Purification and biochemical characterization of beta-hexosaminidase from freshwater cnidarian Hydra vulgaris Ind-Pune

Abstract

Abstract Beta-N-acetylhexosaminidase (Hex) is a vital lysosomal hydrolase ubiquitous in living organisms, that plays a crucial role in cellular homeostasis. Dysfunctions in this enzyme are implicated in severe pathological conditions such as Tay-Sachs and Sandhoff diseases in humans. We report the purification and biochemical characterization of hexosaminidase from the soluble extracts obtained from the polyps of Hydra vulgaris Ind Pune. The Hydra Hex was purified by two-step sequential chromatography (hydrophobic interaction and gel filtration). Our results suggested that the enzyme isoform purified from Hydra is HexB, most likely to be a homodimer with a subunit mass of 65 kDa showing characteristic wobbling specificity. The pH optimum was in the range of 5.0 to 6.0 and the temperature optimum in the range of 50 ºC to 60 ºC. pH stability and temperature stability were found to be 5.0 and 40 ºC respectively. The homology modelling studies corroborated the homodimeric nature of Hydra HexB, and indicated its structural resemblance to human HexB. This first study offers significant insights into Hydra HexB, providing a foundational framework for further extensive investigations on this and other different lysosomal hydrolases in Hydra. The purified enzyme then holds promise for applications in glycomics and glycobiology research, offering new avenues for exploring the intricate roles of carbohydrates in biological systems.