Advisory Committee Chair
Champion Deivanayagam
Document Type
Thesis
Date of Award
2024
Degree Name by School
Master of Science in Biomedical Engineering (MSBME) School of Engineering
Abstract
The Hyaluronate Lyase derived from Group A streptococcal bacteriophage H4489A gene (HylP) is known for cleaving the glycosidic bonds of high molecular weight hyaluronic acid (HMWHA), a substance often overexpressed in certain types of cancer cells. This enzymatic action produces HA oligosaccharides. Previous research, conducted by our team, has demonstrated that these oligosaccharides play a significant role in reducing the invasion, migration, and proliferation of breast cancer cells. This effect occurs through the disruption of Erk1/2 activation and RhoA expression. This bacteriophage hyaluronate lyase (HylP), a 40kDa protein, possesses a unique collagen-like sequence, consisting of 30 amino acids in a Gly-X-Y motif. This motif is absent in HylP1 and HylP2, isoforms of bacteriophage hyaluronate lyase. Interestingly, even after removing this collagen-like motif from HylP1 and HylP2, there was no substantial change in hyaluronidase activity. Consequently, the precise activity of HylP remains enigmatic. To reveal this mystery, our study aimed to identify the active sites by generating triple mutations on the conserved amino acids that form the enzymatic active site of all hyaluronate lyases. This study assessed the wild type and mutated enzyme's activity through various assays and evaluated its enzymatic behavior using Michaelis-Menten kinetics. Furthermore, through kinetics analysis we have established the optimum conditions for hyaluronidase activity under the factors influencing pH, temperature, and substrate concentrations. Future investigation will examine the hyaluronidase activity specifically on breast cancer cells like MCF-7, HS-578T and MD-MBA 231, allowing for a comprehensive understanding of its potential therapeutic implications.
Recommended Citation
Rawal, Urvi Prakash, "Comparative Enzymatic Analysis Of Different Isoforms Of Group A Streptococcal Bacteriophage H4489A Hyaluronidase" (2024). All ETDs from UAB. 3891.
https://digitalcommons.library.uab.edu/etd-collection/3891