Clostridium beijerinckii with high efficiency dechlorination of pentachlorophenol and its construction method and application

A technology of Clostridium beijerinckii and pentachlorophenol, applied in its construction method and application fields in the field of environmental protection, can solve problems such as teratogenicity, high toxicity, and mutagenicity

Active Publication Date: 2020-10-16
南京中环生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, PCP is the most toxic of all phenolic toxic substances, and has strong "three-cause" (carcinogenic, teratogenic, mutagenic) effects

Method used

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  • Clostridium beijerinckii with high efficiency dechlorination of pentachlorophenol and its construction method and application
  • Clostridium beijerinckii with high efficiency dechlorination of pentachlorophenol and its construction method and application
  • Clostridium beijerinckii with high efficiency dechlorination of pentachlorophenol and its construction method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1: Construction of Clostridium beijerinckii Cbei_3304 gene insertion inactivation mutant.

[0027] The principle of constructing Clostridium beijerinckii Cbei_3304 gene insertion inactivation mutant strain is as follows figure 2 As shown, the specific construction process includes the following steps:

[0028] (1) Construction of Cbei_3304 insertion inactivation vector

[0029] 1) Design introns

[0030] According to the Cbei_3304 gene sequence (as shown in SEQ ID No: 1) of Clostridium beijerinckii included in the NCBI database, a suitable insertion gene site is designed by means of software ( http: / / www.clostron.com ), select the insertion between the 101st and 102nd bases, and generate an intron sequence, synthesize the intron sequence S-304 (its sequence is shown in SEQ ID NO: 2), and design the following primers.

[0031] Cloning primers:

[0032] pWJ-OSC-304-S: 5'-ggagtgtcgaggatc ctcgag ataattatccttacacttcgcc-3', its sequence is shown in SEQ ID NO: ...

Embodiment 2

[0050] Example 2: Dechlorination experiment of Clostridium beijerinckii to pentachlorophenol in simulated wastewater.

[0051] (1) Medium formula:

[0052] YPS medium: yeast powder 3g / L, peptone 5g / L, glucose 1g / L, ammonium acetate 2g / L, sodium chloride 3g / L, magnesium sulfate heptahydrate 3g / L, potassium dihydrogen phosphate 1g / L, phosphoric acid Dipotassium hydrogen 1g / L, ferrous sulfate heptahydrate 0.1g / L, pH=6.

[0053] Simulated wastewater: glucose 2g / L, potassium chloride 0.9g / L, ammonium chloride 1.0g / L, sodium dihydrogen phosphate monohydrate 0.6g / L, calcium chloride monohydrate 0.02g / L, yeast extract 0.05g / L L, electron mediator (MV) 0.15g / L, ferric citrate 6.44g / L, pentachlorophenol 3mg / L and trace elements (H 3 BO 3 3g / L, ZnSO 4 ·7H 2 O 0.9g / L, MnCl 2 2H 2 O 2.4g / L, CuSO 4 ·5H 2 O0.6g / L, CoCl 2 ·6H 2 O 3g / L, (NH 4 ) 6 Mo 7 o 24 2H 2 O 0.02g / L, NiCl 2 ·6H 2 (0.1g / L) 5ml / L.

[0054] (2) Activate Clostridium beijerinckii group bacteria M-3304 and i...

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Abstract

The invention discloses a Clostridium beijerinckii recombinant strain for high-efficiency reduction dechlorination of pentachlorophenol. After Cbei_3304 gene undergoes insertional inactivation to make the protein not normally work in NCIMB 8052. The pentachlorophenol reduction dechlorination rate of the Clostridium beijerinckii recombinant strain M-3304 in simulated wastewater containing 3.0 mg / L of pentachlorophenol, 0.15 g / L of methyl viologen (MV) and 6.44 g / L of iron citrate is 90% at 60 h, and the pentachlorophenol reduction dechlorination rate of the original strain Clostridium beijerinckii NCIMB 8052 under the same conditions is only 40%, so the pentachlorophenol reduction dechlorination rate of the Clostridium beijerinckii recombinant strain M-3304 is 2.25 times that of the original strain Clostridium beijerinckii NCIMB 8052. The Clostridium beijerinckii recombinant strain can be applied to the degradation of pentachlorophenol in water, solid garbage, solid wastes and soil, and belongs to the field of environmental protection.

Description

technical field [0001] The invention belongs to the technical field of environmental protection, and in particular relates to a Clostridium beijerinckii recombinant bacterial strain capable of efficiently dechlorinating pentachlorophenol, a construction method thereof and an application in the field of environmental protection. Background technique [0002] Pentachlorophenol (pentachlorophenol, PCP), also known as pentachlorophenol; it often contains a molecule of crystal water, and has a strong pungent odor when slightly heated. Pentachlorophenol can prevent the growth of fungi and inhibit bacteria, and has long been used as herbicides, insecticides, fungicides, and antifungal agents. However, PCP is the most toxic of all phenolic toxic substances, and has strong "three-cause" (carcinogenic, teratogenic, and mutagenic) effects. In the 1970s, the U.S. Environmental Protection Agency included it in the list of priority pollutants (Water Res., 2004, 38: 663-672). In my countr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C02F3/34B09C1/10A62D3/02C12R1/145C02F101/36A62D101/04A62D101/28
CPCA62D3/02A62D2101/04A62D2101/28B09C1/10C02F3/34C02F2101/36C07K14/33
Inventor 郭亭席梓淇朱刚强李永
Owner 南京中环生物科技有限公司
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