Nitrification–denitrification is an environmentally friendly and cost-effective way to treat landfill leachates. Special attention has been given to the nitrification step, usually the limiting one due to its special sensitivity to environmental factors. Here, the effect of the acclimatization of the nitrifying biomass to two different intermediate landfill leachates with different salt concentrations, COD and BOD₅ has been studied. Despite the complete nitrification being successfully performed, the specific nitritation rates were reduced after the biomass adaptation to both landfill leachates caused by the presence of heavy metals and the high salt concentration. NGS analysis of the biomass samples revealed that Proteobacteria (48.5%), Actinobacteriota (14.4%) and Chloroflexi (9.5%) were the dominant phyla in the non-adapted biomass. The leachate feeding led to a decrease in OTU diversity and favored the growth of the phyla Bacteroidetes (27.2%), Euryarchaeota (26.6%) and Proteobacteria (20.0%) accounting for more than 70% of relative abundance. Several OTUs capable of performing the nitritation belong to the Xanthobacteraceae and the Xanthomonadaceae families, the Saccharimonadales order, and the genus Nitrosomonas, Nitrosospira and Paracoccus. In the nitratation process, the Xanthobacteraceae family and Lautropia and Nitrolancea genera were found. Graphical Abstract

Earth’s climate is warming as a result of anthropogenic emissions of greenhouse gases from fossil fuel combustion. Bioenergy, which includes biodiesel, biohydrogen and bioethanol, has emerged as a sustainable alternative fuel source. For this reason, in recent years biodiesel production has become widespread but this industry currently generates a huge amount of glycerol as a by-product, which has become an environmental problem in its own right. A feasible possibility to solve this problem is the use of waste glycerol as a carbon source for microbial transformation into biofuels such as hydrogen and ethanol. For instance, Escherichia coli is a microorganism that can synthesize these compounds under anaerobic conditions. In this work an experimental procedure was established for screening E. coli single mutants to identify strains with enhanced ethanol and/or H2 productions compared to the wild type strain. In an initial screening of 150 single mutants, 12 novel strains (gnd, tdcE, rpiA nanE, tdcB, deoB, sucB, cpsG, frmA, glgC, fumA and gadB) were found to provide enhanced yields for at least one of the target products. The mutations, that improve most significantly the parameters evaluated (gnd and tdcE genes), were combined with other mutations in three engineered E. coli mutant strains in order to further redirect carbon flux towards the desired products. This methodology can be a useful tool to disclose the metabolic pathways that are more susceptible to manipulation in order to obtain higher molar yields of hydrogen and ethanol using glycerol as main carbon source in multiple E. coli mutants.

The development of microalgae uses requires further investigation in cell disruption alternatives to reduce the costs associated to this processing stage. This study aimed to evaluate the main variables affecting an extraction method to obtain protein and amino acids from microalgae. The method was based on a sequential alkaline-enzymatic process, with separate extractions and noncontrolled pH, and was applied to fresh biomass of a resistant species. The processed microalgae were composed of a consortium withNannochloropsissp. as predominant species. After the optimization of the pH of the alkaline reaction, the effect of the time of the alkaline reaction (30–120 min), the time (30–120 min) and temperature (40–60°C) of the enzymatic reaction, and the biomass concentration (50–150 mg·ml⁻¹), on the extraction yields of protein and free amino nitrogen (FAN) and on the final concentration of protein in the extract, was studied using a response surface methodology. Even though all the variables and some interactions among them had a significant effect, the biomass concentration was the most important factor affecting the overall process. The results showed relevant information about the different options in order to maximize not only the response variables individually but also different combinations of them. Assays with optimized values reached maximum yields of 80.3% and 1.07% of protein (% of total protein) and FAN (% of total biomass), respectively, and a protein concentration in the extract of 15.2 mg·ml⁻¹. The study provided the essential information of an alternative approach to obtain protein and amino acids from fresh biomass of resistant microalgae with a high yield, also opening perspectives for further research in particular aspects.