Improving bioreactor production of a recombinant glycoprotein in Escherichia coli: Effect of specific growth rate on protein glycosylation and specific productivity

In the last decades bacterial glycoengineering emerged as a new field as the result of the ability to transfer the Campylobacter jejuni N- glycosylation machinery into Escherichia coli for the production of recombinant glycoproteins that can be used as antigens for diagnosis, vaccines, and therapeutics. However, the identification of critical parameters implicated in the production process and its optimization to jump to a productive scale is still required. In this study, we developed a dual expression glycosylation vector for the production of the recombinant glycoprotein AcrA-O157, a novel antigen that allows the serodiagnosis of the infection with enterohemorrhagic E. coli O157 in humans. Volumetric productivity was studied in different culture media and found that 2xYP had 6.9-fold higher productivity than the extensively used LB. Subsequently, bioreactor batch and exponential-fed-batch cultures were designed to determine the influence of the specific growth rate (μ) on AcrA-O157 glycosylation efficiency, production kinetics, and specific productivity. At μ_max, AcrA glycosylation with O157-polysaccharide and the specific synthesis rate were maximal, constituting the optimal physiological condition for AcrA-O157 production. Our findings should be considered for the design, optimization, and scaling up of AcrA-O157 production and other recombinant glycoproteins attractive for industrial applications.     

Modulation of the central carbon metabolism of Corynebacterium glutamicum improves malonyl-CoA availability and increases plant polyphenol synthesis

In recent years microorganisms have been engineered towards synthesizing interesting plant polyphenols such as flavonoids and stilbenes from glucose. Currently, the low endogenous supply of malonyl-CoA, indispensable for plant polyphenol synthesis, impedes high product titers. Usually, limited malonyl-CoA availability during plant polyphenol production is avoided by supplementing fatty acid synthesis-inhibiting antibiotics such as cerulenin, which are known to increase the intracellular malonyl-CoA pool as a side effect. Motivated by the goal of microbial polyphenol synthesis being independent of such expensive additives, we used rational metabolic engineering approaches to modulate regulation of fatty acid synthesis and flux into the tricarboxylic acid cycle (TCA cycle) in Corynebacterium glutamicum strains capable of flavonoid and stilbene synthesis. Initial experiments showed that sole overexpression of genes coding for the native malonyl-CoA-forming acetyl-CoA carboxylase is not sufficient for increasing polyphenol production in C. glutamicum. Hence, the intracellular acetyl-CoA availability was also increased by reducing the flux into the TCA cycle through reduction of citrate synthase activity. In defined cultivation medium, the constructed C. glutamicum strains accumulated 24 mg·L ⁻¹ (0.088 mM) naringenin or 112 mg·L ⁻¹ (0.49 mM) resveratrol from glucose without supplementation of phenylpropanoid precursor molecules or any inhibitors of fatty acid synthesis.     

Associative learning in immature lacewings (Ceraeochrysa cubana)

It is known that many social insects and arthropod predators and parasitoids can learn the association between a resource and volatile cues. Although there are various studies on the effect of experience in immature arthropods on behavior later in adult life, not much is known about the effects of such experiences on immature behavior. This was investigated here in the lacewing Ceraeochrysa cubana (Hagen) (Neuroptera: Chrysopidae). Whereas adults of this lacewing feed on plant‐provided food and honeydew, larvae are voracious polyphagous predators of several insect pests, and therefore important for biological control. Hence, studying the foraging behavior and the effects of learning in immatures of this species is important. We exposed immatures to the volatile methyl salicylate (MeSA), which was either associated with food or with the absence of food. Subsequently, their response to this volatile was tested in an olfactometer. Immatures that had experienced the association of MeSA with food were attracted to it and immatures that were exposed to MeSA during food deprivation were repelled. Subsequently, predator immatures that had experienced the association between MeSA and food were released on a plant without food and were found to use this volatile in locating patches with food. In contrast, larvae without such experience were found equally on food patches with and without the volatile. We conclude that these immature predators are capable of learning the association between volatiles and food, or the absence of food, and use this during foraging.     

Coastal zone use and migratory behaviour of the southern population of Mugil liza in Brazil

We analysed the ratios Sr:Ca and Ba:Ca in the otoliths of 55 adults of the southern population of Mugil liza in Brazil (Paraná to Rio Grande do Sul) to investigate its coastal zone use and migratory behaviour. All individual M. liza analysed had Sr:Ca and Ba:Ca values indicating that their birth was in the marine environment, which is consistent with the southern population migration to spawn in the ocean,. Juveniles exhibited at least three coastal use and recruitment strategies (contingents): the majority (89%) of M. liza juveniles migrated toward brackish water. They entered the estuary before completing the first year of life (64%) or after (25%) their first year of life. The remaining 11% did not appear to enter brackish or freshwater water as a nursery or at any point in their life cycle. Some adults returned to the estuary after spawning in the ocean but others (of both sexes) never returned to the estuary after spawning, remaining in the marine environment. The pattern of juvenile habitat use in the Brazilian southern population of M. liza seems to be recurrent throughout the extent of its distribution as a consequence of the reproductive spawning aggregation behaviour, which mixes all contingents (with marine or estuarine preferences).     

Diversity of settlement-stage reef fishes captured by light-trap in a tropical south-west Atlantic Ocean coastal reef system

This study reports the results of 5 years of monitoring reef fish post-larvae using light traps in the Bay of Tamandaré, north-east Brazil. An annotated checklist of pre-settlement fish species, their frequency of occurrence and taxonomic characteristics are provided. In total, 4,422 post-larval fishes belonging to 36 families, 56 genera and 76 species were captured. The most species-rich families were Carangidae (7), Lutjanidae (6) and Pomacentridae (4), while the families Gerreidae (30.47%), Holocentridae (16.54%), Blenniidae (12.01%), Labrisomidae (8.36%), Lutjanidae (8.29%) and Acanthuridae (5.95%) were the most abundant. This is the first study of the taxonomic diversity and assemblage structure of settlement-stage reef fishes in the tropical south-west Atlantic Ocean. Although a few common species were not captured due to selectivity of light traps, the composition and taxonomic diversity of this first collection suggests that light traps are useful for studies of the early life history of a wide range of pre-settlement reef fishes.     

Phenotypic plasticity to photon flux density of physiological,anatomical and growth traits in a modern Criollo cocoa clone

Cocoa grows under shade, but some cultivars develop successfully in full sunlight. In order to characterize the response to photosynthetic photon flux density (PPFD) of a Modern Criollo cocoa clone, gas exchange, photochemical activity and leaf traits, and their relation to growth were measured in seedlings growing in a greenhouse at three different values of PPFD, as well as in adults in full sunlight and shade in the field. Plants showed changes in physiological, biochemical, and morpho‐anatomical traits in response to the different light conditions, and in the phenotypic plasticity of these variables. Seedlings subjected to high PPFD in the greenhouse showed decreases in photosynthetic rate, apparent quantum yield of CO₂ fixation and photochemical quenching, and increases in non‐photochemical quenching, suggesting down‐regulation of PSII. In contrast, trees under full sunlight in the field showed a marked reduction in maximum quantum yield of PSII, indicating photoinhibition and supporting that cocoa is a shade tolerant crop. Cocoa showed higher plasticity of physiological and biochemical variables than morpho‐anatomical variables in response to PPFD. Effects of time under treatment in the greenhouse and plant age (greenhouse vs field) on plasticity were observed. The acclimation observed in some of the variables studied after 6 months in high light did not represent a particular advantage to seedlings, since relative growth rate was lower than in low‐ and medium‐light seedlings.     

Impact of metal accumulation on Quercus ilex L. leaf traits

Plant and Soil - The aims of this research were: i) to compare Cr, Cu, Ni and Pb concentrations in Quercus ilex L. leaves collected at urban/industrial and urban areas; ii) to investigate the main...