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Uranyl ion-binding heptapeptide

A technology of uranyl ion and heptapeptide, which is applied in the field of wastewater adsorbent and its preparation and application, can solve the problems of limited types of natural binding peptides, no specific binding peptides of uranyl ions are found, etc., and achieves high affinity and good application prospect. , Improve the effect of adsorption

Active Publication Date: 2020-04-24
EAST CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The metal-binding peptides currently studied are mainly naturally occurring binding peptides, however, the types of natural binding peptides are limited, and no uranyl ion-specific binding peptides have been found

Method used

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  • Uranyl ion-binding heptapeptide

Examples

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Embodiment 1

[0025] A screening method for uranyl ion-binding heptapeptide is provided in this embodiment, comprising the following steps:

[0026] (1) Preparation of uranyl ion chelating resin

[0027] Under sterile conditions, pipette 100 μl of resin into a 1.5mL centrifuge tube, centrifuge at 13000rpm for 5min, and remove the supernatant. Resuspend the resin with 1 mL of sterile water, centrifuge at 13000 rpm for 5 min, and repeat 4 times. Add the treated resin to 150mL sterile uranium standard solution with a concentration of 20μg / mL, shake at 150rpm overnight. 13000rpm, 10min centrifugal, remove supernatant, obtained uranyl ion chelating resin (M U+ ). In the experiment, the resin treated with sterilized ultrapure water was set as the control (M - ).

[0028] (2) The first round of screening of the random heptapeptide library

[0029] Take 100 μl M U+ , M - Add 1000μl 0.1% TBST (pH7.4) to each resin, wash twice. in M U+ , M - 10 μl of phage heptapeptide library (purchased f...

Embodiment 2

[0047] This embodiment provides a method for displaying uranyl ion-binding heptapeptide on the cell surface of Saccharomyces cerevisiae, including the following steps:

[0048] (1) Construction of surface display carrier

[0049] Two DNA sequences encoding uranyl ion-binding heptapeptide were synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd., and connected to the surface display vector pYD1 to form expression vectors pYD-7-4 and pYD-7-5 (such as image 3 shown).

[0050] (2) Transformation of Saccharomyces cerevisiae cells

[0051] Activate the Saccharomyces cerevisiae EBY100 strain, pick 5 single colonies into 50mL YPD liquid medium, and culture overnight at 30°C and 170rpm until OD600 reaches 1.0. Dilute the overnight cultured bacterial solution into a new 50mL YPD liquid medium to make the OD600 0.6, culture at 30°C and 170rpm for 3h, and measure the OD600 to 1.0. Collect the bacteria in a 50mL centrifuge tube, centrifuge at 2500rpm for 5min, discard the supernatan...

Embodiment 3

[0056] This example provides an application of a uranyl ion-binding heptapeptide in the preparation of an adsorption material for treating uranium-containing wastewater, including the following steps: picking single colonies of yeast engineering bacteria EBY100-7-4 and EBY100-7-5 respectively Into 10ml of YNB-CAA medium containing 2% glucose, shake culture at 30°C overnight, until OD600 is between 2~5, centrifuge at 3000~5000rpm at room temperature for 5~10min. The precipitate was washed with double distilled water, centrifuged for 5 min, and repeated twice. Resuspend the cells in YNB-CAA medium containing 2% galactose until the OD600 is 0.5~1. Shake culture at 20°C and 180rpm for 24h, collect the cells, wash with double distilled water, and freeze-dry, then they can be used for the treatment of uranium-containing wastewater. deal with. In the experiment, yeast cells treated with EBY100-pYD and EBY100 were set as controls.

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Abstract

The invention relates to uranyl ion combined heptapeptide, and belongs to the technical field of wastewater adsorbents, and preparation and application thereof. Two kinds of uranyl ion combined heptapeptide with high appetency on uranyl ions in a water solution are obtained through screening a bacteriophage random heptapeptide library; the amino acid sequence is shown as SEQ ID NO:1 or SEQ ID NO:2. Meanwhile, the invention also provides encoding nucleic acid of the uranyl ion combined heptapeptide, a carrier and a cell containing the nucleic acid, and application of the uranyl ion combined heptapeptide. Experiments prove that the uranyl ion combined heptapeptide has high appetency on the uranyl ions in the water solution, and can be specifically combined with the uranyl ions; the combined heptapeptide is shown on the yeast cell surface for preparing an adsorbing material; the adsorption effect of the yeast cell on the uranyl ions in the water solution can be effectively improved; better application prospects are realized.

Description

technical field [0001] The invention belongs to the technical field of wastewater adsorbent and its preparation and application, and relates to a uranyl ion-binding heptapeptide for treating uranium-containing wastewater, a nucleic acid encoding the heptapeptide, a carrier containing the nucleic acid, a cell, and a uranyl ion-binding heptapeptide. Peptide application. Background technique [0002] With the development of the nuclear industry, the demand for natural uranium continues to increase. In the main production links of the uranium mining and metallurgy industry and the application of uranium elements, a large amount of uranium-containing radioactive wastewater is produced. If the uranium-containing wastewater is not treated before discharge, it will cause radioactive pollution to surface water, groundwater and soil, thus posing a great hazard to the environment and human health. How to reduce the radioactive hazards of uranium in wastewater and effectively recycle e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C12N15/11C12N15/81C12N1/19B01J20/24
CPCB01J20/24B01J2220/4856C07K7/06
Inventor 李敏张志宾梁波马腾腾刘云海张卫民高柏王光辉肖化云李小燕肖红伟于梁梁
Owner EAST CHINA UNIV OF TECH
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