Chondroitin lyase enzymes

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/60 (2006.01) A61K 38/51 (2006.01) C12N 9/88 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2194375

The present invention describes a method for the production of two highly purified enzymes capable of degrading chondroitin sulfate polysaccharides. A multi-step purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography, hydroxylapatite chromatography, high resolution ion exchange chromatography and size exclusion is outlined. A 77,000 ~ 5,000 Dalton protein capable of degrading chondroitin sulfates A and C and a 55,000 ~ 2,300 Dalton protein capable of degrading dermatan sulfate were isolated. The genes encoding these enzymes, chondroitinase AC and chondroitinase B, respectively, have been cloned and methods for their use are described.

L'invention concerne un procédé de production de deux enzymes hautement purifiées, capables de dégrader des polysaccharides de chondroïtine-sulfate. Elle concerne également un procédé de purification comportant des étapes multiples, y compris la rupture de cellules, la chromatographie échangeuse de cations, la chromatographie d'affinité, la chromatographie d'hydroxylapatites, la chromatographie échangeuse d'ions à haute résolution et exclusion de tailles. On a isolé une protéine de 77.000 <u>+ </u>5.000 Daltons capable de dégrader des chondroïtines-sulfates A et C, ainsi qu'une protéine de 55.000 <u>+</u> 2.300 Daltons capable de dégrader du sulfate de dermatan. On a cloné les gènes codant lesdites enzymes, respectivement, chondroitinase AC et chondroitinase B et l'invention concerne également leurs procédés d'utilisation.

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