WO2011068297A1 - Variante de peroxydase ayant une stabilité radicalaire améliorée - Google Patents

Variante de peroxydase ayant une stabilité radicalaire améliorée Download PDF

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Publication number
WO2011068297A1
WO2011068297A1 PCT/KR2010/005146 KR2010005146W WO2011068297A1 WO 2011068297 A1 WO2011068297 A1 WO 2011068297A1 KR 2010005146 W KR2010005146 W KR 2010005146W WO 2011068297 A1 WO2011068297 A1 WO 2011068297A1
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WIPO (PCT)
Prior art keywords
cip
variant
seq
native
amino acid
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PCT/KR2010/005146
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English (en)
Inventor
Bong Keun Song
Yong Hwan Kim
Su-Jin Kim
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Korea Research Institute Of Chemical Technology
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Publication date
Application filed by Korea Research Institute Of Chemical Technology filed Critical Korea Research Institute Of Chemical Technology
Publication of WO2011068297A1 publication Critical patent/WO2011068297A1/fr

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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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces

Definitions

  • CiP variant having an improved radical stability.
  • Peak lists from the MS/MS spectra were exported as individual files in data format using Bioworks 3.3 software (Thermo Fisher Scientific, CA, USA) under the following settings: (i) peptide mass range : 500 to 3500 Da; (ii) minimal total ion intensity threshold : 10000; (iii) minimal number of fragment ions : 20; (iv) precursor mass tolerance : 1.4 amu; (v) group scan : 1; and (vi) minimum group count : 1.
  • the energy of the HOMO is related to the potential for oxidation, the higher HOMO energy value, the more susceptible the residue is to oxidization and attack by radicals.
  • the HOMO energy values of alanine, isoleucine and leucine are lower value than those of phenylalanine, histidine, tyrosine and tryptophan.
  • the stability of native CiP and CiP variants was well correlated with the HOMO energy level. This correlation provides support for our hypothesis that the major factor that inactivates CiP during the oxidative reaction was the attack and covalently binding of the phenoxyl radical to the F229 site in CiP.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Botany (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

La présente invention concerne une variante de Coprinus cinereus peroxydase (CiP) ayant une stabilité radicalaire supérieure à celle de la CiP native, un ADN codant pour la variante de CiP, un vecteur d'expression comprenant l'ADN et un microorganisme transformé avec le vecteur d'expression.
PCT/KR2010/005146 2009-12-01 2010-08-05 Variante de peroxydase ayant une stabilité radicalaire améliorée WO2011068297A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020090117904A KR101059873B1 (ko) 2009-12-01 2009-12-01 라디칼 안정성이 증가된 신규한 퍼옥시다아제 변이체
KR10-2009-0117904 2009-12-01

Publications (1)

Publication Number Publication Date
WO2011068297A1 true WO2011068297A1 (fr) 2011-06-09

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PCT/KR2010/005146 WO2011068297A1 (fr) 2009-12-01 2010-08-05 Variante de peroxydase ayant une stabilité radicalaire améliorée

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KR (1) KR101059873B1 (fr)
WO (1) WO2011068297A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101424334B1 (ko) 2012-08-07 2014-07-31 광운대학교 산학협력단 퍼록시다아제 변이형 폴리펩타이드, 이를 코딩하는 핵산분자, 그를 포함하는 재조합벡터, 상기 재조합벡터로 형질전환된 형질전환체, 상기 형질전환체의 제조방법 및 퍼록시다아제 변이형 폴리펩타이드의 제조방법

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DI CERBO, P. ET AL.: "Kinetic evidence for surface residues influencing the a ctive site of Coprinus cinereus peroxidase: analysis of the pH dependence of G154E, P90H and P90H-G154E substrate entrance mutants.", BIOCHIMICA ET BIOPHYSICA ACTA, vol. 1544, no. 1-2, 12 January 2001 (2001-01-12), pages 18 - 27 *
NERI, F. ET AL.: "Mutation of the distal arginine in Coprinus cinereus peroxi dase--structural implications", EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 251, no. 3, 1 February 1998 (1998-02-01), pages 830 - 838 *
SMULEVICH, G. ET AL.: "Versatility of heme coordination demonstrated in a fun gal peroxidase. Absorption and resonance Raman studies of Coprinus cinereus peroxidase and the Asp245-->Asn mutant at various pH values", BIOCHEMISTRY, vol. 35, no. 32, 13 August 1996 (1996-08-13), pages 10576 - 10585 *
VEITCH, N. C. ET AL.: "NMR studies of recombinant Coprinus peroxidase and thr ee site-directed mutants. Implications for peroxidase substrate binding", EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 222, no. 3, 15 June 1994 (1994-06-15), pages 909 - 918 *
VEITCH, N. C. ET AL.: "The Asp245-->Asn mutant of Coprinus cinereus peroxida se. Characterization by 1H-NMR spectroscopy and comparison with the sild-typ e enzyme", BIOCHEMISTRY, vol. 35, no. 45, 12 November 1996 (1996-11-12), pages 14370 - 14380 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023225459A2 (fr) 2022-05-14 2023-11-23 Novozymes A/S Compositions et procédés de prévention, de traitement, de suppression et/ou d'élimination d'infestations et d'infections phytopathogènes

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Publication number Publication date
KR20110061297A (ko) 2011-06-09
KR101059873B1 (ko) 2011-08-29

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