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Preparation of 7-amino-3-desacetoxy cephalosporanic acid by maize milk fermentation-enzyme catalysis method

A technology of deacetoxycephalosporanic acid and enzyme catalysis method is applied in the field of biological enzyme engineering, and can solve the problems that immobilized enzymes depend on imports, the fermentation process technology has not reached industrialization, and the production cost is high.

Inactive Publication Date: 2010-04-07
徐州瑞赛医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Semi-synthetic and biosynthetic methods have also been introduced in China, all of which use penicillin G potassium as a raw material, and the production cost is relatively high. level of industrialization

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0021] Cultivation of seed medium: 1 ml of Streptomyces clavuligerus strain was added to 10 8 Conidia / ml, inoculated in seed medium (g / l) consisting of: glucose 23; (NH 4 ) 2 SO 4 25; KH 2 PO 4 0.7; Lactose 50; CaCO 3 6; corn steep liquor 10; cottonseed flour 10; add water 100ml. Before sterilization, adjust the pH of the medium to 5.3. The seed medium was sealed and cultured with shaking on a rotary shaker at 25°C and 220 rpm for 48 hours.

[0022] Cultivation of fermentation medium: transfer the cultured seed medium to the fermentation medium at an inoculum of 5% (g / l): glucose 10; (NH 4 ) 2 SO 4 16; KH 2 PO 4 0.5; Lactose 60; CaCO 3 9; Corn steep liquor 20; Cottonseed meal 15; Others are water. Before sterilization, adjust the pH of the medium to 6.2. After inoculation, 15% phenylacetic acid solution was added to the fermentation broth at 1:10, and cultured at 25°C and 30 rpm for 168 hours, and the evaporated water was replenished every other day.

[002...

example 2

[0028] Cultivation of the seed medium: the Expansionella strain of Streptomyces clavuligerus (Streptomyces clavuligerus) with 10 9 Conidia / ml, inoculated in the following seed medium (g / l): glucose, 20; (NH 4 ) 2 SO 4 , 18; KH 2 PO 4 , 0.5; Lactose, 30; CaCO 3 , 5; corn steep liquor, 15; Pharmamedia (cotton seed flour), 12; others are water. Before sterilization, the pH of the medium was 5.3. Seal the seed medium and incubate for 42 hours at 27°C and 220 rpm.

[0029] Cultivation of the fermentation medium: the above-mentioned seed medium was inoculated into the production medium (g / l) of the following composition at a volume ratio of 1:18: glucose, 15; (NH 4 ) 2 SO 4 , 11; KH 2 PO 4 , 0.6; lactose, 23; CaCO 3 , 6; corn steep liquor, 25; Pharmamedia (cotton seed flour), 10; others are water. Before sterilization, the pH of the medium was 6.5. After inoculating with the seed medium, 15% phenylacetic acid solution 1:15 was added to the fermentation broth. Airtight...

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Abstract

The invention discloses a corn steep liquor fermentation-enzyme catalysis method, which is used to synthesize phenylacetyl-7-amino-3- desacetoxycephalos-porin acid. Streptomyces clavuligerus with an expandase gene is used to make strain transformation to corn steep liquor to ensure that the corn steep liquor is made into fermentation ring enlargement in a culture medium and is reacted with phenylacetic acid added into the culture medium to generate the phenylacetyl-7-amino-3- desacetoxycephalos-porin acid. Water solution of the phenylacetyl-7-amino-3- desacetoxycephalos-porin acid is obtainedthrough organic solvent extraction and water back extraction. Acetyl transferase and nutrition content are added into the water solution to form an enzyme reaction culture medium and finish a biochemical reaction under appropriate condition, and the phenylacetyl-7-amino-3- desacetoxycephalos-porin acid can be produced after side chains are removed.

Description

technical field [0001] The invention belongs to the technical field of biological enzyme engineering, and relates to a biosynthesis method for preparing and purifying 7-amino-3-deacetoxycephalosporanic acid (7-ADCA). Background technique [0002] Cephalosporins are an important class of β-lactam antibiotics. Originally, it was naturally produced by Acremonium chrysogenum. In recent years, it is mainly obtained by artificial synthesis. 7-Amino-3-deacetoxycephalosporanic acid (7-ADCA) is the key nucleus for synthesizing cephalosporin antibiotics. 7-ADCA is prepared by artificial synthesis, and then cephalexin, cefradine and cephalosporin are further synthesized. A variety of cephalosporin antibiotics such as clorox, ceftazidime and cefadroxil. The synthesis methods of 7-ADCA mainly include chemical synthesis, semi-synthesis and biosynthesis. [0003] At present, chemical synthesis and semi-synthesis are still the main methods for synthesizing 7-ADCA: EP0169144 (1986) intro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/00C12R1/465
Inventor 胡晓王玉成李桂根王静康王理想王友志赵红周伟军王圣良李维泉王健周平江路一飞
Owner 徐州瑞赛医药科技有限公司
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