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Thiamin diphosphate-dependent enzymes and their carboligation activity

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TitleInfo
Title
Thiamin diphosphate-dependent enzymes and their carboligation activity
Name (type = personal)
NamePart (type = family)
Christian
NamePart (type = given)
Abhilash S.
NamePart (type = date)
1992-
DisplayForm
Abhilash S. Christian
Role
RoleTerm (authority = RULIB)
author
Name (type = personal)
NamePart (type = family)
Jordan
NamePart (type = given)
Frank
DisplayForm
Frank Jordan
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
chair
Name (type = corporate)
NamePart
Rutgers University
Role
RoleTerm (authority = RULIB)
degree grantor
Name (type = corporate)
NamePart
Graduate School - Newark
Role
RoleTerm (authority = RULIB)
school
TypeOfResource
Text
Genre (authority = marcgt)
theses
OriginInfo
DateCreated (qualifier = exact)
2018
DateOther (qualifier = exact); (type = degree)
2018-05
CopyrightDate (encoding = w3cdtf); (qualifier = exact)
2018
Place
PlaceTerm (type = code)
xx
Language
LanguageTerm (authority = ISO639-2b); (type = code)
eng
Abstract (type = abstract)
Several enzymes use more than just energy to perform catalysis. Reactions requiring harsh conditions and energy requirement that exceed the cell’s overall energy output are made possible by enzymes. Several of these enzymes are known for using cofactors in their catalysis. One such class of enzyme known as, “thiamin diphosphate dependent enzyme” uses thiamin diphosphate (ThDP) an active form of vitamin B1 to carry out reactions that include cleavage of carbon-carbon, carbon-sulfur, carbon-oxygen and carbon-nitrogen bonds. The understanding of its mechanism has opened a new spectrum of green chemistry involving manipulation of enzymes to give side reactions which create chemical bonds instead of breaking them. Herein, two different ThDP-dependent enzymes and their carboligation reactivity was studied. E1o which is a component of the oxoglutarate dehydrogenase multienzyme complex (OGDHc) and MenD, an enzyme involved in the biosynthesis reaction of vitamin K2. MenD and E1o’s ability to carry out carboligation reaction of bulky aromatic and short aliphatic acceptors with substrate was studied using circular dichroism (CD). The observations using CD were made possible due to an inherent property of the activated nucleophile, “prochirality.” The reaction between the prochiral nucleophile and electrophile led to the formation of chiral products which in turn were observed. To carry out these reactions the plasmid carrying enzyme genes were first over-expressed in E. coli cells and purified using IMAC chromatography (immobilized metal affinity chromatography). The enzymatic reactions were carried out using 2-oxoglutarate (substrate), benzaldehyde (acceptor) and propanal (acceptor) giving their respective products 5-hydroxy-4-oxo-5-phenylpentanoic acid and 5-hydroxy-4-oxo-heptanoic acid. The reactions were also performed using a different 2-oxo-acid (substrate), 2-oxo-5-hexenoic acid with benzaldehyde (acceptor) and propanal (acceptor). The results showed, MenD’s activity was limited due to its mechanism at high substrate levels, leading to little to no product formation attributed to its ping-pong bi-bi mechanism. Moreover, MenD despite having very little sequence similarity to E1o follows a similar reaction mechanism. This opens new opportunities for MenD’s substrate and acceptor spectrum, and its saturation inhibition issues which can be addressed by knocking out or substituting part of its active site residues, or the hydrophobic chains supporting it, using site-directed mutagenesis.
Subject (authority = RUETD)
Topic
Chemistry
Subject (authority = ETD-LCSH)
RelatedItem (type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
Identifier
ETD_8936
PhysicalDescription
Form (authority = gmd)
electronic resource
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
1 online resource (x, 53 p. : ill.)
Note (type = degree)
M.S.
Note (type = bibliography)
Includes bibliographical references
Note (type = statement of responsibility)
by Abhilash S. Christian
RelatedItem (type = host)
TitleInfo
Title
Graduate School - Newark Electronic Theses and Dissertations
Identifier (type = local)
rucore10002600001
Location
PhysicalLocation (authority = marcorg); (displayLabel = Rutgers, The State University of New Jersey)
NjNbRU
Identifier (type = doi)
doi:10.7282/T36Q21NW
Genre (authority = ExL-Esploro)
ETD graduate
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Rights

RightsDeclaration (ID = rulibRdec0006)
The author owns the copyright to this work.
RightsHolder (type = personal)
Name
FamilyName
Christian
GivenName
Abhilash
MiddleName
S.
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime (encoding = w3cdtf); (qualifier = exact); (point = start)
2018-04-20 14:00:55
AssociatedEntity
Name
Abhilash Christian
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - Newark
AssociatedObject
Type
License
Name
Author Agreement License
Detail
I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
Copyright
Status
Copyright protected
Availability
Status
Open
Reason
Permission or license
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Technical

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ETD
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windows xp
DateCreated (point = end); (encoding = w3cdtf); (qualifier = exact)
2018-06-20T10:41:51
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2018-04-29T23:02:37
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