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Alcaligenes faecalis subsp.faecalis strain, enzyme preparation as well as application thereof in degrading pesticide residue

A technology for Alcaligenes faecalis to degrade pesticides, which is applied in the field of biotechnology to degrade pesticide residues. It can solve problems such as mixed pesticide residues that have not been seen, and achieve the effect of removing pesticide residues.

Active Publication Date: 2017-12-12
侨康生物科技(广东)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Improving food safety is of great significance, but so far, there have been no reports on the use of bacterial strains or enzyme preparations made from them to degrade mixed pesticide residues in fruits and vegetables

Method used

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  • Alcaligenes faecalis subsp.faecalis strain, enzyme preparation as well as application thereof in degrading pesticide residue
  • Alcaligenes faecalis subsp.faecalis strain, enzyme preparation as well as application thereof in degrading pesticide residue

Examples

Experimental program
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Effect test

example 1

[0029] Example 1, isolation and identification of Alcaligenes faecalis subspecies fecal strain

[0030] 1. Collection of soil samples

[0031] Collected from the sludge at the discharge outlet of the pesticide factory.

[0032] 2. Isolation and screening of target strains

[0033] Take by weighing sludge 10g in 100mL inorganic salt liquid medium, the composition of inorganic salt liquid medium is as follows:

[0034] MgSO 4 ·7H 2 O 0.2g; K 2 HPO 4 0.1g; (NH 4 ) 2 SO 4 0.1g; CaSO 4 0.04g; FeSO 4 ·7H 2 O0.001g; deionized water 1L; pH 7.0. Sterilize at 121°C for 30min.

[0035] Add chlorpyrifos with a concentration of 100mg / L to the inorganic salt liquid medium to obtain a culture solution, cultivate it on a shaker at 35°C and 180r / min, and after cultivating for one week, insert it into the fresh culture solution with a 10% inoculation amount, and then The turnover is connected once, so the domestication is repeated many times. Finally, isolate and purify by plat...

example 2

[0038] Example 2, Alcaligenes faecalis subsp. faecalis strain induction, domestication

[0039] 1. Chlorpyrifos induction and domestication

[0040]1. The content range of active ingredients prepared by chlorpyrifos: 300mg / L-800mg / L. Tryptone agar containing different concentrations of chlorpyrifos was prepared. Different amounts of chlorpyrifos were added into tryptone agar to make a medium containing chlorpyrifos. Concrete steps: 1) Chlorpyrifos is prepared into a 4500 mg / L pesticide standard solution. Take 13.3mL, 17.8mL, 22.2mL, 26.6mL, 31.1mL, 35.6mL of pesticide standard solutions with a concentration of 4500mg / L and add them to 200mL tryptone agar to prepare chlorpyrifos containing 300mg / L, 400mg / L, 500mg / L , 600mg / L, 700mg / L and 800mg / L tryptone agar medium. Plate production: Pour the prepared tryptone agar containing 300mg / L-800mg / L chlorpyrifos into a plate, 15mL / plate, and use it after cooling and solidifying.

[0041] The components of tryptone agar are: 16g t...

example 3

[0054] Example 3, Utilize Alcaligenes faecalis subspecies FY-2 bacterial strain to prepare biodegradable enzyme preparation

[0055] 1. Strain expansion culture: Inoculate refrigerated strains into 100mL sterile TSB medium, culture at 35°C and 180rpm for 24h, and then they will be the first-grade seeds; transfer the first-grade seeds to 1L with 10% inoculum volume In the TSB medium, cultured at 35°C and 180rpm for 24 hours, it is the secondary seed. The second-grade seeds were transferred to 10 L of industrial medium according to 10% inoculum amount, and cultivated at 35° C. and 180 rpm for 24 hours to obtain the third-grade seeds.

[0056] 2. Inoculate the tertiary seeds into the fermenter, and the inoculation amount is 5% of the fermentation medium. Controlled ventilation volume is 2.5m 3 / h, the temperature is 35°C, the pH is 6.9±0.4, the rotation speed is 180rpm, samples are taken at intervals, and the OD is measured 600 And pH, in the middle according to the foam situa...

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Abstract

The invention discloses an alcaligenes faecalis subsp.faecalis strain, an enzyme preparation as well as application thereof in degrading pesticide residue. The invention provides an alcaligenes faecalis subsp.faecalis FY-2 strain, and a preservation number of the strain is CGMCC (China General Microbiological Culture Collection Center) No. 13518. The invention also provides a method for screening, inducing and domesticating the strain and a preparation method of active enzyme for degrading pesticide residue. Chlorphrifos is used as a carbon source to screen a target bacterium, and the target bacterium is induced and domesticated in single pesticide and mixed pesticide. The strain is fermented, the bacteria are collected, lysed and centrifuged, and the supernatant can be used for preparing the pesticide residue degradation enzyme preparation which can be used for degrading the pesticide residue of vegetables and fruits and has application potential in the aspect of degrading the pesticide residue of the vegetables and fruits.

Description

technical field [0001] The invention relates to the field of biotechnological degradation of pesticide residues, in particular to the application of Alcaligenes faecalis subsp. faecalis strain screening, domestication induction and removal of pesticide residues in vegetables and fruits. Background technique [0002] With the rapid increase in the variety, output and usage of pesticides in recent years, the phenomenon of mixed application of multiple pesticides in the agricultural production process occurs from time to time and is more serious, resulting in multiple pesticides appearing in the same product. Eating vegetables and fruits with excessive pesticide residues will harm the nerve center and cause nerve conduction dysfunction; it can induce mutations, deformity, and affect the health of offspring; it can induce changes in liver enzymes, causing liver enlargement and necrosis; it can invade the kidneys and cause pathological changes. Pesticide residues accumulate in th...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/00A23L5/20C12R1/05
CPCA23L5/25A23L5/28C12N9/00C12N1/205C12R2001/05
Inventor 薛建龙蒋丽娟基特·彦姆常雪妮迈克尔·里奥尼德斯·崎肯德思谢知芳纳泽·米尔冯立雄
Owner 侨康生物科技(广东)有限公司
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