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dc.rights.licenseRestricted to current Rensselaer faculty, staff and students. Access inquiries may be directed to the Rensselaer Libraries.
dc.contributorKoffas, Mattheos A. G.
dc.contributorDordick, Jonathan S.
dc.contributorChakrapani, Vidhya
dc.contributorBarquera, Blanca L.
dc.contributor.authorMorrison, Clifford Steven
dc.date.accessioned2021-11-03T09:18:50Z
dc.date.available2021-11-03T09:18:50Z
dc.date.created2020-08-14T12:20:48Z
dc.date.issued2020-05
dc.identifier.urihttps://hdl.handle.net/20.500.13015/2541
dc.descriptionMay 2020
dc.descriptionSchool of Engineering
dc.description.abstractTherefore, devising engineering strategies for the improved soluble expression of hRen1 is necessary for enabling the recovery of adventitious reduced NAD(P)H cofactor material from electrochemical cofactor regeneration preparations. In this work, the primary sequence of hRen1 has been mutated and fused to various solubility proteins to enhance soluble expression of the enzyme, and their steady-state kinetics parameters are characterized relative to the wild type enzyme. Additionally, engineering strategies for utilizing electrochemical bioreactors for microbial electrosynthesis applications is explored.
dc.description.abstractIndustrial enzymatic reactions that require NAD(P)H to perform redox transformations often require convoluted coupled enzyme regeneration systems to regenerate NAD(P)H from NAD(P) and recycle the cofactor for as many turnovers as possible. Renewed interest in recycling the cofactor via electrochemical means is motivated by the low cost of performing electrochemical reactions, easy monitoring of the reaction progress, and straightforward product recovery. However, direct electrochemical cofactor regeneration methods invariably produce adventitious reduced cofactor side products which results in unproductive loss of NAD(P). Human renalase isoform 1 (hRen1) has been recently identified as an enzyme that is capable of mitigating the cofactor material loss presented by electrochemical cofactor regeneration strategies. The catalytic function of the enzyme is the oxidation of biologically-inactive NAD(P)H isomers. hRen1 is selective for the NAD(P)H isomers and does not oxidize 1,4-NAD(P)H, the biologically-active form of NAD(P)H. Despite the apparent advantage of the addition of hRen1 to an engineered process involving electrochemical cofactor regeneration, recombinant hRen1 has historically been difficult to produce in E. coli, and is also generally not commercially available. hRen1 is well known to exhibit low soluble expression with aggregation into inclusion bodies which results in a total loss of catalytic function even when carefully refolded.
dc.language.isoENG
dc.publisherRensselaer Polytechnic Institute, Troy, NY
dc.relation.ispartofRensselaer Theses and Dissertations Online Collection
dc.subjectChemical engineering
dc.titleEnabling scalable electrochemical cofactor regeneration via improved soluble expression and application of mutant renalase
dc.typeElectronic thesis
dc.typeThesis
dc.digitool.pid180117
dc.digitool.pid180118
dc.digitool.pid180119
dc.rights.holderThis electronic version is a licensed copy owned by Rensselaer Polytechnic Institute, Troy, NY. Copyright of original work retained by author.
dc.description.degreePhD
dc.relation.departmentDept. of Chemical and Biological Engineering


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