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Molecular genetic and physiological studies to unravel the mystery of Sphingomonas wittichii RW1 dibenzo-p-dioxin degradation

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Title
Molecular genetic and physiological studies to unravel the mystery of Sphingomonas wittichii RW1 dibenzo-p-dioxin degradation
Name (type = personal)
NamePart (type = family)
Mutter
NamePart (type = given)
Thamer Yousif
NamePart (type = date)
1978-
DisplayForm
Thamer Yousif Mutter
Role
RoleTerm (authority = RULIB)
author
Name (type = personal)
NamePart (type = family)
Zylstra
NamePart (type = given)
Gerben J
DisplayForm
Gerben J Zylstra
Affiliation
Advisory Committee
Role
RoleTerm (authority = RULIB)
chair
Name (type = corporate)
NamePart
Rutgers University
Role
RoleTerm (authority = RULIB)
degree grantor
Name (type = corporate)
NamePart
School of Graduate Studies
Role
RoleTerm (authority = RULIB)
school
TypeOfResource
Text
Genre (authority = marcgt)
theses
OriginInfo
DateCreated (encoding = w3cdtf); (keyDate = yes); (qualifier = exact)
2019
DateOther (encoding = w3cdtf); (qualifier = exact); (type = degree)
2019-10
Language
LanguageTerm (authority = ISO 639-3:2007); (type = text)
English
Abstract (type = abstract)
Dibenzofuran and dibenzo-p-dioxin are ubiquitous environmental pollutants in soil and sediment. Sphingomonas wittichii RW1 is one of a few strains known for the ability to grow on the related compounds dibenzofuran (DBF) and dibenzo-p-dioxin (DXN) as the sole source of carbon. The genes for the initial steps in the DBF catabolic pathway (ring hydroxylating dioxygenase, ring cleavage dioxygenase, and a hydrolase) which result in the formation of salicylate and a five-carbon fragment have been localized to a mega-plasmid designated pSWIT02 in RW1. Plasmids highly similar to pSWIT02 have been found in other DBF degrading Sphingomonas strains. However, despite having the pSWIT02-encoded DBF degradation pathway these other bacteria are not capable of growth on DXN. This thesis describes involvement of chromosomally encoded genes in dibenzo-p-dioxin degradation by RW1. RW1 lacking the pSWIT02 dbfB gene grows extremely slowly on DBF and accumulates the ring cleavage substrate 2,2',3-trihydroxybiphenyl. The mutant grows normally on DXN as the sole source of carbon indicating that dbfB is not necessary for the DXN catabolic pathway and suggesting involvement of other ring cleavage dioxygenases in DXN pathway. Knockout of gene SWIT3046 resulted in a strain that grows normally on DBF but that does not grow on DXN. The double knockout does not grow on either DBF or DXN. These results prove that separate ring cleavage enzymes are necessary for DBF and DXN degradation.
We then examined the third enzymatic step in RW1, the hydrolase. RW1 lacking the pSWIT02 encoded gene dxnB1 or the chromosome encoded gene dxnB2 grow normally on both DBF and DXN. A double knockout of both genes does not grow on DBF but still grows on DXN. We then examined previously published transcriptomic data that showed that the SWIT0910 encoded hydrolase is up regulated during growth on DBF and DXN. A knockout of SWIT0910 grows normally on DBF but does not grow on DXN. Our results demonstrate that a chromosomally encoded hydrolase, SWIT0910, is absolutely required for growth on DXN and that two different hydrolases (chromosomally and plasmid encoded) contribute equally to growth on DBF.
Genes for three biphenyl ring cleavage dioxygenases from Burkholderia xenovorans LB400, Sphingomonas yanoikuyae B1, and Pseudomonas putida F1 were moved into a mutant lacking the RW1 DBF and DXN ring cleavage genes. All three ring cleavage dioxygenases allowed the mutant RW1 to grow on DBF at different rates. Interestingly, only bphC from Burkholderia xenovorans LB400 allowed RW1 mutant to grow on DXN.
Subject (authority = RUETD)
Topic
Microbial Biology
Subject (authority = local)
Topic
Sphingomonas wittichii
Subject (authority = LCSH)
Topic
Sphingomonas -- Molecular genetics
RelatedItem (type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
Identifier
ETD_10109
PhysicalDescription
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application/pdf
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text/xml
Extent
1 online resource (x, 93 pages) : illustrations
Note (type = degree)
Ph.D.
Note (type = bibliography)
Includes bibliographical references
RelatedItem (type = host)
TitleInfo
Title
School of Graduate Studies Electronic Theses and Dissertations
Identifier (type = local)
rucore10001600001
Location
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NjNbRU
Identifier (type = doi)
doi:10.7282/t3-vc8x-cs80
Genre (authority = ExL-Esploro)
ETD doctoral
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The author owns the copyright to this work.
RightsHolder (type = personal)
Name
FamilyName
Mutter
GivenName
Thamer
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime (encoding = w3cdtf); (qualifier = exact); (point = start)
2019-07-03 22:18:41
AssociatedEntity
Name
Thamer Mutter
Role
Copyright holder
Affiliation
Rutgers University. School of Graduate Studies
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Author Agreement License
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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.
RightsEvent
Type
Embargo
DateTime (encoding = w3cdtf); (qualifier = exact); (point = start)
2019-10-31
DateTime (encoding = w3cdtf); (qualifier = exact); (point = end)
2021-10-30
Detail
Access to this PDF has been restricted at the author's request. It will be publicly available after October 30th, 2021.
Copyright
Status
Copyright protected
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Status
Open
Reason
Permission or license
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windows xp
DateCreated (point = end); (encoding = w3cdtf); (qualifier = exact)
2019-11-11T20:24:09
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