commercial channels; the detection methods used were all existing methods unless otherwise specified. The culture media used were all prepared by the commonly used formulas and methods in the art unless otherwise specified. The culture and separation methods used in the following examples were all commonly used methods in the art unless otherwise specified.

[0031] Example 1 Obtaining Morchella strain ZJYDJ002

[0032] The breeding team isolated and purified the wild Morchella fruiting bodies collected from Ludian Township, Lijiang City, Yunnan Province, with the geographical coordinates: longitude: E099°27.1597', latitude: N27°09.3738', and altitude: 2877m, and obtained the Morchella strain ZJYDJ002.

[0033] The above-mentioned fresh wild Morchella fruiting bodies were collected, and after removing impurities from the surface of the fruiting bodies, they were placed in a ventilated and dry place and dried to a moisture content of 30%. In an ultra-clean workbench, after tissue separation, the separated tissue was inoculated into a slant test tube culture medium containing 1 / 3 of the test tube capacity of a comprehensive PDA culture medium, and after culturing in the dark at 20°C for 3 days, a purification operation commonly used in the field for strain separation was used, and after 4 purifications, a Morchella strain ZJYDJ002 was obtained.

[0034] The comprehensive PDA medium used above is composed of 200 g potato, 20 g glucose, 18 g agar, and 1000 ml distilled water, with natural pH, and is sterilized at 121° C. and 0.1 MPa pressure.

[0035] Example 2 Morphology and molecular identification of Morchella strain ZJYDJ002

[0036] The morphological characteristics of the obtained Morchella strain ZJYDJ002 are as follows:

[0037] 1. Morphological identification

[0038] Morchella strain ZJYDJ002 grew well on PDA medium at 20°C and was sealed after culturing for 4 days. The PDA medium used was composed of 200 g potato, 20 g glucose, 20 g agar, and 1000 ml distilled water, with natural pH.

[0039] Inoculate the inclined test tubes of test tubes 1 to 6. After 2 days of culture at 16-20°C, the mycelium forms light yellow filamentous mycelium. After 4 days of culture, the mycelium turns yellow-brown and begins to produce pigment. After 7 days of culture, yellow sclerotia begin to form. Figure 1 As shown, test tubes 1 to 6 are all parallel treatment samples, and the inoculation amount is 4 pieces / bottle.

[0040] The strain was inoculated on a comprehensive PDA medium plate and cultured under the same conditions for 7 days to obtain the following Figure 2 The plate shown, see Figure 2 It can be seen that the mycelium of strain ZJYDJ002 is thick and no foreign bacteria are found, indicating that the strain has a certain effect of inhibiting foreign bacteria;

[0041] The Morchella strain purchased from the market was selected as a control strain and inoculated with strain ZJYDJ002 on a comprehensive PDA medium plate and cultured for 2 days to obtain the following Figure 3 The comparison diagrams shown in a) and b) show that under the same culture time, Figure 3 a) To show that the existing Morchella strains grow slowly, Figure 3 b) The strain ZJYDJ002 grew faster. The mycelial growth rate was measured to be 2-3 cm / d during the culture, indicating that the strain had a fast germination effect.

[0042] 2. Strain preservation:

[0043] The cultivated strain in the following embodiments is the Morchella strain ZJYDJ002 disclosed in CN202311702383.2, which was deposited in the General Microbiological Center of the China Microbiological Culture Collection Administration Committee on November 10, 2023, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; classified and named Morchella sp., deposit number CGMCC No. 40977. For other undetailed contents, see CN202311702383.2.

[0044] Example 3 Biosorption of lead and cadmium ions in sewage by Morchella strain ZJYDJ002

[0045] 1. Preparation of test tube strains: The strain ZJYDJ002 isolated and purified in Example 1 was inoculated into a slant of a comprehensive PDA culture medium and cultured at 20°C for 4 days to obtain test tube strains. Figure 1 shown.

[0046] The comprehensive PDA medium is composed of 200 parts by weight of potato, 18 parts by weight of glucose, 18 parts by weight of agar, 1 part by weight of potassium dihydrogen phosphate, and 0.5 parts by weight of magnesium sulfate. Water is added to make the volume to 1000 mL, and the pH is natural.

[0047] 2. Liquid culture: 5-6 parts by weight of corn flour, 6-7 parts by weight of soybean meal, 20-21 parts by weight of glucose, 1-2 parts by weight of peptone, 1 part by weight of potassium dihydrogen phosphate, 0.5 parts by weight of magnesium sulfate, and 3 parts by weight of yeast powder. Add water to 1000 mL and adjust the pH to natural to obtain a liquid culture medium.

[0048] Six parallel treatments were set up, each of which contained three conical flasks with 150 mL of liquid culture medium. 75 uL of sewage was added to each conical flask so that the concentrations of lead and cadmium in the culture medium were both 0.5 mg / L. The sewage added in the treatment was prepared by weighing 1.599 g of lead nitrate and 2.032 g of cadmium chloride, dissolving them in 50 ml of 10% nitric acid solution, transferring them to a 1000 mL volumetric flask, adding pure water to the mark, and mixing. The mass concentrations of lead and cadmium in this solution were both 1000 ug / mL for later use.

[0049] Three control treatments were set up at the same time, and three conical flasks containing 150 mL of liquid culture medium were set up for each parallel treatment, and 75 uL of water was added to each conical flask.

[0050] The treatment group and the control group were sterilized at 121°C for 30 minutes, cooled to below 25°C, and inoculated with activated test tubes. The inoculation amount was 4 pieces / bottle, sealed, and cultured at 20°C, 150 rpm, for 7 days. Figure 4 As shown, the six parallel treatments were Figure 4 The conical flasks in the box correspond to each other.

[0051] 3. Take 100 ml of the culture medium after 7 days of cultivation in each treatment to detect the concentration of lead ions and cadmium ions: filter the mycelium of the liquid fermentation culture of Morchella with a 300-mesh sieve or filter cloth, collect the filtered filtrate and centrifuge it at 7000 r / min for 5 min, take the supernatant after centrifugation and pass it through a 0.45 um filter membrane, and immediately acidify the filtrate with nitric acid to pH 1-2, and finally determine it by atomic absorption spectrophotometry.

[0052] The adsorption rate of lead ions and cadmium ions by the strains in each treatment was calculated as follows:

[0053]

[0054] Results: In the high, medium and low-quality sewage as shown in Table 1, the adsorption rates of lead ions and cadmium ions by Morchella strain ZJYDJ002 were 29.40% and 96.60% respectively.

[0055] From the above, it can be seen that the Morchella strain ZJYDJ002 can effectively adsorb the lead ions and cadmium ions contained in the culture medium.

[0056] Example 4 Biosorption of lead and cadmium ions in sewage by Morchella strain ZJYDJ002

[0057] The difference from Example 3 is that 300uL of sewage is added to the conical flask of each treatment group so that the concentrations of lead and cadmium in the culture solution are both 2.0mg / L.

[0058] Add 300uL of water to the conical flask of each control group.

[0059] The treated group and the control group were sterilized at 121°C for 30 min, cooled to below 25°C, inoculated with activated test tube seeds, with the inoculation amount of 4 pieces / bottle, sealed, and cultured at 20°C, 150 rpm, for 7 days.

[0060] According to the method and calculation formula in Example 3, it was obtained that the adsorption rates of lead ions and cadmium ions by Morchella strain ZJYDJ002 in sewage were 64.05% and 96.90%.

[0061] Example 5 Biosorption of lead and cadmium ions in sewage by Morchella strain ZJYDJ002

[0062] The difference from Example 3 is that 600uL of sewage was added to the conical flask of each treatment group, so that the concentrations of lead and cadmium in the culture solution were both 4.0mg / L.

[0063] Add 600uL of water to the conical flask of each control group.

[0064] The treated group and the control group were sterilized at 121°C for 30 min, cooled to below 25°C, inoculated with activated test tube seeds, with the inoculation amount of 4 pieces / bottle, sealed, and cultured at 20°C, 150 rpm, for 7 days.

[0065] According to the method and calculation formula in Example 3, it was obtained that the adsorption rates of lead ions and cadmium ions by Morchella strain ZJYDJ002 in sewage were 82.35% and 92.95%.

[0066] From the results obtained in Examples 3 to 5, it can be seen that strain ZJYDJ002 can significantly reduce the lead ion and cadmium ion concentrations in sewage with a lead ion and cadmium ion concentration of 0.5 to 4 mg / L, and can be used to effectively treat lead ions and cadmium ions in sewage within this concentration range.

[0067] Example 6 Adsorption effect of sewage of different concentrations

[0068] The difference from Example 3 is that after preparing the three kinds of sewage as shown in Table 1, the sewage shown in Table 1 is added, inoculated and cultured according to the treatment group operations of Example 3; at the same time, CK is set as a control, and the only difference between the control and the treatment is that pure water with an equal volume of sewage is added.

[0069] Table 1 Sewage configuration (mg / L)

[0070]

[0071] The concentrations of lead ions and cadmium ions in the culture medium of the treatment group and CK were detected according to the method in Example 3. The results of the detection are shown in Table 2:

[0072] Table 2 Changes in lead and cadmium concentrations in Morchella strain ZJYDJ002 under different treatments

[0073]

[0074] The results in Table 2 show that in high, medium and low sewage concentrations, the strain ZJYDJ002 can decompose or adsorb lead ions and cadmium ions in sewage during its growth, thereby reducing the concentrations of lead ions and cadmium ions in sewage of different concentrations, and the decomposition or adsorption effect is significant compared with the control group CK.

[0075] After inoculation with bacterial suspension, the adsorption rates of lead ions and cadmium ions of strain ZJYDJ002 for high, medium and low concentration sewage are shown in Table 2. This indicates that the strain is suitable for treating sewage in the concentration range in Table 1.

[0076] Example 7

[0077] The difference from Example 3 is that after the bacterial blocks are inoculated in the culture solution, sewage is added to obtain the first culture, and the culture conditions of the first culture are: at 22°C, at a shaking rate of 140 r / min, the culture time is 5 days; the inoculation amount is 5 blocks / 150 mL.

[0078] Specifically, the culture was carried out at 20°C for 3 days to obtain the test tube seeds.

[0079] In the liquid culture medium: 5 parts by weight of corn flour, 6 parts by weight of soybean meal, 20 parts by weight of glucose, and 1 part by weight of peptone.

[0080] Example 8

[0081] The difference from Example 3 is that after the bacterial blocks are inoculated in the culture solution, sewage is added to obtain the first culture, and the culture conditions of the first culture are: at 25°C, at a shaking rate of 180 r / min, the culture time is 6 days; the inoculation amount is 6 blocks / 150 mL.

[0082] Specifically, the culture was carried out at 16°C for 5 days to obtain the test tube seeds.

[0083] In the liquid culture medium: 6 parts by weight of corn flour, 7 parts by weight of soybean meal, 21 parts by weight of glucose, and 2 parts by weight of peptone.

[0084] Example 9

[0085] The difference from Example 3 is:

[0086] Each treatment group inoculated sewage into the second culture; the second culture was a mixture of Morchella ZJYDJ002 mycelium and culture medium after inoculating 4 fungus blocks into 150 ml of liquid culture medium and culturing at 22°C for 8 days. The concentration of lead ions in the sewage used was 3 mg / L; the concentration of cadmium ions was 2 mg / L.

[0087] Example 10

[0088] The difference from Example 3 is:

[0089] Each treatment group inoculated sewage into the second culture; the second culture was a mixture of Morchella ZJYDJ002 mycelium and culture solution after inoculating 4 fungus blocks into 150 ml of liquid culture solution and culturing at 25°C for 7 days. The concentration of lead ions in the sewage used was 2 mg / L; the concentration of cadmium ions was 3 mg / L.

[0090] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for treating sewage containing at least lead ions and / or cadmium ions, characterized in that: The following steps are involved: Inoculating Morchella ZJYDJ002 into a culture solution containing sewage to obtain a first culture and then culturing it; or adding sewage to a culture solution at least including mycelium of Morchella ZJYDJ002 to obtain a second culture and then culturing; Morchella ZJYDJ002, classified as Morchella Morchella sp. ), deposit number CGMCC No .40977; The adsorption rate of lead ions by Morchella strain ZJYDJ002 was 29.40~82.35%; the adsorption rate of cadmium ions by Morchella strain ZJYDJ002 was 92.95~96.90%.

2. The method according to claim 1, characterized in that The amount of sewage added is such that the concentration of lead ions in the mixed solution obtained after adding the sewage is less than 4 mg / L; the concentration of cadmium ions is less than 4 mg / L.

3. The method according to claim 1, characterized in that The amount of sewage added is such that the concentration of lead ions in the mixed solution obtained after adding the sewage is 0.5~4 mg / L; the concentration of cadmium ions is 0.5~4 mg / L.

4. The method according to claim 1, characterized in that The culture conditions of the first culture or the second culture are shaking culture at 20-25°C for at least 7 days; The mycelium culture solution is obtained by inoculating Morchella ZJYDJ002 into the culture solution and shaking and culturing at 20-30° C. for at least 7 days.

5. The method according to claim 1, characterized in that The inoculation amount of Morchella ZJYDJ002 in the culture medium is at least 4 pieces / 150mL.

6. The method according to claim 1, characterized in that The culture solution is 5-6 parts by weight of corn flour, 6-7 parts by weight of soybean meal, 20-21 parts by weight of glucose, 1-2 parts by weight of peptone, 1 part by weight of potassium dihydrogen phosphate, 0.5 parts by weight of magnesium sulfate, and 3 parts by weight of yeast powder, and water is added to 1000 mL, and the pH is adjusted naturally; The inoculation amount of Morchella ZJYDJ002 in the culture medium was 4 to 6 pieces / 150 mL.

7. The method according to claim 1, characterized in that The inoculated Morchella ZJYDJ002 is an activated test tube strain as described above; the test tube strain preparation method is as follows: the strain ZJYDJ002 is inoculated in a slant of a comprehensive PDA medium and cultured at 16-20° C. for at least 3 days to obtain a test tube strain; The comprehensive PDA medium is composed of 200 parts by weight of potato, 18 parts by weight of glucose, 18 parts by weight of agar, 1 part by weight of potassium dihydrogen phosphate, and 0.5 parts by weight of magnesium sulfate. Water is added to make the volume to 1000 mL, and the pH is natural.

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