Date of Graduation


Document Type


Degree Name

Bachelor of Science

Degree Level



Chemistry & Biochemistry


Fan, Chenguang

Committee Member/Reader

Ivey, Mack

Committee Member/Second Reader

Levine, Bill

Committee Member/Third Reader

Zheng, Nan


The contents of this thesis have been modified from the publication “Araujo J, Ottinger S, Venkat S, Gan Q and Fan C (2022) Studying Acetylation of Aconitase Isozymes by Genetic Code Expansion. Front. Chem. 10:862483”. Though studies have found multiple lysine sites in which acetylation takes place in Escherichia Coli aconitase, acetylation’s effects on the enzyme’s activity have yet to be studied. Aconitase is the dehydratase-hydratase found in the citric acid and glyoxylate cycles responsible for the reversible isomerization of citrate to isocitrate via cis-aconitate intermediate. There are two isoforms of aconitase in E. coli: AcnA and AcnB. In this study, the genetic code expansion technique was utilized to generate 14 different site-specifically acetylated aconitases, with both isozymes being represented. Following enzyme assays and kinetic analysis of the variants, it was found that the acetylation of two altered aconitases impacted the level of enzyme kinetics from that of their wild-type counterparts, with AcnA K684 decreasing enzyme activity and AcnB K567 increasing enzyme activity.


lysine, acetylation, aconitase, E. coli, genetic code expansion, enzyme kinetics