Comparative characterization of bovine testicular hyaluronidase and a hyaluronate lyase from Streptococcus agalactiae in pharmaceutical preparations.
Although bovine testicular hyaluronidase (BTH) has been used in several medical fields for many years, these drugs are poorly characterized. We compared pharmaceutical BTH preparations (Neopermease, Hylase "Dessau") and a hyaluronate lyase from Streptococcus agalactiae. The BTH preparations were complex mixtures of proteins (SDS-PAGE, gel filtration) with enzymatic activity in different fractions. In the case of Neopermease the highest specific activity was found in the 58 kDa fraction (optimum at pH 3.6), whereas the 77 and 33 kDa fractions showed markedly lower specific activities at an optimal pH of 6.2. Maximum specific activity of the bacterial enzyme (approx. 1000 micromol min(-1) mg(-1)) was found at pH 5.0, being 410- and 5100-times higher compared to Neopermease and Hylase "Dessau", respectively. The hyaluronate lyase preparation was separated into two main proteins [100 kDa (pI=8.9) and 85 kDa (pI=9.2)] which were enzymatically active in SDS substrate-PAGE. Zymography after limited proteolysis of the bacterial enzyme with trypsin revealed active fragments (75-50 kDa). Our results suggest that hyaluronate lyase is an alternative for BTH, of which there has been a shortage, since companies have stopped the production of BTH preparations due to the risk of BSE.
Oettl
Martin
M
Institut für Pharmazie, Universität Regensburg, Universitätsstrasse 31, D-93040, Regensburg, Germany.
Hoechstetter
Julia
J
Asen
Iris
I
Bernhardt
Günther
G
Buschauer
Armin
A
D000818:Animals; D002417:Cattle; D006821:Hyaluronoglucosaminidase / Q000032:analysis* / Q000378:metabolism; D008297:Male; D004364:Pharmaceutical Preparations; D011133:Polysaccharide-Lyases / Q000032:analysis* / Q000378:metabolism; D013292:Streptococcus agalactiae / Q000201:enzymology*; D013737:Testis / Q000201:enzymology*
D004364:Pharmaceutical Preparations; D006821:Hyaluronoglucosaminidase; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase