The final step in lanthipeptide biosynthesis involves the proteolytic removal of an N-terminal leader peptide. In the class I lanthipeptide epilancin 15X, this step is performed by the subtilisin-like serine peptidase ElxP. Bioinformatic, kinetic, and mass spectrometric analysis revealed that ElxP recognizes the stretch of amino acids DLNPQS located near the proteolytic cleavage site of its substrate, ElxA. When the ElxP recognition motif was inserted into the noncognate lanthipeptide precursor NisA, ElxP was able to proteolytically remove the leader peptide from NisA. Proteolytic removal of the leader peptide by ElxP during the biosynthesis of epilancin 15X exposes an N-terminal dehydroalanine on the core peptide of ElxA that hydrolyzes to a pyruvyl group. The short-chain dehydrogenase ElxO reduces the pyruvyl group to a lactyl moiety in the final step of epilancin 15X maturation. Using synthetic peptides, we also investigated the substrate specificity of ElxO and determined the 1.85 Å resolution X-ray crystal structure of the enzyme.

Airborne bacteria that nucleate ice at relatively warm temperatures (>−10°C) can interact with cloud water droplets, affecting the formation of ice in clouds and the residency time of the cells in the atmosphere. We sampled 65 precipitation events in southeastern Louisiana over 2 years to examine the effect of season, meteorological conditions, storm type, and ecoregion source on the concentration and type of ice-nucleating particles (INPs) deposited. INPs sensitive to heat treatment were inferred to be biological in origin, and the highest concentrations of biological INPs (∼16,000 INPs liter−1 active at ≥–10°C) were observed in snow and sleet samples from wintertime nimbostratus clouds with cloud top temperatures as warm as –7°C. Statistical analysis revealed three temperature classes of biological INPs (INPs active from −5 to –10°C, −11 to –12°C, and −13 to –14°C) and one temperature class of INPs that were sensitive to lysozyme (i.e., bacterial INPs, active from −5 to –10°C). Significant correlations between the INP data and abundances of taxa in the Bacteroidetes, Firmicutes, and unclassified bacterial divisions implied that certain members of these phyla may possess the ice nucleation phenotype. The interrelation between the INP classes and fluorescent dissolved organic matter, major ion concentrations (Na+, Cl−, SO42−, and NO3−), and backward air mass trajectories indicated that the highest concentrations of INPs were sourced from high-latitude North American and Asian continental environments, whereas the lowest values were observed when air was sourced from marine ecoregions. The intra- and extracontinental regions identified as sources of biological INPs in precipitation deposited in the southeastern United States suggests that these bioaerosols can disperse and affect meteorological conditions thousands of kilometers from their terrestrial points of origin. IMPORTANCE The particles most effective at inducing the freezing of water in the atmosphere are microbiological in origin; however, information on the species harboring this phenotype, their environmental distribution, and ecological sources are very limited. Analysis of precipitation collected over 2 years in Louisiana showed that INPs active at the warmest temperatures were sourced from terrestrial ecosystems and displayed behaviors that implicated specific bacterial taxa as the source of the ice nucleation activity. The abundance of biological INPs was highest in precipitation from winter storms and implied that their in-cloud concentrations were sufficient to affect the formation of ice and precipitation in nimbostratus clouds.

Microbes in the atmosphere have broad ecological impacts, including the potential to trigger precipitation through species and strains that act as ice nucleation particles. To characterize spatiotemporal trends of microbial assemblages in precipitation we sequenced 16S (bacterial) and 18S (fungal) rRNA gene amplicon libraries collected from 72 precipitation events in three U.S. states (Idaho, Louisiana, and Virginia) over four seasons. We considered these data from the perspective of a novel metacommunity framework. In agreement with our heuristic, we found evidence for distinct mechanisms underlying the composition and diversity of bacterial and fungal assemblages in precipitation. Specifically, we determined that (1) bacterial operational taxonomic unit (OTU) composition of precipitation was strongly associated with macroscale drivers including season and high-altitude characteristics of storms; (2) fungal OTU composition was strongly correlated with mesoscale drivers including particular spatial locations; (3) b-diversity (heterogeneity of taxa among samples) for both bacteria and fungi was largely maintained by turnover of taxa; however, (4) bacterial assemblages had higher contributions to total b-diversity from nestedness (i.e., lower richness assemblages were largely taxonomic subsets of richer assemblages), due to losses of taxa during dispersal, particularly among potential ice nucleation active bacteria; and (5) fungal assemblages had higher contributions to total b-diversity from turnover due to OTU replacement. Spatiotemporal trends in precipitation-borne metacommunities allowed delineation of a large number of statistically significant indicator taxa for particular sites and seasons, including trends for bacteria that are potentially ice nucleation active. Our findings advance understanding regarding the dispersion of aerosolized microbes via wet deposition, and the development of theory concerning potential assembly rules for bioaerosol assemblages.

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