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Method for continuously synthesizing oligo-isomalto-oligosaccharides

A technology of polysaccharide and chitosan, which is applied in the field of immobilized cell catalysis, can solve the problems of high production cost of IMOs and the inability to obtain α-glucosidase, etc., and achieve improved recycling efficiency, good recycling characteristics, and simple method effective effect

Inactive Publication Date: 2018-08-17
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

my country has a large market demand for isomaltooligosaccharides, but domestic enterprises cannot currently obtain α-glucosidase with high purity. high

Method used

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  • Method for continuously synthesizing oligo-isomalto-oligosaccharides
  • Method for continuously synthesizing oligo-isomalto-oligosaccharides
  • Method for continuously synthesizing oligo-isomalto-oligosaccharides

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Immobilization of Pichia pastoris with α-glucosidase displayed on its surface

[0053] 1. Preparation of immobilized recombinant yeast strains

[0054] The details are as follows: first, 1 g of chitosan is added to 250 mL of 5% (m / v) aqueous acetic acid solution, fully dissolved. Then add 250mL double distilled water in chitosan sol and dilute, add the FeSO of 100mL3.6% (w / v) behind 4 ·7H 2 O aqueous solution. At this time, the solution was acidic, with pH10, and dark green flocs were formed. Take 250mL of the above flocculation solution in a 1000mL beaker, and place it in a microwave oven at 700W for 20min. To obtain black magnetic microspheres coated with chitosan with high paramagnetism, the carrier was washed three times with an aqueous solution with a pH>7.0. After homogenization, paste-like magnetic particles were obtained. Add 20 mL of 5% (v / v) glutaraldehyde solution to every 20 mL of magnetic particle paste, react on a shaker at 90 rpm (4000i con...

Embodiment 2

[0059] Continuous catalytic synthesis of IMOs in the fixed bed of embodiment 2

[0060] 1. Design of fixed bed bioreactor

[0061] A fixed-bed bioreactor was designed and manufactured, and the reactor was made of transparent acrylic tube. The reactor consists of three parts: the upper and lower plugs, the substrate uniform distribution module, and the immobilized cell catalyst filling module. The immobilized cell catalyst filling module is provided with a sampling port. The physical structure of the reactor is as Figure 4 As shown, the inner diameter is 60mm, and the filling layer can be spliced ​​with two specifications of modules with a height of 50mm / 100mm to study the influence of different filling amounts of immobilized bacteria on product distribution and product yield. The upper and lower layers of the bed are filled with 15 g of immobilized bacteria. There are filter membranes between different modules to prevent the leakage of immobilized bacteria. The process f...

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Abstract

The invention discloses a method for continuously synthesizing oligo-isomalto-oligosaccharides. An aldehyde-modified chitosan magnetic carrier is used to further immobilize recombinant Pichia pastorisdisplaying alpha-glucosidase on the surface. Aldehyde groups modifying the surface of the carrier are covalently bound to glycoprotein on the surface of the recombinant Pichia pastoris, and at the same time, through the adsorption effect of chitosan on the recombinant Pichia pastoris, the effect of immobilizing a Pichia pastoris cell catalyst can be achieved. By utilizing the superparamagnetic properties of the carrier, immobilized mycelia and a reaction substrate can be rapidly separated in a magnetic field, and the recycling efficiency of the cell catalyst is improved. The immobilized mycelia have good thermal stability and good recyclability. At the same time, the invention also relates to addition of the immobilized recombinant Pichia pastoris into a fixed bed for direct continuous catalytic synthesis of IMOs, and better production efficiency of the IMOs can be achieved. The immobilization method is simple and effective, and has good operability, the reuse of the mycelia is easy to realize, and the production cost of the IMOs can be reduced.

Description

technical field [0001] The invention belongs to the field of immobilized cell catalysis application, in particular to a method for continuously synthesizing oligomeric isomaltooligosaccharides. Background technique [0002] Isomalto-oligosaccharides (IMOs) are oligosaccharide mixtures containing an α-1,6 glycosidic bond in the molecule, and its functional components mainly include isomaltose, panose and isomaltotriose. IMOs can enrich and grow bifidobacteria in the intestinal tract, making the intestinal system run healthier. At the same time, it has the functions of low calorie, caries prevention, lubricating the intestinal tract, and improving human immunity. It is an important new sugar with nutritional and health functions. source. At present, it has been widely used in health care products industry, food industry and medical industry. In industry, the starch raw material is mainly hydrolyzed into maltose by the amylase system, and then the isomaltose oligosaccharide c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12N11/10C12N15/81C12P19/00C12P19/12C12P19/14C12R1/84
CPCC12N9/2402C12N11/10C12N15/815C12P19/00C12P19/12C12P19/14C12Y302/0102Y02P20/141
Inventor 郑穗平林影韩双艳赵难难王框
Owner SOUTH CHINA UNIV OF TECH
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