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338063

Purification and Biochemical Characterization of Hyaluronidase from Egyptian Echis pyramidum pyramidum Venom

Article

Last updated: 01 Jan 2025

Subjects

-

Tags

Biochemistry

Abstract

A novel hyaluronidase was purified to apparent homogeneity from Egyptian Echis pyramidum pyramidum venom. The enzyme was purified 7.6-fold by Sephacryl S-200 size exclusion followed by CM-Sepharose cation-exchange chromatography with a specific activity of 610 U/mg over the activity of whole venom (80 U/mg). The molecular mass of the purified enzyme was approximately 48 kDa in its native and reduced forms. The maximum activity was estimated at pH 5.5 and 40 oC. The purified hyaluronidase has an absolute activity towards hyaluronic acid with Km and Vmax values of 20µg/ml and 40µg/min, respectively. The Na+ ion enhanced the hyaluronidase activity, whereas Fe+2, Ni+2, Cu+2, Co+2, Zn+2, Mn+2, o-phenanthroline (5 mM) and heparin (10 units) inhibited the enzyme activity by 100%. Antisera against the venoms of Echis pyramidum, Echis coloratus, Cerastes cerastes, Cerastes vipera and Naja haje greatly neutralized E. pyramidum hyaluronidase to a varied extent. In contrast, antisera against Naja nigricollis negatively neutralized the hyaluronidase activity. The study concluded that the purified enzyme shared similar immunogenic properties with venoms of vipers rather than elapids. It also demonstrated the importance of characterization of enzymes from venoms of different species to explore the venomics and antivenomics of Egyptian snake species.

DOI

10.21608/ejchem.2024.257468.9028

Keywords

Snake venom, Enzymes, inhibition, Hyaluronic acid, Neutralization, Antigenic, epitope

Authors

First Name

Walaa

Last Name

Salama

MiddleName

Hussein

Affiliation

Molecular Biology, Biotechnology Research institute, NRC, Egypt

Email

walaahsalama82@gmail.com

City

-

Orcid

0000-0003-1423-5305

Volume

67

Article Issue

13

Related Issue

46555

Issue Date

2024-12-01

Receive Date

2023-12-23

Publish Date

2024-12-01

Page Start

251

Page End

258

Print ISSN

0449-2285

Online ISSN

2357-0245

Link

https://ejchem.journals.ekb.eg/article_338063.html

Detail API

https://ejchem.journals.ekb.eg/service?article_code=338063

Order

338,063

Type

Original Article

Type Code

297

Publication Type

Journal

Publication Title

Egyptian Journal of Chemistry

Publication Link

https://ejchem.journals.ekb.eg/

MainTitle

Purification and Biochemical Characterization of Hyaluronidase from Egyptian Echis pyramidum pyramidum Venom

Details

Type

Article

Created At

30 Dec 2024