Identification of novel human Hyaluronidase Hyal1 inhibitors from saponine derivatives.

Orlando Z, Olschewski I, Brossette T, Hensel, Jose J

Abstract

Hyaluronan (HA) metabolism is very crucial in physiological and pathophysiological processes. It is generally accepted that its degradation to oligomeric chains influences various cellular processes resulting in inflammation, angiogenesis and metastasis [1]. Hyaluronidases are enzymes, which catalyze the degradation of HA. This makes hyaluronidases an interesting pharmaceutical target for a new kind of anti-tumor and anti-inflammation treatment. The ambition of using the model organism Escherichia coli to produce catalytically active human Hyaluronidase Hyal-1 remains a major challenge, yet. Bottlenecks in producing and studying this enzyme comprise the difficylty to avoid the intracellular formation of ‘‘inclusion bodies'' [2]. In this work, we overcome this difficulty by applying the Autodisplay technology. Thereby hHyal-1 was expressed as a part of an artificial autotransporter fusion protein on the cell surface of E. coli. Based on this system we developed a whole cell activity assay using the dye ‘‘stains-all'' [3]. We also evaluated the inhibitory effects of four saponins and 14 plant extracts on the activity of surface displayed hHyal-1.