Preparation and purification of Flavobacterium heparinum chondroitinases AC and B by hydrophobic interaction chromatography

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Preparation and purification of Flavobacterium heparinum chondroitinases AC and B by hydrophobic interaction chromatography.

Flavobacterium heparinum is a soil bacterium that produces several mucopolysaccharidases such as heparinase, heparitinases I and II, and chondroitinases AC, B, C and ABC. The purpose of the present study was to optimize the preparation of F. heparinum chondroitinases, which are very useful tools for the identification and structural characterization of chondroitin and dermatan sulfates. We obse...

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Treatment of bovine nasal cartilage proteoglycan with chondroitinases from Flavobacterium heparinum and Proteus vulgaris.

A calorimetric procedure was developed for assaying the enzyme activity of chondroitinase AC from Flavobacterium heparinum and of chondroitinase ABC from Proteus vulgaris when chondroitin 4-sulfate is used as a substrate. The enzymatic digestion product, 2-acetamido-2-deoxy-3-0-(/3-Dgluco-4-enepyranosyluronic acid)-4-0-s&o-D-galactose, is oxidized with periodic acid to yield, among other produc...

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Heparinase production by Flavobacterium heparinum.

Heparinase production by Flavobacterium heparinum in complex protein digest medium, with heparin employed as the inducer, has been studied and improved. The maximum productivity of heparinase has been increased 156-fold over that achieved by previously published methods to 375 U/liter per h in the complex medium. Rapid deactivation of heparinase activity, both specific and total, was observed a...

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Purification of Plasmid (pVaxLacZ) by Hydrophobic Interaction Chromatography

This paper describes a method for the plasmid DNA purification, which includes an ammonium sulphate precipitation, followed by hydrophobic interaction chromatography (HIC) using Phenyl Sepharose 6 Fast Flow (low sub). The use of HIC took advantage of the more hydrophobic character of single stranded nucleic acid impurities as compared with double-stranded plasmid DNA.

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Purification of Mycobacterium bovis BCG Tokyo antigens by chromatofocusing, lectin-affinity chromatography, and hydrophobic interaction chromatography.

A combination of chromatofocusing, lectin-affinity chromatography, and hydrophobic interaction chromatography resulted in a simple purification of protein antigens of Mycobacterium bovis BCG Tokyo culture filtrate. Identification was established on the basis of chromatographic separation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis determination of molecular weights, and N-termina...

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ژورنال

عنوان ژورنال: Brazilian Journal of Medical and Biological Research

سال: 1999

ISSN: 0100-879X

DOI: 10.1590/s0100-879x1999000500007