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Preparation method of cross-linked beta-lactamase aggregate for treating pharmaceutical wastewater

A technology of lactamase and pharmaceutical wastewater, which is applied in the field of enzyme engineering, can solve the problems of poor versatility, and achieve the effects of good stability, reduced chemical oxygen demand, and high enzyme activity

Active Publication Date: 2016-10-12
UNIV OF ELECTRONICS SCI & TECH OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Although CLEAs technology has obvious advantages, it is not universal. Usually, the preparation conditions optimized for one enzyme may not be suitable for another enzyme or even the same enzyme from different enzyme sources. Therefore, it is still necessary to study the immobilization system of new enzymes. keep exploring

Method used

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  • Preparation method of cross-linked beta-lactamase aggregate for treating pharmaceutical wastewater
  • Preparation method of cross-linked beta-lactamase aggregate for treating pharmaceutical wastewater
  • Preparation method of cross-linked beta-lactamase aggregate for treating pharmaceutical wastewater

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Embodiment Construction

[0028] Specific embodiments of the present invention will be described below in conjunction with the accompanying drawings, so that those skilled in the art can better understand the present invention. It should be noted that in the following description, when detailed descriptions of known functions and designs may dilute the main content of the present invention, these descriptions will be omitted here.

[0029] 1. Embodiment

[0030] 1. Acquisition of recombinant β-lactamase gene

[0031] The present invention is based on the β-lactamase gene on GenBank: AF143804.1 that is resistant to cefotaxime, and restricts the 5' end and 3' end of its coding region (nucleotide sequence) respectively. Endonuclease NcoI is the sequence 5'-ccatgg-3' and EcoRI is 5'-gaattc-3'. Six His tags, namely histidine catcaccatcaccatcac, are inserted before the stop codon taa at the 3' end to obtain the modified β-endo Amidase gene sequence, the process of transforming recombination such as figure...

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Abstract

The invention discloses a preparation method of a cross-linked beta-lactamase aggregate for treating pharmaceutical wastewater. The preparation method comprises the following steps: transferring a recombinant beta-lactamase gene sequence into a host cell E.coli Top10 to obtain recombined bacteria; carrying out induced expression, ultrasonic crushing and centrifugal treatment and taking supernatant to obtain rough enzyme liquid; adding bovine serum albumin into the rough enzyme liquid as an aggregation agent and a protection agent; precipitating and aggregating to form an enzyme aggregate; crossly linking the obtained enzyme aggregate through a bidirectional reagent so as to prepare the cross-linked beta-lactamase aggregate. After a test, the cross-linked beta-lactamase aggregate prepared by the invention is high in enzyme activity and good in stability, can be repeatedly utilized and is used for treating antibiotic wastewater; the chemical oxygen demand is reduced by 72.9 percent; the cross-linked beta-lactamase aggregate has an important application value in the treatment of the antibiotic wastewater in a pharmaceutical industry.

Description

technical field [0001] The invention belongs to the technical field of enzyme engineering, and more specifically relates to a method for preparing cross-linked β-lactamase aggregates for pharmaceutical wastewater treatment. Background technique [0002] In recent years, my country's pharmaceutical industry has developed rapidly. However, a large amount of toxic and harmful wastewater discharged during the pharmaceutical process seriously endangers people's health. Due to the complex composition of pharmaceutical industry wastewater, high organic content, high toxicity, dark color, high salt content, and poor biodegradability, it has a strong inhibitory effect on microbial growth and is difficult to degrade naturally. At the same time, the chemical oxygen demand (Chemical OxygenDemand, COD) of pharmaceutical industrial wastewater can be as high as 6000 mg / L or more. Therefore, it has become a global issue to treat pharmaceutical industrial wastewater and protect people's heal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/86C12N15/70C12N11/02C02F3/34C02F103/34
CPCC02F3/342C02F2103/343C12N9/86C12N11/02C12Y305/02006
Inventor 冯娟吴雪琴汤丽霞邓文凤廖茜仝亚沛
Owner UNIV OF ELECTRONICS SCI & TECH OF CHINA
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