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Penicillin amidase immobilized carrier and its preparing method

A technology of penicillin acylase and immobilized carrier, which is applied in directions such as being immobilized on/in an organic carrier, can solve the problems of difficulty in obtaining preparation cost, complicated preparation of raw materials, etc., and achieves high apparent enzyme activity and low cost of raw materials. Easy-to-obtain and easy-to-operate effects

Inactive Publication Date: 2006-04-26
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved in the present invention is to disclose a penicillin acylase immobilized carrier and its preparation method, so as to overcome the defects of complex preparation of raw materials, difficulty in obtaining and high preparation cost in the prior art, so as to meet the needs of relevant people

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] After mixing 870 milliliters of n-heptane with 280 milliliters of tetrachlorethylene, inject it into a four-necked flask equipped with a thermometer, airway and stirrer, add 0.4 grams of surfactant Span-60 and 50 grams of calcium stearate Heat to 55°C. After mixing evenly, add 160 milliliters of formaldehyde with 32 grams of N, N'-methylenebisacrylamide, 10 grams of acrylamide, 20 grams of glycidyl methacrylate and 0.5 grams of initiator azobisisobutyronitrile. Amide solution. Pure nitrogen was passed through, and the stirring was stopped after 6 hours of polymerization. The product was filtered, washed with acetone, extracted, and then vacuum-dried to constant weight. The apparent enzyme activity of penicillin G potassium salt hydrolysis to prepare 6-APA by immobilized penicillin acylase (IME) on this carrier was measured by p-dimethylaminobenzaldehyde (PDAB) method, which was 110 IU / g.

Embodiment 2

[0024] The difference from Example 1 is that a water separator is installed on the four-necked flask, and the diluent in the monomer phase is a mixed solution of methanol and water instead of formamide. 112 grams of methanol and 40 grams of water were added to the polymerization system, and the amounts and preparation conditions of the remaining components were the same as in Example 1. The corresponding apparent enzyme activity of immobilized penicillin acylase was 125IU / g.

Embodiment 3

[0026] The reaction device is the same as in Example 2. Add 850 ml of cyclohexane into a four-necked bottle, add 0.6 g of Span-60 and 32 g of calcium stearate, and heat to 55°C. After the Span-60 dissolution system reaches uniform mixing, add 156 ml of 60wt% 28 grams of N,N'-methylenebisacrylamide, 20 grams of glycidyl methacrylate and 0.5 grams of azobisisobutyronitrile solution dissolved in methanol aqueous solution were uniformly dispersed and nitrogen gas was introduced. After 6 hours of polymerization, most of the water enters the water separator and is separated from the system, and the reaction is stopped at this time. The aftertreatment of product is with embodiment 1. The corresponding apparent activity of the immobilized penicillin acylase was measured to be 92IU / g.

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Abstract

The invention discloses a preparation method of a penicillin acylase immobilized carrier. In the invention, the surface active agent is used as the dispersant, the vinyl compound is used as the monomer, and the cross-linking polymerization is carried out by adopting the reverse phase suspension technology, so as to obtain the penicillin acylase immobilized carrier. The apparent enzymatic activity of carrier-immobilized penicillin acylase (IME) to hydrolyze penicillin G potassium salt to prepare 6-APA can reach more than 125IU / g. The epoxy group-containing polymer bead carrier obtained by the method of the invention has high apparent enzyme activity, simple process, convenient operation, cheap and easy-to-obtain raw materials, and is easy for industrial scale production.

Description

technical field [0001] The present invention relates to an enzyme immobilization carrier, in particular to a penicillin acylase immobilization carrier, more specifically to a preparation method for a penicillin acylase immobilization carrier of a bead cross-linked polymer containing epoxy groups . Background technique [0002] β-lactam antibiotics, such as semi-synthetic penicillin and semi-synthetic cephalosporin, are currently widely used antibiotics. The traditional semi-synthetic chemical method has many steps, complicated process, low yield and serious pollution. The immobilized penicillin acylase is used for the semi-synthetic modification of antibiotics. Due to the high operational stability of the immobilized enzyme, the production process is easy to achieve continuous operation and controllability. After the reaction, it is easy to separate from the product and can be used repeatedly. Because of the high quality and low production cost of the obtained product, it ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/08C08F20/00C08F2/18
Inventor 卢冠忠薛屏乌云高娃郭杨龙王筠松
Owner EAST CHINA UNIV OF SCI & TECH
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