Chitinophage with aerobic arsenic methylation and volatilization functions and application thereof

A chitin and methylation technology, applied in the field of microorganisms, can solve problems such as unfavorable wide application, excessive arsenic in food, and inability to generate, and achieve the effect of reducing the content of highly toxic inorganic arsenic

Active Publication Date: 2022-04-29
GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Trivalent arsenic is more likely to be absorbed by rice and other crops, causing risk problems such as excessive arsenic in grains
CN201510956127.5 identified an aerobic arsenic-methylating bacterium SM-1 of Cellulophilaceae, but the bacteria can only grow in R2A medium, and cannot be produced in LB and other medium, which has obvious limitations
In addition, the arsenic concentration of the bacteria is low, which is not conducive to wide application

Method used

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  • Chitinophage with aerobic arsenic methylation and volatilization functions and application thereof
  • Chitinophage with aerobic arsenic methylation and volatilization functions and application thereof
  • Chitinophage with aerobic arsenic methylation and volatilization functions and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Isolation and identification of arsenic methylation functional chitinophage YT5

[0038] The samples were collected from arsenic-contaminated paddy soil in a certain place in Hunan. Weigh 100g of the soil sample that has been naturally air-dried and passed through a 10-mesh sieve into a 250ml Erlenmeyer flask, add 100ml of sterilized ultrapure water, seal the bottle mouth with a breathable film, and let it stand at room temperature for 15 days to activate the soil microorganisms. The soil suspension was serially diluted with sterilized ultrapure water, with a concentration gradient of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 and 10 -7 , respectively take 100μl concentration as 10 -4 、10 -5 、10 -6 and 10 -7 Dilutions were applied to ST10 containing 20 μM arsenous acid -1 on solid media plates. ST10 -1The formula of the solid medium is as follows: tryptone 0.5g, yeast extract 0.05g, glucose 0.5g, dilute to 1000ml, and sterilize at 121°C for 20min u...

Embodiment 2

[0045] Example 2: Growth curve and arsenic methylation efficiency of arsenic methylation function chitinophage YT5 in arsenic-containing R2A, LB and TSB media

[0046] R2A, LB and TSB three kinds of liquid medium were used to expand the culture of YT5 and verify the efficiency of arsenic methylation. The details are as follows: Prepare 500ml of sterilized R2A liquid medium, inoculate a single colony in the streaked plate into the culture medium with shaking (30°C, 200rpm) for 10-12 hours, and its OD600 is 0.4-0.5, which is in the middle of the logarithmic phase . The expanded cultured bacterial solution was centrifuged at 5000rpm for 10min to collect the bacterial cells. Take 0.55ml of YT5 bacteria solution (OD600=1.8) and inoculate them in 100ml of sterilized R2A, LB and TSB liquid medium containing 12μM arsenous acid respectively. The formula of LB liquid medium is: 10g / L peptone, 5g / L yeast powder, 10g / L NaCl, and the balance is water; the formula of TSB liquid medium is:...

Embodiment 3

[0049] Example 3: Comparison of efficiency of arsenic methylation functional chitinophage YT5 converting different concentrations of arsenous acid in TSB medium

[0050] Referring to Example 2, 0.55ml of YT5 bacterial solution (OD600=1.8) was inoculated in 100ml of sterilized TSB liquid medium with 3, 6, 12, 24, and 48 μM arsenous acid, respectively. The initial bacterial OD600 of the strain YT5 was 0.01. After the culture solution was cultured at 30°C and 200rpm constant temperature shaking for 72 hours, each sample was taken from each treatment sample, and 1ml of the bacterial solution sample was centrifuged at 12000rpm for 2min, and filtered on a 0.22μm filter membrane. After supernatant, arsenic speciation and quantification were performed by HPLC-ICP-MS.

[0051] Such as Image 6 As shown, the strain YT5 has good arsenic methylation conversion efficiency for different gradients of arsenous acid in TSB medium; within 72 hours, the methylation conversion rate of arsenous aci...

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Abstract

The invention discloses a chitophage strain with aerobic arsenic methylation and volatilization functions and application of the chitophage strain, and belongs to the technical field of microorganisms. The name of the bacterial strain is Chitinophaga filiformes YT5, the bacterial strain is preserved in Guangdong Microbial Culture Collection Center on December 31, 2021 in the building 59, No.100 Courtyard, Xianlie Middle Road, Guangzhou City, Guangdong Province, and the preservation number is GDMCC No: 62178. Chitosan phage YT5 has an obvious methylation effect on arsenious acid and a function of volatilizing methyl arsenic in an arsenic-containing culture medium, main methyl arsenic products are dimethyl arsenic and volatile trimethyl arsenic, the concentrations of the trimethyl arsenic and the dimethyl arsenic are gradually increased along with the increase of culture time, and meanwhile, the concentration of the arsenious acid is reduced along with the increase of time. The chitophage YT5 has important application potential in arsenic-polluted environment remediation, especially arsenic-polluted soil remediation.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a chitinophage strain with aerobic arsenic methylation and volatilization functions and application thereof. Background technique [0002] Arsenic is a toxic and carcinogenic substance ubiquitous in the environment, and it is an environmental problem faced by many countries around the world. In particular, trivalent arsenic (arsenite or arsenite) is more mobile and more toxic in soil and water. Trivalent arsenic is more easily absorbed by rice and other crops, causing risk problems such as excessive arsenic in grains. Therefore, how to effectively control arsenic pollution is the frontier issue of water pollution control and soil remediation, and it is also a major national demand. Studies have shown that microorganisms carrying arsenic methylation functional genes in soil have organic arsine forms such as converting inorganic arsenic into methyl arsenic. On the o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C02F3/34C12R1/01C02F101/20
CPCC12N1/20B09C1/10C02F3/34C02F2101/20
Inventor 方利平李芳柏陈冠虹
Owner GUANGDONG INST OF ECO ENVIRONMENT & SOIL SCI
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