Bacillus of oxidizing glucose, and application

A technology for oxidizing glucose and bacilli, applied in bacteria, fermentation and other directions, can solve the problems of difficult control, difficult separation and purification, and long synthetic route of chemical total synthesis method.

Active Publication Date: 2007-02-21
SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the chemical total synthesis method is greatly restricted in its application due to many unfavorable factors such as long synthetic route, various reagent materials involved, high cost, complicated process and difficult to control, and difficult separation and purification.

Method used

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  • Bacillus of oxidizing glucose, and application
  • Bacillus of oxidizing glucose, and application
  • Bacillus of oxidizing glucose, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Natural isolation of strains and NTG mutagenesis treatment

[0027] culture medium

[0028] Slant / Plate Medium: Glucose 10.0%, Yeast Extract 1.0%, CaCO 3 2.0%, agar 2.4%, pH6.8. This medium is used for the cultivation of starting strains, the natural isolation of strains, and the routine passage of strains.

[0029] natural separation

[0030] Take a fresh slant of the strain and add a small amount of sterile water, gently scrape off the bacterial lawn and pour it into a triangular bottle filled with sterile glass beads and sterile water, shake it fully for 10 minutes, and use a sterile sterile cotton bag filled with absorbent cotton. Filter through a funnel to obtain a bacterial suspension. The bacterial suspension was serially diluted and spread on the plate medium, and cultured at 28°C until a single colony grew.

[0031] NTG mutagenesis

[0032] Replace the above sterile water with sterilized phosphate buffer solution, according to the above natural separatio...

Embodiment 2

[0035] culture medium

[0036] Incline medium: same as embodiment 1

[0037] Liquid medium: D-sorbitol 4.5%, glucose 2.0%, beef extract 1.5%, yeast powder 1.5%, KH 2 PO 4 2.4%, pH 6.5

[0038] After the slant of the strain was cultivated at 28°C to mature, the excavated pieces were inoculated into shake flasks containing liquid medium for liquid propagation and culture at 28°C to measure the OD of the culture medium 600 value to reflect the reproductive status of the strain.

Embodiment 3

[0040] The bacterium liquid cultivated by the procedure of Example 2 is used as the seed liquid, and is inoculated in a 100L fermenter (cultivation liquid formula in the tank: D-sorbitol 5.0%, glucose 0.5%, yeast extract 2.4%, KH 2 PO 4 3.0%, pH 6.0). The temperature is controlled at 28°C, the ventilation rate is 1:1-1.5, and the stirring speed is 300 rpm. After 24 hours of culture, the cells were collected by centrifugation.

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Abstract

This invention relates to Gluconobacter oxydans mutant (CGMCC No. 1184) and its application in preparing 6-(2-hydroxyethyl) amino-6-deoxy-alpha-L-sorbfuranose by microbe conversion. The Gluconobacter oxydans mutant has such advantages as high stability, mild and controllable culture conditions, rapid propagation and short conversion time.

Description

technical field [0001] The invention relates to the technical field of microbial transformation, in particular to a mutant strain of Glucobacterium oxydans CGMCC No.1184, and the strain in 6-(2-hydroxyethyl)-amino-6-deoxy-α-L-sorbofuranose application in preparation. Background technique [0002] Miglitol, as one of the α-glucosidase inhibitors, has been used in the treatment of type II diabetes. According to the literature, the preparation of miglitol can be realized by means of total chemical synthesis or biotransformation combined with chemical synthesis. Among them, the chemical total synthesis method is greatly restricted in its application due to many unfavorable factors such as long synthetic route, various reagent materials involved, high cost, complicated process and difficult to control, and difficult separation and purification. In contrast, the introduction of biotechnology has fundamentally made the preparation of miglitol realistic and feasible. Judging from...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P19/26
Inventor 陈代杰戈梅李治川陶正利陈泉
Owner SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT
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