Cyclodextrin graft-bacteria immobilized cell porous rubber bead and application thereof

A technology for immobilizing cells and cyclodextrins, applied in the directions of immobilization on or in inorganic carriers, immobilization on/in organic carriers, biochemical equipment and methods, etc., to increase the number of cycles and improve porosity , the effect of conducive to circulation

Pending Publication Date: 2020-12-15
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the technology of immobilizing cyclodextrin graft powder and bacterial resting cells by using the blend of graphene oxide and sodium alginate as a carrier for the biotransformation of insoluble compounds has not yet been seen.

Method used

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  • Cyclodextrin graft-bacteria immobilized cell porous rubber bead and application thereof
  • Cyclodextrin graft-bacteria immobilized cell porous rubber bead and application thereof
  • Cyclodextrin graft-bacteria immobilized cell porous rubber bead and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 rubber bead performance measurement

[0044] (1) Preparation of rubber beads

[0045] The preparation method of SA (sodium alginate) bacteria-immobilized cell glue beads is a simple method that accurately weighs 2g of sodium alginate and 3g (wet weight of bacteria collected by centrifugation at 5000rpm, 4°C for 10min, and the following cells are collected by the same method). Arthrobacter resting cells were slowly added to 100 mL of deionized water to form a homogeneous solution under stirring conditions, and then 0.25 mol / L of CaCl was added using a syringe 2 Fix in the solution for 2 hours, and finally wash with deionized water to collect SA bacteria-immobilized cytogel beads.

[0046] The preparation method of β-CD and CMC grafts is as follows: Weigh 5 g of β-cyclodextrin and carboxymethyl cellulose at a mass ratio of 1:1, add 50 mL of 7% sodium hydroxide solution, and stir at 30 ° C Dissolve; add epichlorohydrin according to the volume ratio of epichl...

Embodiment 2

[0065] Example 2: Preparation of β-CD-CMC-GO Bacteria Immobilized Cytogel Beads and Its Application in Cortisone Acetate C1, 2-position Dehydrogenation Reaction

[0066] 1. Preparation of β-CD-CMC-GO bacteria immobilized cytogel beads:

[0067] Resting Cell Preparation:

[0068] The microbial strain is Arthrobacter simplex TCCC 11037, which can realize the dehydrogenation conversion of C1 and 2 positions of steroidal compounds, and convert cortisone acetate (CA) into prednisone acetate;

[0069] Incline medium (g / L): glucose 10, yeast extract 10, agar 20, pH7.2;

[0070] Seed (fermentation) medium (g / L): glucose 10, corn steep liquor 10, peptone 5, potassium dihydrogen phosphate 2.5, pH 7.2;

[0071] Preparation of Arthrobacter simplex resting cells:

[0072] TCCC 11037 was cultured at 32°C, 160r / min, 30 / 250mL seed medium for 18h, then inoculated into a 500mL shake flask containing 120mL fermentation medium at a 5% inoculum size, and cultured at 32°C, 110r / min After 20 hou...

Embodiment 3

[0090] Example 3: Preparation of HP-β-cyclodextrin graft bacteria-immobilized cytogel beads and its application in the dehydrogenation reaction of cortisone acetate C1 and 2

[0091] Except the following content, other is the same as embodiment 2.

[0092] 1. Preparation of HP-β-cyclodextrin graft bacteria-immobilized cell gel beads (HP-β-CD-CMC-GO bacteria-immobilized cell gel beads):

[0093] Prepare carboxymethylcellulose-HP-β-cyclodextrin graft, 5g carboxymethylcellulose is dissolved in 50mL 7% NaOH solution, add 7g HP-β-cyclodextrin, wait for HP-β-cyclodextrin After the essence and carboxymethyl cellulose are completely dissolved, add 8 mL of epichlorohydrin, then continue to stir at 25°C for 1.5h until it becomes a gel, stop stirring, and put it at 70°C for 5h. After the reaction, the product is alternately washed with a large amount of distilled water and ethanol until the eluate is neutral and there is no epichlorohydrin and HP-β-cyclodextrin in the eluate. Then put ...

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Abstract

The present invention belongs to the technical field of biocatalysis and particularly relates to a cyclodextrin graft-bacteria immobilized cell porous rubber bead, a preparation thereof and an application of the cyclodextrin graft-bacteria immobilized cell porous rubber bead in biotransformation. The porous rubber bead is prepared by uniformly mixing sodium alginate, a cyclodextrin-carboxymethyl cellulose graft, bacteria resting cells and graphene oxide in water according to a certain ratio and then dropwise adding the mixture into a metal ion solution for balling. The rubber bead has higher mechanical strength, is beneficial to repeated use, can also efficiently recycle cyclodextrin, reduces losses of the cyclodextrin and cells, and is convenient for separation in a circulation process; the rubber bead is small in pore diameters and regular, has a large number of pores, is higher in porosity, large in specific surface area and small in mass transfer resistance, and beneficial for rapid contact of the cyclodextrin, the cells and a substrate, improves a conversion rate and shortens a reaction period; and moreover, a certain promotion effect is achieved on enzyme activity of the cells, and good cell compatibility is shown.

Description

Technical field: [0001] The invention belongs to the technical field of biocatalysis, and in particular relates to a cyclodextrin graft-bacteria immobilized cell porous glue beads, its preparation and its application in biotransformation. Background technique: [0002] As the second largest class of drugs after antibiotics, steroids have the function of regulating the metabolism and physiological effects of various substances in the living body. Compared with the traditional chemical synthesis method, the method of microbial transformation can form a variety of active intermediates of steroidal drugs with high purity. As a solubilizer, cyclodextrin can increase the solubility of hydrophobic steroids, and its special cavity structure can include steroid substrates, thereby improving the bioavailability and yield of steroid compounds. However, the high price of cyclodextrin restricts its large-scale application in the field of biocatalysis. The cyclodextrin is immobilized on...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10C12N11/04C08G81/00C12P33/00C12P33/02C12R1/06
CPCC12N11/14C12N11/10C12N11/04C08G81/00C12P33/00C12P33/02
Inventor 申雁冰王敏郭俊超李梦涵谢培培骆健美
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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