WO2015145222A3 - Recombinant and stable ssopox enzymes, method of generation thereof and reusable nanobiocatalyst of the same - Google Patents

Recombinant and stable ssopox enzymes, method of generation thereof and reusable nanobiocatalyst of the same Download PDF

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Publication number
WO2015145222A3
WO2015145222A3 PCT/IB2014/064616 IB2014064616W WO2015145222A3 WO 2015145222 A3 WO2015145222 A3 WO 2015145222A3 IB 2014064616 W IB2014064616 W IB 2014064616W WO 2015145222 A3 WO2015145222 A3 WO 2015145222A3
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WO
WIPO (PCT)
Prior art keywords
rssopox
nanobiocatalyst
enzymes
provides
reusable
Prior art date
Application number
PCT/IB2014/064616
Other languages
French (fr)
Other versions
WO2015145222A2 (en
Inventor
Abhay Hariram PANDE
Priyanka BAJAJ
Rajan Kumar TRIPATHY
Ankita JADHAV
Gaurav Shriharishankar CHANDAK
Harsh Dineshchandra PARIKH
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National Institute Of Pharmaceutical Education And Research (Niper)
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Publication of WO2015145222A2 publication Critical patent/WO2015145222A2/en
Publication of WO2015145222A3 publication Critical patent/WO2015145222A3/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/08—Phosphoric triester hydrolases (3.1.8)
    • C12Y301/08001—Aryldialkylphosphatase (3.1.8.1), i.e. paraoxonase
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/30—Organic compounds
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2305/00—Use of specific compounds during water treatment
    • C02F2305/08—Nanoparticles or nanotubes

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Zoology (AREA)
  • Public Health (AREA)
  • Wood Science & Technology (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Water Supply & Treatment (AREA)
  • Environmental & Geological Engineering (AREA)
  • Hydrology & Water Resources (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biotechnology (AREA)
  • Inorganic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Abstract

The present invention provides novel polynucleotide sequences for high level expression in Escherichia coli and encoding rSsoPox(wt) enzyme and its variant having enhanced organophosphate (OP)-hydrolyzing activity. The invention also provides a novel method to produce the recombinant enzymes in highly pure and active form in an unprecedented high yield. The invention also provides novel compositions for long-term storage of purified rSsoPox enzymes. The invention also provides PEGylated-rSsoPox conjugate having decreased protease sensitivity. The invention also provides a method to generate reusable nanobiocatalyst by covalently immobilizing pure r Sso Pox enzyme onto magnetic nanoparticles. The reusable nanobiocatalyst can be recovered from the reaction mixture after its use and can be stored and reused multiple times to degrade the OP-compounds. The rSsoPox enzymes and rSsoPox-immobilized nanobiocatalyst can be used as safe, effective and environmental-friendly OP-compound degrading agent.
PCT/IB2014/064616 2014-03-28 2014-09-18 Recombinant and stable ssopox enzymes, method of generation thereof and reusable nanobiocatalyst of the same WO2015145222A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN902/DEL/2014 2014-03-28
IN902DE2014 IN2014DE00902A (en) 2014-03-28 2014-09-18

Publications (2)

Publication Number Publication Date
WO2015145222A2 WO2015145222A2 (en) 2015-10-01
WO2015145222A3 true WO2015145222A3 (en) 2016-07-14

Family

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PCT/IB2014/064616 WO2015145222A2 (en) 2014-03-28 2014-09-18 Recombinant and stable ssopox enzymes, method of generation thereof and reusable nanobiocatalyst of the same

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IN (1) IN2014DE00902A (en)
WO (1) WO2015145222A2 (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103703128B (en) 2011-03-10 2016-10-05 康奈尔大学 The mesoporous catalyst of the enzyme of magnetic nano-particle and generation free radical and using method thereof
CA2986197C (en) 2015-05-18 2023-10-17 Zymtronix, Llc Magnetically immobilized microbiocidal enzymes
CA2992261A1 (en) 2015-07-15 2017-01-19 Zymtronix, Llc Automated bionanocatalyst production
CA3031802A1 (en) 2016-08-13 2018-02-22 Zymtronix Catalytic Systems, Inc. Magnetically immobilized biocidal enzymes and biocidal chemicals
SE1750143A1 (en) 2017-02-15 2018-08-16 Pharem Biotech Ab Enzyme identification

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100333221A1 (en) * 2007-04-27 2010-12-30 Universite Henri Poincare Nancy 1 Mutated hyperthermophilic phosphotriesterases and uses thereof
WO2014096402A1 (en) * 2012-12-20 2014-06-26 Boumendil Olivier-Georges Enzyme having a nmda receptor antagonist activity and/or an anticholinergic activity

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100333221A1 (en) * 2007-04-27 2010-12-30 Universite Henri Poincare Nancy 1 Mutated hyperthermophilic phosphotriesterases and uses thereof
WO2014096402A1 (en) * 2012-12-20 2014-06-26 Boumendil Olivier-Georges Enzyme having a nmda receptor antagonist activity and/or an anticholinergic activity

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MERONE ET AL.: "A thermostable phosphotriesterase from the archaeon Sulfolobus solfataricus : cloning, overexpression and properties", EXTREMOPHILES, vol. 9, 21 May 2005 (2005-05-21), pages 297 - 305, XP019374100, DOI: doi:10.1007/s00792-005-0445-4 *
NG ET AL.: "Characterization of a Phosphotriesterase-Like Lactonase from Sulfolobus solfataricus and Its Immobilization for Disruption of Quorum Sensing", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 77, no. 4., 23 December 2010 (2010-12-23), pages 1181 - 1186 *

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Publication number Publication date
WO2015145222A2 (en) 2015-10-01
IN2014DE00902A (en) 2015-10-02

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