Applications In Environmental Biotechnology Research Articles

Inoculum source determines the stress resistance of electroactive functional taxa in biofilms: A metagenomic perspective

The inoculum has a crucial impact on bioreactor initialization and performance. However, there is currently a lack of guidance on selecting appropriate inocula for applications in environmental biotechnology. In this study, we applied microbial electrolysis cells (MECs) as models to investigate the differences in the functional potential of electroactive microorganisms (EAMs) within anodic biofilms developed from four different inocula (natural or artificial), using shotgun metagenomic techniques. We specifically focused on extracellular electron transfer (EET) function and stress resistance, which affect the performance and stability of MECs. Community profiling revealed that the family Geobacteraceae was the key EAM taxon in all biofilms, with Geobacter as the dominant genus. The c-type cytochrome gene imcH showed universal importance for Geobacteraceae EET and was utilized as a marker gene to evaluate the EET potential of EAMs. Additionally, stress response functional genes were used to assess the stress resistance potential of Geobacter species. Comparative analysis of imcH gene abundance revealed that EAMs with comparable overall EET potential could be enriched from artificial and natural inocula (P > 0.05). However, quantification of stress response gene copy numbers in the genomes demonstrated that EAMs originating from natural inocula possessed superior stress resistance potential (196 vs. 163). Overall, this study provides novel perspectives on the inoculum effect in bioreactors and offers theoretical guidance for selecting inoculum in environmental engineering applications.

Secretory Production of the Hericium erinaceus Laccase from Saccharomyces cerevisiae.

Mushroom laccases play a crucial role in lignin depolymerization, one of the most critical challenges in lignin utilization. Importantly, laccases can utilize a wide range of substrates, such as toxicants and antibiotics. This study isolated a novel laccase, named HeLac4c, from endophytic white-rot fungi Hericium erinaceus mushrooms. The cDNAs for this enzyme were 1569 bp in length and encoded a protein of 523 amino acids, including a 20 amino-acid signal peptide. Active extracellular production of glycosylated laccases from Saccharomyces cerevisiae was successfully achieved by selecting an optimal translational fusion partner. We observed that 5 and 10 mM Ca2+, Zn2+, and K+ increased laccase activity, whereas 5 mM Fe2+ and Al3+ inhibited laccase activity. The laccase activity was inhibited by the addition of low concentrations of sodium azide and L-cysteine. The optimal pH for the 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt was 4.4. Guaiacylglycerol-β-guaiacyl ether, a lignin model compound, was polymerized by the HeLac4c enzyme. These results indicated that HeLac4c is a novel oxidase biocatalyst for the bioconversion of lignin into value-added products for environmental biotechnological applications.

Graphene hydrogel improves S. putrefaciens' biological treatment of dye wastewater: Impacts of extracellular electron transfer and function of c-type cytochromes

Biocompatible materials and biocarriers have attracted great attention in biological wastewater treatment owing to their excellent performance in improving pollutant removal. Graphene-based material, a biocarrier candidate, with excellent adsorbability and conductivity was increasingly applied in anaerobic digestion due to its exceptional potential in the adsorption and electron transfer process. Nevertheless, the green approach for the formation of bio-graphene complexes and their mechanism in dye removal is limited. The aim of this study is to investigate and assess the performance of biological graphene hydrogel (BGH) formed by Shewanella putrefaciens CN32 on the removal of methyl orange (MO) and methylene blue (MB). The results showed that the formation of BGH is determined by the physicochemical characteristics of graphene oxide, including sheet size, oxidation degree, and interlayer distance. BGHs significantly increased the removal efficiency of dyes in comparison to non-graphene samples, with a 24-h removal rate of MO and MB reaching 92.9% and 91%, respectively. The synergetic mechanism of BGH on the enhanced removal rate of organic dye can be ascribed to GO's ability in accelerating extracellular electron transfer and stimulating biodegradation pathways relating to c-type cytochromes, including MtrA and MtrC. These findings provided an understanding of the relationship between graphene-based nanomaterials and Shewanella, which facilitated their future application in environmental biotechnology.

Thermophilic bacteria from Peruvian hot springs with high potential application in environmental biotechnology

ABSTRACT Hot springs are extreme environments in which well-adapted microorganisms with biotechnological applications can thrive naturally. These thermal environments across Peruvian territory have, until now, remained poorly investigated. In this study, two hot springs, El Tragadero and Quilcate, located in Cajamarca (Peru) were selected in order to investigate the biotechnological potential of indigenous thermophilic bacteria. Enrichment and isolation processes were carried out using microbial mats, sediments, biofilms, and plastic polymers as samples. Screening for biosurfactants and siderophores production, as well as for polyethylene terephthalate (PET) hydrolysis was done using culture-dependent techniques. After molecular identification, Bacillus was found as the most abundant genus in both hot springs. Bacillus velezensis was found producing biosurfactants under high-level temperature. Anoxybacillus species (A. salavatliensis and A. gonensis) are here reported as siderophore-producing bacteria for the first time. Additionally, Brevibacillus and the less-known bacterium Tistrella mobilis were found demonstrating PET hydrolysis activity. Our study provides the first report of thermophilic bacteria isolated from Peruvian hot springs with biotechnological potential for the bioremediation of oil-, metal- and plastic-polluted environments.

ПОЛИФУНКЦИОНАЛЬНЫЕ ШТАММЫ БАКТЕРИЙ-НЕФТЕДЕСТРУКТОРОВ ДЛЯ ЭКОЛОГИЧЕСКОЙ БИОТЕХНОЛОГИИ

The use of polyfunctional strains of degrading bacteria, which are simultaneously capable of decomposing pollutants, stimulating the growth and development of remediant plants, and being resistant to heavy metals, is the most promising way to increase the efficiency of bioremediation of soils contaminated with hydrocarbons. In this work, isolated from oil-contaminated soil and identified up to species using a comparative analysis of the nucleotide sequences of the 16S rRNA gene, bacterial strains of the genus Pseudomonas were able to oxidize hydrocarbons of various classes, oil and diesel fuel to end products (66-138 mg CO2/g of substrate), dissolve inorganic phosphates, and produce IAA (305-1617 ng/ml culture fluid), and are also resistant to lead ions at a concentration of 1,5-2,0 g/l. When barley seeds were inoculated with a diluted liquid culture of bacteria, the length of shoots (by 2,7-18,0%) and roots (total) (by 20,9-33,1%) increased. All strains are recommended for further study of their biotechnologically important properties with a view to possible further application in environmental biotechnology.

A Pesticide Biopurification System: A Source of Biosurfactant-Producing Bacteria with Environmental Biotechnology Applications

Biosurfactants, a wide group of compounds produced by different microorganisms, generally have less toxicity and are more biodegradable than synthetic surfactants. Biosurfactant-producing bacteria can be found in contaminated environments, such as soils receiving pesticide applications constantly, or in pesticides treatment systems where microorganisms are adapted to biodegrading pesticides. Five pesticide-tolerant bacteria previously isolated from a pesticide biopurification system were evaluated as biosurfactant-producers. Pseudomonas rhodesiae C4, Rhodococcus jialingiae C8 and Pseudomonas marginalis C9 strains were positive in qualitative tests. Biosurfactant production by these strains using Bushnell-Haas medium with olive oil at 2% (w/v) was evaluated as emulsification index, oil displacement, droplet collapse test and surface tension. After 144 h, these strains showed a similar emulsification index of >55%. The two Pseudomonas (C4 and C9) strains showed lower superficial tension compared with Rhodococcus strain (C8)—34.47, 37.44 and 47.55 mN/m for strains C4, C9 and C8, respectively. The chemical characterization of the biosurfactants revealed the presence of glycolipids in P. rhodesiae (C4) and glycopeptides in P. marginalis (C9). The degradation of chlorpyrifos increased from 39.2% to 51.6% when biosurfactants produced by P.rhodesiae (C4) were added (10%) with respect to the control. Therefore, biopurification systems are a relevant source of biosurfactant-producing bacteria with environmental biotechnology applications.

Key Enzymes for Anaerobic Lactate Metabolism in Geobacter sulfurreducens.

Growth of Geobacter sulfurreducens PCA on lactate was enhanced by laboratory adaptive evolution. The enhanced growth was considered to be attributed to increased expression of the sucCD genes, encoding a succinyl-coenzyme A (CoA) synthetase. To further investigate the function of the succinyl-CoA synthetase, the sucCD genes were deleted from G. sulfurreducens The mutant showed defective growth on lactate but not on acetate. Introduction of the sucCD genes into the mutant restored the full potential to grow on lactate. These results verify the importance of the succinyl-CoA synthetase in growth on lactate. Genome analysis of Geobacter species identified candidate genes, GSU1623, GSU1624, and GSU1620, for lactate dehydrogenase. Deletion mutants of the identified genes for d-lactate dehydrogenase (ΔGSU1623 ΔGSU1624 mutant) or l-lactate dehydrogenase (ΔGSU1620 mutant) could not grow on d-lactate or l-lactate but could grow on acetate and l- or d-lactate, respectively. Introduction of the respective genes into the mutants allowed growth on the corresponding lactate stereoisomer. These results suggest that the identified genes were essential for d- or l-lactate utilization. The lacZ reporter assay demonstrated that the putative promoter regions were more active during growth on lactate than during growth on acetate, indicating that the genes for the lactate dehydrogenases were expressed more during growth on lactate than during growth on acetate. The gene deletion phenotypes and the expression profiles indicate that there are metabolic switches between lactate and acetate. This study advances the understanding of anaerobic lactate utilization in G. sulfurreducensIMPORTANCE Lactate is a microbial fermentation product as well as a source of carbon and electrons for microorganisms in the environment. Furthermore, lactate is a common amendment for stimulation of microbial growth in environmental biotechnology applications. However, anaerobic metabolism of lactate has been poorly studied for environmentally relevant microorganisms. Geobacter species are found in various environments and environmental biotechnology applications. By employing genomic and genetic approaches, succinyl-CoA synthetase and lactate dehydrogenase were identified as key enzymes in anaerobic metabolism of lactate in Geobacter sulfurreducens, a representative Geobacter species. Differential gene expression during growth on lactate and acetate was observed, demonstrating that G. sulfurreducens could metabolically switch to adapt to available substrates in the environment. The findings provide new insights into basic physiology in lactate metabolism as well as cellular responses to growth conditions in the environment and can be informative for the application of lactate in environmental biotechnology.

Respirometric techniques coupled with laboratory-scale tests for kinetic and stoichiometric characterisation of fungal and bacterial tannin-degrading biofilms.

In environmental biotechnology applications for wastewater treatment, bacterial-based bioprocesses are mostly implemented; on the contrary, the application of fungal-based bioprocesses, is still challenging under non-sterile conditions. In a previous laboratory-scale study, we showed that when specific tannins are used as the sole carbon source, fungi can play a key role in the microbial community, under non-sterile conditions and in the long term. In a previous study, an engineered ecosystem, based on fungal tannin biodegradation, was successfully tested in a laboratory-scale bioreactor under non-sterile conditions. In the present study, a kinetic and stoichiometric characterisation of the biomass developed therein was performed through the application of respirometric techniques applied to the biomass collected from the above-mentioned reactor. To this aim, a respirometric set-up was specifically adapted to obtain valuable information from tannin-degrading fungal biofilms. A mathematical model was also developed and applied to describe both the respirometric profiles and the experimental data collected from the laboratory-scale tests performed in the bioreactor. The microbial growth was described through a Monod-type kinetic equation as a first approach. Substrate inhibition, decay rate and tannin hydrolysis process were included to better describe the behaviour of immobilised biomass selected in the tannin-degrading bioreactor. The model was implemented in AQUASIM using the specific tool Biofilm Compartment to simulate the attached fungal biofilm. Biofilm features and transport parameters were either measured or assumed from the literature. Key kinetic and stoichiometric unknown parameters were successfully estimated, overcoming critical steps for scaling-up a novel fungal-based technology for tannins biodegradation.

Nanotechnology and it’s applications in environmental remediation: an overview

In the last few decades, nanotechnology has come to the fore as a crucial and significant domain in the scientific realm owing to its multidisciplinary nature. The enhanced properties of materials in the nanoscale make them a viable option for different applications in different fields. The conventional method viz. the physical and chemical methods of nanoparticle production, however, pose hazardous risks to the environment. To redress these concerns, researchers have diverted their focus towards the more favourable green method of synthesis which is free from any toxic precursor or strenuous process conditions making it an economical and nature-friendly method. Nanoparticles showed a wide range of application in environmental biotechnology like reduction of pollution, water treatment, remediation, dye degradation and water purification development. This review focuses on the various biogenic precursors for fabrication of nanoparticles and also emphasizes their potential applications in environmental remediation.

Magnetic solid phase extractions of Co(II) and Hg(II) by using magnetized C. micaceus from water and food samples

A new bio-MSPE sorbent based on the use of C. micaceus and γ-Fe2O3 magnetic nanoparticle was prepared for the preconcentrations of Co(II) and Hg(II). Critical parameters including pH, flow rate, quantity of C. micaceus, quantity of γ-Fe2O3 magnetic nanoparticle, eluent (type, concentration and volume), sample volume, and foreign ions were examined. Surface structure and variations after interaction with Co(II) and Hg(II) of bio-MSPE sorbent were investigated by FT-IR, SEM, and EDX. The impact of bio-MSPE column reusage was also tested. The biosorption capacities were determined as 24.7 mg g−1 and 26.2 mg g−1, respectively for Co(II) and Hg(II). Certified reference materials were utilized to find out the accuracy of the prepared bio-MSPE method. This novel bio-MSPE method was accomplished by being applied to real food and water samples. In particular, it will be possible to make use of C. micaceus as new alternatives, in environmental biotechnology applications.

Geomycology: fungi as agents of biogeochemical change

Geomycology is the study of the roles of fungi in geological processes. The biogeochemical importance of fungi is significant in several areas, including nutrient and element cycling, rock, mineral and metal transformations, bioweathering and mycogenic biomineral formation. Such processes can occur in aquatic and terrestrial habitats, but it is the terrestrial environment where fungi probably have the greatest geochemical influence. Of special significance are the mutualistic relationships with phototrophic organisms: lichens (algae, cyanobacteria) and mycorrhizas (plants). Central to many geomycological processes are transformations of metals and minerals, and fungi possess many properties that effect changes in metal speciation, toxicity and mobility, as well as mineral formation or mineral dissolution or deterioration. Some fungal transformations have potential applications in environmental biotechnology, e.g. metal and radionuclide leaching, recovery, detoxification and bioremediation, and in the production or deposition of biominerals or metallic elements with catalytic or other properties. Metal and mineral transformations may also result in adverse effects when these processes result in spoilage and destruction of natural and synthetic materials, rock and mineral-based building materials (e.g. concrete), biocorrosion of metals, alloys and related substances, and adverse effects on radionuclide speciation, mobility and containment.

Glycolipid Biosurfactants, Main Classes, Functional Properties and Related Potential Applications in Environmental Biotechnology

Glycolipids, consisting of a carbohydrate moiety linked to fatty acids, are microbial surface active compounds produced by various microorganisms. Glycolipids are characterized by highly structural diversity and have the ability to decrease the surface and interfacial tension at the surface and interface respectively. It presented initially a detailed classification of glycolipid including rhamnolipids, trehalolipids, mannosylerythritol-lipids, cellobiolipids; along with their producing strain. The review described the main functional properties of glycolipid including emulsification/de-emulsification capacity, foaming and moisturizing, viscosity reduction and hydrocarbon solubilizing and mobilizing capacities. Owing these properties, they can be applied in environmental fields as hydrocarbon emulsifiers, solubilizing and mobilizing agents, for their moisturizing capabilities and ability to reduce viscosity. The review will present a detailed classification of glycolipid biosurfactants, functional properties and the potential related applications in environment and bioremediation.

Characterization of a Marine Microbial Community Used for Enhanced Sulfate Reduction and Copper Precipitation in a Two-Step Process.

Marine microorganisms that are obtained from hydrothermal vent sediments present a great metabolic potential for applications in environmental biotechnology. However, the work done regarding their applications in engineered systems is still scarce. Hence, in this work, the sulfate reduction process carried out by a marine microbial community in an upflow anaerobic sludge blanket (UASB) reactor was investigated for 190days under sequential batch mode. The effects of 1000 to 5500mgL-1 of SO4-2 and the chemical oxygen demand (COD)/SO4-2 ratio were studied along with a kinetic characterization with lactate as the electron donor. Also, the feasibility of using the sulfide produced in the UASB for copper precipitation in a second column was studied under continuous mode. The system presented here is an alternative to sulfidogenesis, particularly when it is necessary to avoid toxicity to sulfide and competition with methanogens. The bioreactor performed better with relatively low concentrations of sulfate (up to 1100mgL-1) and COD/SO4-2 ratios between 1.4 and 3.6. Under the continuous regime, the biogenic sulfide was sufficient to precipitate copper at a removal rate of 234mgL-1day-1. Finally, the identification of the microorganisms in the sludge was carried out; some genera of microorganisms identified were Desulfitobacterium and Clostridium.

Microbial interactions with chromium: basic biological processes and applications in environmental biotechnology.

Chromium (Cr) is a highly toxic metal for microorganisms as well as plants and animal cells. Due to its widespread industrial use, Cr has become a serious pollutant in diverse environmental settings. The hexavalent form of the metal, Cr(VI), is considered a more toxic species than the relatively innocuous and less mobile Cr(III) form. The study of the interactions between microorganisms and Cr has been helpful to unravel the mechanisms allowing organisms to survive in the presence of high concentrations of Cr(VI) and to detoxify and remove the oxyanion. Various mechanisms of interactions with Cr have been identified in diverse species of bacteria and fungi, including biosorption, bioaccumulation, reduction of Cr(VI) to Cr(III), and chromate efflux. Some of these systems have been proposed as potential biotechnological tools for the bioremediation of Cr pollution using bioreactors or by in situ treatments. In this review, the interactions of microorganisms with Cr are summarised, emphasising the importance of new research avenues using advanced methodologies, including proteomic, transcriptomic, and metabolomic analyses, as well as the use of techniques based on X-ray absorption spectroscopy and electron paramagnetic resonance spectroscopy.

Unexplored Brazilian oceanic island host high salt tolerant biosurfactant-producing bacterial strains.

We aimed to isolate biosurfactant-producing bacteria in high salt conditions from uncontaminated soils on the Brazilian oceanic island, Trindade. Blood agar medium was used for the isolation of presumptive biosurfactant-producing bacteria. Confirmation and measurements of biosurfactant production were made using an oil-spreading method. The isolates were identified by fatty acid profiles and partial 16S rRNA gene sequence analysis. A total of 14 isolates obtained from the 12 soil samples were found to produce biosurfactants. Among them, two isolates stood out as being able to produce biosurfactant that is increasingly active in solutions containing up to 175 g L(-1) NaCl. These high salt tolerant biosurfactant producers are affiliated to different species of the genus Bacillus. Soil organic matter showed positive correlation with the number of biosurfactant-producing bacteria isolated from our different sampling sites. The applied approach successfully recovered and identified biosurfactant-producing bacteria from non-contaminated soils. Due to the elevated salt tolerance, as well as their capacity to produce biosurfactants, these isolates are promising for environmental biotechnological applications, especially in the oil production chain.

Ecology and biotechnology of selenium-respiring bacteria.

In nature, selenium is actively cycled between oxic and anoxic habitats, and this cycle plays an important role in carbon and nitrogen mineralization through bacterial anaerobic respiration. Selenium-respiring bacteria (SeRB) are found in geographically diverse, pristine or contaminated environments and play a pivotal role in the selenium cycle. Unlike its structural analogues oxygen and sulfur, the chalcogen selenium and its microbial cycling have received much less attention by the scientific community. This review focuses on microorganisms that use selenate and selenite as terminal electron acceptors, in parallel to the well-studied sulfate-reducing bacteria. It overviews the significant advancements made in recent years on the role of SeRB in the biological selenium cycle and their ecological role, phylogenetic characterization, and metabolism, as well as selenium biomineralization mechanisms and environmental biotechnological applications.

A new two-step screening method for prospecting of trace element accumulating plants

A vulnerable point of the currently used approach to the search for the new species capable of abnormal accumulation (hyperaccumulation) of trace elements is that most studies have been conducted in laboratory conditions and focused on the determination of a limited number of elements. We propose a methodology that enables screening for multi-element accumulating plants. This methodology is based on two analytical steps: a semiquantitative analysis mode by ICP-MS that allows selection of plant samples which are enriched in one or more trace elements, and a quantitative analysis necessary for confirmation of the results derived from the first step. The proposed methodology was tested in the study of 30 plant samples. Ten elements with the highest concentrations obtained in the semiquantitative analyses were determined quantitatively with the following detection limits (in mg/kg): 0.001 for Ag, 0.08 for Ba, 0.002 for Cd, 0.005 for Co, 0.01 for Cr, 0.003 for Cu, 1.4 for Fe, 0.012 for Mn, 0.03 for Ni, 0.006 for Pb, 0.001 for Sc, 0.001 for Tl and 0.06 for Zn. The CRM recovery values obtained were in the range of 80–103 %, and the precision of the measurements (as RSD) was in the range of 0.34–4.05 %. We also propose a simple method for evaluation of typical element concentrations in plants collected for analyses. Our approach provides a novel screening method for both identification of new hyperaccumulators and for studying a larger number of elements accumulated by plants. This method may find its application in environmental biotechnology.

Stabilization of Uranium(VI) at Low pH by Fungal Metabolites: Applications in Environmental Biotechnology

Uranium contamination of soils and water is a worldwide problem due to geology or anthropogenic release such as mining, or use of inorganic fertilizers. In situ remediation of low and moderately contaminated sites is a complicated procedure due to the complex chemistry of uranium. This study demonstrates that at pH 3.5, a fungal strain isolated from unprocessed uranium bearing shale creates hydrochemical conditions that immobilize 97% of a total of 10mg L-1 dissolved uranium in a 0.20μm pore system. The redistribution occurred within 10minutes and remained for five weeks and just 12% of the inventory was retrieved in the biomass. Size exclusion chromatography of the dissolved phase identified organic substances in the range of more than 60kD down to 100 D as a response to time of incubation. Geochemical modeling indicates formation of uranium-organic complexes where ligand size, coordination chemistry and their tendency to agglomerate determine the redistribution.

Enhancing the Laccase Production and Laccase Gene Expression in the White-Rot Fungus Trametes velutina 5930 with Great Potential for Biotechnological Applications by Different Metal Ions and Aromatic Compounds

Laccase is useful for various biotechnological and industrial applications. The white-rot fungus Trametes velutina 5930 and its laccase, isolated from the Shennongjia Nature Reserve in China by our laboratory, has great potential for practical application in environmental biotechnology. However, the original level of laccase produced by Trametes velutina 5930 was relatively low in the absence of any inducer. Therefore, in order to enhance the laccase production by Trametes velutina 5930 and make better use of this fungus in the field of environmental biotechnology, the regulation of laccase production and laccase gene expression in Trametes velutina 5930 were investigated in this study. Different metal ions such as Cu2+ and Fe2+ could stimulate the laccase synthesis and laccase gene transcription in Trametes velutina 5930. Some aromatic compounds structurally related to lignin, such as tannic acid, syringic acid, cinnamic acid, gallic acid and guaiacol, could also enhance the level of laccase activity and laccase gene transcription. We also found that there existed a positive synergistic effect of aromatic compound and metal ion on the laccase production and laccase gene transcription in Trametes velutina 5930. Taken together, our study may contribute to the improvement of laccase productivity by Trametes velutina 5930.

Geomycology: Fungi as Agents of Biogeochemical Change

Geomycology is the study of the roles of fungi in geological processes. The biogeochemical importance of fungi is significant in several areas, including nutrient and element cycling, rock, mineral and metal transformations, bioweathering and mycogenic biomineral formation. Such processes can occur in aquatic and terrestrial habitats, but it is the terrestrial environment where fungi probably have the greatest geochemical influence. Of special significance are the mutualistic relationships with photo trop hie organisms: lichens (algae, cyanobacteria) and mycorrhizas (plants). Central to many geomycological processes are transformations of metals and minerals, and fungi possess many properties that effect changes in metal speciation, toxicity and mobility, as well as mineral formation or mineral dissolution or deterioration. Some fungal transformations have potential applications in environmental biotechnology, e.g. metal and radionuclide leaching, recovery, detoxification and bioremediation, and in the production or deposition of biominerals or metallic elements with catalytic or other properties. Metal and mineral transformations may also result in adverse effects when these processes result in spoilage and destruction of natural and synthetic materials, rock and mineral-based building materials (e.g. concrete), biocorrosion of metals, alloys and related substances, and adverse effects on radionuclide speciation, mobility and containment.