Search Results (2,338)

Human liver subcellular fractions, including liver microsomes (HLM), liver cytosol fractions, and S9 fractions, are extensively utilized in in vitro assays to predict liver metabolism. The S9 fractions are supernatants of human liver homogenates that contain both microsomes and cytosol, which include most cytochrome P450 (CYP) enzymes and soluble phase II enzymes such as glucuronosyltransferases and sulfotransferases. This study reports on the direct electrochemistry and biocatalytic features of redox-active enzymes in S9 fractions for the first time. We investigated the electrochemical properties of S9 films by immobilizing them onto a high-purity graphite (HPG) electrode and performing cyclic voltammetry under anaerobic (Ar-saturated) and aerobic (O₂-saturated) conditions. The heterogeneous electron transfer rate between the S9 film and the HPG electrode was found to be 14 ± 3 s⁻¹, with a formal potential of −0.451 V vs. Ag/AgCl reference electrode, which confirmed the electrochemical activation of the FAD/FMN cofactor containing CYP450-reductase (CPR) as the electron receiver from the electrode. The S9 films have also demonstrated catalytic oxygen reduction under aerobic conditions, identical to HLM films attached to similar electrodes. Additionally, we investigated CYP activity in the S9 biofilm for phase I metabolism using diclofenac hydroxylation as a probe reaction and identified metabolic products using liquid chromatography–mass spectrometry (LC-MS). Investigating the feasibility of utilizing liver S9 fractions in such electrochemical assays offers significant advantages for pharmacological and toxicological evaluations of new drugs in development while providing valuable insights for the development of efficient biosensor and bioreactor platforms. Full article

A novel, user-friendly bioreactor for the cultivation of cellularised membranes for tissue engineering has been successfully designed, manufactured, and validated. This bioreactor features a culture vessel and a cover, the latter equipped with one or more sidewalls to ensure airtightness in two distinct zones, thereby maintaining sterile conditions. The cover, designed to integrate seamlessly with the culture vessel, includes several ports compatible with commercial connectors. This design allows the introduction of cells and culture medium without requiring the opening of the cover, thus preserving sterility. Additionally, the cover is equipped with flanges that effectively press the membrane against the bottom surface of the culture vessel, preventing it from shrinking or shifting. This ensures that cells can properly adhere to the membrane and proliferate. Manufactured under Good Manufacturing Practice (GMP) conditions, the bioreactor supports cultivation in optimal aseptic environments, thereby preventing external contamination. This feature is critical for the safe transportation of cultivated tissue to clinical settings. Validation tests have confirmed the bioreactor’s excellent performance, endorsing its suitability for intended applications in tissue engineering. Full article

The swine industry generates millions of gallons (thousands of cubic meters) of wastewater every day, posing significant environmental risk due to high concentrations of organics and nutrients. This study aims to investigate the effectiveness of attached-growth anaerobic bioreactors for treating swine wastewater by utilizing sulfate-reducing bacteria, focusing on the impact of chemical oxygen demand (COD)/sulfate mass ratios on organics degradation. A series of lab-scale anaerobic bioreactors were employed to treat swine wastewater for a 14-day period. The study evaluated changes in pH, acidity, alkalinity, COD, sulfate, and various nutrients along with total suspended solids (TSS) and volatile suspended solids (VSS) before and after treatment. At a COD/sulfate mass ratio of 2:1, the bioreactors achieved optimum removal efficiencies of 80% for TSS, 83% for VSS, 86–88% for COD, 82–87% for sulfate, 73% for sulfide, and 73% for sulfite. The nutrient removal efficiency was 67% for nitrate and 72% for nitrite. The acidity and alkalinity were effectively controlled, with alkalinity values reaching up to 2161 ± 92.5 mg/L and pH within the range of 7–7.24. The findings demonstrated that anaerobic bioreactor at a COD/sulfate mass ratio of 2:1 significantly enhanced the degradation of organic matter coupling with sulfate reduction in swine wastewater, providing an efficient and sustainable treatment method. Full article

In the green alga Chlamydomonas reinhardtii, hydrogen production is catalyzed via the [FeFe]-hydrogenases HydA1 and HydA2. The electrons required for the catalysis are transferred from ferredoxin (FDX) towards the hydrogenases. In the light, ferredoxin receives its electrons from photosystem I (PSI) so that H₂ production becomes a fully light-driven process. HydA1 and HydA2 are highly O₂ sensitive; consequently, the formation of H₂ occurs mainly under anoxic conditions. Yet, photo-H₂ production is tightly coupled to the efficiency of photosynthetic electron transport and linked to the photosynthetic control via the Cyt b₆f complex, the control of electron transfer at the level of photosystem II (PSII) and the structural remodeling of photosystem I (PSI). These processes also determine the efficiency of linear (LEF) and cyclic electron flow (CEF). The latter is competitive with H₂ photoproduction. Additionally, the CBB cycle competes with H₂ photoproduction. Consequently, an in-depth understanding of light-driven H₂ production via photosynthetic electron transfer and its competition with CO₂ fixation is essential for improving photo-H₂ production. At the same time, the smart design of photo-H₂ production schemes and photo-H₂ bioreactors are challenges for efficient up-scaling of light-driven photo-H₂ production. Full article

Waste cooking oil is a common byproduct in the culinary industry, often posing disposal challenges. This study explores its conversion into the valuable bioplastic material, medium-chain-length polyhydroxyalkanoate (mcl-PHA), through microbial biosynthesis in controlled bioreactor conditions. Twenty-four bacterial isolates were obtained from oil-contaminated soil and waste materials in Mahd Ad-Dahab, Saudi Arabia. The best PHA-producing isolates were identified via 16S rDNA analysis as Neobacillus niacini and Metabacillus niabensis, with the sequences deposited in GenBank (accession numbers: PP346270 and PP346271). This study evaluated the effects of various carbon and nitrogen sources, as well as environmental factors, such as pH, temperature, and shaking speed, on the PHA production titer. Neobacillus niacini favored waste cooking oil and yeast extract, achieving a PHA production titer of 1.13 g/L, while Metabacillus niabensis preferred waste olive oil and urea, with a PHA production titer of 0.85 g/L. Both strains exhibited optimal growth at a neutral pH of 7, under optimal shaking -flask conditions. The bioreactor performance showed improved PHA production under controlled pH conditions, with a final titer of 9.75 g/L for Neobacillus niacini and 4.78 g/L for Metabacillus niabensis. Fourier transform infrared (FT-IR) spectroscopy and gas chromatography–mass spectrometry (GC-MS) confirmed the biosynthesized polymer as mcl-PHA. This research not only offers a sustainable method for transforming waste into valuable materials, but also provides insights into the optimal conditions for microbial PHA production, advancing environmental science and materials engineering. Full article

This study investigates the densification/granulation of activated sludge with poor settleability, treating real industrial wastewater from a tank truck cleaning company. The wastewater is low in nutrients, acidic in nature, and high and variable in chemical oxygen demand (COD, ranging from 2770 mg·L⁻¹ to 14,050 mg·L⁻¹). A microbial selection strategy was applied to promote slow-growing glycogen-accumulating microorganisms (GAO) by the implementation of an anaerobic feast/aerobic famine strategy in a sequencing batch reactor (SBR). After 60 to 70 days, the uptake of carbon during the anaerobic phase exceeded 80%, the sludge morphology improved, and the sludge volume index (SVI) dropped below 50 mL·g⁻¹. 16S rRNA gene sequencing showed the enrichment of the GAOs Defluviicoccus and Candidatus Competibacter. Stable sludge densification was maintained when using a constant organic loading rate (OLR) of 0.85 ± 0.05 gCOD·(L·d)⁻¹, but the sludge quality deteriorated when switching to a variable OLR. In view of the integration of densified/granular sludge in a membrane bioreactor configuration, the filtration properties of the densified SBR sludge were compared to the seed sludge from the full-scale plant. The densified sludge showed a significantly lower resistance due to pore blockage and a significantly higher sustainable flux (45 vs. 15 L·(m²·h)⁻¹). Full article

Corn steep liquor is a waste product from the process of treating corn grain for starch extraction. It is used as a substrate in anaerobic digestion with simultaneous hydrogen and methane production in a cascade of two anaerobic bioreactors. For process research and optimisation, adequate mathematical models are required. So, the authors aim to present a high-quality model of the corn steep liquor process for the sequential production of H₂ and CH₄. This paper proposes a technique for identifying the best mathematical model of the process using the metaheuristics crow search algorithm (CSA). The CSA was applied for the first time to mathematical modelling of the considered two-stage anaerobic digestion process, using real experimental data. Based on the analysis of the numerical data from the model parameter identification procedures, the influence of the main CSA parameters—the flight length, fl, and the awareness probability, AP—was investigated. Applying classical statistical tests and an innovative approach, InterCriteria Analysis, recommendations about the optimal CSA parameter tuning were proposed. The best CSA algorithm performance was achieved for the AP = 0.05, fl = 3.0, followed by AP = 0.10, fl = 2.5, and AP = 0.15, fl = 3.0. The optimal tuning of the CSA parameters resulted in a 29% improvement in solution accuracy. As a result, a mathematical model of the considered two-stage anaerobic digestion process with a high degree of accuracy was developed. Full article

Microbial degradation represents an eco-friendly alternative to traditional physicochemical treatments in removing persistent and toxic environmental pollutants, including synthetic dyes (i.e., methylene blue, MB) employed in different industries. The exploitation of thermophilic bacilli, such as those isolated from the shallow hydrothermal vents of the Eolian Islands (Italy), could provide valuable resources for the treatment of warm, dye-containing wastewater. In this study, we evaluated the ability of preformed biofilms on polypropylene perforated balls (BBs) of fifteen thermophilic bacilli, to decolor, degrade, and detoxify MB in aqueous solutions. Among them, BBs of Bacillus licheniformis B3-15 and Bacillus sp. s7s-1 were able to decolorize MB more than 50% in saline solution (NaCl 2%), incubated in static conditions at 45 °C for 48 h. At optimized initial conditions (10 mg L⁻¹ MB, pH 5.2 for B3-15 or pH 4 for s7s-1), the two strains enhanced their decolorization potential, reaching 96% and 67%, respectively. As indicated by ATR-FTIR spectroscopy, the treatment with BB B3-15 was the most efficient in degrading the Cl–C and –NH groups of MB. This degraded solution was 40% less toxic than undegraded MB, and it has no impact on the bioluminescence of Vibrio harveyi, nor the growth of the marine diatom Phaeodactylum tricornutum. Biofilm formed by strain B3-15 on polypropylene perforated balls could be proposed as a component of bioreactors in the treatment of warm, dye-containing wastewater to concomitantly remediate MB pollution and simultaneously counteract harmful effects in aquatic environments. Full article

Weeds cause significant agricultural losses worldwide, and herbicides have traditionally been the main solution to this problem. However, the extensive use of herbicides has led to multiple cases of weed resistance, which could generate an increase in the application concentration and consequently a higher persistence in the environment, hindering natural degradation processes. Consequently, more environmentally friendly alternatives, such as microbial bioherbicides, have been sought. Although these bioherbicides are promising, their efficacy remains a challenge, as evidenced by their limited commercial and industrial production. This article reviews the current status of microbial-based bioherbicides and highlights the potential of cell-free metabolites to improve their efficacy and commercial attractiveness. Stirred tank bioreactors are identified as the most widely used for production-scale submerged fermentation. In addition, the use of alternative carbon and nitrogen sources, such as industrial waste, supports the circular economy. Furthermore, this article discusses the optimization of downstream processes using bioprospecting and in silico technologies to identify target metabolites, which leads to more precise and efficient production strategies. Bacterial bioherbicides, particularly those derived from Pseudomonas and Xanthomonas, and fungal bioherbicides from genera such as Alternaria, Colletotrichum, Trichoderma and Phoma, show significant potential. Nevertheless, limitations such as their restricted range of action, their persistence in the environment, and regulatory issues restrict their commercial availability. The utilization of cell-free microbial metabolites is proposed as a promising solution due to their simpler handling and application. In addition, modern technologies, including encapsulation and integrated management with chemical herbicides, are investigated to enhance the efficacy and sustainability of bioherbicides. Full article

Tanneries generate copious amounts of potentially toxic sludge and effluent from the processing of skins and hides to leather. The effluent requires remediation before discharge to protect the receiving environment. A range of physicochemical methods are used for pre- and post-treatment, but biological secondary remediation remains the most popular choice for the reduction of the organic and macronutrient fraction of tannery effluent. This review provides an update and critical discussion of biological systems used to remediate tannery effluent. While the conventional activated sludge process and similar technologies are widely used by tanneries, they have inherent problems related to poor sludge settling, low removal efficiencies, and high energy requirements. Treatment wetlands are recommended for the passive polishing step of beamhouse effluent. Hybrid systems that incorporate anoxic and/or anaerobic zones with sludge and/or effluent recycling have been shown to be effective for the removal of organics and nitrogenous species at laboratory scale, and some have been piloted. Novel systems have also been proposed for the removal and recovery of elemental sulfur and/or energy and/or process water in support of a circular economy. Full-scale studies showing successful long-term operation of such systems are now required to convince tanneries to modernize and invest in new infrastructure. Full article

This article considers the task of developing mathematical models and their computer implementation that would establish the dependence of pH (acidity of the environment) on the volume and structure of raw materials for daily loading, as well as on the operating parameters of temperature and humidity based on the interval analysis of experimental data obtained during BGP research of a given type. In the process of research, based on the developed interval models, it was established that this indicator depends on the volume and structure of raw materials, as well as on the temperature and humidity of the substrate in the bioreactor. To build this mathematical model, it is proposed to use the method of interval data analysis and the method of identification of model parameters based on multidimensional optimization. The results of experimental studies for a specific type of biogas plant are given, and interval models with guaranteed prognostic properties that characterize the pH of the environment depending on the specific type of bio-raw material of solid and liquid fractions, temperature, and humidity are obtained. Based on the use of different types of raw materials, the developed models, based on experimental data, describe different configurations of the structure and volumes of raw materials for daily loading. The obtained mathematical models are an algebraic nonlinear equation that can be applied to control the level of pH of the environment in the bioreactor by determining the optimal volumes of raw materials of each type during the loading period depending on the temperature and humidity of the substrate in the bioreactor. Full article

Antibiotic Resistance Genes (ARGs) are contaminants of emerging concern with marked potential to impact public and environmental health. This review focusses on factors that influence the presence, abundance, and dissemination of ARGs within Wastewater Treatment Plants (WWTPs) and associated effluents. Antibiotic-Resistant Bacteria (ARB) and ARGs have been detected in the influent and the effluent of WWTPs worldwide. Different levels of wastewater treatment (primary, secondary, and tertiary) show different degrees of removal efficiency of ARGs, with further differences being observed when ARGs are captured as intracellular or extracellular forms. Furthermore, routinely used molecular methodologies such as quantitative polymerase chain reaction or whole genome sequencing may also vary in resistome identification and in quantifying ARG removal efficiencies from WWTP effluents. Additionally, we provide an overview of the One Health risk assessment framework, as well as future strategies on how WWTPs can be assessed for environmental and public health impact. Full article

The aim of the study was to compare the variable costs of planting material production using the example of vitro cultures of Pennisetum × advena ‘Rubrum’. In the study, temporary immersion system (TIS)- and agar-based methods were used in innovative workday organisation. The workday structure involved a six-hour passaging period followed by a two-hour break for medium preparation, autoclaving, and maintenance tasks. The TIS was found to be more cost-effective than the agar cultures, with lower labour costs and comparable growing expenses. The most expensive element of agar production was labour which was 43% of the costs. The second biggest cost was materials and reagents which represented 25%. In a TIS, production materials and reagents are the most expensive part of production (44%), while labour represents 24% of costs. A TIS offers a much faster multiplication of plants than agar cultures. Plants obtained in the multiplication phase are two times cheaper using a TIS. Rooting accounted for a significant portion of production costs in both methods. Overall, the TIS demonstrated superior efficiency and cost-effectiveness compared to agar cultures in producing Pennisetum × advena ‘Rubrum’ plants. Full article

This article summarizes the authors’ experiences in the development and application of the General Dynamical Model Approach related to adaptive linearizing control of biotechnological processes. Special attention has been given to some original, innovative solutions in model-based process control theory: new formalization of biotechnological process kinetics, derivation and tuning of the general software sensor of the full kinetics of biotechnological processes, and a general algorithm for fully adaptive linearizing control with software sensors. These theoretical solutions are the basis of three control strategies—fully adaptive control of the main substrate, partially adaptive control of intermediate metabolite, and recognition and stabilization of the desired physiological state based on the proposed theoretical solutions. Each strategy is illustrated in different case studies. The advantages and limitations of each of them are identified and discussed. The derived algorithms for monitoring and controlling the considered biotechnological processes are realized and included in a software platform named Interactive System for Education in Modelling and Control of Bioprocesses (InSEMCoBio). The InSEMCoBio modules and their main functions are discussed. The effectiveness of the proposed control strategies (achieving maximum productivity) has been proven through a series of simulation investigations of the considered case studies. Full article

Cellobionic acid (CBA) can be obtained through the oxidation of cellobiose, the monomer of cellulose. CBA serves as a plant-based alternative to its stereoisomer lactobionic acid, which is used in the pharmaceutical, cosmetic, and food industries. Gluconobacter oxydans is a well-established whole-cell biocatalyst with membrane-bound dehydrogenases (mDH) for regio-specific oxidations. As G. oxydans wildtype cells show low cellobiose oxidation activities, the glucose mDH from Pseudomonas taetrolens was overexpressed in G. oxydans BP9, a multi mDH deletion strain. Whole-cell biotransformation studies were performed with resting cells of the engineered G. oxydans in stirred tank bioreactors. Initial biomass specific cellobionate formation rates increased with increasing cellobiose concentrations up to 190 g L⁻¹, and were constant until the solubility limit. The maximal volumetric CBA formation rates and the oxygen uptake rates increased linearly with the concentration of engineered G. oxydans. This enables the estimation of the maximum biocatalyst concentration limited by the maximum oxygen transfer rate of any bioreactor. Thus, 5.2 g L⁻¹ G. oxydans was sufficient to produce 502 g L⁻¹ CBA with >99% yield in a simple aerobic batch process. The highly concentrated CBA will reduce downstream processing costs considerably after cell separation. Full article