Immobilization alters heparin cleaving properties of heparinase I

Abstract

We report here a novel observation that immobilization of heparinase I on CNBr-activated Sepharose results in heparin degradation properties that are different from heparinase I in the free solution form. Studies over a range of pHs (5-8) and temperatures (5-50°C) as well as under batch and flow conditions show that immobilized heparinase 1 displays altered pH and temperature optima, and a higher propensity for generation of longer chains (hexa- and octa-) with variable sulfation as compared to that in the free form, which is known to yield disaccharides. The immobilized enzyme retained good eliminase activity over at least five cycles of reuse. In combination, results suggest that heparinase I immobilization may offer a more productive route to longer, variably sulfated sequences.

Polymeric fluorescent heparin as one-step FRET substrate of human heparanase

“…Hexylamine (15 mM) was used as an ion-pairing agent and hexafluoroisopropanol (100 mM) was used as an organic modifier in both solvent systems. The ESI-MS peaks were acquired in the positive ionization mode with a capillary voltage of 3.2 kV, extractor voltage of 1 V, cone voltage between 20 and 180 V, the radio frequency of 0.1 V, source temperature of 150 °C, and desolvation temperature of 350 °C (Bhushan, Alabbas, Kuberan, Gupta, & Desai, 2017).…”