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Lactococcus lactis subsp. lactic acid strain, enzyme preparation and its application in degrading pesticide residues

A technology of Lactococcus lactis and lactic acid subsp. is applied in the field of biotechnology degradation of pesticide residues, which 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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  • Lactococcus lactis subsp. lactic acid strain, enzyme preparation and its application in degrading pesticide residues
  • Lactococcus lactis subsp. lactic acid strain, enzyme preparation and its application in degrading pesticide residues

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0029] Example 1, isolation and identification of Lactococcus lactis subsp.

[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 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 purify by plate dilution method, inocula...

example 2

[0038] Example 2, Lactococcus lactis subsp. lactis 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: 15g trypt...

example 3

[0055] Example 3, utilize Lactococcus lactis subsp. lactis strain FY-5 to prepare biodegradation enzyme preparation

[0056] 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.

[0057] 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 6.2±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 situatio...

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Abstract

The invention discloses a strain of Lactococcus lactis subsp. lactis, an enzyme preparation and an application thereof for degrading pesticide residues. The present invention provides Lactococcus lactis subsp. lactis FY-5 strain, and its preservation number is CGMCC No.13521. The invention also provides a method for strain screening, induction, domestication and active enzyme preparation for pesticide residue degradation, that is, using chlorpyrifos as a carbon source to screen target bacteria, and subject the target bacteria to a single pesticide or mixed pesticides for induction and domestication. The bacterial strain is fermented, the bacteria are collected, the cells are broken, and centrifuged, and the supernatant can be used to prepare a pesticide residue-degrading enzyme preparation, which can be used for the degradation of pesticide residues in fruits and vegetables, and has application potential in degrading pesticide residues in fruits and vegetables.

Description

technical field [0001] The invention relates to the field of biotechnological degradation of pesticide residues, in particular to the application of Lactococcus lactis subsp. 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 the human body, and if they exceed a certain limit, they will cause some chronic diseases, such as muscle numbness,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/00C11D7/42C11D3/386C12R1/46
CPCC11D3/386C12N9/00C12N1/205C12R2001/46
Inventor 蒋丽娟迈克尔·里奥尼德斯·崎肯德思谢知芳纳泽·米尔冯立雄常雪妮基特·彦姆薛建龙
Owner 侨康生物科技(广东)有限公司
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