Type
Text
Type
Thesis
Advisor
Sampson, Nicole S
Date
2016-12-01
Keywords
Chemistry
Department
Department of Chemistry
Language
en_US
Source
This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.
Identifier
http://hdl.handle.net/11401/77029
Publisher
The Graduate School, Stony Brook University: Stony Brook, NY.
Format
application/pdf
Abstract
Mycobacterium tuberculosis (Mtb) is the bacterium that causes tuberculosis (TB) in humans. TB is one of the deadliest infectious diseases in the world. Mtb resides in the lipid- enriched environment of the host and metabolizes cholesterol as a carbon source using its own enzymes. Among the total 4000 genes of Mtb, 250 genes correlate to the enzymatic reactions of lipid utilization. The degradation of cholesterol side chain is characterized by three cycles of β-oxidation. Each of the β-oxidation cycles involves hydration of the dehydrogenated cholesterol intermediate substrate. Rv3538 and EchA19 are expected to function as an enoyl-CoA hydratase in the cholesterol metabolism of Mtb. In this project, the hydratase candidates Rv3538 and EchA19 were expressed and tested with the dehydrogenated cholesterol intermediates of the β-oxidation cycles. Rv3538, echA19, and chsH1-chsH2 are controlled by same regulator of KstR1. The enoyl-CoA hydratase candidates of Rv3538, EchA19, and ChsH1-ChsH2 are predicted to be involved in similar enzymatic reactions. The enoyl-CoA hydratase, ChsH1-ChsH2, was previously identified to hydrate the cholesterol intermediate of the 3rd β-oxidation cycle. The substrate 3-oxo-chol-4-en-24-oyl-CoA (3-OCO-CoA) is involved in the 2nd β-oxidation cycle. The target protein of Rv3538 was investigated for the hydration of the 2nd β-oxidation. The 1st β- oxidation cycle contains 3-oxo-cholest-4-en-26-oyl-CoA (3-OCS-CoA) as a substrate. The enzymatic reaction of EchA19 and dehydrogenated 3-OCS-CoA was monitored using MALDI TOF mass spectroscopy. EchA13, fadE17, and fadE18 are members of the Mce3R regulon. The substrate of EchA13 is predicted to be a product of the enzymatic reaction of FadE17 or FadE18. The investigation about EchA13 would give valuable information to identify FadE17 and FadE18 in the future. EchA13 is expected to function as an enoyl-CoA hydratase for Mtb. EchA13 was expressed and tested with the substrate of C12-enoyl-CoA. The enzymatic reaction was monitored by MALDI TOF mass spectroscopy. | Mycobacterium tuberculosis (Mtb) is the bacterium that causes tuberculosis (TB) in humans. TB is one of the deadliest infectious diseases in the world. Mtb resides in the lipid- enriched environment of the host and metabolizes cholesterol as a carbon source using its own enzymes. Among the total 4000 genes of Mtb, 250 genes correlate to the enzymatic reactions of lipid utilization. The degradation of cholesterol side chain is characterized by three cycles of β-oxidation. Each of the β-oxidation cycles involves hydration of the dehydrogenated cholesterol intermediate substrate. Rv3538 and EchA19 are expected to function as an enoyl-CoA hydratase in the cholesterol metabolism of Mtb. In this project, the hydratase candidates Rv3538 and EchA19 were expressed and tested with the dehydrogenated cholesterol intermediates of the β-oxidation cycles. Rv3538, echA19, and chsH1-chsH2 are controlled by same regulator of KstR1. The enoyl-CoA hydratase candidates of Rv3538, EchA19, and ChsH1-ChsH2 are predicted to be involved in similar enzymatic reactions. The enoyl-CoA hydratase, ChsH1-ChsH2, was previously identified to hydrate the cholesterol intermediate of the 3rd β-oxidation cycle. The substrate 3-oxo-chol-4-en-24-oyl-CoA (3-OCO-CoA) is involved in the 2nd β-oxidation cycle. The target protein of Rv3538 was investigated for the hydration of the 2nd β-oxidation. The 1st β- oxidation cycle contains 3-oxo-cholest-4-en-26-oyl-CoA (3-OCS-CoA) as a substrate. The enzymatic reaction of EchA19 and dehydrogenated 3-OCS-CoA was monitored using MALDI TOF mass spectroscopy. EchA13, fadE17, and fadE18 are members of the Mce3R regulon. The substrate of EchA13 is predicted to be a product of the enzymatic reaction of FadE17 or FadE18. The investigation about EchA13 would give valuable information to identify FadE17 and FadE18 in the future. EchA13 is expected to function as an enoyl-CoA hydratase for Mtb. EchA13 was expressed and tested with the substrate of C12-enoyl-CoA. The enzymatic reaction was monitored by MALDI TOF mass spectroscopy. | 77 pages
Recommended Citation
Jang, Jungwon, "The investigation of enoyl-CoA hydratases in cholesterol metabolism of Mycobacterium tuberculosis" (2016). Stony Brook Theses and Dissertations Collection, 2006-2020 (closed to submissions). 2883.
https://commons.library.stonybrook.edu/stony-brook-theses-and-dissertations-collection/2883