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Chondroitinase AC - Research Grade PN 50-013

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Chondroitinase AC - Research Grade Datasheet

Synonyms

Chondroitin sulfate lyase; chondroitin sulfate eliminase

 

Source

Flavobacterium heparinum (recombinant)

 

EC Number

CAS Number

4.2.2.5

9047-57-8

Catalyzed Reaction

The enzyme cleaves, via an elimination mechanism, sulfated and non-sulfated polysaccharide chains containing 1-4 linkages between hexosamines and glucuronic acid residues.The reaction yields oligosaccharide products (mainly disaccharides) containing unsaturated uronic acids which can be detected by UV spectroscopy at 232 nm. The enzyme is active on chondroitin sulfates A and C, chondroitin and hyaluronic acid, but is not active on dermatan sulfate (chondroitin sulfate B).

 

Substrate Specificity

Chondroitin sulfates A and C, chondroitin, hyaluronic acid. (The specific activity with chondroitin sulfate A is approx. 1.5 times higher than the specific activity with chondroitin sulfate C).

 

Properties

  • Molecular weight: 79,557 Da
  • Isoelectric point: 9.0 – 9.1
  • pH optimum for activity: 4.5 – 6 with chondroitin sulfate A , and 6 – 7 with chondroitin sulfate C
  • pH range for activity: 3.5 – 9 with chondroitin sulfate A, and 4.5 to 9 with chondroitin sulfate C
  • Optima testingl temperature range: 20 ° C – 37 ° C
  • Optimal storage temperature: –70 ° C
  • Crystal structure has been determined and published (see references).

Purity

≥ 90 % by reversed phase HPLC analysis.

 

Chondroitinase AC Datasheet graphic

Specific Activity

≥ 200 IU/mg (substrate: chondroitin sulfate A)

One international unit (IU) is defined as the amount of enzyme that will liberate 1.0 µmole unsaturated oligosaccharides from chondroitin sulfates A and C and hyaluronic acid per minute at 30 ° C and pH 8.0.

 

Stability

  • PN 50-013 (vial of 0.5 IU): Expiration is 30 months from manufacturing date, frozen at – 70 ° C in aqueous buffers containing Sodium Chloride, Sodium Phosphate and Sucrose 5%.

Applications

  • As research reagent (glycosaminoglycan degradation).
  • For the preparation of di- and oligo-saccharides of chondroitin sulfates and the preparation of oligosaccharide libraries.
  • Degradation of hyaluronic acid.

 

Availability

A proprietary expression system for F. heparinum and the fermentation and isolation processes developed by IBEX Pharmaceuticals allow the production of large quantities of high purity product.

 

References

  • Review: "Enzymatic Degradation of Glycosaminoglycans". S. Ernst et al. in Critical Reviews in Biochemistry and Molecular Biology (1995), 30(5): 387-444.·
  • "Isolation and Expression in Escherichia coli of cslA and cslB, Genes Coding for the Chondroitin Sulfate-Degrading Enzymes Chondroitinase AC and Chondroitinase B, Respectively, from Flavobacterium heparinum". A.L. Tkalec, D. Fink, F. Blain, G. Zhang-Sun, M. Laliberté, D.C. Bennett, K. Gu, J.J.F. Zimmermann and H. Su, in Applied and Environmental Microbiology (2000) 66(1): 29-35.·
  • "Purification, Characterization and Specificity of Chondroitin Lyases and Glycuronidase from Flavobacterium heparinum". K. Gu, R.J. Linhardt, M. Laliberté, K. Gu and J. Zimmermann, in Biochem. J. (1995) 312: 569-577.·
  • "A comparative Study Between a Chondroitinase B and a Chondroitinase AC from Flavobacterium heparinum". M.Y.M. Michelacci and D.C.P. Dietrich, in Biochemical Journal (1975) 151: 121-129.·
  • "Crystal Structure of Chondroitin AC Lyase, a Representative of a Family of Glycosaminoglycan Degrading Enzymes". J. Féthière, B. Eggimann and M. Cygler, in J. Mol. Biol. (1999) 288: 635-647.