Lysinibacillus fusiformis strain and enzyme preparation, and application of same to degradation of pesticide residues

A lysinic bacillus and spindle-shaped technology, which is applied in the field of biotechnology to degrade pesticide residues, can solve the problems of mixed pesticide residues that have not been seen, and achieve the effect of removing pesticide residues

Inactive Publication Date: 2017-11-24
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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  • Lysinibacillus fusiformis strain and enzyme preparation, and application of same to degradation of pesticide residues
  • Lysinibacillus fusiformis strain and enzyme preparation, and application of same to degradation of pesticide residues
  • Lysinibacillus fusiformis strain and enzyme preparation, and application of same to degradation of pesticide residues

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0030] Example 1, the isolation and identification of spindle-shaped lysine bacillus strain

[0031] 1. Collection of soil samples

[0032] Collected from the sludge at the discharge outlet of the pesticide factory.

[0033] 2. Isolation and screening of strains

[0034] Take by weighing sludge 10g in 100mL inorganic salt liquid medium, the composition of inorganic salt liquid medium is as follows:

[0035] 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.

[0036] 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 purify by plate dilution...

example 2

[0039] Example 2, spindle-shaped lysine bacillus strain induction, domestication

[0040] 1. Chlorpyrifos induction and domestication

[0041]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.

[0042] The components of tryptone agar are: 18g trypton...

example 3

[0056] Example 3, utilize spindle-shaped lysine bacillus strain FY-7 to prepare biodegradable enzyme preparation

[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, cultured at 37°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 37° C. and 180 rpm for 24 hours to obtain the third-grade seeds.

[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 the middle according to the foam situation...

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Abstract

The invention discloses a Lysinibacillus fusiformis strain and an enzyme preparation, and application of the same to degradation of pesticide residues. The invention provides the Lysinibacillus fusiformis strain FY-7, with an accession number of CGMCC No. 13523. The invention also provides screening, induction and domestication of the strain and a preparation method for a pesticide residue-degrading active enzyme. Chlopyrifos is used as a carbon source for screening of the target strain, and then the target strain is subjected to induction and domestication with a single pesticide and a mixed pesticide. The strain is subjected to fermentation, then bacterial cells are collected, broken and centrifuged, and the obtained supernatant can be used for preparing the pesticide residue-degrading enzyme preparation. The prepared pesticide residue-degrading enzyme preparation can be used for degrading pesticide residues on vegetable and fruit and has application potential in degradation of pesticide residues on vegetable and fruit.

Description

technical field [0001] The invention relates to the field of biotechnological degradation of pesticide residues, in particular to the application of Lysinibacillus fusiformis strain screening, inducing domestication and removing pesticide residues in fruits and vegetables. 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 body, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/00C11D7/42C11D3/386C12R1/01
CPCC11D3/386C12N9/00C12N1/205C12R2001/01
Inventor 常雪妮纳泽·米尔冯立雄蒋丽娟基特·彦姆薛建龙迈克尔·里奥尼德斯·崎肯德思谢知芳
Owner 侨康生物科技(广东)有限公司
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