Physiological aspects of l-lysine production: effect of nutritional limitations on a producing strain of Corynebacterium glutamicum

The effect of nutritional limitations, such as phosphorus and carbon, on the production of l-lysine by Corynebacterium glutamicum was studied in continuous culture. We observed that phosphate-limited cultures at low growth rates were favourable to l-lysine production. l-Lysine was produced when a culture at low dilution rate (0.03 h^–1) was established. A dilution rate of about 0.04 h^–1 should be maintained in order to assure good productivity and an l-lysine yield of 0.53 g/g. Under carbon-limiting conditions the maintenance energy and growth yield of 0.03 g/g·g^–1·h^–1 and 0.41 g/g, respectively, have been obtained. Under these limiting conditions the l-lysine production was not favoured even at lower dilution rates.Correspondence to: N. Coello     

PDZ/PDZ interaction between PSD‐95 and nNOS neuronal proteins

N‐Methyl‐D‐aspartate (NMDA) receptors are key components in synaptic communication and are highly relevant in central nervous disorders, where they trigger excessive calcium entry into the neuronal cells causing harmful overproduction of nitric oxide by the neuronal nitric oxide synthase (nNOS) protein. Remarkably, NMDA receptor activation is aided by a second protein, postsynaptic density of 95 kDa (PSD95), forming the ternary protein complex NMDA/PSD95/nNOS. To minimize the potential side effects derived from blocking this ternary complex or either of its protein components, a promising approach points to the disruption of the PSD‐95/nNOS interaction which is mediated by a PDZ/PDZ domain complex. Since the rational development of molecules targeting such protein‐protein interaction relies on energetic and structural information herein, we include a thermodynamic and structural analysis of the PSD95‐PDZ2/nNOS‐PDZ. Two energetically relevant events are structurally linked to a “two‐faced” or two areas of recognition between both domains. First, the assembly of a four‐stranded antiparallel β‐sheet between the β hairpins of nNOS and of PSD95‐PDZ2, mainly enthalpic in nature, contributes 80% to the affinity. Second, binding is entropically reinforced by the hydrophobic interaction between side chains of the same nNOS β‐hairpin with the side chains of α2‐helix at the binding site of PSD95‐PDZ2, contributing the remaining 20% of the total affinity. These results suggest strategies for the future rational design of molecules able to disrupt this complex and constitute the first exhaustive thermodynamic analysis of a PDZ/PDZ interaction.     

Length-weight relationship of 74 fish species caught in the continental coast of Ecuador

This work analysis the length-weight relationships for fish species captured by artisanal fishery along the continental coast of Ecuador. Fishermen used the following fishing gear: beach seine nets (5.1–31.8 mm), shrimp trawl nets (21.8–31.8 mm) and bottom gill nets (54.1–203.2 mm). Sampling took place during the years 2012 and 2014. Frequency of sampling was monthly, and it was completed systematically from Southern toward Northern Provinces. Length-weight relationship (LWR) was realized by the power model (y = a × x^b), for species with a bivariate dataset >70. A total of 59 commercial and 15 non-commercial species were registered. For 74 species, our analysis including a higher total length not reported in LWR estimates previously. The determination coefficient (r²) value was always above 0.95.     

Kluyveromyces lactis sexual pheromones. Gene structures and cellular responses to alpha-factor

The Kluyveromyces lactis genes for sexual pheromones have been analyzed. The alpha-factor gene encodes a predicted polypeptide of 187 amino acid residues containing four tridecapeptide repeats (WSWITLRPGQPIF). A nucleotide blast search of the entire K. lactis genome sequence allowed the identification of the nonannotated putative a-pheromone gene that encodes a predicted protein of 33 residues containing one copy of the dodecapeptide a-factor (WIIPGFVWVPQC). The role of the K. lactis structural genes KlMFalpha1 and KlMFA1 in mating has been investigated by the construction of disruption mutations that totally eliminate gene functions. Mutants of both alleles showed sex-dependent sterility, indicating that these are single-copy genes and essential for mating. MATalpha, Klsst2 mutants, which, by analogy to Saccharomyces cerevisiae, are defective in Galpha-GTPase activity, showed increased sensitivity to synthetic alpha-factor and increased capacity to mate. Additionally, Klbar1 mutants (putatively defective in alpha-pheromone proteolysis) showed delay in mating but sensitivity to alpha-pheromone. From these results, it can be deduced that the K. lactis MATa cell produces the homolog of the S. cerevisiaealpha-pheromone, whereas the MATalpha cell produces the a-pheromone.     

The role of abscisic acid in controlling leaf water loss, survival and growth of micropropagated Tagetes erecta plants when transferred directly to the field

Plants of Tagetes erecta L. (marigold) cultivated in vitro in ventilated containers exhibited greater control of leaf water loss and increased survival in the field than plants cultivated in sealed containers. Increased field survival of plants cultivated in ventilated containers was attributed to higher levels of endogenous abscisic acid (ABA). Therefore, ABA was supplied exogenously to plants in sealed or ventilated containers by adding ABA (10(-6), 10(-5), 10(-4) M) to the in vitro culture media in order to evaluate control of leaf water loss, growth and field survival. The addition of 10(-4) M ABA to the culture media in sealed containers produced plants that had similar control of leaf water loss and were morphologically similar to plants cultivated in ventilated containers without the addition of ABA. Field survival of 10(-4) M ABA plants (75%) was increased compared to plants cultivated in sealed containers without ABA (31%), with survival being closer to that of plants cultivated in ventilated containers (90-100%). Plants cultivated with 10(-4) M ABA (sealed and ventilated) also exhibited increased plant vigour and leaf area in the field compared to plants cultivated without ABA. The results suggest that the limited field survival and growth of plants cultured in vitro are related to the limited ABA concentrations they accumulate while in vitro. Consequently, conditions that increase the endogenous ABA concentrations of in vitro plants (like ventilation or ABA addition to the medium) would improve the control of leaf water loss, field survival and plant vigour.     

Use of microbial activity parameters for determination of a biosolid stability index

Variations in microbial activity during the aerobic digestion of sludge generated at wastewater treatment plants were studied. Results obtained by the measurement of enzymatic activity and microbiological parameters were compared with those determined by traditional methods (COD, suspended solids, etc.). Their variation with digestion time was monitored for batch digestion over a period of 135 days. The relationship between these measurements and control parameters of the sludge was also investigated. It was found that the traditional physicochemical and microbiological parameters present a series of problems which detract from their usefulness. The enzymatic parameters dehydrogenase activity (primary metabolism) and esterase activity (secondary metabolism) are better able to characterise the process, and the ratio of these two variables may be used to estimate the degree of endogenesis and, consequently, the degree of stability of the aerobic sludge digestion. In addition, these techniques are swift and simple to employ.     

Feather degradation by Kocuria rosea in submerged culture

A strain of Kocuria rosea with keratinolytic activity was studied. In batch culture, the optimum temperature for feather degradation, bacterial growth and protease secretion was at 40 °C. A specific growth rate of 0.17 h⁻¹ was attained in basal medium with feathers as fermentation substrate. Under these conditions, after 36 h of incubation, biomass and caseinolytic activity reached 3.2 g/l and 0.15 U/ml, respectively. Extracellular protease secretion was associated with the exponential growth phase. In batch fermentation, feather degradation up to 51% in 72 h was obtained with a conversion yield in biomass of 0.32 g/g. No organic acids were detected in the fermentation broth in significant amount. This revised version was published online in July 2006 with corrections to the Cover Date.