Sulfur Oxidation in Chlorobium tepidum (syn. Chlorobaculum tepidum): Genetic and Proteomic Analyses

2008 ◽  
pp. 117-126 ◽  
Author(s):  
Leong-Keat Chan ◽  
Rachael Morgan-Kiss ◽  
Thomas E. Hanson
2016 ◽  
Vol 82 (21) ◽  
pp. 6431-6439 ◽  
Author(s):  
Amalie T. Levy ◽  
Kelvin H. Lee ◽  
Thomas E. Hanson

ABSTRACTMicrobial sulfur metabolism, particularly the formation and consumption of insoluble elemental sulfur (S0), is an important biogeochemical engine that has been harnessed for applications ranging from bioleaching and biomining to remediation of waste streams.Chlorobaculum tepidum, a low-light-adapted photoautolithotrophic sulfur-oxidizing bacterium, oxidizes multiple sulfur species and displays a preference for more reduced electron donors: sulfide > S0> thiosulfate. To understand this preference in the context of light energy availability, an “energy landscape” of phototrophic sulfur oxidation was constructed by varying electron donor identity, light flux, and culture duration. Biomass and cellular parameters ofC. tepidumcultures grown across this landscape were analyzed. From these data, a correction factor for colorimetric protein assays was developed, enabling more accurate biomass measurements forC. tepidum, as well as other organisms.C. tepidum's bulk amino acid composition correlated with energy landscape parameters, including a tendency toward less energetically expensive amino acids under reduced light flux. This correlation, paired with an observation of increased cell size and storage carbon production under electron-rich growth conditions, suggests thatC. tepidumhas evolved to cope with changing energy availability by tuning its proteome for energetic efficiency and storing compounds for leaner times.IMPORTANCEHow microbes cope with and adapt to varying energy availability is an important factor in understanding microbial ecology and in designing efficient biotechnological processes. We explored the response of a model phototrophic organism,Chlorobaculum tepidum, across a factorial experimental design that enabled simultaneous variation and analysis of multiple growth conditions, what we term the “energy landscape.”C. tepidumbiomass composition shifted toward less energetically expensive amino acids at low light levels. This observation provides experimental evidence for evolved efficiencies in microbial proteomes and emphasizes the role that energy flux may play in the adaptive responses of organisms. From a practical standpoint, our data suggest that bulk biomass amino acid composition could provide a simple proxy to monitor and identify energy stress in microbial systems.


2012 ◽  
Vol 195 (2) ◽  
pp. 399-408 ◽  
Author(s):  
Brian J. Eddie ◽  
Thomas E. Hanson

ABSTRACTChlorobaculum tepidumis a green sulfur bacterium (GSB) that is a model system for phototrophic sulfur oxidation. Despite over 2 decades of research, conspicuous gaps exist in our understanding of its electron donor metabolism and regulation. RNA sequencing (RNA-seq) was used to provide a global picture of theC. tepidumtranscriptome during growth on thiosulfate as the sole electron donor and at time points following the addition of sulfide to such a culture. Following sulfide addition, 121 to 150 protein-coding genes displayed significant changes in expression depending upon the time point. These changes included a rapid decrease in expression of thiosulfate and elemental sulfur oxidation genes. Genes and gene loci with increased expression included CT1087, encoding a sulfide:quinone oxidoreductase required for growth in high sulfide concentrations; a polysulfide reductase-like complex operon,psrABC(CT0496 to CT0494); and, surprisingly, a large cluster of genes involved in iron acquisition. Finally, two genes that are conserved as a cassette in anaerobic bacteria and archaea, CT1276 and CT1277, displayed a strong increase in expression. The CT1277 gene product contains a DNA-binding domain, suggesting a role for it in sulfide-dependent gene expression changes.


2010 ◽  
Vol 104 (2-3) ◽  
pp. 153-162 ◽  
Author(s):  
Kalliopi Kouyianou ◽  
Michalis Aivaliotis ◽  
Kris Gevaert ◽  
Michael Karas ◽  
Georgios Tsiotis

2008 ◽  
Vol 191 (3) ◽  
pp. 1026-1034 ◽  
Author(s):  
Leong-Keat Chan ◽  
Rachael M. Morgan-Kiss ◽  
Thomas E. Hanson

ABSTRACT Sulfide:quinone oxidoreductase (SQR) catalyzes sulfide oxidation during sulfide-dependent chemo- and phototrophic growth in bacteria. The green sulfur bacterium Chlorobaculum tepidum (formerly Chlorobium tepidum) can grow on sulfide as the sole electron donor and sulfur source. C. tepidum contains genes encoding three SQR homologs: CT0117, CT0876, and CT1087. This study examined which, if any, of the SQR homologs possess sulfide-dependent ubiquinone reduction activity and are required for growth on sulfide. In contrast to CT0117 and CT0876, transcripts of CT1087 were detected only when cells actively oxidized sulfide. Mutation of CT0117 or CT1087 in C. tepidum decreased SQR activity in membrane fractions, and the CT1087 mutant could not grow with ≥6 mM sulfide. Mutation of both CT0117 and CT1087 in C. tepidum completely abolished SQR activity, and the double mutant failed to grow with ≥4 mM sulfide. A C-terminal His6-tagged CT1087 protein was membrane localized, as was SQR activity. Epitope-tagged CT1087 was detected only when sulfide was actively consumed by cells. Recombinantly produced CT1087 and CT0117 proteins had SQR activity, while CT0876 did not. In summary, we conclude that, under the conditions tested, both CT0117 and CT1087 function as SQR proteins in C. tepidum. CT0876 may support the growth of C. tepidum at low sulfide concentrations, but no evidence was found for SQR activity associated with this protein.


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