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Carrier for immobilization as well as preparation method thereof and immobilized beta-glucosaccharase

A technology of immobilized carrier and glucosidase, which is applied in the field of solid-phase enzyme preparation, can solve the problems of easy shedding of biological enzymes, low activity of immobilized enzymes, low operational stability, etc., and achieve short reaction time and high reaction efficiency High and guaranteed catalytic efficiency

Inactive Publication Date: 2014-04-09
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the above deficiencies or improvement needs of the prior art, the present invention provides an immobilized carrier and its preparation method and immobilized β-glucosidase, the purpose of which is to use metal ion coordination bonds such as β-glucosidase The combination of biological enzymes and magnetic nanoparticles forms immobilized biological enzymes, thereby solving the current technical problems of low enzyme activity, easy fall-off of biological enzymes, and low operational stability of immobilized enzymes on magnetic nanoparticles

Method used

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  • Carrier for immobilization as well as preparation method thereof and immobilized beta-glucosaccharase
  • Carrier for immobilization as well as preparation method thereof and immobilized beta-glucosaccharase
  • Carrier for immobilization as well as preparation method thereof and immobilized beta-glucosaccharase

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preparation example Construction

[0041] The preparation method of immobilized carrier provided by the invention comprises the following steps:

[0042] (1) Preparation of magnetic nanoparticles.

[0043] (2) Preparation of magnetic nanoparticles coated with polyhydric sol-gel compounds.

[0044] The polyhydroxy sol-gel compound is dissolved in water and heated to form a sol. The magnetic nanoparticles are added to the polyhydroxy sol-gel compound sol, and ultrasonically dispersed to obtain the magnetic nano particles and the polyhydroxy sol-gel compound suspension. Add the suspension into an emulsifier composed of surfactants and organic solvents, and stir vigorously at high temperature to obtain a uniform dispersion system. The uniformly dispersed system is continuously stirred, cooled at normal temperature, and after the temperature is lowered to room temperature, magnetic nanoparticles coated with polyhydric sol-gel compounds are obtained. Magnetic nanoparticles coated with polyhydric sol-gel compounds ...

Embodiment 1

[0055] An immobilization carrier includes magnetic nanoparticles, carboxyl-rich composite materials and transition metal ions.

[0056] The magnetic nanoparticles are Fe 3 o 4 , with an average particle size of 5 nm.

[0057] The carboxyl-rich composite material is wrapped on the surface of the magnetic nanoparticles to form a carboxyl-rich shell with an average thickness of 30 nanometers. The carboxyl-rich composite material includes three layers, which are agarose, epichlorohydrin and iminodiacetic acid in sequence from inside to outside.

[0058] The transition metal ion is Co 2+ , containing 50 micromoles of transition metal ions per gram of support.

[0059] The carboxyl-rich composite material is wrapped on the surface of the magnetic nanoparticles to form a carboxyl-rich shell, and the surface carboxyl groups are complexed with transition metal ions, and the transition metal ions are combined with the enzyme to be immobilized through metal ion coordination bonds.

...

Embodiment 2

[0074] An immobilization carrier includes magnetic nanoparticles, carboxyl-rich composite materials and transition metal ions.

[0075] The magnetic nanoparticles are Fe 3 o 4 , with an average particle size of 11 nm.

[0076] The carboxyl-rich composite material is wrapped on the surface of the magnetic nanoparticles to form a carboxyl-rich shell with an average thickness of 20 nanometers. The carboxyl-rich composite material includes three layers, which are agarose, epichlorohydrin and nitrilotriacetic acid in sequence from inside to outside.

[0077] The transition metal ion is Ni 2+ , containing 100 micromoles of transition metal ions per gram of support.

[0078] The carboxyl-rich composite material is wrapped on the surface of the magnetic nanoparticles to form a carboxyl-rich shell, and the surface carboxyl groups are complexed with transition metal ions, and the transition metal ions are combined with the enzyme to be immobilized through metal ion coordination bond...

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Abstract

The invention discloses a carrier for immobilization as well as a preparation method thereof and immobilized beta-glucosaccharase. The carrier for the immobilization comprises magnetic nano particles, a carboxyl-rich composite material and transitional metal ions, wherein the carboxyl-rich composite material wraps the surface of the magnetic nano particles to form a carboxyl-rich shell, carboxyl on the surface is complexed with the transitional metal ions, and the transitional metal ions are combined with to-be-immobilized enzyme through a metal ion coordination bond. The preparation method comprises the following steps of (1) preparing the magnetic nano particles; (2) carrying out hydroxyl functional modification for the particles; (3) carrying out epoxy functional modification for the particles; (4) carrying out carboxyl functional modification for the particles; (5) complexing the particles with the transitional metal ions. The functional carrier is combined with beta-glucosaccharase through the metal coordination bond to obtain immobilized beta-glucosaccharase. The biological enzyme is firm in combination, high in enzyme activity, strong in operation stability and suitable for the mass production.

Description

technical field [0001] The invention belongs to the field of solid-phase enzyme preparation, and more specifically relates to an immobilized carrier, a preparation method thereof and an immobilized β-glucosidase. Background technique [0002] Enzyme immobilization refers to an enzyme engineering technology that uses carriers to restrict or bind enzymes in a certain area through physical and chemical methods, so that the enzyme can perform catalytic reactions in this specific area. Enzyme immobilization technology has become one of the most active research focuses in the field of enzyme engineering in recent years because of its advantages of enabling enzyme recovery and continuous operation and reducing production costs. [0003] As an immobilization carrier, magnetic nanoparticles are widely used because they facilitate separation and reuse. At present, the manufacturing method of the magnetic nanoparticle carrier mainly wraps the magnetic nano core with chitosan, and cros...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/08C12N11/04C12N9/42
Inventor 闫云君陈亭亭张后今徐莉黄霜霜
Owner HUAZHONG UNIV OF SCI & TECH
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