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Penicillin acylation enzyme-fixing carrier preparation method and carrying method

A penicillin acylase, carrier technology, applied in the direction of immobilization on/in organic carriers, hydrolase, etc., can solve the problems of unsatisfactory mechanical properties and unsuitable reaction conditions, etc.

Inactive Publication Date: 2007-07-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Biomaterials, due to their better biocompatibility, will provide some help to immobilized enzymes, such as improving their chemical stability and reducing the inactivation of enzymes during immobilization, but often this type of carrier The mechanical properties are not very ideal, and it cannot be well adapted to the harsh reaction conditions in industry

Method used

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  • Penicillin acylation enzyme-fixing carrier preparation method and carrying method
  • Penicillin acylation enzyme-fixing carrier preparation method and carrying method
  • Penicillin acylation enzyme-fixing carrier preparation method and carrying method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 5.0 g of polymethyl methacrylate spherical porous resin was ground and sieved to obtain resin particles with a particle size of 0.5 mm. Dissolve 1.0 g of chitosan in 50 ml of 2% acetic acid solution, and then add 0.5 g of PEG20000 to form a transparent and clear solution. Mix the resin and chitosan solution evenly, and dry at 40°C. Transfer the mixed solids into a round-bottomed flask, add 50ml of water, and stir mechanically at 200r / min. After 30 minutes, adjust the pH value to 10 with NaOH aqueous solution, add 0.5g of 2-chloroethylamine hydrochloride, and control the temperature at 40°C. After reacting for 5 hours, filter with suction, wash, and dry at 40°C.

[0045] Take 1.0 g of the prepared carrier in a Erlenmeyer flask, add 1 mol / L sodium bicarbonate solution, react at 50°C for 12 hours, separate, then add 10 ml of 2.0% glyoxal solution, shake on a shaking table, 200r / min, the temperature is controlled at 40°C. Take it out after 6 hours, age at 30°C for 24 ho...

Embodiment 2

[0047] 20.0 g of poly(p-styrene alkylhydroxyl) spherical porous resin was ground and sieved to obtain resin particles with a particle size of 0.5 mm. Dissolve 1.0 g of chitosan in 50 ml of 1% hydrochloric acid solution, and then add 0.5 g of PEG2000 to form a transparent and clear solution. Mix the resin with the chitosan solution and dry at 40°C. Transfer the mixed solids into a round bottom flask, add 50ml DMSO, and after 30 minutes, mechanically stir at 500r / min, adjust the pH value to 9 with NaOH aqueous solution, add 5ml of trimethoxychloromethylsilane, control the temperature at 60°C, and react After 4 hours, filter with suction, wash, and dry at 40°C.

[0048] Take 1.0 g of the prepared carrier in an Erlenmeyer flask, add 1 mol / L ammonia water, react at 60°C for 15 hours, separate, then add 10 ml of 2.0% glutaraldehyde solution, vibrate on a shaker, 200r / min, temperature Controlled at 40°C. Take it out after 4 hours, age at 30°C for 36 hours, add free penicillin acyl...

Embodiment 3

[0050] 10.0 g of polyacrylic acid tertiary amino porous resin was ground and sieved to obtain resin particles with a particle size of 0.5 mm. Dissolve 1.0 g of chitosan in 50 ml of 1% hydrochloric acid solution, then add 0.5 g of glucose to form a transparent and clear solution. Mix the resin and chitosan solution evenly, and dry at 40°C. Transfer the mixed solids into a round bottom flask, add 25ml DMF and 25ml ethanol, after 30 minutes, mechanically stir at 500r / min, adjust the pH value to 11 with NaOH aqueous solution, add 5ml of epichlorohydrin, control the temperature at 90°C, and react After 8 hours, filter with suction, wash, and dry at 50°C.

[0051] Take 1.0g of the prepared carrier in a Erlenmeyer flask, add 1mol / L sodium polyphosphate solution, react at 30°C for 12 hours, separate, then add 10ml of 2.0% glyoxal solution, oscillate on a shaking table, 200r / min, the temperature is controlled at 40°C. Take it out after 6 hours, age at 30°C for 48 hours, add free pe...

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Abstract

The invention discloses a making method of fixed penicillin acylated enzyme, which comprises the following steps: (1) grinding the porous resin with lateral functional group; sieving; drying; activating; blending with chitose solution; drying; obtaining the solid; (2) adding solid in the step (1) in the polar hydrophilic solvent; adding adjuvant to suck after decorating; washing; drying to obtain the porous resin carrier; (3) loading acylated penicillin on the porous resin carrier; obtaining the product.

Description

technical field [0001] The invention relates to a preparation and loading method of a carrier for immobilizing penicillin acylase, which is mainly used in the industrial preparation and synthesis process of β-lactam antibiotics and their mother nuclei. Background technique [0002] In the early 1960s, the semi-synthetic industry of β-lactam antibiotics gradually emerged, which made 6-aminopenicillin alkanoic acid (6-APA), the mother nucleus of β-lactam, an important drug synthesis intermediate. Therefore, people first started to synthesize 6-APA by cleaving penicillin G by penicillin acylase (PGA). Subsequently, due to the unique selectivity of PGA, people studied the selective catalysis of PGA, using 6-APA or 7-amino deacetoxycephalosporanic acid (7-ADCA) as the mother nucleus of β-lactam, plus Appropriate side chains for the semisynthesis of β-lactam antibiotics. [0003] For more than 40 years, PGA has become more and more stable through continuous screening and genetic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12N9/78
Inventor 林贤福金鑫吴起吕德水陈志春
Owner ZHEJIANG UNIV
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