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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

Signal Word

Danger

Hazard Statements

Hazard Classifications

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

Target Organs

Liver,Kidney

Storage Class Code

10 - Combustible liquids


Certificates of Analysis (COA)

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Postharvest application of a novel chitinase cloned from Metschnikowia fructicola and overexpressed in Pichia pastoris to control brown rot of peaches
Banani H, et al.
International Journal of Food Microbiology, 199, 54-61 (2015)
Flávio Augusto Cardozo et al.
World journal of microbiology & biotechnology, 35(8), 114-114 (2019-07-25)
N-acetyl-D-glucosamine (GlcNAc) is an important amino-monosaccharide with great potential for biotechnological applications. It has traditionally been produced by the chemical hydrolysis of chitin, despite certain industrial and environmental drawbacks, including acidic wastes, low yields and high costs. Therefore, enzymatic production
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
Patima Permpoonpattana et al.
Journal of bacteriology, 193(23), 6461-6470 (2011-09-29)
Clostridium difficile is an important human pathogen and one where the primary cause of disease is due to the transmission of spores. We have investigated the proteins found in the outer coat layers of C. difficile spores of pathogenic strain
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)

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