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Bacillus elongatus lysinus strain, enzyme preparation and its application in degrading pesticide residues

A technology of Bacillus lysine to degrade pesticides, applied in the field of biotechnology degradation of pesticide residues, can solve problems such as mixed pesticide residues that have not yet been seen, and achieve the effect of removing pesticide residues

Inactive Publication Date: 2020-05-22
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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  • Bacillus elongatus lysinus strain, enzyme preparation and its application in degrading pesticide residues

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0028] Example 1, isolation and identification of lysine bacillus strain

[0029] 1. Acquisition of strains

[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 37°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 ...

example 2

[0038] Example 2, induction and acclimatization of elongated lysine bacillus strain

[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: 17g try...

example 3

[0055] Example 3, utilize elongated lysine bacillus strain FY-3 to prepare biodegradation enzyme preparation

[0056] 1. Obtaining the fermentation broth

[0057] 1. Strain expansion culture: Inoculate refrigerated strains into 100mL sterile TSB medium, cultivate at 37°C and 180rpm for 24 hours, and then they will become first-class seeds; transfer the first-class seeds to 1L with 10% inoculum volume In the TSB medium, cultivated at 37°C and 180rpm for 24 hours, it is the second-grade seed; transfer the second-grade seed to 10L industrial medium according to 10% inoculation amount, and cultivate it at 37°C and 180rpm for 24 hours, which is the third-grade seed seed.

[0058] 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 37°C, the pH is 7.2±0.4, the rotation speed is 180rpm, samples are taken at intervals, and the OD is measured 600 And pH, in th...

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Abstract

The invention discloses a Bacillus elongatus strain, an enzyme preparation and an application for degrading pesticide residues. The strain of Lysinibacillus macroides FY-3 provided by the present invention has a deposit number of CGMCC No.13519. The invention also provides a method for strain screening, induction, domestication and preparation of a pesticide residue degrading active enzyme, that is, using chlorpyrifos as a carbon source to screen a target strain, and inducing and domesticating the target strain with a single pesticide and a mixed pesticide. The strains are fermented, the cells are collected, the cells are disrupted, centrifuged, and the supernatant can be used to prepare a pesticide residue-degrading enzyme preparation, which can degrade pesticide residues in vegetables and fruits, and has application potential in the degradation of pesticide residues in vegetables and fruits.

Description

technical field [0001] The invention relates to the field of biotechnological degradation of pesticide residues, in particular to the screening, induction, domestication and application of Lysinibacillus macroides strains in degrading pesticide residues in vegetables and fruits. Background technique [0002] With the sharp 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 the human ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/00A23L5/20C12R1/01
CPCA23L5/25A23L5/28C12N9/00C12N1/205C12R2001/01
Inventor 常雪妮基特·彦姆薛建龙迈克尔·里奥尼德斯·崎肯德思谢知芳纳泽·米尔冯立雄蒋丽娟
Owner 侨康生物科技(广东)有限公司
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