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Zivkovic DaVena PDF A APPROVED.pdf (1.66 MB)
ETD Abstract Container
Abstract Header
Enzymatic Characterization of Aldose Reductase and Its Inhibitors
Author Info
Zivkovic, DaVena
Permalink:
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1472069987
Abstract Details
Year and Degree
2016, Master of Science in Chemistry, Youngstown State University, Department of Chemistry.
Abstract
Human aldose reductase (hAR) is a NADPH-dependent oxidoreductase that reduces aldehydes to their corresponding alcohols with the oxidation of NADPH. It has been shown that two forms of this enzyme can occur. When Cysteine-298 in the active site is mutated, different kinetic variables are experienced that correlate to the form of enzyme being obtained. Mutant C298A kinetic studies showed that the Michaelis-Menten constant,
K
m
, had an 18-fold increase, the turnover number, kcat, decreased about 83%, and the catalytic efficency,
k
cat
/
K
m
, decreased about 100% for the mutant enzyme compared to the wildtype for the forward reaction using DL-glyceraldehyde as substrate and NADPH as cofactor. Mutant C298A kinetic studies showed that the Michaelis-Menten constant,
K
m
, had an 4-fold decrease, the turnover number, kcat, decreased about 60%, and the catalytic efficency,
k
cat
/
K
m
, increased about 39% for the mutant enzyme compared to the wildtype for the reverse reaction using benzyl alcohol as substrate and NADP+ as cofactor. The mutation proved beneficial for the enzyme, shown to have been the targeted native form of the enzyme. Thermal analysis of clofibric acid samples in sodium chloride mixtures of 10% (w/w) ratios were investigated by thermogravimetric analysis and mass spectroscopy. Kinetic parameters like activation energy
E
a
, pre-exponential factor
A
, and order of reaction
n
were calculated for each sample's decomposition. Trends within the kinetic parameters occurred with increase of clofibric acid percentage. Mass spectroscopy data shows a detection of CO
2
with decomposition of clofibric acid samples. This data can provide insight on the possible mechanisms in clofibric acid decomposition.
Committee
Ganesaratnam Balendiran, PhD (Advisor)
Brian Leskiw, PhD (Committee Member)
Russell Moser, PhD (Committee Member)
Pages
107 p.
Subject Headings
Analytical Chemistry
;
Biochemistry
Keywords
Aldose Reductase
;
Enzyme Kinetics
;
Thermogravimetric Analysis
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Citations
Zivkovic, D. (2016).
Enzymatic Characterization of Aldose Reductase and Its Inhibitors
[Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1472069987
APA Style (7th edition)
Zivkovic, DaVena .
Enzymatic Characterization of Aldose Reductase and Its Inhibitors .
2016. Youngstown State University, Master's thesis.
OhioLINK Electronic Theses and Dissertations Center
, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1472069987.
MLA Style (8th edition)
Zivkovic, DaVena . "Enzymatic Characterization of Aldose Reductase and Its Inhibitors ." Master's thesis, Youngstown State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1472069987
Chicago Manual of Style (17th edition)
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Document number:
ysu1472069987
Download Count:
119
Copyright Info
© 2016, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.