Anaerobic degradation of limonene and p-xylene in freshwater enrichment cultures

155 Pages·2009·6.99 MB·English

Anaerobic degradation of limonene and
p
-xylene in freshwater enrichment
cultures
Amelia Rotaru
2009

Anaerobic degradation of limonene and p-xylene in
fresh-water enrichment cultures
Dissertation
zur
ErlangungdesGradeseines
DoktorsderNaturwissenschaften
Dr. rer. nat.
FachbereichBiologie/Chemie
derUniversitätBremen
vorgelegtvon
Amelia-ElenaRotaru
aus
RamnicuValcea
Romania
Bremen,2009
3
DieUntersuchungenzurvorliegendenDoktorarbeitwurdenamMax-Planck-InstitutfürMa-
rineMikrobiologieinBremendurchgeführt.
Gutachter1: Prof. Dr. FriedrichWiddel,UniversitätBremen
Gutachter2: PDJensHarder,MaxPlanckInstitutfürMarineMikrobiologie
4
Aceastacarteestededicataparintilormeicarem-ausustinutcu
dragostesiincredere,de-alungulacestei„calatorii“innecunoscut.
5
6
Contents
I Anaerobic aliphatic hydrocarbon degradation 15
1 Anaerobicdegradationofsaturatedaliphatichydrocarbons 17
2 Anaerobicdegradationofunsaturatedaliphatichydrocarbons 21
3 Anaerobiclimonenedegradation: resultsanddiscussions 25
3.1 Scopeofthestudy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
3.2 Degradationoflimoneneundermethanogenicconditions . . . . . . . . . . . 26
3.3 Microbialcommunitycomposition . . . . . . . . . . . . . . . . . . . . . . . . . 30
3.4 IsolationofnovelDeltaproteobacteria . . . . . . . . . . . . . . . . . . . . . . 37
3.5 Conclusionsandoutlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
4 References 41
II Anaerobic aromatic hydrocarbon degradation 49
5 Monoaromatichydrocarbondegradation 51
6 Polyaromatichydrocarbondegradation 59
7 Anaerobicp-xylenedegradation: resultsanddiscussions 61
7.1 Scopeofthestudy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
7.2 Cultivationofp-xylenedegradingmicroorganismsunderdenitrifyingconditions 62
7.2.1 Enrichment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
7.2.2 Isolationattempts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
7.2.3 Growthtestsonotherhydrocarbons . . . . . . . . . . . . . . . . . . . 63
7.3 Dominance of novel Betaproteobacteria in p-xylene degrading enrichment
cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
7.4 Quantificationofp-xylenedegradation . . . . . . . . . . . . . . . . . . . . . . 66
7.5 Mechanismofp-Xyleneactivation . . . . . . . . . . . . . . . . . . . . . . . . 67
7.6 Conclusionsandoutlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
8 References 73
7
Contents
9 Explanationofcontribution 83
III Manuscripts 85
10 Manuscript1 87
11 Manuscript2 113
12 Manuscript3 129
13 Annexes 151
14 Acknowledgments 153
8
List of abbreviations
BssA α-subunitofbenzylsuccinatesynthase
bssA geneencodingtheα-subunitofbenzylsuccinatesynthase
BTEX benzene,toluene,ethylbenzene,xylenes
CARD-FISH catalyzedreporterdeposition-fluorescenceinsitu hybridization
DAPI (4’,6-diamidino)2-phenylindol
ΔG0’ deltaG(freeenergy)understandardconditions(25°C,1atm,pH7.00)
FISH fluorescentinsitu hybridization
GC gaschromatography
PAH polyaromatichydrocarbons
RNA ribonucleicacid
rRNA ribosomalRNA
SDS-PAGE sodiumdodecylsulfate-polyacrylamidegelelectrophoresis
SR sulfatereduction
SRB sulfatereducingbacteria
TEA terminalelectronacceptor
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Contents
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