Project description:Cyanobacterial blooms associated with the benthic mat have been rising. Besides the ongoing concern about toxins production, cyanobacteria are actively involved in marine biofilms, representing several economic and environmental impacts. Proteomic studies on cyanobacterial biofilms could be an effective approach to establish metabolic pathways that affect these fouling organisms and, consequently, obtain novel control strategies against them. Currently, there are few studies in this field on filamentous cyanobacteria. Thus, standard methodologies for following cyanobacterial biofilm development for a long-term assay and a quantitative proteomics analysis were performed in this work. Biofilm development from unidentified filamentous Synechococcales LEGE 06021 was evaluated on different surfaces, glass and perspex, and at two significant shear rates for marine environments (4 s-1 and 40 s-1). Higher biofilm development was observed at 4 s-1, and these biofilms showed a lower roughness coefficient value than those formed at higher shear. Overall, about 1,877 proteins were identified, and differences in proteome were more noticeable between the two hydrodynamic conditions than those found between the two surfaces. 20 Differentially Expressed Proteins (DEPs) were found between 4 s-1 vs. 40 s-1, of which 15 DEPs were found on glass, whereas five DEPs were found on perspex. On the glass, some of these DEPs include phage tail proteins, orange carotenoid protein, enzymes like cyanophynase, glutathione-dependent formaldehyde dehydrogenase, and MoaD/ThiS family protein, while on perspex, the DEPs include enzymes such as transketolase, dihydroxy-acid dehydratase, iron ABC transporter substrate-binding protein or transcription termination/antitermination protein NusG. In summary, the biofilm structure, chlorophyll a content, total biomass, and proteomic profile are more affected by the hydrodynamic conditions than by the surfaces employed. These findings suggest that most of the metabolic changes could be produced to counterbalance the different shear rates. However, the differential expression of some proteins could be associated with the surfaces used. This study helps to consolidate the knowledge of the main factors affecting biofilm development, and sheds new lights on putative targets to address new antimicrobial strategies.
Project description:Increasing atmospheric CO2 concentrations are causing decreased pH over vast expanses of the ocean. This decreasing pH may alter biogeochemical cycling of carbon and nitrogen via the microbial process of nitrification, a key process that couples these cycles in the ocean, but which is often sensitive to acidic conditions. Recent reports indicate a decrease in oceanic nitrification rates under experimentally lowered pH. How composition and abundance of ammonia oxidizing bacteria (AOB) and archaea (AOA) assemblages respond to decreasing oceanic pH, however, is unknown. We sampled microbes from two different acidification experiments and used a combination of qPCR and functional gene microarrays for the ammonia monooxygenase gene (amoA) to assess how acidification alters the structure of ammonia oxidizer assemblages. We show that despite widely different experimental conditions, acidification consistently altered the community composition of AOB by increasing the relative abundance of taxa related to the Nitrosomonas ureae clade. In one experiment this increase was sufficient to cause an increase in the overall abundance of AOB. There were no systematic shifts in the community structure or abundance of AOA in either experiment. These different responses to acidification underscore the important role of microbial community structure in the resiliency of marine ecosystems. SUBMITTER_CITATION: Title: Acidification alters the composition of ammonia oxidizing microbial assemblages in marine mesocosms Journal: Marine Ecology Progress Series Issue: 492 Pages: 1-8 DOI: 10.3354/meps 10526 Authors: Jennifer L Bowen Patrick J Kearns Michael Holcomb Bess B Ward
Project description:Many chemically complex cyanobacterial polyketides and nonribosomal peptides are of great pharmaceutical interest, but the levels required for exploitation are difficult to achieve from native sources. Here we develop a framework for the expression of these multifunctional cyanobacterial assembly lines in Escherichia coli using the lyngbyatoxin biosynthetic pathway, derived from a marine microbial assemblage dominated by the cyanobacterium Moorea producens. Heterologous expression of this pathway afforded high titers of both lyngbyatoxin A (25.6 mg L(-1)) and its precursor indolactam-V (150 mg L(-1)). Production, isolation, and identification of all expected chemical intermediates of lyngbyatoxin biosynthesis in E. coli also confirmed the previously proposed biosynthetic route, setting a solid chemical foundation for future pathway engineering. The successful production of the nonribosomal peptide lyngbyatoxin A in E. coli also opens the possibility for future heterologous expression, characterization, and exploitation of other cyanobacterial natural product pathways.