Preparation method of glutaryl-7-aminocefaphytanic acid acyltransferase

A technology of aminocephalosporanic acid acylase and glutaryl, which is applied in the field of preparation of glutaryl-7-aminocephalosporanic acid acylase, and can solve the problems of expensive reagents, poor quality, complicated process and the like

Inactive Publication Date: 2003-07-09
HUNAN FLAG BIOTECHNOLOGY CO LTD
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Problems solved by technology

And the key intermediate of semi-synthetic cephalosporin is 7-aminocephalosporanic acid (7-ACA), the method of traditional production in the past mainly is by ...

Method used

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Embodiment

[0024] GL-7-ACA acylase producing bacteria Escherichia coli genetically engineered bacteria (code name: E.coli-GLAA2002), purchased from South Korea Jinmei Co., Ltd., producing bacteria RecombinantEscherichia coli BL21 (DE3), GL-7-ACA acylase The source of enzyme gene is Pseudomonas sp. The bacterium was inoculated into a 500ml Erlenmeyer flask equipped with 100ml shake flask seed medium, which contained 1.0 grams of tryptone, 0.5 grams of yeast extract, and 1.0 grams of sodium chloride. The culture temperature was controlled at 25° C., the pH value was 6.5, and cultured for 12 hours. Then 400ml shake flask mycelium was moved into a fermenter equipped with 100 liters of medium for fermented seed culture, wherein 1500 grams of yeast extract, 240 grams of ammonium chloride, 220 grams of malic acid, and 680 grams of potassium dihydrogen phosphate. The culture temperature is 25° C., the pH value is 6.5, the air flow rate is 1:1 V / V / M, and the culture is performed for 12 hours...

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Abstract

The method for preparing glutaryl-7-aminocephaphylanic acid acylase is characterized by that it utilizes colibacillus gene engineering bacterium to make shake-flask strain culture in culture medium, fermentation strain culture and fermentation culture, then utilizes centrifuge to collect thallus, soakes to extract enzyme liquor, makes the enzyme liquor undergo the processes of plate-frame filtration, ultrafiltration concentration, purification and fixation so as to obtain the invented product GL-7-ACA ACY.

Description

Technical field: [0001] The invention relates to a preparation method of glutaryl-7-aminocephalosporanic acid acylase used for the key intermediate 7-aminocephalosporanic acid in the synthesis of semi-synthetic cephalosporins. Background technique: [0002] Cephalosporins are antibiotics with important efficacy, while semi-synthetic cephalosporins have stronger antibacterial ability or can improve pharmacokinetic properties. And the key intermediate of semi-synthetic cephalosporin is 7-aminocephalosporanic acid (7-ACA), the method of traditional production in the past mainly is by chemical method, has four major steps, and process is complicated, and used reagent is expensive, and pollution is serious, And the yield is low and the quality is poor. The biotransformation method is a better method. Studies have shown that cephalosporin C can be oxidatively deaminated by D-amino acid oxidase, and further decarboxylated to generate GL-7-ACA, while the 7-side chain of GL-7-ACA c...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N9/80C12N15/55
Inventor 许岗朱敏
Owner HUNAN FLAG BIOTECHNOLOGY CO LTD
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