Lignocellulolytic Potential of Microbial Consortia Isolated from a Local Biogas Plant: The Case of Thermostable Xylanases Secreted by Mesophilic Bacteria.

Lignocellulose biomasses (LCB), including spent mushroom substrate (SMS), pose environmental challenges if not properly managed. At the same time, these renewable resources hold immense potential for biofuel and chemicals production. With the mushroom market growth expected to amplify SMS quantities, repurposing or disposal strategies are critical. This study explores the use of SMS for cultivating microbial communities to produce carbohydrate-active enzymes (CAZymes). Addressing a research gap in using anaerobic digesters for enriching microbiomes feeding on SMS, this study investigates microbial diversity and secreted CAZymes under varied temperatures (37 °C, 50 °C, and 70 °C) and substrates (SMS as well as pure carboxymethylcellulose, and xylan). Enriched microbiomes demonstrated temperature-dependent preferences for cellulose, hemicellulose, and lignin degradation, supported by thermal and elemental analyses. Enzyme assays confirmed lignocellulolytic enzyme secretion correlating with substrate degradation trends. Notably, thermogravimetric analysis (TGA), coupled with differential scanning calorimetry (TGA-DSC), emerged as a rapid approach for saccharification potential determination of LCB. Microbiomes isolated at mesophilic temperature secreted thermophilic hemicellulases exhibiting robust stability and superior enzymatic activity compared to commercial enzymes, aligning with biorefinery conditions. PCR-DGGE and metagenomic analyses showcased dynamic shifts in microbiome composition and functional potential based on environmental conditions, impacting CAZyme abundance and diversity. The meta-functional analysis emphasised the role of CAZymes in biomass transformation, indicating microbial strategies for lignocellulose degradation. Temperature and substrate specificity influenced the degradative potential, highlighting the complexity of environmental-microbial interactions. This study demonstrates a temperature-driven microbial selection for lignocellulose degradation, unveiling thermophilic xylanases with industrial promise. Insights gained contribute to optimizing enzyme production and formulating efficient biomass conversion strategies. Understanding microbial consortia responses to temperature and substrate variations elucidates bioconversion dynamics, emphasizing tailored strategies for harnessing their biotechnological potential.

Hormesis of black soldier fly larva: Influence and interactions in livestock manure recycling.

Black soldier fly larvae (BSFL) are considered important organisms, utilized as tools to transform waste including manure into valuable products. The growth and cultivation of BSFL are influenced by various factors, such as the presence of toxic substances in the feed and parasites. These factors play a crucial role in hormesis, and contributing to regulate these contaminants hermetic doses to get sustainable byproducts. This review aims to understand the effects on BSFL growth and activities in the presence of compounds like organic and inorganic pollutants. It also assesses the impact of microbes on BSFL growth and explores the bioaccumulation of pharmaceutical compounds, specifically focusing on heavy metals, pesticides, pharmaceuticals, indigenous bacteria, insects, and nematodes. The review concludes by addressing knowledge gaps, proposing future biorefineries, and offering recommendations for further research.

Metabolic engineering of Corynebacterium glutamicum for the high-level production of valerolactam, a nylon-5 monomer.

Valerolactam (VL) is an important precursor chemical for nylon-5 and nylon 6,5. It has been produced by petroleum-based route involving harsh reaction conditions and generating toxic wastes. Here, we report the complete biosynthesis of VL by metabolically engineered Corynebacterium glutamicum overproducing L-lysine. The pathway comprising L-lysine monooxygenase (davB) and 5-aminovaleramide amidohydrolase (davA) from Pseudomonas putida, and ß-alanine CoA transferase (act) from Clostridium propionicum was introduced into the C. glutamicum GA16 strain. To increase the VL flux, competitive pathways predicted from sRNA knockdown target screening were deleted. This engineered C. glutamicum strain produced VL as a major product, but still secreted significant amount of its precursor, 5-aminovaleric acid (5AVA). To circumvent this problem, putative 5AVA transporter genes were screened and engineered in the genome, thereby reuptaking 5AVA excreted. Also, multiple copies of the act gene were integrated into the genome to strengthen the conversion of 5AVA to VL. The final VL10 (pVL1) strain was constructed by enhancing glucose uptake system, which produced 9.68 g/L of VL in flask culture. Fed-batch fermentation of the VL10 (pVL1) strain produced 76.1 g/L of VL from glucose with the yield and productivity of 0.28 g/g and 0.99 g/L/h, respectively, showcasing a high potential for bio-based production of VL from renewable resources.

Arabinose as an overlooked sugar for microbial bioproduction of chemical building blocks.

The circular economy is anticipated to bring a disruptive transformation in manufacturing technologies. Robust and industrial scalable microbial strains that can simultaneously assimilate and valorize multiple carbon substrates are highly desirable, as waste bioresources contain substantial amounts of renewable and fermentable carbon, which is diverse. Lignocellulosic biomass (LCB) is identified as an inexhaustible and alternative resource to reduce global dependence on oil. Glucose, xylose, and arabinose are the major monomeric sugars in LCB. However, primary research has focused on the use of glucose. On the other hand, the valorization of pentose sugars, xylose, and arabinose, has been mainly overlooked, despite possible assimilation by vast microbial communities. The present review highlights the research efforts that have explicitly proven the suitability of arabinose as the starting feedstock for producing various chemical building blocks via biological routes. It begins by analyzing the availability of various arabinose-rich biorenewable sources that can serve as potential feedstocks for biorefineries. The subsequent section outlines the current understanding of arabinose metabolism, biochemical routes prevalent in prokaryotic and eukaryotic systems, and possible products that can be derived from this sugar. Further, currently, exemplar products from arabinose, including arabitol, 2,3-butanediol, 1,2,3-butanetriol, ethanol, lactic acid, and xylitol are discussed, which have been produced by native and non-native microbial strains using metabolic engineering and genome editing tools. The final section deals with the challenges and obstacles associated with arabinose-based production, followed by concluding remarks and prospects.

Microalgae are not an umbrella solution for power industry waste abatement but could play a role in their valorization.

Microalgae have long been regarded as a promising solution for biological carbon abatement from the power industry, offering renewable biomass without competing for land or water resources used for food crops. In this study, we extensively examined the application of photosynthetic microorganisms for closing carbon, nitrogen, and micronutrient loops in the power industry. Subsequently, we explored the bottom-up integration of algal biorefineries into power industry waste streams for increased economic benefits and reduced environmental impacts. Analysis of the available data indicated that microalgae integration with the power industry is primarily performed using flue-gas-assisted cultivation. This approach allows for carbon sequestration typically below one gram per liter per day, too low to significantly impact carbon abatement at achievable scales of microalgae cultivation. Alternative approaches are also being explored. For example, soluble bicarbonate platforms allow for higher biomass productivity and temporary carbon storage. Meanwhile, the use of ashes and waste heat and thermophilic strains can result in lower cultivation costs and better control of cultivation conditions. These approaches offer further incremental improvement to microalgae-based carbon abatement systems in the power industry but are unlikely to be an umbrella solution for carbon reduction. Consequently, in the near term, microalgae-based carbon valorization systems are likely to be limited to niche applications involving the synthesis of high-value products. For microalgae to truly transform carbon abatement processes radical improvements in both biology and engineering approaches are urgently needed.

Analysis of Single-Step Pretreatments for Lignocellulosic Platform Isolation as the Basis of Biorefinery Design.

Biorefinery feasibility is highly influenced by the early design of the best feedstock transformation pathway to obtain value-added products. Pretreatment has been identified as the critical stage in biorefinery design since proper pretreatment influences subsequent reaction, separation, and purification processes. However, many pretreatment analyses have focused on preserving and valorizing six-carbon sugars for future use in bioconversion processes, leaving aside fractions such as hemicellulose and lignin. To date, there has been no pretreatment systematization for the removal of lignocellulosic fractions. This work defines pretreatment efficacy through operational, economic, environmental, and social indicators. Thus, using the data reported in the literature, as well as the results of the simulation schemes, a multi-criteria weighting of the best-performing schemes for the isolation or removal of cellulose, hemicellulose, and lignin was carried out. As a main result, it was concluded that dilute acid is the most effective for cellulose isolation and hemicellulose removal for producing platform products based on six- and five-carbon sugars, respectively. Additionally, the kraft process is the best methodology for lignin removal and its future use in biorefineries. The results of this work help to elucidate a methodological systematization of the pretreatment efficacy in the design of biorefineries as an early feasibility stage considering sustainability aspects.

Seaweed for climate mitigation, wastewater treatment, bioenergy, bioplastic, biochar, food, pharmaceuticals, and cosmetics: a review.

The development and recycling of biomass production can partly solve issues of energy, climate change, population growth, food and feed shortages, and environmental pollution. For instance, the use of seaweeds as feedstocks can reduce our reliance on fossil fuel resources, ensure the synthesis of cost-effective and eco-friendly products and biofuels, and develop sustainable biorefinery processes. Nonetheless, seaweeds use in several biorefineries is still in the infancy stage compared to terrestrial plants-based lignocellulosic biomass. Therefore, here we review seaweed biorefineries with focus on seaweed production, economical benefits, and seaweed use as feedstock for anaerobic digestion, biochar, bioplastics, crop health, food, livestock feed, pharmaceuticals and cosmetics. Globally, seaweeds could sequester between 61 and 268 megatonnes of carbon per year, with an average of 173 megatonnes. Nearly 90% of carbon is sequestered by exporting biomass to deep water, while the remaining 10% is buried in coastal sediments. 500 gigatonnes of seaweeds could replace nearly 40% of the current soy protein production. Seaweeds contain valuable bioactive molecules that could be applied as antimicrobial, antioxidant, antiviral, antifungal, anticancer, contraceptive, anti-inflammatory, anti-coagulants, and in other cosmetics and skincare products.

Advances in sustainable approaches utilizing orange peel waste to produce highly value-added bioproducts.

Orange peel waste (OPW), a discarded part of orange fruit, is a rich source of essential constituents that can be transformed into highly value-added bioproducts. OPW is being generated in million tonnes globally and returns to the environment without complete benefit. Thus, a high volume of annually produced OPW in the industry requires effective valorization. In this regard, limited data is available that summarizes the broader spectrum for the sustainable fate of OPW to produce value-added bioproducts. The main objective of this treatise is to explore the sustainable production of bioproducts from OPW. Therefore, this review covers all the aspects of OPW, from its production to complete valorization. The review encompasses the extraction technologies employed for extracting different valuable bioactive compounds, such as: essential oil (EO), pectin, and carotenoids, from OPW. Furthermore, the suitability of bioconversion technologies (digestion/fermentation) in transforming OPW to other useful bioproducts, such as: biochemicals (lactic acid and succinic acid), biopolysaccharides (xanthan and curdlan gum), and bioenergy (biomethane and bioethanol) is discussed. Also, it includes the concept of OPW-based biorefineries and their development that shall play a definite role in future to cover demands for: food, chemicals, materials, fuels, power, and heat. Lastly, this review focuses on OPW-supplemented functional food products such as: beverages, yogurts, and extruded products. In conclusion, insights provided in this review maximize the potential of OPW for commercial purposes, leading to a safe, and waste-free environment.

Valorisation of residues from municipal wastewater sieving through anaerobic (co-)digestion with biological sludge.

The Circular and Green Economy principles is inspiring new approaches to municipal wastewater treatment plants (MWWTPs) design and operation. Recently, an ever-growing interest is devoted to exploring the alternatives for switching the WWTPs from being able to 'simply' removing contaminants from water to biorefinery-like plants where energy and material can be recovered. In this perspective, both wastewater and residues from process can be valorised for recovering nutrients (N and P), producing value added products (i.e. biopolymers), energy vectors and biofuels (i.e. bio-H2, bio-CH4 and bioethanol). As an additional benefit, changing the approach for WWTPs design and operation will decrease the overall amount of landfilled residues. In this context, the present research is aimed at evaluating the CH4 production potential of MWW screening units' residues. While such a stream is typically landfilled, the expected progressive increase of biodegradable matter content due to the ban on single-use plastic along with the boost of bioplastics makes the investigation of different biochemical valorisation routes more and more interesting from an environmental and economical perspective. Thus, a full-scale data collection campaign was performed to gain information on screening residues amount and properties and to analyse the relationship with influent flowrate. The most relevant residue properties were measured, and lab-scale tests were carried out to evaluate the bio-CH4 potential.

Role of extremophiles and their extremozymes in biorefinery process of lignocellulose degradation.

Technological advances in the field of life sciences have led to discovery of organisms that live in harsh environmental conditions referred to as extremophiles. These organisms have adapted themselves to thrive in extreme habitat giving these organisms an advantage over conventional mesophilic organisms in various industrial applications. Extremozymes produced by these extremophiles have high tolerance to inhospitable environmental conditions making them an ideal enzyme system for various industrial processes. A notable application of these extremophiles and extremozymes is their use in the degradation of recalcitrant lignocellulosic biomass and application in biorefineries. For maximum utilization of the trapped carbon source from this obstinate biomass, pretreatment is a necessary step that requires various physiochemical and enzymatic treatments. From search for novel extremophiles and extremozymes to development of various genetic and protein engineering techniques, investigation on extremozymes with enhanced stability and efficiency is been done. Since extremozymes are easily calibrated to work under such conditions, they have become an emerging topic in the research field of biofuel production. The review discusses the various extremozymes that play an important role in lignocellulose degradation along with recent studies on their molecular and genetic evolution for industrial application and production of biofuels and various value-added products.

Residential Proximity to Biorefinery Sources of Air Pollution and Respiratory Diseases in New York State.

Understanding potential health risks associated with biofuel production is critical to sustainably combating energy insecurity and climate change. However, the specific health impacts associated with biorefinery-related emissions are not yet well characterized. We evaluated the relationship between respiratory emergency department (ED) visits (2011-2015) and residential exposure to biorefineries by comparing 15 biorefinery sites to 15 control areas across New York (NY) State. We further examined these associations by biorefinery types (e.g., corn, wood, or soybean), seasons, and lower respiratory disease subtypes. We measured biorefinery exposure using residential proximity in a cross-sectional study and estimation of biorefinery emission via AERMOD-simulated modeling. After controlling for multiple confounders, we consistently found that respiratory ED visit rates among residents living within 10 km of biorefineries were significantly higher (rate ratios (RRs) range from 1.03 to 3.64) than those in control areas across our two types of exposure indices. This relationship held across biorefinery types (higher in corn and soybean biorefineries), seasons (higher in spring and winter), air pollutant types (highest for NO2), and respiratory subtypes (highest for emphysema). Further research is needed to confirm our findings.

New (and Old) Monomers from Biorefineries to Make Polymer Chemistry More Sustainable.

This opinion article describes recent approaches to use the "biorefinery" concept to lower the carbon footprint of typical mass polymers, by replacing parts of the fossil monomers with similar or even the same monomer made from regrowing dendritic biomass. Herein, the new and green catalytic synthetic routes are for lactic acid (LA), isosorbide (IS), 2,5-furandicarboxylic acid (FDCA), and p-xylene (pXL). Furthermore, the synthesis of two unconventional lignocellulosic biomass derivable monomers, i.e., α-methylene-γ-valerolactone (MeGVL) and levoglucosenol (LG), are presented. All those have the potential to enter in a cost-effective way, also the mass market and thereby recover lost areas for polymer materials. The differences of catalytic unit operations of the biorefinery are also discussed and the challenges that must be addressed along the synthesis path of each monomers.

Loops around the Heme Pocket Have a Critical Role in the Function and Stability of BsDyP from Bacillus subtilis.

Bacillus subtilis BsDyP belongs to class I of the dye-decolorizing peroxidase (DyP) family of enzymes and is an interesting biocatalyst due to its high redox potential, broad substrate spectrum and thermostability. This work reports the optimization of BsDyP using directed evolution for improved oxidation of 2,6-dimethoxyphenol, a model lignin-derived phenolic. After three rounds of evolution, one variant was identified displaying 7-fold higher catalytic rates and higher production yields as compared to the wild-type enzyme. The analysis of X-ray structures of the wild type and the evolved variant showed that the heme pocket is delimited by three long conserved loop regions and a small α helix where, incidentally, the mutations were inserted in the course of evolution. One loop in the proximal side of the heme pocket becomes more flexible in the evolved variant and the size of the active site cavity is increased, as well as the width of its mouth, resulting in an enhanced exposure of the heme to solvent. These conformational changes have a positive functional role in facilitating electron transfer from the substrate to the enzyme. However, they concomitantly resulted in decreasing the enzyme's overall stability by 2 kcal mol-1, indicating a trade-off between functionality and stability. Furthermore, the evolved variant exhibited slightly reduced thermal stability compared to the wild type. The obtained data indicate that understanding the role of loops close to the heme pocket in the catalysis and stability of DyPs is critical for the development of new and more powerful biocatalysts: loops can be modulated for tuning important DyP properties such as activity, specificity and stability.

A review on modern and smart technologies for efficient waste disposal and management.

In the current scenario, the word waste management holds much importance in every individual's life. Pollution and the generation of vast waste quantities with no proper waste management process have become one of humanity's biggest threats. This review article provides a complete review of the innovative technologies currently employed to handle and dispose of the waste successfully. This work aims to include the different solid, liquid, gaseous, and radioactive waste management processes. The novel and improved plasma gasification concepts, transmutation, incineration, bio-refineries, microbial fuel cells (MFC) have been thoroughly explained. In addition, some new techniques like Mr. Trash Wheel and the Smart bin approach provide much hope of adequately managing waste. The work's novelty lies in adopting several successful methods of various countries for waste disposal and management. To incorporate or improve India'sIndia's same techniques and processes, we have to tackle the ever-increasing waste disposal problems and find economic and eco-friendly ways of waste management.

Added-Value Chemicals from Lignin Oxidation.

Lignin is the second most abundant component, next to cellulose, in lignocellulosic biomass. Large amounts of this polymer are produced annually in the pulp and paper industries as a coproduct from the cooking process-most of it burned as fuel for energy. Strategies regarding lignin valorization have attracted significant attention over the recent decades due to lignin's aromatic structure. Oxidative depolymerization allows converting lignin into added-value compounds, as phenolic monomers and/or dicarboxylic acids, which could be an excellent alternative to aromatic petrochemicals. However, the major challenge is to enhance the reactivity and selectivity of the lignin structure towards depolymerization and prevent condensation reactions. This review includes a comprehensive overview of the main contributions of lignin valorization through oxidative depolymerization to produce added-value compounds (vanillin and syringaldehyde) that have been developed over the recent decades in the LSRE group. An evaluation of the valuable products obtained from oxidation in an alkaline medium with oxygen of lignins and liquors from different sources and delignification processes is also provided. A review of C4 dicarboxylic acids obtained from lignin oxidation is also included, emphasizing catalytic conversion by O2 or H2O2 oxidation.

Whey Utilization: Sustainable Uses and Environmental Approach.

The dairy industry produces large amounts of whey as a by- or co-product, which has led to considerable environmental problems due to its high organic matter content. Over the past decades, possibilities of more environmentally and economically efficient whey utilisation have been studied, primarily to convert unwanted end products into a valuable raw material. Sustainable whey management is mostly oriented to biotechnological and food applications for the development of value-added products such as whey powders, whey proteins, functional food and beverages, edible films and coatings, lactic acid and other biochemicals, bioplastic, biofuels and similar valuable bioproducts. This paper provides an overview of the sustainable utilization of whey and its constituents, considering new refining approaches and integrated processes to convert whey, or lactose and whey proteins to high value-added whey-based products.

Toward Replacing Ethylene Oxide in a Sustainable World: Glycolaldehyde as a Bio-Based C2 Platform Molecule.

Fossil-based platform molecules such as ethylene and ethylene oxide currently serve as the primary feedstock for the C2 -based chemical industry. However, in the search for a more sustainable chemical industry, fossil-based resources may preferentially be replaced by renewable alternatives, provided there is realistic economic feasibility. This Review compares and critically discusses several production routes toward bio-based structural analogues of ethylene oxide and the required adaptations for their implementation in state-of-the-art C2 -based chemical processes. For example, glycolaldehyde, a structural analogue obtainable from carbohydrates by atom-economic retro-aldol reactions, may replace ethylene oxide's leading role. This alternative chemical route may not only allow the carbon footprint of conventional chemicals production to be lowered, but the introduction of a bio-based pathway may also contribute to safer production processes. Where possible, challenges, drawbacks, and prospects are highlighted.

C/N ratio and carbon source-dependent lipid production profiling in Rhodotorula toruloides.

Microbial oils are lipids produced by oleaginous microorganisms, which can be used as a potential feedstock for oleochemical production. The oleaginous yeast Rhodotorula toruloides can co-produce microbial oils and high-value compounds from low-cost substrates, such as xylose and acetic acid (from hemicellulosic hydrolysates) and raw glycerol (a byproduct of biodiesel production). One step towards economic viability is identifying the best conditions for lipid production, primarily the most suitable carbon-to-nitrogen ratio (C/N). Here, we aimed to identify the best conditions and cultivation mode for lipid production by R. toruloides using various low-cost substrates and a range of C/N ratios (60, 80, 100, and 120). Turbidostat mode was used to achieve a steady state at the maximal specific growth rate and to avoid continuously changing environmental conditions (i.e., C/N ratio) that inherently occur in batch mode. Regardless of the carbon source, higher C/N ratios increased lipid yields (up to 60% on xylose at a C/N of 120) but decreased the specific growth rate. Growth on glycerol resulted in the highest specific growth and lipid production (0.085 g lipids/gDW*h) rates at C/Ns between 60 and 100. We went on to study lipid production using glycerol in both batch and fed-batch modes, which resulted in lower specific lipid production rates compared with turbisdostat, however, fed batch is superior in terms of biomass production and lipid titers. By combining the data we obtained in these experiments with a genome-scale metabolic model of R. toruloides, we identified targets for improvements in lipid production that could be carried out either by metabolic engineering or process optimization.

Sugar Production from Hybrid Poplar Sawdust: Optimization of Enzymatic Hydrolysis and Wet Explosion Pretreatment.

Wet explosion pretreatment of hybrid poplar sawdust (PSD) for the production of fermentable sugar was carried out in the pilot-scale. The effects of pretreatment conditions, such as temperature (170-190 °C), oxygen dosage (0.5-7.5% of dry matter (DM), w/w), residence time (10-30 min), on cellulose and hemicellulose digestibility after enzymatic hydrolysis were ascertained with a central composite design of the experiment. Further, enzymatic hydrolysis was optimized in terms of temperature, pH, and a mixture of CTec2 and HTec2 enzymes (Novozymes). Predictive modeling showed that cellulose and hemicellulose digestibility of 75.1% and 83.1%, respectively, could be achieved with a pretreatment at 177 °C with 7.5% O2 and a retention time of 30 min. An increased cellulose digestibility of 87.1% ± 0.1 could be achieved by pretreating at 190 °C; however, the hemicellulose yield would be significantly reduced. It was evident that more severe conditions were required for maximal cellulose digestibility than that of hemicellulose digestibility and that an optimal sugar yield demanded a set of conditions, which overall resulted in the maximum sugar yield.

Integrating Value Considerations in the Decision Making for the Design of Biorefineries.

Biobased production has been promoted as a sustainable alternative to fossil resources. However, controversies over its impact on sustainability highlight societal concerns, value tensions and uncertainties that have not been taken into account during its development. In this work, the consideration of stakeholders' values in a biorefinery design project is investigated. Value sensitive design (VSD) is a promising approach to the design of technologies with consideration of stakeholders' values, however, it is not directly applicable for complex systems like biorefineries. Therefore, some elements of VSD, such as the identification of relevant values and their connection to a technology's features, are brought into biorefinery design practice. Midstream modulation (MM), an approach to promoting the consideration of societal aspects during research and development activities, is applied to promote reflection and value considerations during the design decision making. As result, it is shown that MM interventions during the design process led to new design alternatives in support of stakeholders' values, and allowed to recognize and respond to emerging value tensions within the scope of the project. In this way, the present work shows a novel approach for the technical investigation of VSD, especially for biorefineries. Also, based on this work it is argued that not only reflection, but also flexibility and openness are important for the application of VSD in the context of biorefinery design.