A new affinity method for purification of bovine testicular hyaluronidase enzyme and an investigation of the effects of some compounds on this enzyme


KAYA M. O. , ARSLAN O., ÖZENSOY GÜLER Ö.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY, vol.30, no.4, pp.524-527, 2015 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 30 Issue: 4
  • Publication Date: 2015
  • Doi Number: 10.3109/14756366.2014.949253
  • Title of Journal : JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
  • Page Numbers: pp.524-527

Abstract

In this study, a new affinity gel for the purification of bovine testicular hyaluronidase (BTH) was synthesized. L-Tyrosine was added as the extension arm to the Sepharose-4B activated with cyanogen bromide. m-Anisidine is a specific inhibitor of BTH enzyme. m-Anisidine was clamped to the newly formed Sepharose-4B-L-tyrosine as a ligand. As a result, an affinity gel having the chemical structure of Sepharose-4B-L-tyrosine-m-anisidine was obtained. BTH purified by ammonium sulfate precipitation and affinity chromatography was obtained with a 16.95% yield and 881.78 degree of purity. The kinetic constants K-M and V-Max for BTH were determined by using hyaluronic acid as a substrate. K-M and V-Max values obtained from the Lineweaver-Burk graph were found to be 2.23mM and 19.85 U/mL, respectively. In vitro effects of some chemicals were determined on purified BTH enzyme. Some chemically active ingredients were 1,1-dimethyl piperidinium chloride, beta-naphthoxyacetic acid and gibberellic acid. Gibberellic acid showed the best inhibition effect on BTH.