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Compositions and Methods for Pesticide Degradation

a technology of pesticide degradation and composition, applied in the field of compositions and methods concerning pesticide degradation, can solve the problems of affecting the appearance and quality of crops and fruits sprayed with massive amounts of pesticide, affecting the health of people, so as to reduce the half-life of pesticides

Inactive Publication Date: 2020-12-24
UNIV OF VIRGINIA ALUMNI PATENTS FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides methods for reducing the half-life of a pesticide called malathion, which can be applied to crops or agricultural products. The methods involve using a composition described in the invention, which can decrease the half-life of malathion to about 1, 2, 3, or 4 hours, depending on the desired outcome. This can help to improve the effectiveness and safety of the pesticide.

Problems solved by technology

While the widespread use of pesticides has had several benefits for the agricultural industry in terms of combating the adverse economic impact of pest-related crop loss, excessive pesticide use poses significant problems to the environment and health.
For example, the appearance and quality of crops and fruits sprayed with massive amounts of pesticide are significantly compromised.
Further, excessive pesticide exposure leads to detrimental health effects such as nausea, seizures, and even death.
However, biosafety concerns hinder synthetic biology applications because microorganisms (e.g., bacteria) rapidly replicate and may contain one or more antibiotic resistance genes.

Method used

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  • Compositions and Methods for Pesticide Degradation
  • Compositions and Methods for Pesticide Degradation
  • Compositions and Methods for Pesticide Degradation

Examples

Experimental program
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example 1

[0052]Construction of an IPTG-Inducible ftsZ Operon for the Production of E. coli Minicells

[0053]Despite many attractive qualities for synthetic biology, Escherichia coli and other engineering-amenable bacteria suffer from a number of inherent biosafety issues that limit their practical application. A safer alternative exists in the little-known nano-chassis called a minicell—the anucleoid product of an aberrant, asymmetric cell division. To introduce minicells to synthetic biology, a minicell-inducing operon from standardized parts was engineered. Through IPTG-tunable overexpression of the tubulin-homolog FtsZ, this operon enables optimization of E. coli minicell production. Whereas a spatially oscillating “Z ring” composed of FtsZ protein marks cleavage at the cell midline in wild type E. coli, overexpression of FtsZ initiates Z-ring assembly distal to the midline to form a minicell (FIG. 1). These engineered minicells are achromosomal bacterial particles that are unable to replic...

example 2

[0061]Plasmid Construct for Surface Expression of Organophosphate Hydrolase (OPH) on Minicells

[0062]An exemplary plasmid construct for surface expression of organophosphate hydrolase (OPH) on minicells is shown in FIGS. 7A-7B. The Knr designation represents the kanamycin resistance gene used in order to ascertain whether isolated minicells expressed OPH; any minicell lacking the plasmid would die upon exposure to sufficient levels of kanamycin. The PAllacO-1 designation represents the lac promoter that regulates the expression of our INPNC-OPH protein. In the absence of IPTG, expression of the INPNC-OPH protein will be actively repressed until IPTG is in solution. The p15A represents the origin of replication of the plasmid. This allows for the plasmid to be copied within the cell in order to ensure that maximal expression of the desired protein—INPNC-OPH—upon induction with IPTG. The INPNC-OPH gene codes for organophosphate hydrolase fused to an ice nucleation protein. The ice nucl...

example 3

[0063]Induction and Purification of Minicells

[0064]Bacterial strains that naturally produce minicells (commonly at a 2:1 ratio (2 bacterial cells for every one minicell) were used to obtain OPH-expressing minicells (produced using the construct shown in FIGS. 7A-7B and described in Example 2). Minicells were purified by differential centrifugation at 2,000 g for 10 min at 4° C. to pellet the parent bacteria followed by centrifugation of the supernatant at 10,000 g for 10 min at 4° C. to pellet minicells. The minicell pellet was then re-suspended in 50 mL LB and incubated at 37° C., 180 rpm for 90 min with 100 mg / L ceftriaxone. This dose of ceftriaxone is sufficient to cause cell lysis without having any detrimental effect on minicell integrity. The resulting minicell preparation was filtered at room temperature through a 0.45 μm dead end filter (Millipore SE1M003M00) to remove any remaining parent cells, followed by additional filtration with a 0.22 μm cross-flow filter (Millipore G...

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Abstract

Compositions and methods related to anucelate cells (e.g., bacterial minicells) for pesticide degradation applications including related cells, polypeptides, and vectors.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a Continuation of U.S. application Ser. No. 16 / 092,697, filed Oct. 10, 2018, which is a 371 national phase of International Application No. PCT / US2017 / 027048, filed Apr. 11, 2017 and published as WO 2017 / 0180650, which claims priority to U.S. Provisional Patent Application No. 62 / 320,790 filed on Apr. 11, 2016, each of which are incorporated herein by reference in its entirety.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY VIA EFS-WEB[0002]The entire content of the following electronic submission of the sequence listing submitted via the USPTO EFS-WEB server is incorporated herein by reference in its entirety for all purposes. The sequence listing is within the electronically filed text file that is identified as follows:File Name: UVA_SEQL_ST25.txtDate of Creation: 8 Oct. 2018[0003]Size (bytes): 1,055 bytesTECHNICAL FIELD OF THE INVENTION[0004]The invention relates generally to compositions and methods concern...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N9/16C07K14/195A62D3/02
CPCC12N9/16A62D3/02C12Y301/08002A62D2101/26C07K2319/00A62D2101/04C07K14/195
Inventor POURTAHERI, PAYAMZOMORODI, SEPEHRDAVIS, ZACHERY GEORGESHAKEEL, AMEER HAMZAFRANK, JOSEPHMOSHASHA, SHAUN RAFIEKHOKHLACHEV, ANDREIKESTER, MARK
Owner UNIV OF VIRGINIA ALUMNI PATENTS FOUND
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