Chlorpyrifos degrading bacteria and application thereof

A technology for degrading bacteria and colonies of chlorpyrifos, which can be applied to bacteria, microorganisms, biochemical equipment and methods, etc., and can solve problems such as limited means, low efficiency, and secondary pollution.

Inactive Publication Date: 2014-12-31
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional physical and chemical treatment methods are not only inefficient and have limited means, but also produce secondary pollution during the treatment process. Bioremediation methods that use biological or enzy...

Method used

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  • Chlorpyrifos degrading bacteria and application thereof
  • Chlorpyrifos degrading bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment one: Erwinia xishuaiensis SCU-B244 T screening and separation

[0022] The cricket samples used in the experiment are adults of Gryllus chinensis, which are mostly distributed in farmland. They were captured in batches in August 2012 from the wasteland of Jiang’an Campus, Sichuan University, Shuangliu County, Chengdu, Sichuan (30°33’ 38.04”North 104°00 '20.17” East, 481 meters above sea level). After the microorganisms were eluted from the crickets with sterile water, they were spread on TSB, Chase, Gao’s No. 1, PDA, beef extract peptone and parasite inorganic salt medium by the dilution plate method, and cultivated at 37°C until visible Colonies appeared, and were identified after purification.

[0023] Description of strains:

[0024] Gram-negative bacteria, facultative aerobic, cream-colored colonies. Oxidase negative, catalase positive. The pH growth range is 6 to 9, and the optimum growth salt concentration is 1% to 2%. The bacteria has alkaline ...

Embodiment 2

[0025] Embodiment two: Erwinia xishuaiensis SCU-B244 T multiphase taxonomic identification of

[0026] the strain Erwinia xishuaiensis SCU-B244 T Biolog automatic bacterial identification system was used for carbon source utilization, fatty acid composition was determined by GC-MS analysis, enzymatic properties were identified by API ZYM reagent strips, acid production was identified by API50CHE reagent strips, and the reference strain was Erwinia oleae DSM 23398 T . Carbon source utilization was determined by Biolog automatic bacterial identification system. The result is as follows:

[0027] Available carbon sources: dextrin, N-acetyl-D-glucosamine, L-arabinose, D-fructose, D-galactose, α-D-glucose, maltose, D-mannitol, D-mannose, D-melibiose, β-methyl-D-glucoside, allulose L-raffinose, D-trehalose, turanose, methylpyruvate, citric acid, formic acid, D-lactobionic acid Esters, D-galacturonic acid, D-gluconic acid, p-hydroxyphenylacetic acid, α-ketoglutaric acid, ...

Embodiment 3

[0035] Example 3: 16S rDNA sequencing and evolutionary tree construction

[0036] The total bacterial DNA was extracted by SDS method, the 16S rRNA gene sequence of the strain was amplified by PCR method, and the DNA sequencing was completed by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Go to the GenBank (http: / / www.ncbi.nlm.nih.gov / ) database to submit the partial sequence of the 16S rRNA gene measured by each strain for registration to obtain the sequence number. Then perform BLAST comparison on NCBI (National Center for Biotechnology Information) to find the strain with the closest relationship. The sequences of the corresponding strains with the closest relatives were compared by BioEdit, and then the phylogenetic tree was constructed using the Neighbor-Joinin (N-J) method on the software MEGA 5.0. The corresponding primers, reaction system and conditions for PCR are as follows:

[0037] Primer PA8: 5'-CCGTCGACGAGCTCAGAGTTTGATCCTGGCTCAG-3' and PB1: 5'-CC...

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Abstract

The invention discloses a chlorpyrifos degrading bacteria Erwiniaxishuaiensis SCU-B244<T> = CGMCCNo.1.12772<T> = KCTC42022<T> (Figure) (i). Through the separation from the cricket body surface, screening, physiological and biochemical identification, the bacteria is identified as gram negative and facultative aerobic bacteria, which has cream bacterium colony, and is oxidase negative and catalase positive; the cells mainly comprises fatty acids of C16:0, C16:1delta<9>, C18:1delta<9>, C11:0<3-OH> and C14:0<3-OH>, GC content in cells is 49.3%-49.5%; and the bacteria has alkaline phosphatase, esterase (C4), lipase (C8), acid phosphatase, beta-galactosidase and beta-glucosidase activities. The bacteria has one day degradation rate on chlorpyrifos of 44.64%, and can be widely used in the fields of development of chlopyrifos degrading enzyme and various types of biological engineering enzyme preparations. Figure instructions: the figure is photos of Erwinia xishuaiensis SCU-B244<T> under 30000 times scanning electron microscope.

Description

technical field [0001] The invention relates to the field of microbes and their applications, in particular to the technical field of chlorpyrifos-degrading Enterobacteriaceae and its applications. Background technique [0002] Organophosphorus pesticides are widely used in agricultural production because of their low residue, high efficiency and low cost of use. On the one hand, they increase the yield of crops, but at the same time, they also bring many problems. The most important thing is the pollution of the environment and the destruction of the ecology by pesticides, which directly or indirectly affect human health. Studies have shown that organophosphorus pesticides are mutagenic and teratogenic, and nervous and immune system diseases have also been proved to be related to organophosphorus pesticides. How to eliminate the harm of organophosphorus pesticides to the environment and humans has become a matter of human health and national economy. developmental issues. ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02C12R1/18A62D101/04A62D101/26
Inventor 田永强刘波王磊邱鹏龙秀锋
Owner SICHUAN UNIV
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