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dc.contributor.authorGao, Jie
dc.contributor.otherQueen's University (Kingston, Ont.). Theses (Queen's University (Kingston, Ont.))en
dc.date2015-12-14 11:34:37.309en
dc.date2015-12-15 10:11:43.51en
dc.date.accessioned2015-12-16T15:56:40Z
dc.date.issued2015-12-16
dc.identifier.urihttp://hdl.handle.net/1974/13874
dc.descriptionThesis (Master, Chemical Engineering) -- Queen's University, 2015-12-15 10:11:43.51en
dc.description.abstractThe main goal of the thesis is to develop strategies for the fed-batch production of medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) by Pseudomonas putida KT2440 from decanoic acid (DA) co-feeding with glucose (G). Since decanoic acid is poorly water soluble and a solid at room temperature, two methods were tested to maintain it as a liquid by either mixing it with acetic acid (AA) or heating it to feed into the bioreactor. Exponentially feeding the carbon substrates at a ratio of 3:3:4 of DA:AA:G at a specific growth rate (µ) of 0.15 h-1 resulted in 26 g L-1 biomass containing 57% PHA. The final PHA composition was 74 mol% 3-hydroxydecanoate (3HD), 14 mol% 3-hydroxyoctanoate (3HO) and 12 mol% 3-hydroxyhexanoate (3HHx). As the ratio of DA:AA:G increased from 3:3:4 to 5:1:4, 50% higher biomass and 73% higher PHA were obtained. An even higher biomass concentration of 75 g L-1 containing 74% PHA was achieved by applying the same specific growth rate followed by constant substrate feeding strategy. Heating decanoic acid without acetic acid was performed as a control experiment at feeding ratio of DA:G (1:1) resulting in a final higher biomass of 41 g L-1 but containing only 58% PHA. A fadB and fadA knockout mutant of P. putida KT2440 named P. putida DBA-F1was constructed to increase the fraction of the dominant monomer (3HD) in PHA by limiting the fatty acid β-oxidation activity. Using a feeding ratio of decanoic acid and acetic acid (DAA) to glucose (G) (6:4) at specific growth rate of 0.15 h-1, significantly different monomeric composition (97% 3HD:3% 3HO) but a lower biomass (8.5 g L-1) and PHA concentrations (3.5 g L-1) were obtained compared to the wild type. The performance could be improved at a feeding ratio of DAA:G of 2:8, leading to 18 g L-1 biomass containing 100% poly(3-hydroxydecanoate) (PHD). MCL-PHA with enriched 3HD monomers could also be obtained using acrylic acid (Aa) as a chemical inhibitor of the β-oxidation pathway by P. putida KT2440. At a ratio of DAA:G:Aa (6:4:0.2), 21 g L-1 MCL-PHA with 84 mol% 3HD was produced.en_US
dc.languageenen
dc.language.isoenen_US
dc.relation.ispartofseriesCanadian thesesen
dc.rightsQueen's University's Thesis/Dissertation Non-Exclusive License for Deposit to QSpace and Library and Archives Canadaen
dc.rightsProQuest PhD and Master's Theses International Dissemination Agreementen
dc.rightsIntellectual Property Guidelines at Queen's Universityen
dc.rightsCopying and Preserving Your Thesisen
dc.rightsThis publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner.en
dc.rightsThis publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner.en
dc.subjectβ-oxidation inhibitionen_US
dc.subjectDecanoic Aciden_US
dc.subjectMCL-PHAen_US
dc.subjectPoly-hydroxydecanoateen_US
dc.subjectFed-batchen_US
dc.subjectFermentationen_US
dc.subjectMedium-chain-length Poly-3-hydroxyalkanoateen_US
dc.subjectPseudomonas putidaen_US
dc.titleProcess development for fed-batch production of medium-chain-length poly(3-hydroxyalkanoate) from decanoic aciden_US
dc.typeThesisen_US
dc.description.restricted-thesispatent applicationen
dc.description.degreeMasteren
dc.contributor.supervisorRamsay, Juliana A.en
dc.contributor.supervisorRamsay, Bruce A.en
dc.contributor.departmentChemical Engineeringen
dc.embargo.terms1825en
dc.embargo.liftdate2020-12-14


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