A method for degrading tetrabutylphosphonium bromide using tetrahymena

Abstract

The application discloses a method for degrading tetrabutylphosphonium bromide by using tetrahymena. The method is for degrading tetrabutylphosphonium bromide by using tetrahymena in a starvation medium under aerobic conditions. The application first proposes the purpose of eliminating tetrabutylphosphonium bromide through biological degradation of tetrahymena pyriformis. The application provides a new idea for degradation of tetrabutylphosphonium bromide, and has potential to be further optimized and popularized. The method has the advantages of green environmental protection and no energy consumption.

Description

Technical Field

[0001] The invention belongs to the field of biodegradation of ionic liquids, and particularly relates to a method for degrading tetrabutylphosphonium bromide by utilizing Tetrahymena. Background Art

[0002] Ionic liquids generally refer to molten salts composed of organic cations and organic / inorganic anions with a melting temperature below 100°C. As alternatives to organic solvents, ionic liquids hold significant promise in catalysis, food extraction, and food analysis. Quaternary phosphine ionic liquids, such as tetrabutylphosphonium bromide, are common. However, their difficulty in rapid biodegradation poses significant environmental risks, a critical issue that must be addressed before their widespread application.

[0003] Biodegradation primarily utilizes the metabolic activity of microorganisms to convert organic pollutants into non-toxic carbon dioxide and water. Since the 19th century, biodegradation technology has played a vital role in water treatment and has been widely used for its environmentally friendly, energy-saving properties. Tetrahymena piriformis, a protozoan long used as a model organism in molecular biology and toxicology, is found in freshwater systems worldwide and can be cultivated at low cost, making it well-suited for studying the degradation of tetrabutylphosphonium bromide. Summary of the Invention

[0004] The present invention provides a method for biodegrading tetrabutylphosphonium bromide, specifically using Tetrahymena pyriformis. Tetrabutylphosphonium bromide's carbon source is of little use when Tetrahymena is in a nutrient-rich medium, so the present invention performs the degradation process in a starvation medium.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] A method for degrading tetrabutylphosphonium bromide by using Tetrahymena is to degrade tetrabutylphosphonium bromide by using Tetrahymena in a starvation culture medium under aerobic conditions.

[0007] Preferably, the Tetrahymena is Tetrahymena piriformis.

[0008] Preferably, the Tetrahymena is cultured in a nutrient-rich medium and then enriched and transferred to a starvation medium.

[0009] More preferably, the Tetrahymena is cultured in large quantities using SSP medium at 26-30° C. and 160 rpm, and then enriched and washed by centrifugation at room temperature and 1500 rpm for 3 minutes.

[0010] Preferably, the starvation medium is Dryl's buffer or 10 mM Tris-HCl (pH 7.4) buffer.

[0011] Preferably, the Tetrahymena concentration is 200,000-3,000,000 / mL, and the count is performed using a hemocytometer.

[0012] Preferably, the added concentration of tetrabutylphosphonium bromide is 0.1-10 mg / mL.

[0013] Preferably, the Tetrahymena degradation temperature is 20-30° C. and the rotation speed is 0-220 rpm.

[0014] The present invention has the following beneficial effects:

[0015] The present invention provides a method for biodegrading tetrabutylphosphonium bromide for the first time, specifically utilizing Tetrahymena piriformis to biodegrade tetrabutylphosphonium bromide in a starvation culture medium. The method of the present invention has the advantages of being environmentally friendly and energy-saving. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is the secondary mass spectrum of the degradation intermediate product of tetrabutylphosphonium bromide in Example 1.

[0017] Figure 2 The degradation pathway of tetrabutylphosphonium bromide in Example 1 is DETAILED DESCRIPTION

[0018] In order to make the above-mentioned purpose of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are further described in detail below in conjunction with specific examples, but the protection scope of the present invention is not limited to the following examples.

[0019] Unless otherwise specified, the experimental reagents used in the following examples are all conventional biochemical reagents.

[0020] Example 1

[0021] SSP medium ( 10g of peptone, 1g of yeast extract powder, 2g of glucose, 0.03g of sodium ferric ethylenediaminetetraacetic acid, 1000mL of distilled water, mixed and sterilized) was cultured at 28°C and 160rpm for 24 hours. The mixture was collected by centrifugation at 1500rpm for 3min at room temperature and washed three times with Dryl's buffer (2mM sodium citrate, 1mM NaH2PO4, 1mM Na2HPO4·12H2O, 1000mL of distilled water, mixed and sterilized, and then 1.5mM CaCl2 was added). The Tetrahymena in Dryl's buffer was counted using a hemocytometer until the concentration of Tetrahymena in the Dryl's buffer reached 1 million / mL. 1mg / mL of tetrabutylphosphonium bromide ([BPy]Cl) was added and the LC-MS results after degradation at 28°C and 160rpm for 13 days were as follows: Figure 1As shown in Figure 2, the degradation efficiency reached 64.76±30.80%, and the secondary mass spectrometry showed the degradation intermediates. Figure 2 shown.

[0022] Example 2

[0023] SSP medium ( Tetrahymena pyriformis was cultured with 10 g of peptone, 1 g of yeast extract powder, 2 g of glucose, 0.03 g of sodium ferric ethylenediaminetetraacetic acid, and 1000 mL of distilled water (mixed and sterilized) at 28°C and 160 rpm for 24 hours. The cells were collected by centrifugation at 1500 rpm for 3 minutes at room temperature and washed three times with Dryl's buffer (2 mM sodium citrate, 1 mM NaH2PO4, 1 mM Na2HPO4·12H2O, 1000 mL of distilled water, mixed and sterilized, and then supplemented with 1.5 mM CaCl2). The Tetrahymena cells were counted using a hemocytometer until the concentration of Tetrahymena in Dryl's buffer reached 3 million cells / mL. After 24 days of degradation at 20°C and 220 rpm, the ionic liquid degradation efficiency reached 56.11 ± 9.20%.

[0024] Example 3

[0025] SSP medium ( Peptone (10 g), yeast extract (1 g), glucose (2 g), sodium ferric ethylenediaminetetraacetic acid (EDTA) (0.03 g), and distilled water (1000 mL, mixed and sterilized) were cultured at 28°C and 160 rpm for 24 hours. The cells were collected by centrifugation at 1500 rpm for 3 minutes at room temperature and washed three times with 10 mM Tris-HCl (pH 7.4) buffer. The cells were counted using a hemocytometer to a concentration of 200,000 Tetrahymena / mL in 10 mM Tris-HCl (pH 7.4). After addition of 0.1 mg / mL [BPy]Cl and degradation at 30°C for 24 days, the ionic liquid degradation efficiency reached 30.56 ± 3.37%.

[0026] The above are merely preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be construed as limiting the present invention, and the scope of protection of the present invention should be determined by the scope defined in the claims. Persons skilled in the art will appreciate that improvements and modifications may be made without departing from the spirit and scope of the present invention, and such improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for degrading tetrabutylphosphonium bromide using Tetrahymena, characterized in that: The method comprises the following steps: degrading tetrabutylphosphonium bromide by using Tetrahymena in a starvation culture medium under aerobic conditions; the Tetrahymena is Tetrahymena pear-shaped.

2. The method according to claim 1, characterized in that The starvation medium is Dryl's buffer or 10 mM Tris-HCl buffer, and the pH value of the Tris-HCl buffer is 7.

4.

3. The method according to claim 1, characterized in that The concentration of the Tetrahymena is 200,000 to 3,000,000 / mL.

4. The method according to claim 1, wherein The added concentration of the tetrabutylphosphonium bromide is 0.1-10 mg / mL.

5. The method according to claim 1, wherein The degradation temperature is 20-30° C. and the rotation speed is 0-220 rpm.

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