One-step enzymatic method for preparing 7-aminocephalosporanic acid
A technology of aminocephalosporanic acid and amino, which is applied in the field of one-step enzymatic method for preparing 7-aminocephalosporanic acid, and can solve the problem of insufficient yield, purity and cost of the two-step enzymatic method, insufficient description of preparation conditions, and product yield. and purity effects, to achieve the effect of mild, safe and fast reaction conditions, fewer steps, and simple equipment
Inactive Publication Date: 2011-08-17
HARBIN PHARMA GRP CO LTD GENERAL PHARMA FACTORY
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Problems solved by technology
Although the two-step enzymatic method solves the environmental pollution problem of the chemical method, the two-step enzymatic method is somewhat insufficient in terms of yield, purity and cost, and the process of the latter step GL-7-ACA is difficult to control
[0009] Although the one-step enzymatic method is superior to the two-step enzymatic method for preparing 7-aminocephalosporanic acid, the efficiency of different enzymes is very different, and its specific treatment method will also have an impact on the yield and purity of the product.
[0010] The prior art discloses a method for preparing 7-aminocephalosporanic acid by one-step enzymatic method, but the preparation conditions are not fully explained. The present invention finds the best production conditions through screening, and obtains a method that combines The advantages of the chemical method (high yield and high purity) and the two-step enzymatic method (environmental protection and high yield), while being simpler than the existing one-step enzymatic method, the product yield is higher, the purity is higher, and the reaction conditions are milder and faster. Significant savings in production costs
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Embodiment 1
[0032] The pretreatment of embodiment 1. cephalosporin C (CPC) liquid:
[0033] Take 1 liter of cephalosporin C (CPC) fermentation filtrate (75mM, equivalent to 31.1 grams of pure CPC), filter through a 1 μm membrane, and adjust the pH to 10 with ammonia water.
Embodiment 2
[0034] Embodiment 2 reaction conditions control:
[0035] Add 167 grams of (NRB-103) enzyme into the reactor, wash twice with water, add CPC solution, under rapid stirring, control the pH to 10 with ammonia water, react for 30 minutes, and the 7-ACA conversion rate is 98%.
Embodiment 3
[0036] The processing of embodiment 3 lysate (7-ACA liquid):
[0037] To the resulting 2 liters of 7-ACA solution was added 10 g of Na 2 S 2 o 4 , adjust the pH to 7 with ammonia water, add Tween 80, 1 ml, add 200 ml of dichloromethane, stir for 10 minutes, separate phases, add 2.0 grams of activated carbon to the water phase, add 1.0 grams of EDTA, stir for 15 minutes, filter Remove carbon to obtain 7-ACA treatment solution.
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The invention relates to a one-step enzymatic method for preparing 7-aminocephalosporanic acid. The method for preparing the 7-aminocephalosporanic acid comprises four steps of: 1, treating fermentation liquid of cephalosporin C; 2, performing enzymolysis of the cephalosporin C under the action of acyltransferase of the cephalosporin C to obtain the 7-aminocephalosporanic acid; 3, treating the lysate of 7-aminocephalosporanic acid obtained through the enzymolysis; and 4, crystallizing and drying the 7-aminocephalosporanic acid.
Description
Technical field: [0001] The present invention relates to the application of an enzymatic cleavage method, in particular to a one-step enzymatic application for preparing 7-aminocephalosporanic acid. Background technique: [0002] 7-aminocephalosporanic acid, also known as: 7-aminocephalosporanic acid (7-aminocephalosporanic acid, 7-ACA), structural formula: [0003] [0004] It is an important basic raw material for semi-synthetic cephalosporins. At present, it is usually prepared from cephalosporin C (Cephalosporin C, CPC) by chemical method or two-step enzymatic cleavage, and the 7-position D-α-aminohexyl is removed. The diacid side chain gives 7-ACA. [0005] The production of 7-ACA by chemical method has the fatal defect of excreting a large amount of environmental pollutants, which is incompatible with today's concept of environmental protection. Later, a two-step enzymatic method was developed to produce 7-ACA, using cephalosporin C to pass through two enzymes, nam...
Claims
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IPC IPC(8): C12P35/02
Inventor 赵玉新黄宇红韩东胜邹国利刘治林户巧芬杨佳宁李国峰徐丹
Owner HARBIN PHARMA GRP CO LTD GENERAL PHARMA FACTORY
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