ES2186560A1 - Method for determining chitinase activity based on usage of fluorescent substrates involves incubation of sample of enzyme with chitinase activity or of preparation containing such enzyme in presence of appropriate substrate - Google Patents

Method for determining chitinase activity based on usage of fluorescent substrates involves incubation of sample of enzyme with chitinase activity or of preparation containing such enzyme in presence of appropriate substrate

Info

Publication number
ES2186560A1
ES2186560A1 ES200101809A ES200101809A ES2186560A1 ES 2186560 A1 ES2186560 A1 ES 2186560A1 ES 200101809 A ES200101809 A ES 200101809A ES 200101809 A ES200101809 A ES 200101809A ES 2186560 A1 ES2186560 A1 ES 2186560A1
Authority
ES
Spain
Prior art keywords
enzyme
chitinase activity
usage
sample
determining
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
ES200101809A
Other languages
Spanish (es)
Other versions
ES2186560B1 (en
Inventor
Vladimir Tikhonov
Llorca Luis Vicente Lopez
Calvete Jesus Salinas
Prieto Elena Monfort
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universidad de Alicante
Original Assignee
Universidad de Alicante
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Universidad de Alicante filed Critical Universidad de Alicante
Priority to ES200101809A priority Critical patent/ES2186560B1/en
Publication of ES2186560A1 publication Critical patent/ES2186560A1/en
Application granted granted Critical
Publication of ES2186560B1 publication Critical patent/ES2186560B1/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • 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)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/42—
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Optics & Photonics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

The method for determining chitinase activity based on the usage of fluorescent substrates involves the incubation of a sample of an enzyme with chitinase activity or of a preparation containing such an enzyme in the presence of an appropriate substrate. The substrate is marked with a fluorochrome, under conditions in which are formed the products of the reaction catalyzed by the chitinase activity. The emission of fluorescence is then measured.
ES200101809A 2001-08-01 2001-08-01 METHOD FOR THE DETERMINATION OF CHITINASE ACTIVITY BASED ON THE USE OF FLUORESCENT SUBSTRATES. Expired - Fee Related ES2186560B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
ES200101809A ES2186560B1 (en) 2001-08-01 2001-08-01 METHOD FOR THE DETERMINATION OF CHITINASE ACTIVITY BASED ON THE USE OF FLUORESCENT SUBSTRATES.

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES200101809A ES2186560B1 (en) 2001-08-01 2001-08-01 METHOD FOR THE DETERMINATION OF CHITINASE ACTIVITY BASED ON THE USE OF FLUORESCENT SUBSTRATES.

Publications (2)

Publication Number Publication Date
ES2186560A1 true ES2186560A1 (en) 2003-05-01
ES2186560B1 ES2186560B1 (en) 2004-09-16

Family

ID=8498603

Family Applications (1)

Application Number Title Priority Date Filing Date
ES200101809A Expired - Fee Related ES2186560B1 (en) 2001-08-01 2001-08-01 METHOD FOR THE DETERMINATION OF CHITINASE ACTIVITY BASED ON THE USE OF FLUORESCENT SUBSTRATES.

Country Status (1)

Country Link
ES (1) ES2186560B1 (en)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5587292A (en) * 1987-05-22 1996-12-24 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Diagnosis of fungal infections with a chitinase

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5587292A (en) * 1987-05-22 1996-12-24 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Diagnosis of fungal infections with a chitinase

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
HONDA, Y. et al.: "Chemo- and enzymatic synthesis of partially and fully N-deacetylated 4-methylumbelliferyl chitobiosides: fluorogenic substrates for chitinase", Bioorganic & Medicinal Chemistry Letters, (2000), Vol. 10, pßginas 827-829, todo el documento. *
HONDA, Y. et al.: "Kinetic analysis of the reaction catalyzed by chitinase A1 from Bacillus circulans WL-12 toward the novel substrates, partially N-deacetylated 4-methylumbelliferyl chitobiosides", FEBS Lett., (2000), Vol. 476, pßginas 194-197, todo el documento. *

Also Published As

Publication number Publication date
ES2186560B1 (en) 2004-09-16

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