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A method of obtaining halophilic bacteria with high-salt wastewater treatment function

A high-salt wastewater and functional technology, applied in the direction of biological water/sewage treatment, water/sludge/sewage treatment, bacteria, etc., can solve the problems of microbial growth, toxicity, high toxicity, threat and impact on human life and production, and achieve The effect of stable performance of single bacteria

Active Publication Date: 2020-07-28
宜兴国际环保城科技发展有限公司
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Problems solved by technology

[0002] In recent years, with the vigorous development of my country's industrial industry, the discharge of industrial sewage has increased rapidly. With the diversification of chemical synthesis processes, most of the high-salt and high-chlorine sewage has been added to the industrial sewage. The growth of microorganisms can be inhibited or poisonous, and the discharge of a large amount of standard sewage will inevitably pollute the surrounding environment and receiving water bodies, endangering the surrounding sea areas, thus posing a serious threat and impact on human life and production

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  • A method of obtaining halophilic bacteria with high-salt wastewater treatment function

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Embodiment 1

[0020] Example 1: Acquisition of total halophilic genes

[0021] Saline-alkali lakes are high-saline-alkali environments in nature, mostly inland lakes, which can form a relatively closed microbial ecological environment. The microbial community of saline-alkali lakes includes algae, cyanobacteria, aerobic bacteria, anaerobic bacteria, facultative aerobic bacteria, Archaea, etc. Archaea are an important component of the microbial community in saline-alkali lakes. They are adapted to high-salt-alkali environments. Most of them belong to the type of chemical energy organic nutrition. Because halophilic archaea have different osmotic pressure, cell membrane ion distribution, The characteristics of halophilicity and anaphylaxis are difficult to cultivate under conventional conditions. It is precisely because of the specificity of the above-mentioned physicochemical properties of archaea that they have different metabolic pathways from conventional microorganisms and have the potential...

Embodiment 2

[0024] Example 2: Construction of a halophilic gene library

[0025] The purified DNA was sequenced and the salt lake halophilic gene library was constructed using the E.coli expression system (E.coli DH5a), and the genomic DNA was randomly interrupted to 100bp, 150bp, 200bp, 250bp, 500bp, 1.0kb, 1.5kb, 2.5kb After ligating the DNA fragment with the adaptor, PCR amplified 100bp, 150bp, 200bp, 250bp, 500bp sequences and sequenced on the computer to obtain short sequence libraries. The 1.0kb and 1.5kb large fragment libraries were constructed using Cre-Lox library construction technology. After connecting both ends of the large DNA fragment with LoxP adapters, circularization and sequencing were performed; according to the genome construction steps, insert fragments of 250 bp, 500 bp, 1.0 kb, and 1.5 kb were constructed to insert into the library, and the genome sequencing was commissioned by Nanjing GenScript Company through SOLEXA The platform is complete. Inoculate the above-me...

Embodiment 3

[0026] Example 3: Transfer of halophilic gene into Bacillus subtilis

[0027] The codons of SAL1, SAL5, and SAL6 were modified to adapt to the Bacillus subtilis coding system, and the nucleotide sequence of the modified gene SAL1c was shown in SEQ ID NO:1, and the nucleotide sequence of SAL5c was shown in SEQ ID NO: 2. As shown, the nucleotide sequence of SAL6c is shown in SEQ ID NO: 3. The nucleotide sequences of SAL1c, SAL5c, and SAL6c were synthesized by Nanjing GenScript Biotechnology Co., Ltd. The synthesized sequence is also connected to NcoI at the 5'end. The restriction site, the 3'end is also connected with a SwaI restriction site. The synthesized nucleotide sequences of SAL1c, SAL5c, and SAL6c were respectively connected to the cloning vector pGEM-T (purchased from Promega), and the operation steps were carried out according to the vector instructions to obtain recombinant cloning vectors pGEM-SALc1, pGEM-SALc5, pGEM-SALc6 ( Vector structure: Amp represents the ampicil...

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Abstract

The invention provides a method for obtaining halophilic bacteria with a high-salt wastewater treatment function. The method particularly includes transferring a halophilic related gene into a specific microorganism. After the obtained halophilic bacteria are added to a deep biochemical pool, the microbial life in background sludge in the deep biochemical pool is not affected. The halophilic bacteria are resistant and degradable to high-salt high-chlorine and high-alkali wastewater, and can treat the wastewater with poor biodegradability and have stable single bacteria performance. The performance can be subjected to passage by means of genetic modification, and the method is especially suitable for industrial scenes with deep sewage treatment demand.

Description

Technical field [0001] The invention relates to the field of sewage treatment, in particular to a method for obtaining halophilic bacteria with high-salt wastewater treatment functions. Background technique [0002] In recent years, with the vigorous development of my country’s industrial industry, the discharge of industrial wastewater has increased rapidly. With the diversified development of chemical synthesis processes, most of the high-salt and high-chlorine wastewater has been added to industrial wastewater. The growth of microorganisms has inhibitory or toxic effects, and the discharge of a large amount of standard sewage is bound to pollute the surrounding environment and receiving water bodies, endanger the surrounding seas, and cause serious threats and impacts to human life and production. [0003] Bio-enhancement technology belongs to biological treatment technology. Bio-enhancement technology refers to adding microorganisms with specific functions to the biological trea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34
Inventor 周春松周佳玮贾建洪王浩孙坚
Owner 宜兴国际环保城科技发展有限公司
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