Properties of Chitinase from <i>Vibrio alginolyticus</i>, as Assayed with the Chromogenic Substrate 3,4-Dinitrophenyl Tetra-<i>N</i>-acetylchitotetraoside

Aribisala, O. A.; Gooday, Graham W.

doi:10.1042/bst0060568

Cited by 27 publications

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“…In the former category, examples include measurement of release of reducing sugars or N-acetylglucosamine (requiring N-acetylglucosaminidase together with chitinase) (Ulhoa and Peberdy 1991;Vasseur et al 1990); the use of [3H]_ or [14C)-chitin (Molano et al 1977;Cabib 1988;Rast et al 1991); visco metric measurements of soluble chitin derivatives (Ohtakara 1988;Lindsay and Gooday 1985b); and release of soluble dye-labelled products from dyed chitin derivatives (Wirth and Wolf 1990;Evrall et al 1990). In the latter category, chromogenic soluble model substrates have provided the basis for useful assays, notably 3,4-dinitrophenyl tetra-N-acetyl-f3 chitotetraose (Aribisala and Gooday 1978;Rast et al 1991). More versatile, however, are assays following the hydrolysis of glycosides of the fluorophore, 4-methylumbelliferone.…”

Section: Identification and Assay Of Chitinolytic Activitiesmentioning

confidence: 99%

Physiology of microbial degradation of chitin and chitosan

Gooday

1994

Biochemistry of Microbial Degradation

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101

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Chi tin, the (1-4 )-J3-linked homopolymer of N-acetyl-D-glucosamine (Fig. 1), is produced in enormous amounts in the biosphere. A recent working estimate for both annual production and steady-state amount is of the order of lO lD to 1011 tons (Gooday 1990a). Chitin is utilized as a structural component by most species alive today. Its phylogenetic distribution is clearly defined: (a) Prokaryotes. Despite its chemical similarity to the polysaccharide backbone of peptidoglycan, chitin has only been reported as a possible component of streptomycete spores and of the stalks of some prosthecate bacteria. (b) Protista. Chitin provides the tough structural material for many protists; in cyst walls of some ciliates and amoebae; in the lorica walls of some ciliates and chrysophyte algae; in the flotation spines of centric diatoms; and in the walls of some chlorophyte algae and oomycete fungi (Gooday 1990a). (c) Fungi. Chitin appears to be ubiquitous in the fungi (Bartnicki-Garcia and Lippman 1982). Reported exceptions, such as Schizosaccharomyces, prove to have small but essential amounts of chitin. Pneumocystis carinii, of uncertain affinity, has chitin in the walls of its cysts and trophozoites (Walker et a1. 1990). (d) Animals. Chitin is the characteristic tough material playing a range of structural roles among most invertebrates (Jeuniaux 1963(Jeuniaux , 1982. It is absent from vertebrates. (e) Plants. Chitin sensu stricto is probably absent from plants, but polymers rich in (1-4)-J3-linked N-acetylglucosamine have been reported (Benhamou and Asselin 1989). Chitin occurs in a wide variety of manners. Three hydrogen-bonded crystalline forms have been characterised: a-chitin with antiparallel chains, 13chitin with parallel chains and )I-chitin with a three-chain unit cell, two "up"one "down" (Blackwell 1988). a-Chitin is by far the most common, being the form found in fungi and most protistan and invertebrate exoskeletons. The importance of physical form to biological function is indicated by squid, Loligo,

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Section: Identification and Assay Of Chitinolytic Activitiesmentioning

confidence: 99%

Physiology of microbial degradation of chitin and chitosan

Gooday

1994

Biochemistry of Microbial Degradation

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101

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“…The K,,, value was determined by the Lineweaver-Burke method using colloidal chitin as substrate. The substrate specificity of the purified enzyme was tested against various substances : carboxymethylchitin (0.6 mg ml-') and chitosan (0.6 mg ml-I), obtained as described by Reyes et al (1989), chitin from P. oxalicum cell wall (1 mg ml-') (Copa -Patiiio et al 1990b) and 3,4dinitrophenyl-tetra-Nacetyl-/3-mhitotetraoside (0.5 mg ml -I) (Koch-Light) as described by Aribisala and Gooday (1978).…”

Section: Enzyme Preparation and Purificationmentioning

confidence: 99%

Purification and properties of a chitinase from Penicillium oxalicum autolysates

Rodríguez

Copa-Patiño

Pérez-Leblic

1995

Lett Appl Microbiol

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A chitinase (EC. 3.2.1.14) from autolysed culture filtrate of Penicillium oxalicum was purified by precipitation with ammonium sulphate, gel filtration and ion exchange chromatographies. The purified enzyme showed a single protein band in SDS gel electrophoresis. The enzyme is an acidic protein with a pI of 4.5 and has a molecular weight of 54,900 as estimated from SDS gel electrophoresis and 21,500 from gel filtration. The optimum pH and temperature were 5.0 and 35 degrees C, respectively. The enzyme was stable at temperatures up to 45 degrees C and in a pH range between 4.0 and 6.0. The Km was 2.5 mg ml-1 for colloidal chitin, Hg2+ and Ag+ were effective inhibitors. The viscosimetric study carried out using carboxymethyl chitin as substrate revealed the endotype action of this enzyme.

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“…The soluble chitinase substrate 3,4-dinitrophenyl-N-acetylchitotetraoside consists of an N-acetylglucosamine (GlcNAc) tetramer linked 0-( 1-4) to 3,4-dinitrophenol. It is specific for chitinase and allows simple, rapid and accurate determinations (Aribisala & Gooday, 1978). Assays were carried out by adding 0.1 ml of a 0.5mgml-' aqueous solution of DNP(GlcNAc), to 0.1 ml citric acid: NaOH (Ssrensen) buffer and equilibrating to 37" C in a semi-micro cuvette; then, to this, 0.1 ml of test solution was added and the absorbance at 400 nm of the reaction mixture monitored in a spectrophotometer connected to a chart recorder for 3-5 min at 37" C and the activity quantified by reference to molar co-efficient data for 3,4-dinitrophenol.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Seasonal activities of chitinase and chitobiase in the digestive tract and serum of cod, Gadus morhua (L.)

Lindsay

1987

Journal of Fish Biology

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No statistically significant seasonal variations of chitinase and chitobiase in cod, Gadus morhua, stomach, intestine or serum could be detected. Enzyme activity often varied considerably (up to six-fold) between fish captured at the same time, but no correlation could be detected between the presence of chitin or the amount of prey in the stomach and the activities of chitinase and chitobiase. Lysozyme was absent from all the tissues tested.

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Properties of Chitinase from Vibrio alginolyticus, as Assayed with the Chromogenic Substrate 3,4-Dinitrophenyl Tetra-N-acetylchitotetraoside

Cited by 27 publications

References 0 publications

Physiology of microbial degradation of chitin and chitosan

Physiology of microbial degradation of chitin and chitosan

Purification and properties of a chitinase from Penicillium oxalicum autolysates

Seasonal activities of chitinase and chitobiase in the digestive tract and serum of cod, Gadus morhua (L.)

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