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Application of algal toxin-degrading enzymes in inhibiting cyanobacteria and degrading algal toxins

A technology for degrading algal toxins and algal toxins, applied in the directions of enzymes, enzymes, chemical instruments and methods, etc., can solve the problems of degradation, cumbersome operation, strong killing ability of oxidants, etc., achieve efficient inhibition or killing, and solve the problems of water pollution. Effect

Active Publication Date: 2021-10-12
HUAZHONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although coagulation, activated carbon absorption, and membrane treatment technologies can partially reduce the content of algal toxins in water, these methods can only transfer algal toxins to other places and cannot effectively degrade algal toxins, and consume time and materials. Disadvantages of high cost
Although the method of oxidative degradation of algal toxins has high degradation efficiency and low price, the oxidant has strong biocidal ability and poor specificity, and it is easy to react with organic pollutants to produce substances harmful to human health
(2) Bacterial degradation method: This method uses fine sand filtration and biofilms containing selective biodegradable bacteria to treat water samples. Need to consume nutrients, easily affect the stability of the ecosystem, and may produce unknown secondary metabolic toxic substances, causing secondary pollution
That is, algae removal and degradation of algae toxins are usually two independent steps, which must be achieved by certain means (or agents), and it is difficult to comprehensively control algicide and degradation of algae toxins at the same time.

Method used

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  • Application of algal toxin-degrading enzymes in inhibiting cyanobacteria and degrading algal toxins
  • Application of algal toxin-degrading enzymes in inhibiting cyanobacteria and degrading algal toxins
  • Application of algal toxin-degrading enzymes in inhibiting cyanobacteria and degrading algal toxins

Examples

Experimental program
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Effect test

preparation example 1

[0048] This preparation example is used to illustrate the preparation method of cyanotoxin degrading enzyme I.

[0049] The MlrA coding sequence (Accession Number: AF411068.1) was constructed on the prokaryotic expression vector pMAL-C2X (inserted between the BamHI and HindIII restriction sites) to obtain the expression vector pMAL-C2X-MlrA expressing MlrA. For the specific construction method, see " Chapter 1 and Chapter 15 of Molecular Cloning Laboratory Guide, Third Edition (J. Sambrook et al., Science Press, 2002).

[0050] Take the prepared competent DH5α out of the refrigerator and freeze-thaw on ice, add 10 μL of the ligation product (expression vector pMAL-C2X-MlrA), reset the EP tube in ice for 30 minutes, and then put it in a 42°C Place in a water bath for 90 seconds, then put it in ice and let it stand for 5 minutes. Add 600 μL of LB medium to each tube, incubate in a shaker at 37°C and 180 rpm for 70 min; then centrifuge at 4000 rpm, take 600 μL of supernatant, mi...

preparation example 2

[0058] This preparation example is used to illustrate the preparation method of algae toxin degrading enzyme II.

[0059] The MlrA coding sequence (Accession Number: AB468058.1) was constructed on the prokaryotic expression vector pET22b (inserted between the BamHI and HindIII restriction sites) to obtain the expression vector pET22b-MlrA expressing MlrA. For the specific construction method, see "Molecular Cloning Experiment Guide" "Third Edition (J. Sambrook et al., Science Press, 2002) Chapters 1 and 15.

[0060] Take the prepared competent DH5α out of the refrigerator and put it on ice to freeze-thaw, add 10 μL of the ligation product (expression vector pET22b-MlrA), reset the EP tube in ice for 30 minutes, and then put it in a water bath at 42°C , placed for 90s, then placed in ice, and allowed to stand for 5min. Add 600 μL of LB medium to each tube, incubate in a shaker at 37°C and 180 rpm for 70 min; then centrifuge at 4000 rpm, take 600 μL of supernatant, mix the prec...

Embodiment 1

[0069] Experiments on Inhibition of Growth of Cyanobacteria and Degradation of Algal Toxin by Algal Toxin Degrading Enzyme

[0070] (1) Add the enzyme preparation to 100mL containing the lower initial algae cell concentration (about 1.0×10 5 algae cells / mL), including Synechocystis 6803, Microcystis aeruginosa 905, and Microcystis aeruginosa 912, were treated continuously for 2, 4, and 6 days to observe and detect the inhibition of different algae. Among them, the inhibition of Microcystis aeruginosa 905 by enzyme preparation 3 with different concentrations is shown in figure 1 And table 3, from which it can be seen that the enzyme preparation has a good inhibitory effect on cyanobacteria when it contains 50-400mg in 1L algae liquid. In addition, measure the OD of the algae liquor (enzyme dosage is 200mg) after continuous treatment 6 days 730 value, and start with "(initial OD 730 Value - OD after treatment 730 value) / initial OD 730 The calculation result of "value × 100%...

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Abstract

The invention relates to the fields of agriculture, fishery production and environmental protection, and discloses the application of algae toxin degrading enzymes in inhibiting cyanobacteria and degrading algae toxins. It specifically discloses the application of algae toxin degrading enzyme in inhibiting or killing cyanobacteria and degrading algae toxin while inhibiting or killing cyanobacteria. The invention also discloses a composition for inhibiting or killing cyanobacteria while degrading algae toxin and a preparation method thereof. The present invention can not only obtain the effect of inhibiting or killing cyanobacteria, but also degrade algal toxins through the use of algal toxin degrading enzymes. Therefore, the algal toxin degrading enzymes of the present invention can be widely used in agriculture, fishery (including aquaculture, etc.) production and Water environment pollution treatment.

Description

technical field [0001] The present invention relates to the fields of agriculture, fishery production and environmental protection, in particular to the application of microcystins degrading enzyme (MDE) in inhibiting or killing cyanobacteria, the application of MDE in inhibiting or killing cyanobacteria while degrading algae toxins, Composition for inhibiting or killing cyanobacteria while degrading algal toxin, its preparation method and application. Background technique [0002] In recent years, there have been large-scale outbreaks of cyanobacteria blooms in various places. The algae damage has directly affected the domestic and industrial water in the surrounding areas, seriously damaged the ecological environment, and brought huge economic losses to the development of agriculture, fishery production and tourism. Bring great hidden danger to public health, cause serious social public safety problem. [0003] In the comprehensive management of algae hazards, two key iss...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C02F3/34C12N9/00C12N15/70
CPCC02F3/342C02F2103/007C02F2303/04C12N9/00C12N15/70
Inventor 冯玲玲刘红林郭小亮巩小涵万坚冯江涛廖琴
Owner HUAZHONG NORMAL UNIV
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