RUcore: Rutgers University Community Repository
Identification and characterization of monooxygenase enzymes involved in 1,4-dioxane degradation in Pseudonocardia sp. strain ENV478, Mycobacterium sp. strain ENV421, and Nocardia sp. strain ENV425
Descriptive
TypeOfResource
Text
TitleInfo
(ID = T-1)
Title
Identification and characterization of monooxygenase enzymes involved in 1,4-dioxane degradation in Pseudonocardia sp. strain ENV478, Mycobacterium sp. strain ENV421, and Nocardia sp. strain ENV425
SubTitle
PartName
PartNumber
NonSort
Identifier
ETD_1641
Identifier
(type = hdl)
http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.000051379
Language
(objectPart = )
LanguageTerm
(authority = ISO639-2);
(type = code)
eng
Genre
(authority = marcgt)
theses
Subject
(ID = SBJ-1);
(authority = RUETD)
Topic
Microbiology and Molecular Genetics
Subject
(ID = SBJ-2);
(authority = ETD-LCSH)
Topic
Monooxygenases
Abstract
The first part of this dissertation deals with the identification and analysis of oxygenases possibly involved in a biodegradation of a possible human carcinogen, 1, 4-dioxane.
The cometabolic oxidations of 1, 4-dioxane in three gram positive bacteria were analyzed. In Mycobacterium sp. ENV421 and Nocardia sp. ENV425, 1, 4-dioxane is oxidized during growth on propane. Three putative propane oxidizing enzymes (alkane monooxygenase, soluble diiron monooxygenase, and cytochrome P450) were identified in both strains. While in strain ENV425 only soluble diiron monooxygenase was expressed after growth on propane, in strain ENV421 all three genes were expressed. Although 1,4-dioxane oxidation activity could not be detected in heterologous host gene expression studies, aliphatic oxidation activity was observed with cytochrome P450 CYP153 from strain ENV421.
In strain Pseudonocardia sp. ENV478, 1, 4-dioxane oxidation activity was induced in the presence of tetrahydrofuran (THF). A THF-inducible soluble diiron monooxygenase was identified in strain ENV478. The oxygenase gene was transcribed in a complex operonic fashion yielding multiple transcripts with varying lengths. A gene knockdown experiment using antisense technology was employed to show that this gene is essential in THF and 1,4-dioxane degradation.
The second part of this dissertation dealt with the isolation and characterization of a novel solid alkane-utilizing β-Proteobacterium, Aquabacterium njensis sp. nov.
A novel liquid and solid alkane-degrading bacterium was isolated from hydrocarbon-contaminated soil in New Jersey, USA. The morphological, genomic, and metabolic properties of this organism were examined, including, observation with scanning electron microscopy, DNA-DNA hybridization, and nutritional screening of mainly aliphatic compounds as the sole source of carbon for growth. The preliminary screen of known alkane oxidation enzymes identified two alkane monooxygenase (AlkB) genes in the HMGZ-01 strain.
Sequence analysis of 16S rRNA showed that it has the highest similarity to the members of the genus Aquabacterium. The ability to hydrolyze urea and gelatin, NaCl tolerance, substrate utilization pattern and anaerobic growth distinguish the isolated strain from other members of this genus. We proposed the new species, Aquabacterium njensis, to be created with a single member and a type strain (HMGZ-01T).
The cometabolic oxidations of 1, 4-dioxane in three gram positive bacteria were analyzed. In Mycobacterium sp. ENV421 and Nocardia sp. ENV425, 1, 4-dioxane is oxidized during growth on propane. Three putative propane oxidizing enzymes (alkane monooxygenase, soluble diiron monooxygenase, and cytochrome P450) were identified in both strains. While in strain ENV425 only soluble diiron monooxygenase was expressed after growth on propane, in strain ENV421 all three genes were expressed. Although 1,4-dioxane oxidation activity could not be detected in heterologous host gene expression studies, aliphatic oxidation activity was observed with cytochrome P450 CYP153 from strain ENV421.
In strain Pseudonocardia sp. ENV478, 1, 4-dioxane oxidation activity was induced in the presence of tetrahydrofuran (THF). A THF-inducible soluble diiron monooxygenase was identified in strain ENV478. The oxygenase gene was transcribed in a complex operonic fashion yielding multiple transcripts with varying lengths. A gene knockdown experiment using antisense technology was employed to show that this gene is essential in THF and 1,4-dioxane degradation.
The second part of this dissertation dealt with the isolation and characterization of a novel solid alkane-utilizing β-Proteobacterium, Aquabacterium njensis sp. nov.
A novel liquid and solid alkane-degrading bacterium was isolated from hydrocarbon-contaminated soil in New Jersey, USA. The morphological, genomic, and metabolic properties of this organism were examined, including, observation with scanning electron microscopy, DNA-DNA hybridization, and nutritional screening of mainly aliphatic compounds as the sole source of carbon for growth. The preliminary screen of known alkane oxidation enzymes identified two alkane monooxygenase (AlkB) genes in the HMGZ-01 strain.
Sequence analysis of 16S rRNA showed that it has the highest similarity to the members of the genus Aquabacterium. The ability to hydrolyze urea and gelatin, NaCl tolerance, substrate utilization pattern and anaerobic growth distinguish the isolated strain from other members of this genus. We proposed the new species, Aquabacterium njensis, to be created with a single member and a type strain (HMGZ-01T).
PhysicalDescription
Form
(authority = gmd)
electronic resource
Extent
xix, 162 p. : ill.
InternetMediaType
application/pdf
InternetMediaType
text/xml
Note
(type = degree)
Ph.D.
Note
(type = bibliography)
Includes bibliographical references (p. 151-161)
Note
(type = statement of responsibility)
by Hisako Masuda
Name
(ID = NAME-1);
(type = personal)
NamePart
(type = family)
Masuda
NamePart
(type = given)
Hisako
NamePart
(type = date)
1980
Role
RoleTerm
(authority = RULIB);
(type = )
author
DisplayForm
Hisako Masuda
Name
(ID = NAME-2);
(type = personal)
NamePart
(type = family)
Zylstra
NamePart
(type = given)
Gerben
Role
RoleTerm
(authority = RULIB);
(type = )
chair
Affiliation
Advisory Committee
DisplayForm
Gerben J Zylstra
Name
(ID = NAME-3);
(type = personal)
NamePart
(type = family)
Chase, Jr
NamePart
(type = given)
Theodore
Role
RoleTerm
(authority = RULIB);
(type = )
internal member
Affiliation
Advisory Committee
DisplayForm
Theodore Chase, Jr
Name
(ID = NAME-4);
(type = personal)
NamePart
(type = family)
Vetriani
NamePart
(type = given)
Constantino
Role
RoleTerm
(authority = RULIB);
(type = )
internal member
Affiliation
Advisory Committee
DisplayForm
Constantino Vetriani
Name
(ID = NAME-5);
(type = personal)
NamePart
(type = family)
Bennett
NamePart
(type = given)
Joan
Role
RoleTerm
(authority = RULIB);
(type = )
outside member
Affiliation
Advisory Committee
DisplayForm
Joan W Bennett
Name
(ID = NAME-1);
(type = corporate)
NamePart
Rutgers University
Role
RoleTerm
(authority = RULIB);
(type = )
degree grantor
Name
(ID = NAME-2);
(type = corporate)
NamePart
Graduate School - New Brunswick
Role
RoleTerm
(authority = RULIB);
(type = )
school
OriginInfo
DateCreated
(point = );
(qualifier = exact)
2009
DateOther
(qualifier = exact);
(type = degree)
2009-05
Place
PlaceTerm
(type = code)
xx
Location
PhysicalLocation
(authority = marcorg)
NjNbRU
RelatedItem
(type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier
(type = RULIB)
ETD
RelatedItem
(type = host)
TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier
(type = local)
rucore19991600001
Identifier
(type = doi)
doi:10.7282/T31N81CB
Genre
(authority = ExL-Esploro)
ETD doctoral
Back to the top
Rights
RightsDeclaration
(AUTHORITY = GS);
(ID = rulibRdec0006)
The author owns the copyright to this work.
Copyright
Status
Copyright protected
Availability
Status
Open
RightsEvent
(AUTHORITY = rulib);
(ID = 1)
Type
Permission or license
Detail
Non-exclusive ETD license
AssociatedObject
(AUTHORITY = rulib);
(ID = 1)
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.
Back to the top
Technical
ContentModel
ETD
MimeType
(TYPE = file)
application/pdf
MimeType
(TYPE = container)
application/x-tar
FileSize
(UNIT = bytes)
5109760
Checksum
(METHOD = SHA1)
07781d32792dd0d5094d88e6b74a3d32eeea5af0
Back to the top
Statistical Profile