Main Variables Affecting a Chemical-Enzymatic Method to Obtain Protein and Amino Acids from Resistant Microalgae

he 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 with Nannochloropsis sp. as predominant species. After the optimization of the pH of the alkaline reaction, the effect of the time of the alkaline reaction (30-120min), the time (30-120min) and temperature (40-60 degrees C) of the enzymatic reaction, and the biomass concentration (50-150mgml(-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 15.2mgml(-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

Feedback and Feedforward Control of a Biotrickling Filter for H2S Desulfurization with Nitrite as Electron Acceptor

Biotrickling filters’ control for H2S removal has special challenges because of complexity of the systems. Feedback and feedforward control were implemented in an anoxic biotrickling filter, operated in co-current flow mode and using nitrite as an electron acceptor. The feedback controller was tuned by three methods—two based on Ziegler-Nichols’ rules (step-response and maintained oscillation) and the third using the Approximate M-constrained Integral Gain Optimization (AMIGO). Inlet H2S staircase step perturbations were studied using a feedforward control and the e ect of EBRT considered by feedback control. The tuning method by maintained oscillation shows the lower errors. The selected controller was a PI, because unstable behavior at the lowest H2S inlet loading was found under a PID controller. The PI control was able to maintain an outlet H2S concentration of 14.7 0.45 ppmV at three EBRT, studied at 117 s, 92 s and 67 s. Therefore, desulfurized biogas could be used to feed a fuel cell. Feedforward control enhances BTF performance compared to the system without control. The maximum outlet H2S concentration was reduced by 26.18%, although sulfur selectivity did not exceed 55%, as elemental sulfur was the main oxidation product

Influence of Leachate and Nitrifying Bacteria on Photosynthetic Biogas Upgrading in a Two-Stage System

Photosynthetic biogas upgrading using two-stage systems allows the absorption of carbon dioxide (CO2) in an absorption unit and its subsequent assimilation by microalgae. The production of microalgae requires large amounts of nutrients, thus making scale-up difficult and reducing economic feasibility. The photosynthetic process produces oxygen (O-2) (1 mol per mol of CO2 consumed), which can be desorbed into purified biogas. Two-stage systems reduce its impact but do not eliminate it. In this study, we test the use of landfill leachate as a nutrient source and propose a viable and economical strategy for reducing the O-2 concentration. First, the liquid/gas (L/G) ratio and flow mode of the absorber were optimized for 20% and 40% CO2 with COMBO medium, then landfill leachate was used as a nutrient source. Finally, the system was inoculated with nitrifying bacteria. Leachate was found to be suitable as a nutrient source and to result in a significant improvement in CO2 absorption, with outlet concentrations of 0.01% and 0.6% for 20% and 40% CO2, respectively, being obtained. The use of nitrifying bacteria allowed a reduction in dissolved oxygen (DO) concentration, although it also resulted in a lower pH, thus making CO2 uptake slightly more difficult

Integration of a nitrification bioreactor and an anoxic biotrickling filter for simultaneous ammonium-rich water treatment and biogas desulfurization

A preliminary assessment has been carried out on the integration of an anoxic biotrickling filter and a nitrification bioreactor for the simultaneous treatment of ammonium-rich water and H2S contained in a biogas stream. The nutrient consumption in the biotrickling filter was as follows (mol−1 NO3−-N): 6.3·10−4 ± 1.2·10−4 mol PO43−-P, 0.04 ± 0.05 mol NH4+-N and 0.04 ± 0.03 mol K+-K. Furthermore, it was possible to supply a mixture of biogenic NO3− and NO2− into the biotrickling filter from the nitrification bioreactor to obtain a maximum elimination capacity of 152 gH2S–S m−3 h−1. The equivalence between the two compounds was 1 mol NO3−-N equal to 1.6 mol NO2−-N. The biotrickling filter was also operated under a stepped variable inlet load (30–100 gH2S–S m−3 h−1) and outlet H2S concentrations of less than 150 ppmV were obtained. It was also possible to maintain the outlet H2S concentration close to 15 ppmV with a feedback controller by manipulating the feed flow (in the nitrification bioreactor). Two stepped variable inlet loads were tested (60–111 and 16–102 gH2S–S m−3 h−1) under this type of control. The implementation of feedback control could enable the exploitation of biogas in a fuel cell, since the H2S concentrations were 15.1 ± 4.3 and 15.0 ± 3.4 ppmV. Finally, the anoxic biotrickling filter experienced partial denitrification and this implied a loss of the desulfurization effectiveness related to SO42− production. © 2021 The AuthorsThe Spanish Government ( Ministerio de Economía y Competitividad ) and European FEDER funds provided financial support through the project CTM2012-37927-C03 ‘Monitoring, modelling and control towards the optimization of anoxic and aerobic desulfurizing biotrickling filters’

Recent advances in biological technologies for anoxic biogas desulfurization.

Recovery of the energy contained in biogas will be essential in coming years to reduce greenhouse gas emissions and our current dependence on fossil fuels. The elimination of H2S is a priority to avoid equipment corrosion, poisoning of catalytic systems and SO2 emissions in combustion engines. This review describes the advances made in this technology using fixed biomass bioreactors (FBB) and suspended growth bioreactors (SGB) since the first studies in this field in 2008. Anoxic desulfurization has been studied mainly in biotrickling filters (BTF). Elimination capacities (EC) up to 287 gS m-3 h-1 have been achieved, with a removal efficiency (RE) of 99%. Both nitrate and nitrite have been successfully used as electron acceptor. SGBs can solve some operational problems present in FBBs, such as clogging or nutrient distribution issues. However, they present greater difficulties in gas-liquid mass transfer, although ECs of up to 194 gS m-3 h-1 have been reported in both gas-lift and stirred tank reactors. One of the major disadvantages of using anoxic biodesulfurization compared to aerobic biodesulfurization is the need to provide reagents (nitrates and/or nitrites), with the consequent increase in operating costs. A solution proposed in this respect is the use of nitrified effluents, some ammonium-rich effluents nitrified include landfill leachate and digested effluent from the anaerobic digester have been tested successfully. Among the microbial diversity found in the bioreactors, the genera Thiobacillus, Sulfurimonas and Sedimenticola play a key role in anoxic removal of H2S. Finally, a summary of future trends in technology is provided

Optimization of culture media for etanol production from glicerol by Escherichia coli

http://dx.doi.org/10.1016/j.biombioe.2011.12.002The culture media for glycerol to ethanol biotransformation by Escherichia coli under anaerobic conditions was optimized. A Plackett-Burman screening design was used to determine which factors were significant with a 95% confidence interval. A full factorial 22 and response surface model were employed to determine the optimum conditions for the selected response variable. The response variable considered in this work was biomass productivity (QX). Profiles for biomass growth, glycerol consumption and ethanol production were obtained at optimum conditions and fermentation parameters were calculated. Glycerol to ethanol yield and ethanol specific productivity were determined to be 845 g kg 1 of glycerol and 212 g kg 1 h 1 of cell mass respectively. Optimized culture media is presented along with the main results and experimental profiles obtained from this work

Methane concentration and bacterial communities' dynamics during the anoxic desulfurization of landfill biogas under diverse nitrate sources and hydraulic residence times

Landfill biogas contains certain amounts of H2S that must be removed in order to prevent both equipment corrosion and SO2 emissions to the atmosphere when burnt. Anoxic desulfurization has been proven to be an eco-friendly and cost-efficient method to remove H2S from biogas. Nevertheless, and despite all the reported benefits, the potential consumption of methane (CH4) during the anoxic desulfurization of landfill biogas is a factor that has not yet been thoroughly investigated. The present study evaluates the microbial composition and methane assimilation activity of three microbial samples obtained when feeding different nitrate sources, namely nitrified landfill leachate (M1) or chemical nitrate (M2, M3) with 10 days (M2) and 1.5 days (M3) hydraulic residence times. The samples were characterized by the prevalence of sulfide oxidizing bacteria [Thiomicrospira (11.4-25.5 %), Family Rhodobacteraceae (9.9-14.3 %), Sulfurimonas (0.34-17.9 %), Thioclava (0-23.5 %) and Arcobacter (0-11.5 %)], as well as the presence of methane oxidizing bacteria [Halomonas (0.2-16.0 %), Methylophaga (0-0.2 %) and Methylophilacea (0-0.1 %)] and heterotrophic bacteria [Lentimicrobium (0.1-9.7 %) and Roseovarius (0.1-1.2 %)]. The highest CH4 assimilation levels were reached under anoxic conditions at 34.0 and 50.1 g CH4 m-3 h-1 using nitrate and nitrite, respectively. The oxygen present in the landfill biogas itself had a detrimental effect on the anoxic bioreactor nitrate removal efficiency. The presence of organic matter in the nitrified influent gave rise to CH4 inside the anoxic desulfurization bioreactors, which resulted in the offsetting of the CH4 oxidation caused by methane-oxidizing bacteria (MOB). © 2023 The Authors

Characterization of Bacterial and Archaeal Communities by DGGE and Next Generation Sequencing (NGS) of Nitrification Bioreactors Using Two Different Intermediate Landfill Leachates as Ammonium Substrate

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.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. The Spanish Government (Ministry of Economy and Competitiveness) and the Vice-rectorate for Research of the University of Cadiz provided financial support through the project CTM2016-79089-R "Enhancement of landfill gas by an integrated biological system (Biointegra3)" and UCA/REC01VI/2017 (Universidad de Cadiz) respectivel

Study of the role played by NfsA, NfsB nitroreductase and NemA flavin reductase from Escherichia coli in the conversion of ethyl 2-(2′-nitrophenoxy)acetate to 4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one (D-DIBOA), a benzohydroxamic acid with interesting biological properties

Benzohydroxamic acids, such as 4-hydroxy-(2H)- 1,4-benzoxazin-3(4H)-one (D-DIBOA), exhibit interesting herbicidal, fungicidal and bactericidal properties. Recently, the chemical synthesis of D-DIBOA has been simplified to only two steps. In a previous paper, we demonstrated that the second step could be replaced by a biotransformation using Escherichia coli to reduce the nitro group of the precursor, ethyl 2-(2′-nitrophenoxy)acetate and obtain D-DIBOA. The NfsA and NfsB nitroreductases and the NemA xenobiotic reductase of E. coli have the capacity to reduce one or two nitro groups from a wide variety of nitroaromatic compounds, which are similar to the precursor. By this reason, we hypothesised that these three enzymes could be involved in this biotransformation. We have analysed the biotransformation yield (BY) of mutant strains in which one, two or three of these genes were knocked out, showing that only in the double nfsA/nfsB and in the triple nfsA/nfsB/nemA mutants, the BY was 0%. These results suggested that NfsA and NfsB are responsible for the biotransformation in the tested conditions. To confirm this, the nfsA and nfsB open reading frames were cloned into the pBAD expression vector and transformed into the nfsA and nfsB single mutants, respectively. In both cases, the biotransformation capacity of the strains was recovered (6.09±0.06% as in the wild-type strain) and incremented considerably when NfsA and NfsB were overexpressed (40.33%±9.42% and 59.68%±2.0% respectively)

Anoxic biogas biodesulfurization promoting elemental sulfur production in a Continuous Stirred Tank Bioreactor

Biological desulfurization of biogas has been extensively studied using biotrickling filters (BTFs). However, the accumulation of elemental sulfur (S0) on the packing material limits the use of this technology. To overcome this issue, the use of a continuous stirred tank bioreactor (CSTBR) under anoxic conditions for biogas desulfurization and S0 production is proposed in the present study. The effect of the main parameters (stirring speed, N/S molar ratio, hydraulic residence time (HRT) and gas residence time (GRT)) on the bioreactor performance was studied. Under an inlet load (IL) of 100 g S-H2S m–3 h–1 and a GRT of 119 s, the CSTBR optimal operating conditions were 60 rpm, N/S molar ratio of 1.1 and a HRT of 42 h, in which a removal efficiency (RE) and S 0 production of 98.6 ± 0.4% and 88% were obtained, respectively. Under a GRT of 41s and an IL of 232 g S-H2S m–3 h–1 the maximum elimination capacity (EC) of 166.0 ± 7.2 g S-H2S m–3 h–1 (RE = 71.7 ± 3.1%) was obtained. A proportional-integral feedback control strategy was successfully applied to the bioreactor operated under a stepped variable IL