Characterization and Immobilization of A Novel Hyaluronidase Produced By Streptomyces roseofulvus

Abstract

M AXIMUM hyaluronidase production by Streptomyces roseofulvus S10 (LC314796) was attained when it was cultured in submerged fermentation process under favorable conditions, pH 5 at 40ºC for 6 days. Hyaluronidase was purified to its homogeneity by 9.2 fold with molecular weight of 97kDa under denaturing SDS-PAGE. Mg +2 exerted highly stimulatory effect on S. roseofulvus S10 hyaluronidase activity and was significantly reduced in presence of Mn +2 , Zn +2 , and EDTA. Optimum reaction was attained at pH 9 and the pH stability of enzyme ranged between 9-10 at 35°C. To protect the intrinsic activity and halftime of hyaluronidase, several carriers and immobilization of hyaluronidase were investigated. The immobilized enzyme had higher thermal stability than free one with T m values; 46.1°C and 24.7°C, respectively. Maximum affinity of free and immobilized hyaluronidase was for hyaluronic acid followed by bovine albumin. Free enzyme had a high catalytic affinity of hyaluronic acid compared with immobilized enzyme. Our results demonstrated that S. roseofulvus S10 hyaluronidase was highly stable to pH and high temperature. These properties of long-term stability facilitate its wide range of applications.

Continuous Ethanol Production from Molasses via Immobilized Saccharomyces cerevisiae on Different Carriers on Pilot Scale

“…It well known that alginate present as gel beads in the medium but during fermentation carbon dioxide which produced by yeast cells caused the formation of cracks and capillaries in the alginate hydrogel, thus reducing the diffusion limit and so the transport of nutrients and metabolites is facilitated through the created structures using capillary forces (Poreda et al, 2011). In contrast Reda & Shanawany (2020) revealed that the maximum hyaluronidase enzyme immobilization yield (67.2%) was observed with Ca-alginate followed by agar-agar and Silica gel respectively. inoculum size of S. cerevisiae immobilized on wheat straw in continuous fermentation culture using a mixture of Egyptian cane and beet molasses with 16% initial sugar at 30 °C and pH4.5.…”

Purification and characterization of the produced hyaluronidase by Brucella Intermedia MEFS for antioxidant and anticancer applications

“…In order to purify the hyaluronidase from crude filtrate, the enzyme filtrate was firstly precipitated using 70% saturation ammonium sulfate solution as recommended by Reda & El-Shanawany [ 16 ]. The protein precipitate was dissolved using a 50 mM phosphate buffer (pH 7) and dialyzed three times using the same buffer.…”

Prospective bacterial and fungal sources of hyaluronic acid: A review

“…The co-expression of udgA and hasA genes from S. zooepidemicus results in a sharp decrease in the amount of product [143] . Unlike the hyaluronidase from streptococci, the activity of the hyaluronidase from Streptomyces roseofulvus S10 (LC314796) rises markedly in the presence of Mg 2+ , but decreases in the presence of Mn 2+ and Zn 2+ ions [290] .…”