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High tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles and antibacterial application of high tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles

A technology of high tellurite and synthetic biology, applied in the direction of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., to achieve the effects of simple preservation, safe and controllable acquisition methods, and stable biological functions

Active Publication Date: 2022-05-03
HUBEI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] It has been reported in the literature that many microorganisms can reduce Te(IV) to tellurium nanostructures extracellularly or intracellularly through enzymatic or non-enzymatic reactions, but there are few studies on the synthesis of tellurium nanostructures using fungi. In addition, this study also used Biosynthesized tellurium nanomaterials are used as antibacterial materials, on the one hand, to alleviate the high toxicity of Te(IV) in the environment, on the other hand, the produced biological tellurium nanoparticles have a unique antibacterial function, which has dual significance for alleviating environmental problems, This invention fills the technical gap in this direction

Method used

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  • High tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles and antibacterial application of high tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles
  • High tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles and antibacterial application of high tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles
  • High tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles and antibacterial application of high tellurite tolerant bacterium-mediated synthesized biological tellurium nano-particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Isolation, screening and identification of tellurite-tolerant strains

[0038] (1) Select 1g of waste dump soil sample, add 10mL of sterilized normal saline with a mass volume ratio of 0.85%, and mix in a shaker at 37°C and 200rpm for 30min;

[0039] (2) Heat to dissolve 100mL solid LB medium (tryptone 10g, yeast extract 5g, sodium chloride 10g, agar 20g, add dd H 2 (2) Dilute to 1000mL, 121°C, use after 20min sterilization), add potassium tellurite (KT) with a final concentration of 0.1mmol / L 2 TeO 3 ), pour the plate, take 200 μL of the above-mentioned dissolved soil sample, spread it evenly on the surface of the plate, and incubate at 37°C for 16 hours;

[0040] (3) Select black mycelium, inoculate to solid PDA plate (potato 200g, glucose 20g, agar 20g, add ddH 2 (2) Dilute to 1000mL, 121°C, use after 20min sterilization), and cultivate at 28°C for 6-7 days;

[0041] (4) Cut small pieces of solid PDA (containing mycelium), add it to 100mL liquid PDA, an...

Embodiment 2

[0048] Example 2 Tellurite tolerance test

[0049] (1) Optimizing culture medium: the primary shake flask uses corn flour medium (corn flour 40g / L, KNO 3 2 g / L, NaHPO 4 1g / L, MgSO 4 ·7H 2 O 0.3g / L), seal the medium after preparation, and sterilize at 121°C for 20min. The secondary shake flask uses improved PDA medium (potato 200g / L, KNO 3 2g / L, NaHPO 4 1g / L, MgSO 4 ·7H 2 (0.3g / L), the culture medium was prepared and sealed, and sterilized at 121°C for 20 minutes;

[0050] (2) Using the slant strain in Example 1, cut small pieces of solid PDA (containing mycelia), add it to a 100mL first-stage shaker flask, and cultivate it for 3 to 5 days at 28°C and 200r / min. , to obtain an appropriate amount of liquid bacteria;

[0051] (3) Take 5mL of the liquid strains in the primary shake flask, add them to the 100mL secondary shake flask, and cultivate them for 3 to 5 days at 28°C and 200r / min to obtain a large amount of liquid strains;

[0052] (4) Use the culture medium form...

Embodiment 3

[0057] Example 3 Tellurite reduction test

[0058] (1) Make a standard curve: a. Prepare 5 mg / mL sodium borohydride. b. Prepare 10mM K 2 TeO 3 , and carried out 8 times of serial dilution, leaving 0 value. c. Prepare 300 mM PBS (pH 7). d. Add 50 μL sodium borohydride, 50 μL potassium tellurite and 200 μL PBS into a 500 μL centrifuge tube, mix well, and react at 60°C for 10 minutes. e. Take 200 μL of the reaction solution into a 96-well plate, at OD 500 Measure the absorbance and make a standard curve;

[0059] (2) Using the secondary shake flask liquid strain in Example 2, take 5 mL of the bacterial liquid and add it to 100 mL of the improved PDA liquid medium (containing potassium tellurite with a final concentration of 0.5 mM);

[0060] (3) Take 500 μL of the culture medium every 24 hours, centrifuge at 12,000 rpm for 10 minutes, take 50 μL of the sample, mix with 50 μL of sodium borohydride and 200 μL of PBS, and react at 60°C for 10 minutes, then take 200 μL of the r...

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Abstract

The invention provides a high tellurite tolerant bacterium mediated synthesized biological tellurium nanoparticle, which is prepared by the following steps: 1) obtaining a strain mortierella sp.AB1 (Mortierella sp.AB1), and preserving in the China Center for Type Culture Collection (CCTCC) with the preservation number of CCTCC M 20211177); 2) fermenting; 3) adding a tellurite solution into the improved PDA for continuous growth; 4) collecting thalli, performing suction filtration, washing, performing suction filtration, freezing, grinding and resuspending; and 5) washing with Tris-HCl and n-caprylic alcohol, washing with deionized water in sequence, and re-suspending. The invention further provides antibacterial application of the biological tellurium nano-particles. The strain is stable in biological function, and the tellurite reducing capacity and the biological tellurium nano-particle synthesis capacity are high; the product is low in production raw material price, simple in synthesis process condition, safe and controllable, simple in preservation mode, high in stability and strong in antibacterial ability.

Description

technical field [0001] The present invention relates to the technical field of biological nanomaterials, more specifically to a high tellurite tolerant bacteria-mediated synthesis of biological tellurium nanoparticles and its antibacterial application. Background technique [0002] In the past research, tellurium and its compounds have been widely used in various industries such as metallurgy, electronics and applied chemical industry, and the existence of tellurium oxides in the environment has aroused people's attention to water and soil pollution. Studies have shown that even at low concentrations, tellurium oxides are toxic to most microorganisms, and elemental tellurium is less toxic, less soluble, and more widely used than tellurium oxides. Using microbial detoxification mechanism to reduce highly toxic Te(IV) to less toxic elemental tellurium is an important way to facilitate the bioremediation of Te(IV) pollution and the rational utilization of tellurium resources. ...

Claims

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Application Information

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IPC IPC(8): C12P3/00C12N1/14A61K33/04A61P31/04C12R1/645
CPCC12P3/00C12N1/14A61K33/04A61P31/04Y02A50/30
Inventor 夏险涂俊铭谭峥敖波严镇钧吴金
Owner HUBEI NORMAL UNIV
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