Noninvasive tool for optical online monitoring of individual biomass concentrations in a defined coculture

Cocultures bear great potential in the conversion of complex substrates and process intensification, as well as, in the formation of unique components only available due to inter-species interactions. Dynamic data of coculture composition is necessary for understanding and optimizing coculture systems. However, most standard online determined parameters measure the sum of all species in the reactor system. The kinetic behavior of the individual species remains unknown. Up to now, different offline methods are available to determine the culture composition, as well as the online measurement of fluorescence of genetically modified organisms. To avoid any genetic modification, a noninvasive online monitoring tool based on the scattered light spectrum was developed for microtiter plate cultivations. To demonstrate the potential, a coculture consisting of the bacterium Lactococcus lactis and the yeast Kluyveromyces marxianus was cultivated. Via partial least squares regression of scattered light spectra, the online determination of the individual biomass concentrations without further sampling and analyses is possible. The results were successfully validated by a Coulter counter-analysis, taking advantage of the different cell sizes of both organisms. The findings prove the applicability of the new method to follow in detail the dynamics of a coculture.     

Engineering Escherichia coli for propionic acid production through the Wood–Werkman cycle

Native to propionibacteria, the Wood–Werkman cycle enables propionate production via succinate decarboxylation. Current limitations in engineering propionibacteria strains have redirected attention toward the heterologous production in model organisms. Here, we report the functional expression of the Wood–Werkman cycle in Escherichia coli to enable propionate and 1-propanol production. The initial proof-of-concept attempt showed that the cycle can be used for production. However, production levels were low (0.17 mM). In silico optimization of the expression system by operon rearrangement and ribosomal-binding site tuning improved performance by fivefold. Adaptive laboratory evolution further improved performance redirecting almost 30% of total carbon through the Wood–Werkman cycle, achieving propionate and propanol titers of 9 and 5 mM, respectively. Rational engineering to reduce the generation of byproducts showed that lactate (∆ldhA) and formate (∆pflB) knockout strains exhibit an improved propionate and 1-propanol production, while the ethanol (∆adhE) knockout strain only showed improved propionate production.     

Multiple RNA interactions position Mrd1 at the site of the small subunit pseudoknot within the 90S pre-ribosome

Ribosomal subunit biogenesis in eukaryotes is a complex multistep process. Mrd1 is an essential and conserved small (40S) ribosomal subunit synthesis factor that is required for early cleavages in the 35S pre-ribosomal RNA (rRNA). Yeast Mrd1 contains five RNA-binding domains (RBDs), all of which are necessary for optimal function of the protein. Proteomic data showed that Mrd1 is part of the early pre-ribosomal complexes, and deletion of individual RBDs perturbs the pre-ribosomal structure. In vivo ultraviolet cross-linking showed that Mrd1 binds to the pre-rRNA at two sites within the 18S region, in helix 27 (h27) and helix 28. The major binding site lies in h27, and mutational analyses shows that this interaction requires the RBD1-3 region of Mrd1. RBD2 plays the dominant role in h27 binding, but other RBDs also contribute directly. h27 and helix 28 are located close to the sequences that form the central pseudoknot, a key structural feature of the mature 40S subunit. We speculate that the modular structure of Mrd1 coordinates pseudoknot formation with pre-rRNA processing and subunit assembly.     

Calcium and iron as key drivers of brown moss composition through differential effects on phosphorus availability

Brown moss-dominated rich fens are characterized by minerotrophic conditions, in which calcium (Ca) and iron (Fe) concentrations show large variations. We examined the relative importance of Ca and Fe in relation to the occurrence of three typical brown moss species: Scorpidium scorpioides, Scorpidium cossonii, and Hamatocaulis vernicosus. Peat chemistry was examined in 24 stands of brown moss-dominated rich fens: 12 in the Netherlands and 12 in central Sweden. Ca and Fe turned out to be important drivers of brown moss composition. Fens dominated by Scorpidium scorpioides or Scorpidium cossonii were characterized by high pore water Ca-concentrations and total soil Ca-contents, but low P-availability. In these Ca-rich, but Fe-poor fens, foliar N:P ratios of vascular vegetation exceeded 20?g?g^?1, indicating phosphorus (P)-limitation due to Ca-P precipitation or low P-sorption capacity due to low Fe-levels. In contrast, fens dominated by Hamatocaulis vernicosus were characterized by high pore water Fe-concentrations and total soil Fe-contents, but also relatively high P-availability. N:P ratios in these fens were below 13.5?g?g^?1, indicating potential nitrogen (N)-limitation. We conclude that the relative roles of Ca and Fe, as related to the geohydrological conditions present, strongly determine the brown moss composition in rich fens through their differential effects on plant P-availability.     

Conservation status of the two cryptic species of Hamatocaulis vernicosus (Bryophyta) in Sweden

Cryptic species are rarer than their combined, morphologically recognisable species. Each cryptic species may have its own habitat requirements and distribution, and each should be considered separately in biodiversity conservation. This investigation explores how well the two cryptic species of the wetland moss Hamatocaulis vernicosus (Mitt.) Hedenäs s.l., included in Annex II of the EU Habitat Directive, are safeguarded in existing protected sites in Sweden. Further, the northern distribution limit of the southern of the two cryptic species is explored. The distributions of the two cryptic species and their intraspecific variation are judged by the nuclear ITS1?+?2 and the two chloroplast markers rpl16 and trnL-trnF for a set of 89 specimens. The genetic differences between the two cryptic species are significant, but there are no differences between the protected and non-protected subsets within the respective species. The protected areas therefore represent these two species’ genetic variation well. The populations of both cryptic species appear stable, according to their genetic signals. One of the two cryptic species occurs almost throughout Sweden, whereas the other occurs only to the south of the southern limit of the southern boreal zone, except for two finds slightly further north in climatically mild areas.