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Bacillus licheniformis UTM104 for producing pyrethroid hydrolase and application of bacillus licheniformis UTM104

A technology of Bacillus licheniformis and UTM104, which is applied in the field of fermentation, can solve problems such as pesticide residues and health threats, and achieve the effects of excellent strain performance, food safety protection, good social and economic benefits

Active Publication Date: 2015-04-08
大地绿源环保科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the other hand, while the use of pesticides increases the yield of crops, it also causes pesticide residues in food and soil, causing various food safety problems and posing a huge threat to people's health.

Method used

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  • Bacillus licheniformis UTM104 for producing pyrethroid hydrolase and application of bacillus licheniformis UTM104
  • Bacillus licheniformis UTM104 for producing pyrethroid hydrolase and application of bacillus licheniformis UTM104
  • Bacillus licheniformis UTM104 for producing pyrethroid hydrolase and application of bacillus licheniformis UTM104

Examples

Experimental program
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Effect test

Embodiment 1

[0032] The isolation and identification of embodiment 1 Bacillus licheniformis UTM104

[0033] Take about 1 gram of Tengchong hot spring bottom mud sample and place it in a 250ml Erlenmeyer flask filled with multiple small glass beads and 100ml sterile water, and incubate with shaking at 40°C for 1 hour at a speed of 160 rpm, then let it stand for 30 minutes. Draw 1ml of the supernatant under sterile conditions and insert it into 100ml of sterilized enrichment medium (15.0g colloidal chitin, 5.0g yeast powder, (NH 4 ) 2 SO 4 1.0g, MgSO 4 ·5H 2 O 0.3g, KH 2 PO 4 1.36 g, 1000 ml of water, pH 4.5), shake culture at 40° C. for 3 days, and rotate at 160 rpm. Take 1ml of the bacterial solution cultured for 3 days, add it to a test tube filled with 9ml sterile water, and prepare 10% by 10-fold dilution method. -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7the dilution. Take 0.1ml and spread on three solid medium plates containing colloidal chitin (15.0g colloidal ch...

Embodiment 2

[0037] Example 2 Detection of Chitinase Activity of Bacterial Strain UTM104 and its Gene Sequence

[0038] The UTM104 strain was inoculated on the colloidal chitin solid medium plate of Example 1, and cultured at 37° C. for 3 days. It was observed to have a transparent chitinase hydrolysis circle. According to the primer chitinase-f of chitinase gene design: 5'-TCA TCG GCT ACT ATC C-3' and chitinase-r: 5'-TCA CCG GAT TGA TCA G-3', and use bacterial strain UTM104DNA of the present invention as Template for PCR amplification reaction, PCR reaction program: 95°C pre-denaturation for 5 minutes, 95°C denaturation for 30 seconds, 56°C annealing for 45 seconds, 72°C extension for 90 seconds, 35 cycles, 72°C extension for 10 minutes, electrophoresis A nucleotide fragment with a size of about 1.7kb was detected, sequenced and sequenced in the NCBI GenBank database using the Blast program to obtain the sequence fragment encoding the chitinase gene, the gene sequence of which is shown i...

Embodiment 3

[0039] Example 3 Detection of cellulase activity of bacterial strain UTM104 and related gene sequences

[0040] The UTM104 strain was planted in sodium carboxymethyl cellulose medium (sodium carboxymethyl cellulose 5g, KH 2 PO 4 lg, agar 17g, NaNO 3 3g, KCL 0.5g, MgSO 4 0.5g, FeSO 4 0.01g, 1000ml of distilled water, pH 5.5~6.0), cultured at 40°C for 48 hours, stained with 0.2% Congo red for 30 minutes, washed the dye solution with distilled water, soaked in NaCl with a concentration of 1mol / L for 1 hour, and finally used 5% acetic acid fixes the color. The formation of a colorless transparent circle around the colony indicates that the bacterium secretes cellulase. Cellulase is a multi-enzyme mixture consisting of cellobiase, endo-β-glucanase and the like.

[0041] Primer betaglu-f: 5'-AAC GGT CCG CAT CAT C-3' and betaglu-r: 5'-CGA GGA AGA TGC CGA T-3' were designed according to the endo-β-glucanase gene, with the strain of the present invention UTM104DNA was used as a ...

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Abstract

The invention provides a bacillus licheniformis UTM104 strain for producing pyrethroid hydrolase and application of the bacillus licheniformis UTM104 strain. The biological preservation number of the bacillus licheniformis UTM104 strain is CGMCC No.9509. The UTM104 strain can be subjected to aerobic composting fermentation in organic solid wastes such as sewage sludge, household garbage, animal carcass, livestock manure and crop straws, can form a stable ecological system of a bacterial colony in an organic material, and can massively and rapidly breed, so that the organic matters in the organic solid wastes can be effectively degraded, and thus the reduction and harmless targets are rapidly achieved. Meanwhile, a microbial agent prepared by the bacillus licheniformis UTM104 strain is capable of transforming the organic solid wastes into a bio-fertilizer; and the UTM104 strain is capable of degrading pyrethroid pesticide residues on soil and plants, so that the food safety is ensured; and the strain disclosed by the invention is excellent in property, and has the advantages of relatively good social benefits and economic benefits.

Description

technical field [0001] The invention relates to the field of fermentation, in particular to a Bacillus licheniformis UTM104 strain carrying a pyrethroid hydrolase gene and its application in degrading urban and rural organic solid waste and preparing biological fertilizers. Background technique [0002] At present, the organic solid waste produced by agricultural and animal husbandry production has become an important source of urban and rural environmental pollution. The annual large-scale production of crop straw and livestock manure and other waste not only causes farmland occupation, soil and groundwater pollution, but also produces odor pollution. Air. On the other hand, while the use of pesticides increases the yield of crops, it also causes pesticide residues in food and soil, causing various food safety problems and posing a huge threat to people's health. How to effectively remove agricultural products and pesticide residues in soil has become a widely concerned is...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F17/00C12R1/10
CPCC05F11/08C05F17/00C12N1/20C12N1/205C12R2001/10Y02W30/40
Inventor 刘永跃何璧梅许宜北汪涌
Owner 大地绿源环保科技(北京)有限公司
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