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Dissertation/Thesis Abstract

Mechanistic Studies of Lysozyme Amyloid Fibril Formation and Pilot Studies of Chitinase Extraction from Chitinimonas taiwanesis
by Fulton, Alexandrya, M.S., Southern Illinois University at Edwardsville, 2020, 69; 27957011
Abstract (Summary)

I. Mechanism of Amyloid Formation of lysozyme: Lysozyme is an antibacterial enzyme that catalyzes the hydrolysis of 1,4-beta-linkages between N-acetylmuramic acid (NAM) and N-acetyl-D-glucosamine (NAG). Human variants of lysozyme are involved in fibrillogenic amyloid formation through beta-sheet aggregation which underlies a range of fatal diseases. However, the mechanism for such a formation, including kinetics and potential partial misfolded or oligomerized intermediates, is not clear. Here we studied the lysozyme amyloid formation in 50mM glycine at different pH values at 70oC and monitored the aggregation by dynamic light scattering (DSL). Differential scanning calorimetry (DSC) measurements indicates that the aggregation is correlated with the thermostability of lysozyme determined by differential scanning calorimetry (DSC). Ions and chemicals, such as Ca2+, Mg2+, and polymers, interacting with lysozyme, evidenced by fluorescence spectroscopy, either speed or inhibit amyloid formation. Here we elucidated the amyloid formation of lysozyme to its stability.

II. Isolation and Characterization of Novel Chitinase from Chitinimonas taiwanesis: Chitinase a highly conserved glycosyl hydrolase enzyme that catalyzes the hydrolysis of chitin. Chitinase has many isoforms which can vary in sequence and length of amino acid residues. The chitinase from the bacterial strain Chitimonas taiwanesis has not been characterized by its enzymatic activity and substrate specificity. Here we specifically cultured the bacterial strand using a minimized buffer. The excreted chitinase protein and the hydrolyzed product(s) of chitin were isolated for electrophoresis and thin layer chromatography.

Indexing (document details)
Advisor: Wei, Chin-Chuan
Commitee: Lu, Yun, Fowler, Thomas J.
School: Southern Illinois University at Edwardsville
Department: Chemistry
School Location: United States -- Illinois
Source: MAI 81/12(E), Masters Abstracts International
Subjects: Biophysics, Biochemistry, Molecular biology
Keywords: Amyloid, Characterization, Chitinase, Extraction, Lysozyme, Protein
Publication Number: 27957011
ISBN: 9798645497408
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