High-temperature-resistant salt-tolerant microbial inoculant culture nutrient solution and preparation method thereof

Abstract

The invention discloses a high-temperature-resistant salt-tolerant microbial inoculant culture nutrient solution and a preparation method thereof. The high-temperature-resistant salt-tolerant microbial inoculant culture nutrient solution comprises the following components: 1.0% (w / v) cane sugar or 1.0% (w / v) glucose; 0.5% (w / v) potassium nitrate (KNO3); 0.3% (w / v) of monopotassium phosphate (KH2PO4); 2-3% (w / v) sodium chloride (NaCl); 0.05% (w / v) of nano silicate; 0.1% (w / v) of a seaweed extract; 0.001% (w / v) of carbon nanotubes; 0.002% (w / v) of amino acid chelated iron; 0.2% (w / v) of agarose or 0.2% (w / v) of xanthan gum; 0.01% (w / v) of a mixture of vitamin C and vitamin E; 0.0005% (w / v) of gibberellin and cytokinin; and 0.005% (w / v) HEPES buffer solution. By adding the nano silicate and the carbon nanotubes, the thermal stability is enhanced, microbial cells are protected to survive and breed in a high-temperature environment, the salinity adaptability is improved by the sodium chloride and the seaweed extract, the growth of microorganisms in a high-salt environment is enhanced, the microorganisms can be effectively protected to survive and breed in the high-temperature and high-salt environment, and the survival rate of the microorganisms in the high-temperature and high-salt environment is improved. Sufficient nutritional ingredients are provided to promote rapid growth of microorganisms, meanwhile, the pH value of the nutrient solution is kept stable, and smooth proceeding of the culture process is ensured.

Description

Technical Field

[0001] The invention relates to the technical field of microbial nutrient solution, and in particular to a high temperature resistant and salt tolerant microbial agent culture nutrient solution and a preparation method thereof. Background Art

[0002] High temperature and salt resistant microbial culture nutrient solution plays an important role in practical applications, especially in microbial research and industrial production in extreme environments. For example, in the fields of oil extraction, seawater desalination, salt lake resource development, environmental remediation, etc., it is necessary to cultivate specific microorganisms under high temperature and high salt conditions to achieve efficient biotransformation and pollutant degradation. The survival and reproduction ability of these microorganisms in extreme environments directly affects the process effect and economic benefits.

[0003] However, the current microbial culture nutrient solution has some significant disadvantages. First, ordinary nutrient solution has poor stability under high temperature conditions, which can easily lead to the decomposition of nutrients and affect the normal growth of microorganisms. Second, the insufficient osmotic pressure regulation ability in a high-salt environment makes it difficult for microorganisms to adapt and survive, reducing the culture efficiency.

[0004] In view of the above problems, we have introduced a high temperature resistant and salt tolerant microbial culture nutrient solution and a preparation method thereof. Summary of the invention

[0005] The invention discloses a high temperature resistant and salt resistant microbial agent culture nutrient solution and a preparation method thereof, aiming to solve the technical problems in the background technology.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A high temperature resistant and salt tolerant microbial agent culture nutrient solution, comprising the following components:

[0008] 1.0% (w / v) sucrose or 1.0% (w / v) glucose;

[0009] 0.5% (w / v) potassium nitrate (KNO3);

[0010] 0.3% (w / v) potassium dihydrogen phosphate (KH2PO4);

[0011] 2-3% (w / v) sodium chloride (NaCl);

[0012] 0.05% (w / v) nanosilicate;

[0013] 0.1% (w / v) seaweed extract;

[0014] 0.001% (w / v) carbon nanotubes;

[0015] 0.002% (w / v) amino acid chelated iron;

[0016] 0.2% (w / v) agarose or 0.2% (w / v) xanthan gum;

[0017] 0.01% (w / v) vitamin C and E mixture;

[0018] 0.0005% (w / v) gibberellins and cytokinins;

[0019] 0.005% (w / v) HEPES buffer.

[0020] In a preferred embodiment, the particle size of the nano-silicate is 10-100 nanometers.

[0021] In a preferred embodiment, the carbon nanotubes are multi-walled carbon nanotubes and have been sterilized and dispersed.

[0022] In a preferred embodiment, the seaweed extract is derived from red algae or green algae.

[0023] In a preferred embodiment, the source of the agarose or xanthan gum is microbial fermentation or plant extraction.

[0024] In a preferred embodiment, the ratio of vitamin C to vitamin E in the vitamin C and E mixture is 1:1.

[0025] In a preferred embodiment, the gibberellins and cytokinins are derived from plant extraction or chemical synthesis and are aseptically processed.

[0026] In a preferred embodiment, the pH value of the HEPES buffer is 7.0-7.5.

[0027] A method for preparing a high temperature resistant and salt tolerant microbial agent culture nutrient solution comprises the following steps:

[0028] S1. Add 1.0% (w / v) sucrose or glucose, 0.5% (w / v) potassium nitrate (KNO3), 0.3% (w / v) potassium dihydrogen phosphate (KH2PO4) and 2-3% (w / v) sodium chloride (NaCl) to 90% deionized water preheated to 60-70°C, and stir until completely dissolved;

[0029] S2, sequentially add 0.05% (w / v) nanosilicate, 0.1% (w / v) seaweed extract, 0.001% (w / v) carbon nanotubes, 0.002% (w / v) amino acid chelated iron, 0.2% (w / v) agarose or xanthan gum, 0.01% (w / v) vitamin C and E mixture, 0.0005% (w / v) gibberellin and cytokinin, 0.005% (w / v) HEPES buffer, and gently stir after adding each ingredient to ensure uniform distribution;

[0030] S3, using sodium hydroxide (NaOH) or hydrochloric acid (HCl) to adjust the pH value to 7.0-7.5;

[0031] S4. Add deionized water or distilled water to the final volume to ensure that the concentration of all ingredients meets the ratio requirements;

[0032] S5. Place the prepared nutrient solution into an autoclave and sterilize at 121°C for 15-20 minutes;

[0033] S6. After sterilization, cool the nutrient solution to room temperature and store it in a clean and dry environment for future use.

[0034] The high temperature resistant and salt tolerant microbial agent culture nutrient solution and the preparation method thereof provided by the present invention have the following advantages:

[0035] The invention enhances thermal stability and protects microbial cells from surviving and reproducing in a high-temperature environment by adding nano-silicate and carbon nanotubes; sodium chloride and seaweed extract improve salinity adaptability and enhance the growth of microorganisms in a high-salt environment; sucrose or glucose provides sufficient carbon source; potassium nitrate and potassium dihydrogen phosphate provide necessary nitrogen source and phosphorus source; the invention can effectively protect microorganisms from surviving and reproducing in a high-temperature and high-salt environment, provide sufficient nutrients to promote rapid growth of microorganisms, and simultaneously maintain the pH value of the nutrient solution stable, thereby ensuring the smooth progress of the cultivation process. DETAILED DESCRIPTION

[0036] The invention discloses a high temperature resistant and salt tolerant microbial agent culture nutrient solution and a preparation method thereof.

[0037] A high temperature resistant and salt tolerant microbial agent culture nutrient solution, the composition and concentration of which are as follows:

[0038] 1.0% (w / v) sucrose or glucose, 0.5% (w / v) potassium nitrate (KNO3), 0.3% (w / v) potassium dihydrogen phosphate (KH2PO4), 2-3% (w / v) sodium chloride (NaCl), 0.05% (w / v) nanosilicate, the particle size of the nanosilicate is 10-100 nanometers, 0.1% (w / v) seaweed extract, the seaweed extract is derived from red algae or green algae, 0.001% (w / v) carbon nanotubes, the carbon nanotubes are multi-walled carbon nanotubes, and have been sterilized and dispersed, 0.0 0.2% (w / v) amino acid chelated iron, 0.2% (w / v) agarose or xanthan gum, the agarose or xanthan gum is derived from microbial fermentation or plant extraction, 0.01% (w / v) vitamin C and E mixture, the ratio of vitamin C and E is 1:1, 0.0005% (w / v) gibberellins and cytokinins, the gibberellins and cytokinins are derived from plant extraction or chemical synthesis and are aseptically treated, 0.005% (w / v) HEPES buffer, the pH value of the HEPES buffer is 7.0-7.5;

[0039] Place 90% deionized water or distilled water preheated to 60-70°C in a clean container, add 1.0% (w / v) sucrose or glucose, 0.5% (w / v) potassium nitrate (KNO3), 0.3% (w / v) potassium dihydrogen phosphate (KH2PO4) and 2-3% (w / v) sodium chloride (NaCl) in sequence, stir gently until completely dissolved, ensure that all ingredients are completely dissolved and no particles remain, then add 0.05% (w / v) nanosilicate, 0.1% (w / v) seaweed extract, 0.001% (w / v) carbon nanotubes, 0.002% (w / v) amino acid chelated iron, 0.2% (w / v) agarose or xanthan gum, 0.01% (w / v) Vitamin C and E mixture, 0.0005% (w / v) gibberellin and cytokinin, 0.005% (w / v) HEPES buffer, gently stir after adding each ingredient to ensure uniform distribution, use a pH meter to test the pH value of the nutrient solution, use sodium hydroxide (NaOH) or hydrochloric acid (HCl) to adjust to between 7.0 and 7.5 when necessary, ensure that the pH value is within the range suitable for microbial growth, add deionized water or distilled water to the final volume, ensure that the concentration of all ingredients meets the requirements of the formula, transfer the prepared nutrient solution to an autoclave, sterilize at 121°C for 15-20 minutes, and after sterilization, cool the nutrient solution to room temperature and store it in a clean and dry environment for use;

[0040] Things to note during operation:

[0041] The carbon nanotubes need to be sterilized and dispersed in advance to avoid condensation in the nutrient solution;

[0042] Seaweed extracts and polysaccharides (such as agarose or xanthan gum) may increase the viscosity of the culture medium, and the stirring speed and time need to be adjusted appropriately to ensure uniform distribution of the ingredients;

[0043] All added components must be sterile to avoid contamination of the culture medium;

[0044] The pH value should be adjusted carefully to avoid the pH value being too high or too low, which may affect the growth of microorganisms.

[0045] A method for preparing a high temperature resistant and salt tolerant microbial agent culture nutrient solution comprises the following steps:

[0046] S1. Add 1.0% (w / v) sucrose or glucose, 0.5% (w / v) potassium nitrate (KNO3), 0.3% (w / v) potassium dihydrogen phosphate (KH2PO4) and 2-3% (w / v) sodium chloride (NaCl) to 90% deionized water preheated to 60-70°C, and stir until completely dissolved;

[0047] S2, sequentially add 0.05% (w / v) nanosilicate, 0.1% (w / v) seaweed extract, 0.001% (w / v) carbon nanotubes, 0.002% (w / v) amino acid chelated iron, 0.2% (w / v) agarose or xanthan gum, 0.01% (w / v) vitamin C and E mixture, 0.0005% (w / v) gibberellin and cytokinin, 0.005% (w / v) HEPES buffer, and gently stir after adding each ingredient to ensure uniform distribution;

[0048] S3, using sodium hydroxide (NaOH) or hydrochloric acid (HCl) to adjust the pH value to 7.0-7.5;

[0049] S4. Add deionized water or distilled water to the final volume to ensure that the concentration of all ingredients meets the ratio requirements;

[0050] S5. Place the prepared nutrient solution into an autoclave and sterilize at 121°C for 15-20 minutes;

[0051] S6. After sterilization, cool the nutrient solution to room temperature and store it in a clean and dry environment for future use.

[0052] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. The replacement may be a replacement of a part of the structure, device, method step, or a complete technical solution. Any equivalent replacement or change according to the technical solution and the inventive concept of the present invention shall be covered within the protection scope of the present invention.

Claims

1. A high temperature resistant and salt tolerant microbial agent culture nutrient solution, characterized in that: Includes the following ingredients: 1.0% (w / v) sucrose or 1.0% (w / v) glucose; 0.5% (w / v) potassium nitrate (KNO3); 0.3% (w / v) potassium dihydrogen phosphate (KH2PO4); 2-3% (w / v) sodium chloride (NaCl); 0.05% (w / v) nanosilicate; 0.1% (w / v) seaweed extract; 0.001% (w / v) carbon nanotubes; 0.002% (w / v) amino acid chelated iron; 0.2% (w / v) agarose or 0.2% (w / v) xanthan gum; 0.01% (w / v) vitamin C and E mixture; 0.0005% (w / v) gibberellins and cytokinins; 0.005% (w / v) HEPES buffer.

2. The high temperature resistant and salt tolerant microbial agent culture nutrient solution according to claim 1, characterized in that: The particle size of the nano silicate is 10-100 nanometers.

3. The high temperature resistant and salt tolerant microbial agent culture nutrient solution according to claim 1, characterized in that: The carbon nanotubes are multi-walled carbon nanotubes and have been sterilized and dispersed.

4. The high temperature resistant and salt tolerant microbial agent culture nutrient solution according to claim 1, characterized in that: The seaweed extract is derived from red algae or green algae.

5. The high temperature resistant and salt tolerant microbial agent culture nutrient solution according to claim 1, characterized in that: The agarose or xanthan gum is obtained from microbial fermentation or plant extraction.

6. The high temperature resistant and salt tolerant microbial agent culture nutrient solution according to claim 1, characterized in that: The ratio of vitamin C to vitamin E in the vitamin C and E mixture is 1:

1.

7. The high temperature resistant and salt tolerant microbial agent culture nutrient solution according to claim 1, characterized in that: The gibberellins and cytokinins are derived from plant extraction or chemical synthesis and are aseptically processed.

8. The high temperature resistant and salt tolerant microbial agent culture nutrient solution according to claim 1, characterized in that: The pH value of the HEPES buffer is 7.0-7.

5.

9. The method for preparing a high temperature resistant and salt tolerant microbial agent culture nutrient solution according to any one of claims 1 to 8, characterized in that: The following steps are involved: S1. Add 1.0% (w / v) sucrose or glucose, 0.5% (w / v) potassium nitrate (KNO3), 0.3% (w / v) potassium dihydrogen phosphate (KH2PO4) and 2-3% (w / v) sodium chloride (NaCl) to 90% deionized water preheated to 60-70°C, and stir until completely dissolved; S2, sequentially add 0.05% (w / v) nanosilicate, 0.1% (w / v) seaweed extract, 0.001% (w / v) carbon nanotubes, 0.002% (w / v) amino acid chelated iron, 0.2% (w / v) agarose or xanthan gum, 0.01% (w / v) vitamin C and E mixture, 0.0005% (w / v) gibberellin and cytokinin, 0.005% (w / v) HEPES buffer, and gently stir after adding each ingredient to ensure uniform distribution; S3, using sodium hydroxide (NaOH) or hydrochloric acid (HCl) to adjust the pH value to 7.0-7.5; S4. Add deionized water or distilled water to the final volume to ensure that the concentration of all ingredients meets the ratio requirements; S5. Place the prepared nutrient solution into an autoclave and sterilize at 121°C for 15-20 minutes; S6. After sterilization, cool the nutrient solution to room temperature and store it in a clean and dry environment for future use.