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035 a| (Sirsi) o1268150269;

035 a| (OCoLC)1268150269;

049 a| EREE;

090 a| QK495.C9;

100 1 a| Pahl, Matthew,e| author.?| UNAUTHORIZED;

245 10 a| Investigating the role of conserved cysteine residues in arabidopsis thaliana diacylglycerol acyltransferase 1 /c| by Matthew Pahl.;

264 1 a| [Greenville, N.C.] :b| [East Carolina University],c| 2021.;

300 a| 1 online resource (71 pages) :b| color illustrations;

336 a| textb| txt2| rdacontent;

337 a| computerb| c2| rdamedia;

338 a| online resourceb| cr2| rdacarrier;

347 a| text file;

347 b| PDF;

347 c| 1.839 MB;

538 a| System requirements: Adobe Reader.;

538 a| Mode of access: World Wide Web.;

502 b| M.S.c| East Carolina Universityd| 2021;

500 a| Presented to the faculty of the Department of Biology;

500 a| Advisor: Patrick Horn;

500 a| Title from PDF t.p. (viewed September 14, 2022).;

520 3 a| Triacylglycerols (TAGs) are biomolecules found in all organisms comprised of three fatty acid chains esterified to a glycerol backbone. In humans, TAGs are important energy storage reserves whose misregulation are associated with diseases such as obesity, diabetes, and heart disease. TAGs are also important for plant growth and development, in particular for seed lipid (or oil) reserves, as decreased TAG production is associated with poor quality seeds incapable of germination. As extractable chemical feedstocks, TAGs are a major component of biodiesel production, a green fuel source that is domestically produced and burns cleaner than typical petroleum-based fuels. Biochemically, the most common and rate-limiting final step for generating TAGs requires a diacylglycerol acyltransferase (DGAT) enzyme. This enzyme esterifies a fatty acyl-CoA to a diacylglycerol molecule. Despite its importance, the membrane-spanning topology and hydrophobicity of DGATs has prevented determination of its three-dimensional structure until recently. Plant DGAT enzymes contain cysteine-associated motifs (e.g. CXXXXC) that are reminiscent of redox-regulated switches and may be important for DGAT structure/activity (and therefore TAG accumulation). We have performed site-directed mutagenesis on several highly conserved cysteines and tested the associated lipid production levels in the model systems yeast, tobacco, and Arabidopsis thaliana. The lipid amounts and compositions produced from the mutated enzymes, along with protein amounts from western blots, were compared to the wild-type to determine how the cysteine mutations affect DGAT-catalyzed TAG production. This study will inform the strategic bioengineering of DGATs that could lead to improvements in biodiesel production, animal feed, and specialized dietary supplements produced by oleaginous plants or microorganisms.;

504 a| Includes bibliographical references.;

650 0 a| Arabidopsis.=| ^A689221;

650 0 a| Triglycerides.=| ^A11627;

650 0 a| Esters.=| ^A50633;

650 0 a| Cysteine.=| ^A1448742;

653 a| Plant;

653 a| Lipid;

653 a| Yeast;

655 7 a| Academic theses.2| fast0| (OCoLC)fst01726453;

655 7 a| Academic theses.2| lcgft;

655 7 a| Thèses et écrits académiques.2| rvmgf0| (CaQQLa)RVMGF-000001173;

655 2 a| Academic Dissertation.0| (DNLM)D019478?| UNAUTHORIZED;

700 1 a| Horn, Patrick,e| degree supervisor.?| UNAUTHORIZED;

710 2 a| East Carolina University.b| Department of Biology.=| ^A637467;

856 40 u| http://hdl.handle.net/10342/9413z| Access via ScholarShip;

949 o| wjh;

994 a| C0b| ERE;

596 a| 1 4;

998 a| 5668828;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 5668828-1001l| JNETm| JOYNERr| Ys| Yt| JNE3ETDu| 9/14/2021x| ETDz| JERESOURCE;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 5668828-2001l| HSLELECm| HSLr| Ys| Yt| HEETDu| 9/14/2021x| ETDz| HERESOURCE;

Investigating the role of conserved cysteine residues in arabidopsis thaliana diacylglycerol acyltransferase 1 / by Matthew Pahl.

Author/creator	Pahl, Matthew author.
Other author	Horn, Patrick, degree supervisor.
Other author	East Carolina University. Department of Biology.
Format	Theses and dissertations
Publication	[Greenville, N.C.] : [East Carolina University], 2021.
Description	1 online resource (71 pages) : color illustrations
Supplemental Content	Access via ScholarShip
Subjects	Arabidopsis. Triglycerides. Esters. Cysteine.

Summary	Triacylglycerols (TAGs) are biomolecules found in all organisms comprised of three fatty acid chains esterified to a glycerol backbone. In humans, TAGs are important energy storage reserves whose misregulation are associated with diseases such as obesity, diabetes, and heart disease. TAGs are also important for plant growth and development, in particular for seed lipid (or oil) reserves, as decreased TAG production is associated with poor quality seeds incapable of germination. As extractable chemical feedstocks, TAGs are a major component of biodiesel production, a green fuel source that is domestically produced and burns cleaner than typical petroleum-based fuels. Biochemically, the most common and rate-limiting final step for generating TAGs requires a diacylglycerol acyltransferase (DGAT) enzyme. This enzyme esterifies a fatty acyl-CoA to a diacylglycerol molecule. Despite its importance, the membrane-spanning topology and hydrophobicity of DGATs has prevented determination of its three-dimensional structure until recently. Plant DGAT enzymes contain cysteine-associated motifs (e.g. CXXXXC) that are reminiscent of redox-regulated switches and may be important for DGAT structure/activity (and therefore TAG accumulation). We have performed site-directed mutagenesis on several highly conserved cysteines and tested the associated lipid production levels in the model systems yeast, tobacco, and Arabidopsis thaliana. The lipid amounts and compositions produced from the mutated enzymes, along with protein amounts from western blots, were compared to the wild-type to determine how the cysteine mutations affect DGAT-catalyzed TAG production. This study will inform the strategic bioengineering of DGATs that could lead to improvements in biodiesel production, animal feed, and specialized dietary supplements produced by oleaginous plants or microorganisms.
General note	Presented to the faculty of the Department of Biology
General note	Advisor: Patrick Horn
General note	Title from PDF t.p. (viewed September 14, 2022).
Dissertation note	M.S. East Carolina University 2021
Bibliography note	Includes bibliographical references.
Technical details	System requirements: Adobe Reader.
Technical details	Mode of access: World Wide Web.
Genre/form	Academic theses.
Genre/form	Academic theses.
Genre/form	Thèses et écrits académiques.

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Investigating the role of conserved cysteine residues in arabidopsis thaliana diacylglycerol acyltransferase 1 / by Matthew Pahl.

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