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Small-grain hemoglobin/hydrotalcite-like compound nano-composite and preparation method and application thereof

A nanocomposite and hemoglobin technology, which is applied in the field of small particle size hemoglobin/hydrotalcite-like nanocomposite, can solve the problems of complicated operation and complicated steps, and achieve the effects of low preparation cost, simple operation and mild conditions.

Active Publication Date: 2015-09-30
滁州市南方黑芝麻食品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods either add new raw materials, or the steps are cumbersome and the operation is complicated, and there is still a long way from the requirements of green chemistry.
However, there have been no reports on the preparation of small particle size protein / hydrotalcite-like nanocomposites and their biocatalysis using redox proteins as biomolecular templates by co-precipitation in T-shaped microreactors.

Method used

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  • Small-grain hemoglobin/hydrotalcite-like compound nano-composite and preparation method and application thereof
  • Small-grain hemoglobin/hydrotalcite-like compound nano-composite and preparation method and application thereof
  • Small-grain hemoglobin/hydrotalcite-like compound nano-composite and preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0024] Weigh MgCl separately 2 ·6H 2 O and AlCl 3 ·9H 2 O, add deionized water to prepare MgCl with a total metal ion concentration of 6mmol / L 2 And AlCl 3 Mix the salt solution, add hemoglobin to it to make the concentration 0.2g / L, and record it as solution A after mixing; prepare the sodium hydroxide solution with the concentration of 0.02mol / L, record it as solution B; open the T-type microreactor, Solution A and solution B were added to the salt tube and lye tube of the T-type microreactor respectively, and the flow rate was controlled to keep the pH of the reaction mixture at about 9.0, and the reaction product was received at the outlet of the mixture; the reaction mixture was at N 2 Under protective conditions, stir at low temperature for 24 hours, after the end, centrifuge the mixture at 12000 rpm for 10 minutes, and wash the precipitate with ultrapure water 3 times to obtain hemoglobin / hydrotalcite-like nanocomposite gel, which is recorded as Hb-LDH C6 .

Embodiment 2

[0026] Weigh MgCl separately 2 ·6H 2 O and AlCl 3 ·9H 2 O, add deionized water to prepare MgCl with a total metal ion concentration of 12mmol / L 2 And AlCl 3 Mix the salt solution, add hemoglobin to it to make the concentration 0.2g / L, and record it as solution A after mixing; prepare a sodium hydroxide solution with a concentration of 0.02mol / L, record it as solution B; open the T-type microreactor, Solution A and solution B were added into the salt tube and lye tube of the T-type microreactor respectively, and the flow rate was controlled to keep the pH of the reaction mixture around 9.0. The reaction product was received at the outlet of the mixture; the reaction mixture was at N 2 Under protective conditions, stir at low temperature for 24 hours, after the end, centrifuge the mixture at 12000 rpm for 10 minutes, and wash the precipitate with ultrapure water 3 times to obtain hemoglobin / hydrotalcite-like nanocomposite gel, which is recorded as Hb-LDH C12 .

Embodiment 3

[0028] Weigh MgCl separately 2 ·6H 2 O and AlCl 3 ·9H 2 O, add deionized water to prepare MgCl with a total metal ion concentration of 24mmol / L 2 And AlCl 3 Mix the salt solution, add hemoglobin to it to make the concentration 0.2g / L, and record it as solution A after mixing; prepare a sodium hydroxide solution with a concentration of 0.02mol / L, record it as solution B; open the T-type microreactor, Solution A and solution B were added to the salt tube and lye tube of the T-type microreactor respectively, and the flow rate was controlled to keep the pH of the reaction mixture at about 9.0, and the reaction product was received at the outlet of the mixture; the reaction mixture was at N 2 Under protective conditions, stir at low temperature for 24 hours, after the end, centrifuge the mixture at 12000 rpm for 10 minutes, and wash the precipitate with ultrapure water 3 times to obtain hemoglobin / hydrotalcite-like nanocomposite gel, which is recorded as Hb-LDH C24 .

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Abstract

The invention provides a small-grain hemoglobin / hydrotalcite-like compound nano-composite. The small-grain hemoglobin / hydrotalcite-like compound nano-composite is prepared by using a T-type microreactor through a coprecipitation method under the induction of hemoglobin template molecules with hemoglobin as a biomolecule source, a divalent metal salt Mg (NO3) and trivalent metal salt Al (NO3) mixed solution as a metal ion source and a sodium hydroxide solution as an alkali source. In the nano-composite, hemoglobin molecules are evenly adsorbed to the surfaces of hydrotalcite-like particles, and an original secondary structure of protein of the nano-composite is maintained. The nano-composite is shaped like sponge-shaped amorphous small grains and has a large specific surface area. The average grain diameter is smaller than 40 nm, and the thickness is smaller than 5 nm; the Zeta potential ranges from +16 mV to +30 mV; the hemoglobin / hydrotalcite-like compound nano-composite has high very thermal stability, the enzyme activity retention rate of the nano-composite ranges from 31% to 63.5% when catalyzing o-phenylenediamine on a 90-DEG C high temperature condition and is far higher than the enzyme activity retention rate 7.28% of free hemoglobin and the enzyme activity retention rate 21.44% of a big-grain hydrotalcite-like immobilized hemoglobin composite.

Description

Technical field: [0001] The invention relates to a small particle size hemoglobin / hydrotalcite-like nanocomposite; the invention also relates to a preparation method of the nanocomposite and its application to catalyze o-phenylenediamine. Background technique: [0002] The assembly of protein and inorganic nanoparticles has extensive basic and applied research value in the fields of biocatalysis, biomedical diagnosis and biosensors. The assembly of bio-inorganic interfaces involves van der Waals forces, hydrophobic affinity, electrostatic interaction and hydrogen bonding. Although inorganic nanoparticles will improve their performance after immobilizing proteins, the immobilized proteins may be denatured and inactivated due to improper interface assembly conditions. In order to make the fixed protein maintain its original secondary structure and biological activity, it is particularly important to find inorganic nanomaterials with good biocompatibility and many active sites. [0...

Claims

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Application Information

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IPC IPC(8): C07K17/14B01J31/38
Inventor 詹天荣芦思思谭正伟杨琦
Owner 滁州市南方黑芝麻食品有限公司
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