Peroxidase and preparation of degradable malachite green from mangrove sediments

A technology of peroxidase and peroxidase, applied in biochemical equipment and methods, botany equipment and methods, enzymes, etc., can solve problems such as rare environmental restoration

Active Publication Date: 2017-08-08
XIAMEN UNIV
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Problems solved by technology

In terms of patents, peroxidases from various sources have been used in the fields of quantification, color development and chemical modification, but peroxidases obtained through metagenomic sequencing of mangrove sediments have not been used in malachite green degradation and environmental restoration. rare

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  • Peroxidase and preparation of degradable malachite green from mangrove sediments
  • Peroxidase and preparation of degradable malachite green from mangrove sediments
  • Peroxidase and preparation of degradable malachite green from mangrove sediments

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[0055] The present invention will be further described below in conjunction with specific embodiments and accompanying drawings. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific condition in the following examples, generally according to conventional conditions such as the experimental conditions described in the Molecular Cloning Laboratory Handbook (New York: Cold Spring Harbor Laboratory Press, 2002), or according to the reagent or instrument manufacturer's instructions suggested conditions.

[0056] 1. Extraction of environmental microbial total DNA (eDNA) from mangrove sediments

[0057]The mangrove sediments stored in the -80°C refrigerator were taken out and melted at 4°C, and the Mo Bio soil microbial extraction kit ( Soil DNA Isolation Kit (purchased from Mo Bio Company) was used to extract environmen...

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Abstract

The invention discloses peroxidase of degradable malachite green from mangrove sediments and a preparation method for the peroxidase, and relates to peroxidase. A peroxidase gene is from a microbial metagenome sequence of the mangrove sediments. The preparation method comprises the following steps of cloning the peroxidase gene, inserting an expression vector and constructing a recombinant expression vector carrying with the peroxidase; converting the recombinant expression vector into E.coli BL21(DE3); selecting positive clone in converted E.coli BL21(DE3) to perform fermented cultivation in a culture medium; centrifuging and collecting the fermented E.coli BL21(DE3) cells, re-suspending the E.coli BL21(DE3) cells in lysis buffer to be lysed, centrifuging, collecting liquid supernatant, then, uniformly mixing the liquid supernatant with R10-Flammable which is pre-combined with NiSO4, and performing imidazole elution to obtain the superoxide Mgv-rPOD. The obtained peroxidase has remarkable degradation activities on malachite green.

Description

technical field [0001] The invention relates to a peroxidase capable of degrading malachite green, in particular to a peroxidase capable of degrading malachite green derived from mangrove deposits and its preparation. Background technique [0002] Malachite Green (MG) is a triphenylmethane dye used in paper, textile and leather industries. And it is also widely used in aquaculture because of its high-efficiency antibacterial effect. However, studies in recent years have shown that malachite green is extremely carcinogenic and mutagenic to mammalian cells (Fessard et al.1999; Srivastava et al.2004), so it has been banned by many countries. However, due to the effectiveness of malachite green in antagonizing aquatic pathogens and the lack of suitable substitutes, it is still widely used, thus posing a great threat to the offshore ecological environment and human health. [0003] Peroxidase (Peroxidase, EC 1.11.1) is a class of oxidative enzymes that use organic matter as a s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/53C12N9/08C12N15/70A62D3/02A62D101/26
Inventor 赵晶曲武
Owner XIAMEN UNIV
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