Environment sensitive gel crosslinked with metalloprotein, and applications thereof

A metalloprotein and sensitive technology, applied in the direction of alkali metal compounds, alkali metal oxides/hydroxides, and other chemical processes, can solve the problems of poor metal selectivity and low sensitivity

Active Publication Date: 2019-11-12
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, Xiaojie Ju et al. used NIPAM and BCAm to prepare hydrogels with temperature-sensitive properties, which can adsorb and desorb Pb using temperature changes. 2+ , but Pb 2+ The concentration is as high as 4mM, and cannot remove Pb at low concentrations (such as μM or nM levels) 2+ Ions (Ju XJ,Zhang SB,Zhou MY,Xie R,Chu LY,Novel heavy-metaladsorption materials:ion-recognition P(NIPAM-co-BCAm)hydrogels for removal oflead(II)ions.J Hazard Mater,2009,167 :114-118.); Neeshma Dave et al. used acrylamide and DNA-prepared hydrogels to detect and remove 10nM Hg 2+ , but the gel can only be regenerated by acid treatment (Dave N, Chan MY, Huang PJ, Smith BD, Liu JW, Regenerale DNAfunctionalized hydrogels for ultra

Method used

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  • Environment sensitive gel crosslinked with metalloprotein, and applications thereof
  • Environment sensitive gel crosslinked with metalloprotein, and applications thereof
  • Environment sensitive gel crosslinked with metalloprotein, and applications thereof

Examples

Experimental program
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Example Embodiment

[0057] Example 1

[0058] 1. The modified cadmium ion regulatory protein CadR of the present invention can be expressed, purified and modified using the following methods:

[0059] (1) Transform the pet28a plasmid encoding the modified cadmium ion regulatory protein obtained by total gene synthesis into E.coli BL21 (DE3), use 0.5mM IPTG to induce expression at 37°C for 4h, and use a high-speed centrifuge to harvest and Store at -80°C until use. Resuspend in 30mL buffer and lyse with ultrasound (buffer is 20mM Tris-HCl, pH 6.8, 100mM NaCl, 5mM 2-mercaptoethanol (β-ME), 1mM PMSF, 10% glycerol), and centrifuge the lysate at 15,000 rpm in a high-speed centrifuge , Use a 0.45μm filter membrane to filter the supernatant, and then load the sample on a 5mL nickel column. After 10% buffer B is equilibrated, 10%-100% buffer B is used to elute 5 column volumes. After concentrating the collected samples, use a desalting column to replace the solution with buffer A, and then use HRV3C enzyme ...

Example Embodiment

[0064] Example 2

[0065] Dissolve 220mg isopropylacrylamide monomer, 2.4mg N,N-methylenebisacrylamide, 1mg anhydrous sodium sulfite in 2ml secondary water, stir magnetically for 10min under nitrogen protection, and add 300μL dropwise (4mg dissolved Use 300μL of secondary water) potassium persulfate as the initiator. Under the protection of nitrogen, continue magnetic stirring for 1 min. Add 1 mL (0.33 mg / mL) of modified modified cadmium ion-regulated protein CadR, and place it at 20°C for about 20 min under a nitrogen atmosphere. Will polymerize into a gel. Add secondary water soaking at room temperature for 12 hours to elute uncrosslinked monomers, stand at 37°C for about 1 hour, centrifuge at 8000 rpm for 5 minutes to remove uncrosslinked proteins. After immersing in secondary water at 37°C for 4 hours to reduce the volume, replace the secondary water with 10mM Bis-Tris (pH 7.0), eluting with 0.1mM TCEP buffer for 3 times, and then desorb at 4-25°C using a mixer For 48h, use...

Example Embodiment

[0066] Example 3

[0067] The gel 1 before and after the release of metal ions was nitrated with 6 mL of concentrated nitric acid, and the Cd ion concentration in it was measured by inductively coupled plasma mass spectrometry (ICP-MS). The results showed that the total amount of Cd ions in the gel before metal release was 63.7nmol, the total amount of Cd after desorption at 4℃ was 0.25nmol, and the total amount of Cd ions after desorption at 25℃ was 0.32nmol. It shows that 99.5% of the metal can be completely released by reducing the temperature to swell the gel. The appearance of the gel before and after swelling of gel 1 is as Figure 7 Shown.

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Abstract

The invention discloses a special large molecular gel with environment sensitivity. According to a preparation method, metalloprotein is embedded into a gel three dimensional network through chemicalcovalent bonds. The gel is prepared through copolymerization of an environment sensitive gel monomer or a skeleton formed through crosslinking of the environment sensitive gel monomer with a metalloprotein containing reaction groups or a complex of the metalloprotein with metal ions. Shrinkage and swelling of the environment sensitive gel with changing of environment can be realized, so that the chelating capacity of the metalloprotein on metal ions is adjusted, and high efficiency and selective capturing and release of metal ions are realized. The special large molecular gel with environmentsensitivity is capable of realizing high efficiency, stable, circulation capturing of metal ions at extremely concentration in liquid.

Description

technical field [0001] The invention belongs to the field of functional materials, and relates to an environment-sensitive gel of cross-linked metalloproteins and an application thereof. Utilize the properties of environment-sensitive gels that can shrink and swell with environmental changes to adjust the ability of metalloproteins and / or metal ion-binding domains of metalloproteins to chelate metal ions, so as to achieve efficient and selective capture of metal ions and freed. Background technique [0002] Heavy metal ion pollution in the environment will have serious impacts and harms on the human body and other organisms. These heavy metal ions are different from the pollution of other organic compounds. Most organic compounds can be purified or degraded by nature itself, while heavy metal ions can only be used as a One form is transformed into another form, which is easily enriched in living organisms, and then produces severe toxicity. For example, it will cause irrev...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/28C02F1/28B01D15/00C08F289/00C08F222/38C08F220/34C08F220/54C02F101/20
CPCB01D15/00B01J20/264B01J20/28047C02F1/285C02F2101/20C08F222/385C08F289/00C08F220/34C08F220/54Y02C20/40
Inventor 陈浩吴奇黄善青胡青原魏天彪
Owner NANJING UNIV
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