Hmelo Laura R.
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ArticleKinetic constraints on acylated homoserine lactone-based quorum sensing in marine environments(Inter-Research, 2009-02-04) Hmelo, Laura R. ; Van Mooy, Benjamin A. S.Quorum sensing (QS) via acylated homoserine lactones (AHLs) was discovered in the ocean, yet AHLs are expected to be very short-lived at seawater pH due to rapid abiotic degradation. Quorum quenching, the enzymatic degradation of AHLs, is also likely. To better understand the potential for QS to regulate behaviors of marine bacteria, we investigated the degradation of a variety of AHL molecules in several types of seawater media. We did this by incubating AHLs and tracking their concentration using HPLC/electrospray-ionization mass-spectrometry (HPLC/ESI-MS). AHL concentrations decreased with time, and degradation rate coefficients were calculated by applying a first-order rate law. The rate of abiotic degradation showed strong dependence on acyl chain length and the presence of 3-oxo substitutions on the acyl chain. We found that the rate of abiotic degradation of AHLs in artificial seawater was much slower than that predicted by an oft-cited equation for non-marine media that takes only pH into account. However, AHLs degraded more rapidly in natural seawater than in artificial seawater, an observation we found to be due to quorum quenching enzyme activity. By applying calculated degradation rates in a simple steady-state calculation, we suggest that despite the observed quorum quenching activity, AHLs are likely to be viable signals in organic particles and in other microbial ‘hotpsots’ in marine environments.
ArticleProchlorococcus extracellular vesicles: molecular composition and adsorption to diverse microbes(Society for Applied Microbiology, 2021-11-12) Biller, Steven J. ; Lundeen, Rachel A. ; Hmelo, Laura R. ; Becker, Kevin W. ; Arellano, Aldo A. ; Dooley, Keven ; Heal, Katherine R. ; Carlson, Laura Truxal ; Van Mooy, Benjamin A. S. ; Ingalls, Anitra ; Chisholm, Sallie W.Extracellular vesicles are small (~50–200 nm diameter) membrane-bound structures released by cells from all domains of life. While vesicles are abundant in the oceans, their functions, both for cells themselves and the emergent ecosystem, remain a mystery. To better characterize these particles – a prerequisite for determining function – we analysed the lipid, protein, and metabolite content of vesicles produced by the marine cyanobacterium Prochlorococcus. We show that Prochlorococcus exports a diverse array of cellular compounds into the surrounding seawater enclosed within discrete vesicles. Vesicles produced by two different strains contain some materials in common, but also display numerous strain-specific differences, reflecting functional complexity within vesicle populations. The vesicles contain active enzymes, indicating that they can mediate extracellular biogeochemical reactions in the ocean. We further demonstrate that vesicles from Prochlorococcus and other bacteria associate with diverse microbes including the most abundant marine bacterium, Pelagibacter. Together, our data point toward hypotheses concerning the functional roles of vesicles in marine ecosystems including, but not limited to, possibly mediating energy and nutrient transfers, catalysing extracellular biochemical reactions, and mitigating toxicity of reactive oxygen species.
ThesisMicrobial interactions associated with biofilms attached to Trichodesmium spp. and detrital particles in the ocean(Massachusetts Institute of Technology and Woods Hole Oceanographic Institution, 2010-06) Hmelo, Laura R.Quorum sensing (QS) via acylated homoserine lactones (AHLs) was discovered in the ocean, yet little is known about its role in the ocean beyond its involvement in certain symbiotic interactions. The objectives of this thesis were to constrain the chemical stability of AHLs in seawater, explore the production of AHLs in marine particulate environments, and probe selected behaviors which might be controlled by AHL-QS. I established that AHLs are more stable in seawater than previously expected and are likely to accumulate within biofilms. Based on this result, I chose to study AHL-QS in the bacterial communities inhabiting biofilms attached to Trichodesmium spp. and detrital (photosynthetically-derived sinking particulate organic carbon, POC) particles. These hot spots of microbial activity are primary sites of interaction between marine primary producers and heterotrophs and crucial components of the biological pump. Biofilm communities associated with Trichodesmium thiebautii colonies in the Sargasso Sea differed considerably from seawater microbial communities. In addition, there was no overlap between the communities associated with tuft and puff colonies. These results suggest that bacterial communities associated with Trichodesmium are not random; rather, Trichodesmium selects for specific microbial flora. Novel 16S rRNA gene sequences are present both in clone libraries constructed from DNA extracted from colonies of Trichodesmium spp. and in culture collections derived from wild and laboratory cultivated Trichodesmium spp., supporting the idea that the phycosphere of Trichodesmium is a unique microenvironment. Using high performance liquid chromatography-mass spectrometry, I demonstrated that bacteria isolated from Trichodesmium synthesize AHLs. In addition, I detected AHLs in sinking particles collected from a site off of Vancouver Island, Canada. AHLs were subsequently added to laboratory cultures of non-axenic Trichodesmium colonies and sinking POC samples. This is the first time AHLs have been detected in POC and indicates that AHL-QS was occurring in POC. Further, I showed that AHLs enhanced certain organic-matter degrading hydrolytic enzyme activities. My results suggest that AHL-QS is a factor regulating biogeochemically relevant enzyme activities on sinking POC and within the biofilms attached to Trichodesmium colonies and thereby may impact the timing and magnitude of biogeochemical fluxes in the ocean.