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Immobilization method of alcaligenes faecalis penicillin G acylase

A technology of Alcaligenes faecalis and penicillin, which is applied in the field of immobilizing Alcaligenes faecalis penicillin G acylase, which can solve the problems of undisclosed process details, high cost of reagents and carriers, and low immobilization efficiency, and achieve immobilization Effects of reduced time, improved stability and efficiency, and improved pH stability

Inactive Publication Date: 2009-03-11
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After decades of research and development, a variety of immobilization carriers and methods with various properties have been developed successively. Immobilization technology has made great progress, but it still cannot meet all the requirements of an ideal biocatalyst. There are not many enzymes, the main reason is the high cost of reagents and carriers used for immobilization, followed by low immobilization efficiency and poor stability; and due to commercial confidentiality, many process details are not disclosed

Method used

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  • Immobilization method of alcaligenes faecalis penicillin G acylase
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  • Immobilization method of alcaligenes faecalis penicillin G acylase

Examples

Experimental program
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Effect test

Embodiment 1

[0054] Example 1 Immobilization with carrier Eupergit C

[0055] Alcaligenes faecalis penicillin G acylase with a specific activity of 334U / g protein is used, the enzyme is dissolved in phosphate buffer solution with pH 8.0, and the enzyme solution is concentrated or diluted to 120mg / ml. Add 1g of wet carrier Eupergit C to 5ml of enzyme solution, and react with uniform mixing for 260 hours at room temperature. Suction filtration with a sand core funnel, the obtained immobilized product was washed 3-5 times with phosphate buffer solution, and the immobilization was completed.

Embodiment 2

[0056] Example 2 Immobilization of Alcaligenes faecalis penicillin G acylase with carrier ZH-EP

[0057] Alcaligenes faecalis penicillin G acylase with a specific activity of 334U / g protein is used, the enzyme is dissolved in phosphate buffer solution with pH 9.0, and the enzyme solution is concentrated or diluted to 120mg / ml. Add 1 g of wet carrier ZH-EP to 5 ml of enzyme solution, and react for 116 hours at room temperature with uniform mixing. Suction filtration with a sand core funnel, the obtained immobilized product was washed 3-5 times with phosphate buffer solution, and the immobilization was completed.

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Abstract

The invention relates to a process for immobilizing an enzyme in biological chemistry, in particular to a method for immobilizing a bacillus foecalis alkaligenes penicillin G acylase. The carrier for immobilizing the bacillus foecalis alkaligenes penicillin G acylase is ZH-EP. In a buffer solution system, when the ZH-EP is used to immobilize, the concentration of the enzyme is between 100 and 160 mg / ml, the immobilizing time is between 106 and 120 hours, the pH value of the buffer solution is between 8.5 and 10.0, the concentration of salt in the reaction system is between 1.0 and 1.5M, and the weight volume ratio of the carrier to the buffer solution is 1g : 4 to 6ml. The method improves the temperature and the pH stability of the immobilized enzyme compared with a free enzyme. Through continuous batches of hydrolysis of penicillin G kali salt, the immobilized bacillus foecalis alkaligenes penicillin G acylase shows the excellent operation stability and the superiority to the free enzyme.

Description

technical field [0001] The invention relates to a process for immobilizing enzymes in biochemical industry, in particular to a method for immobilizing Alcaligenes faecalis penicillin G acylase with a novel epoxy carrier ZH-EP. Background technique [0002] Since Fleming discovered penicillin in 1929, penicillin has made great contributions to human health care. However, with the use and abuse, bacterial resistance to penicillin is also increasing. The development of new antibiotics is an effective measure to solve the problem. In the 1960s, researchers discovered penicillin G acylase (PGA; EC 3.5.1.11), and developed immobilized enzyme technology to hydrolyze penicillin and cephalosporin G to produce 6-aminopenicillanic acid (6-APA ) and 7-amino deacetyl cephalosporanic acid (7-ADCA). 6-APA and 7-ADCA are important pharmaceutical intermediates of semi-synthetic antibiotics. Immobilized penicillin acylase is an important means for the production of semi-synthetic β-lactam...

Claims

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Application Information

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IPC IPC(8): C12N11/00C12N9/78
Inventor 许国旺孙健袁中一
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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