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CS0980

Sigma-Aldrich

Chitinase Assay Kit

sufficient for 100 multiwell tests

Synonym(s):

Chitinase Activity Detection Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.54

usage

sufficient for 100 multiwell tests

Quality Level

absorbance ratio

405 nm (Absorbance)

shipped in

wet ice

storage temp.

2-8°C

Gene Information

General description

The Chitinase Assay Kit is based on the enzymatic hydrolysis of chitinase substrates. The hydrolysis releases p-nitrophenol, which can be measured colormetrically at 405 nm.

Application

Chitinase Assay Kit has been used in screening chitinase activity of recombinant chitinase from yeast, Metschnikowia fructicola, bacterial endophyte 3A12 and Clostridium difficile spores.

The kit provides all the reagents required for efficient detection of chitinase activity in fungal and bacterial growth media, macrophage lysates, and purified enzyme preparations. In addition, the kit provides three different substrates for the detection of the various types of the chitinolytic activity:
  • 4-Nitrophenyl N,N′-diacetyl-β-D-chitobioside - substrate suitable for exochitinase activity detection (chitobiosidase activity)
  • 4-Nitrophenyl N-acetyl-β-D-glucosaminide - substrate suitable for exochitinase activity detection (β-N-acetylglucosaminidase activity)
4-Nitrophenyl β-D-N,N′,N′′-triacetylchitotriose - substrate suitable for endochitinase activity detection

Biochem/physiol Actions

Chitinase catalyzes the hydrolytic cleavage of the β-1→4-glycoside bond present in biopolymers of N-acetylglucosamine, primarily in chitin. Chitinases are widely distributed in living organisms and are found in fungi, bacteria, parasites, plants, and animals. They are classified in families based on amino acid sequence similarities.

The chitinolytic enzymes are also categorized based on their enzymatic action on chitin substrates. Endochitinases are defined as the enzymes catalyzing the random cleavage at internal points in the chitin chain. Exochitinases catalyze the progressive release of acetylchitobiose or N-acetylglucosamine from the non-reducing end of chitin, and are referred to as chitobiosidase and β-N-acetylglucosaminidase, respectively.

Chitinases perform different functions in different organisms. In bacteria, they are mainly involved in nutritional processes. In yeast and various fungi, these enzymes participate in morphogenesis. In animals and plants, chitinases primarily play a role in the defense of the organism against pathogen attack.

Analysis Note

The kit assay is based on the enzymatic hydrolysis of chitinase substrates. This hydrolysis releases p-nitrophenol (4-nitrophenol), which upon ionization in basic pH can be measured colorimetrically at 405 nm.

Kit Components Only

Product No.
Description

  • Assay Buffer 20 mL

  • 4-Nitrophenyl N-acetyl-β-D-glucosaminide 10 mg

  • 4-Nitrophenyl N,N′-diacetyl-β-D-chitobioside 5 mg

  • 4-Nitrophenyl β-D-N,N′N′′-triacetylchitotriose 1 mg

  • Chitinase from Trichoderma viride 1 mg

  • p-Nitrophenol Solution, 10 mM 1 mL

  • Sodium Carbonate 1 g

Related product

Product No.
Description
Pricing

pictograms

Health hazard

signalword

Danger

Hazard Classifications

Carc. 2 - Eye Irrit. 2 - Resp. Sens. 1 - STOT RE 2 Oral

target_organs

Liver,Kidney

Storage Class

10 - Combustible liquids


Certificates of Analysis (COA)

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Ahmed Akrem et al.
Acta crystallographica. Section F, Structural biology and crystallization communications, 67(Pt 3), 340-343 (2011-03-12)
A chitinase has been isolated and purified from Crocus vernus corms. N-terminal amino-acid sequence analysis of the approximately 30 kDa protein showed 33% identity to narbonin, a seed protein from Vicia narbonensis L. The C. vernus chitinase was crystallized by the
Functional characterization of Clostridium difficile spore coat proteins
Permpoonpattana P, et al.
Journal of Bacteriology, 195, 1492?1503-1492?1503 (2013)
E Patrick Fuerst et al.
Frontiers in plant science, 8, 2259-2259 (2018-02-08)
Seeds have well-established passive physical and chemical defense mechanisms that protect their food reserves from decay-inducing organisms and herbivores. However, there are few studies evaluating potential biochemical defenses of dormant seeds against pathogens. Caryopsis decay by the pathogenic Fusarium avenaceum
Isolation, purification, crystallization and preliminary crystallographic studies of a chitinase from Crocus vernus
Akrem,A et al.
Acta Crystallographica Section A, Foundations of Crystallography, F67, 340-343 (2011)
Ante Jerkovic et al.
Plant physiology, 152(3), 1459-1470 (2010-01-12)
Bran from bread wheat (Triticum aestivum 'Babbler') grain is composed of many outer layers of dead maternal tissues that overlie living aleurone cells. The dead cell layers function as a barrier resistant to degradation, whereas the aleurone layer is involved

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