Staff View
Identification and characterization of a novel monooxygenase from Burkholderia xenovorans LB400

Descriptive

TitleInfo (displayLabel = Citation Title); (type = uniform)
Title
Identification and characterization of a novel monooxygenase from Burkholderia xenovorans LB400
Name (ID = NAME001); (type = personal)
NamePart (type = family)
Montes-Matias
NamePart (type = given)
Marie Carmen
DisplayForm
Marie Carmen Montes-Matias
Role
RoleTerm (authority = RUETD)
author
Name (ID = NAME002); (type = personal)
NamePart (type = family)
Zylstra
NamePart (type = given)
Gerben
Affiliation
Advisory Committee
DisplayForm
Gerben Zylstra
Role
RoleTerm (authority = RULIB)
chair
Name (ID = NAME003); (type = personal)
NamePart (type = family)
Chase
NamePart (type = given)
Theodore
Affiliation
Advisory Committee
DisplayForm
Theodore Chase
Role
RoleTerm (authority = RULIB)
internal member
Name (ID = NAME004); (type = personal)
NamePart (type = family)
Bini
NamePart (type = given)
Elisabetta
Affiliation
Advisory Committee
DisplayForm
Elisabetta Bini
Role
RoleTerm (authority = RULIB)
internal member
Name (ID = NAME005); (type = personal)
NamePart (type = family)
Kobayashi
NamePart (type = given)
Donald
Affiliation
Advisory Committee
DisplayForm
Donald Kobayashi
Role
RoleTerm (authority = RULIB)
outside member
Name (ID = NAME006); (type = corporate)
NamePart
Rutgers University
Role
RoleTerm (authority = RULIB)
degree grantor
Name (ID = NAME007); (type = corporate)
NamePart
Graduate School - New Brunswick
Role
RoleTerm (authority = RULIB)
school
TypeOfResource
Text
Genre (authority = marcgt)
theses
OriginInfo
DateCreated (qualifier = exact)
2008
DateOther (qualifier = exact); (type = degree)
2008-10
Language
LanguageTerm
English
PhysicalDescription
Form (authority = marcform)
electronic
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
xi, 128 pages
Abstract
Whole genome sequences have been key elements in discovering new genes and in predicting protein function. A gene found in Burkholderia xenovorans LB400 encodes a protein that is 77% identical and 85% similar to the p-cymene monooxygenase from Pseudomonas putida F1. Phylogenetic analysis of this enzyme reveals that it is closely related to cymene, xylene and alkane monooxygenase. Protein alignment of these sequences identified 10 conserved histidine residues, of which 8 were reported to be essential for catalysis in alkane monooxygenase. The identified consensus sequence for these enzymes is [HX(3)HX(25)HX(3)HHX(139)HX(2)HH], which is characteristic of membrane-bound hydroxylases with an active site that require iron and the activation of molecular oxygen for their catalytic cycle. In order to analyze the B. xenovorans LB400 monooxygenase enzyme in more detail, we cloned the cognate genes into the expression vector pQE.30 and performed biotransformation assays to determine the substrate specificity of the CymA1A2-like enzyme. The novel monooxygenase found in LB400 hydroxylates p-cymene to the product p-cymen-8-ol. The cymene monooxygenase found in P. putida F1 hydroxylated the methyl group of p-cymene, while the novel monooxygenase catalyzed the hydroxylation of the carbon of the isopropyl group adjacent to the benzene ring. The enzyme also oxidized the substrates cumene, isobutylbenzene, n-butylbenzene, propylbenzene, 1, 4-diisopropylbenzene, tert-butyltoluene, sec-butylbenzene, and 4-ethyltolunene in addition to p-cymene. The collected data indicate that the LB400 monooxygenase described above is able to catalyze an interesting and novel reaction, but its physiological role is still unidentified. Steric hindrance and chain length definitely play an important role in determining the enzyme specificity. Metabolite analysis and RT-PCR data indicated that there is no correlation between the novel enzyme activity and growth of the strain on p-cymene, propylbenzene, 1,4-diisopropylbenzene, 4-ethyltoluene, n-butylbenzene, isobutylbenzene, sec-butylbenzene, tert-butylbenzene, and tert-butyltoluene. Other pathways and enzymes are involved in the degradation of these substrates. This data indicates that although bioinformatic analysis is a valuable tool in protein analysis and function prediction, it cannot always accurately predict the catalytic activity and physiological role of an enzyme.
Note (type = degree)
Ph.D.
Note (type = bibliography)
Includes bibliographical references (p. 120-127).
Subject (ID = SUBJ1); (authority = RUETD)
Topic
Microbiology and Molecular Genetics
Subject (ID = SUBJ2); (authority = ETD-LCSH)
Topic
Monooxygenases
RelatedItem (type = host)
TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier (type = local)
rucore19991600001
Identifier (type = hdl)
http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.17537
Identifier
ETD_1200
Location
PhysicalLocation (authority = marcorg); (displayLabel = Rutgers, The State University of New Jersey)
NjNbRU
Identifier (type = doi)
doi:10.7282/T36T0MXS
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
AssociatedEntity (AUTHORITY = rulib); (ID = 1)
Name
Marie Montes-Matias
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
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

Format (TYPE = mime); (VERSION = )
application/x-tar
FileSize (UNIT = bytes)
2990080
Checksum (METHOD = SHA1)
6cbb16e615c4af8f54749ed7982d8677bb0c94b9
ContentModel
ETD
CompressionScheme
other
OperatingSystem (VERSION = 5.1)
windows xp
Format (TYPE = mime); (VERSION = NULL)
application/x-tar
Back to the top
Version 8.3.13
Rutgers University Libraries - Copyright ©2021