The Tat system can transport folded, signal peptide-containing proteins (Tat substrates) across energized membranes of prokaryotes and plant plastids. A twin-arginine motif in the signal peptide of Tat substrates is recognized by TatC-containing complexes, and TatA permits the membrane passage. Often, as in the model Tat systems of Escherichia coli and plant plastids, a third component – TatB – is involved that resembles TatA but has a higher affinity to TatC. It is not known why most TatA dissociates from TatBC complexes in vivo and distributes more evenly in the membrane. Here we show a TatBC-independent substrate-binding to TatA from Escherichia coli, and we provide evidence that this binding enhances Tat transport. First hints came from in vivo cross-linking data, which could be confirmed by affinity co-purification of TatA with the natural Tat substrates HiPIP and NrfC. Two positions on the surface of HiPIP could be identified that are important for the TatA interaction and transport efficiency, indicating physiological relevance of the interaction. Distributed TatA thus may serve to accompany membrane-interacting Tat substrates to the few TatBC spots in the cells.

The hypothesis was that forage species and ensiling conditions have an impact on the formation of biogenic amines and the feed choice of goats in short‐term preference trials. At ensiling, lucerne (Medicago sativa L., first cut), red clover (Trifolium pratense L., first cut) and Italian ryegrass (Lolium multiflorum Lam., second cut) were treated differently to obtain a range of fermentation qualities. Six treatments of each forage species were prepared and included different dry‐matter concentrations, chemical and biological silage additives, and additions of soil. Silages were sampled for chemical analyses (proximate constituents, fermentation products and other volatile compounds, crude protein fractions and biogenic amines) and stored anaerobically in vacuum‐sealed plastic bags for use in preference trials (one trial for each forage species) with Saanen‐type wethers (n = 8 or 6). Each possible two‐way combination of the six silage treatments and a standard hay (n = 21 combinations) was offered for ad libitum intake for 3 hr. Data were analysed using multidimensional scaling, analysis of variance and correlation analysis between silage characteristics and dry‐matter intake (DMI). For each forage species, fermentation characteristics and crude protein fractions revealed only small differences among treatments. Although the degree of proteolysis, as measured by non‐protein nitrogen, of all silages was high, biogenic amine and butyric acid concentrations were low. The different treatments apparently had no direct influence on the formation of biogenic amines and feed choice. The preference behaviour within one forage species was strongly divergent, but DMI rankings of the three species were very similar.

Tat systems translocate folded proteins across biological membranes of prokaryotes and plant plastids. TatBC complexes recognize N-terminal Tat signal peptides that contain a sequence motif with two conserved arginines (RR-motif), and transport takes place after a recruitment of TatA. Unfolded Tat substrate domains lower translocation efficiency and too long linkers lead to translocation arrest. To identify the components that interact with transported proteins during their passage through the translocon, we used a Tat substrate that arrests translocation at a long unfolded linker region, and we chose in vivo site-directed photo cross-linking to specifically detect the interactions of this linker region. For comparison, we included the interactions of the signal peptide and of the folded domain at the C-terminus of this construct. The data show that the linker contacts only two, structurally similar Tat components, namely TatA and TatB. These contacts depend on the recognition of the Tat-specific signal peptide. Only when membrane translocation of the globular domain was allowed--i.e., in the absence of the linker--we observed the same TatAB-contacts also to the globular domain. The data thus suggest that mature protein domains are translocated through a TatAB environment.

Hermetia illucens larvae are able to convert a broad range of organic substrates into animal protein and are believed to have the potential to help solving the supply problem for animal protein in the future. Many farmed animals such as poultry or pigs need an adequate amount of essential amino acids like lysine for optimal growth and performance. In most diets for farmed animals, essential amino acids are supplemented as feed additive to avoid excessive protein intake while ensuring a sufficient supply of essential amino acids. There is no information on the requirements of lysine for the growth of H. illucens larvae. Furthermore, it remains open whether lysine may have an extra effect on growth and development in the cultivation of such larvae. The aim of the study was to evaluate, if an addition of lysine to their feed substrate will affect these parameters. Therefore, in the present study up to 3.0% crystalline lysine was added to a basic substrate and survival, growth, and development of larvae were recorded. Furthermore, larval nutritional values like crude fat, crude protein, and lysine content were analysed. The addition of lysine did not improve growth, development or nutritional values of H. illucens larvae. In contrast, the addition of lysine led to reduced larval size, survival and prepupal rates, especially when high amounts of lysine were provided. The results of this study indicate that a lysine content of 0.3% in the substrate is sufficient for optimal larval growth. Using H. illucens larvae for animal protein production could, therefore, be accomplished with less lysine compared to many other farmed animals.

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