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With regard to social and environmental sustainability, second-generation biofuel and biogas production from lignocellulosic material provides considerable potential, since lignocellulose represents an inexhaustible, ubiquitous natural resource, and is therefore one important step towards independence from fossil fuel combustion. However, the highly heterogeneous structure and recalcitrant nature of lignocellulose restricts its commercial utilization in biogas plants. Improvements therefore rely on effective pretreatment methods to overcome structural impediments, thus facilitating the accessibility and digestibility of (ligno)cellulosic substrates during anaerobic digestion. While chemical and physical pretreatment strategies exhibit inherent drawbacks including the formation of inhibitory products, biological pretreatment is increasingly being advocated as an environmentally friendly process with low energy input, low disposal costs, and milder operating conditions. Nevertheless, the promising potential of biological pretreatment techniques is not yet fully exploited. Hence, we intended to provide a detailed insight into currently applied pretreatment techniques, with a special focus on biological ones for downstream processing of lignocellulosic biomass in anaerobic digestion.

Bioethanol is recognized as a valuable substitute for renewable energy sources to meet the fuel and energy demand of the nation, considered an environmentally friendly resource obtained from agricultural residues such as sugarcane bagasse, rice straw, husk, wheat straw and corn stover. The energy demand is sustained using lignocellulosic biomass to produce bioethanol. Lignocellulosic biomass (LCBs) is the point of attention in replacing the dependence on fossil fuels. The recalcitrant structure of the lignocellulosic biomass is disrupted using effective pretreatment techniques that separate complex interlinked structures among cellulose, hemicellulose, and lignin. Pretreatment of biomass involves various physical, chemical, biological, and physiochemical protocols which are of importance, dependent upon their individual or combined dissolution effect. Physical pretreatment involves a reduction in the size of the biomass using mechanical, extrusion, irradiation, and sonification methods while chemical pretreatment involves the breaking of various bonds present in the LCB structure. This can be obtained by using an acidic, alkaline, ionic liquid, and organosolvent methods. Biological pretreatment is considered an environment-friendly and safe process involving various bacterial and fungal microorganisms. Distinct pretreatment methods, when combined and utilized in synchronization lead to more effective disruption of LCB, making biomass more accessible for further processing. These could be utilized in terms of their effectiveness for a particular type of cellulosic fiber and are namely steam explosion, liquid hot water, ammonia fibre explosion, CO 2 explosion, and wet air oxidation methods. The present review encircles various distinct and integrated pretreatment processes developed till now and their advancement according to the current trend and future aspects to make lignocellulosic biomass available for further hydrolysis and fermentation.

There is a growing need of substrate flexibility for biobased production of energy and value-added products that allows the application of variable biodegradable residues within a circular economy. It can be used to balance fluctuating energy provision of other renewable sources. Hydrolysis presents one of the biggest limitations during anaerobic digestion. Methods to improve it will result in broader process applicability and improved integration into regional material cycles. Recently, one focus of anaerobic digestion research has been directed to systems with a separate hydrolysis–acidogenesis stage as it might be promised to improve process performance. Conditions can be adjusted to each class of microorganisms individually without harming methanogenic microorganisms. Extensive research of separate biomass pretreatment via biological, chemical, physical or mixed methods has been conducted. Nevertheless, several methods lack economic efficiency, have a high environmental impact or focus on specific substrates. Pretreatment via a separate hydrolysis stage as cell-driven biotransformation in a suspension might be an alternative that enables high yields, flexible feeding and production, and a better process control. In this review, we summarize existing technologies for microbial hydrolytic biotransformation in a separate reactor stage and the impacts of substrate, operational parameters, combined methods and process design as well as remaining challenges.

Lignocellulosic biomass is the most abundant, low-cost, bio-renewable resource that holds enormous importance as alternative source for production of biofuels and other biochemicals that can be utilized as building blocks for production of new materials. Enzymatic hydrolysis is an essential step involved in the bioconversion of lignocellulose to produce fermentable monosaccharides. However, to allow the enzymatic hydrolysis, a pretreatment step is needed in order to remove the lignin barrier and break down the crystalline structure of cellulose. The present manuscript is dedicated to reviewing the most commonly applied “green” pretreatment processes used in bioconversion of lignocellulosic biomasses within the “biorefinery” concept. In this frame, the effects of different pretreatment methods on lignocellulosic biomass are described along with an in-depth discussion on the benefits and drawbacks of each method, including generation of potentially inhibitory compounds for enzymatic hydrolysis, effect on cellulose digestibility, and generation of compounds toxic for the environment, and energy and economic demand.

Background Energy is the basis and assurance for a world's stable development; however, as traditional non-renewable energy sources deplete, the development and study of renewable clean energy have emerged. Using microalgae as a carbon source for anaerobic bacteria to generate biohydrogen is a clean energy generation system that both local and global peers see as promising. Results Klebsiella pneumonia , Enterobacter cloacae , and their coculture were used to synthesize biohydrogen using Oscillatoria acuminata biomass via dark fermentation. The total carbohydrate content in O. acuminata was 237.39 mg/L. To enhance the content of fermentable reducing sugars, thermochemical, biological, and biological with magnesium zinc ferrite nanoparticles (Mg-Zn Fe 2 O 4 -NPs) pretreatments were applied. Crude hydrolytic enzymes extracted from Trichoderma harzianum of biological pretreatment were enhanced by Mg-Zn Fe 2 O 4 -NPs and significantly increased reducing sugars (230.48 mg/g) four times than thermochemical pretreatment (45.34 mg/g). K. pneumonia demonstrated a greater accumulated hydrogen level (1022 mLH 2 /L) than E. cloacae (813 mLH 2 /L), while their coculture showed superior results (1520 mLH 2 /L) and shortened the production time to 48 h instead of 72 h in single culture pretreatments. Biological pretreatment + Mg-Zn Fe 2 O 4 NPs using coculture significantly stimulated hydrogen yield (3254 mLH 2 /L), hydrogen efficiency)216.9 mL H 2 /g reducing sugar( and hydrogen production rate (67.7 mL/L/h) to the maximum among all pretreatments. Conclusion These results confirm the effectiveness of biological treatments + Mg-Zn Fe 2 O 4 -NPs and coculture dark fermentation in upregulating biohydrogen production.

The pretreatment of lignocellulosic biomass (LC biomass) prior to the anaerobic digestion (AD) process is a mandatory step to improve feedstock biodegradability and biogas production. An important potential is provided by lignocellulosic materials since lignocellulose represents a major source for biogas production, thus contributing to the environmental sustainability. The main limitation of LC biomass for use is its resistant structure. Lately, biological pretreatment (BP) gained popularity because they are eco-friendly methods that do not require chemical or energy input. A large number of bacteria and fungi possess great ability to convert high molecular weight compounds from the substrate into lower mass compounds due to the synthesis of microbial extracellular enzymes. Microbial strains isolated from various sources are used singly or in combination to break down the recalcitrant polymeric structures and thus increase biogasgeneration. Enzymatic treatment of LC biomass depends mainly on enzymes like hemicellulases and cellulases generated by microorganisms. The articles main purpose is to provide an overview regarding the enzymatic/biological pretreatment as one of the most potent techniques for enhancing biogas production.

The rising global population and worldwide industrialization have led to unprecedented energy demand that is causing fast depletion of fossil reserves. This has led to search for alternative energy sources that are renewable and environment friendly. Use of lignocellulosic biomass for energy generation is considered a promising approach as it does not compete with food supply. However, the lignin component of the biomass acts as a natural barrier that prevents its efficient utilization. In order to remove the lignin and increase the amount of fermentable sugars, the lignocellulosic biomass is pretreated using physical and chemical methods which are costly and hazardous for environment. Moreover, during the traditional pretreatment process, numerous inhibitory compounds are generated that adversely affect the growth of fermentative microbes. Alternatively, biological methods that use microbes and their enzymes disrupt lignin polymers and increase the accessibility of the carbohydrates for the sugar generation. Microbial laccases have been considered as an efficient biocatalyst for delignification and detoxification offering a green initiative for energy generation process. The present review aims to bring together recent studies in bioenergy generation using laccase biocatalyst in the pretreatment processes. The work provides an overview of the sustainable and eco-friendly approach of biological delignification and detoxification through whole-cell and enzymatic methods, use of laccase-mediator system, and immobilized laccases for this purpose. It also summarizes the advantages, associated challenges, and potential prospects to overcome the limitations.

Landfill leachates from municipal landfills are usually heavily contaminated and thus require treatments before direct discharge into natural waters. Selecting the appropriate technology for leachate treatment is still a major challenge for operations in municipal landfills. Biodegradation is effective for treating young leachates, whereas old leachates require processes such as chemical oxidation, coagulation–flocculation, chemical precipitation, ozonation, activated carbon adsorption, and reverse osmosis. Recently, the combination of biological pretreatments followed by physico-chemical processes has been shown to be very efficient. Here we review the efficiency of biological treatment in combination with reverse osmosis to clean landfill leachates. We studied in particular processes including a membrane bioreactor, activated sludge, a rotating biological contactor, and up-flow anaerobic sludge blanket treatments, followed by reverse osmosis. We found a 99–99.5% removal of the chemical oxygen demand (COD), and a 99–99.8% removal of N–NH4+ using reverse osmosis and activated sludge. Using reverse osmosis with a rotating biological contactor, we observed 99% removal of COD, biochemical oxygen demand and N–NH4+. The combination of reverse osmosis, activated sludge and rotating biological contactor removed 98–99.2% of Cl− and 99–99.7% of Pb. Total suspended solids are best removed, up to 99%, by either a combination of reverse osmosis with membrane bioreactor, or reverse osmosis with activated sludge.

Rugulopteryx okamurae is an invasive brown alga causing severe environmental and economic problems on the western Mediterranean coasts. Thus, in addition to the difficulties caused to the fishing and tourism sectors, there is a need to manage its accumulation on the beaches. This work aims to valorise this waste by using it as raw material for producing monosaccharides through a two-stage sequential process. These sugars could be used for different fermentative processes to obtain high-value-added bioproducts. In this work, biological pretreatment of the previously conditioned seaweed with the fungus Aspergillus awamori in solid-state fermentation (SSF), followed by enzymatic hydrolysis with a commercial enzyme cocktail, was performed. The effect of the extension of the biological pretreatment (2, 5, 8 and 12 days) on the subsequent release of total reducing sugars (TRS) in the enzymatic hydrolysis stage was studied. To analyse this effect, experimental data of TRS produced along the hydrolysis were fitted to simple first-order kinetics. Also, the secretion of cellulase and alginate lyase by the fungus, along with the biological pretreatment, was determined. The results suggest that 5 days of biological pretreatment of the macroalgae with A. awamori followed by enzymatic saccharification for 24 h with Cellic CTec2® (112 FP units/g of dry biomass) are the best conditions tested, allowing the production of around 240 g of TRS per kg of dried biomass. The main sugars obtained were glucose (95.8 %) and mannitol (1.5 %), followed by galactose (1 %), arabinose (0.9 %) and fucose (0.5 %).Key points• Five-day SSF by A. awamori was the best condition to pretreat R. okamurae.• Five-day SSF was optimal for alginate lyase production (1.63 ±0.011 IU/g biomass).• A maximum yield of 239 mg TRS/g biomass was obtained (with 95.8 % glucose).

Lignocellulosic agricultural residues such as coconut husk are abundant but recalcitrant substrates that limit efficient biogas production during anaerobic digestion. We evaluated the effect of biological pretreatment of coconut husk on archaea, volatile fatty acid (VFA) profiles, and biogas production during anaerobic digestion. Coconut husk was pretreated for 50 days using a consortium of cellulolytic bacteria isolated from poultry feces and subsequently digested alongside untreated husk (control) in batch reactors. Cellulose-degrading bacteria were presumptively characterized based on morphological, cultural, and biochemical characteristics and included Bacillus megaterium , Bacillus licheniformis , Lactobacillus plantarum , Bacillus cereus , and Bacillus subtilis based on morphological, cultural, and biochemical characteristics. Ten archaeal genera were detected across treatments, including Methanobrevibacter , Methanomassiliicoccus , Methanobacterium , Methanoculleus , Methanosarcina , Methanomicrobium , Desulfurococcus , Thermosphaera , Methanogenium , and Methanoregula . The control digestate exhibited higher archaeal diversity with a low methane production. The pretreated reactor (Ch + Cdb) produced a cumulative biogas yield of 88,355 mL gVS⁻ 1 , compared with 67,105 mL gVS⁻ 1 in the control, representing a significant 1.32-fold increase ( p  ≤ 0.05). Methane production commenced on day 3 in Ch + Cdb relative to day 15 in the control indicating a shortened lag phase and enhanced breakdown of substrate. Mean methane concentration increased from 44.52 ± 25.4% in the control (Ch) to 56.38 ± 16.54% in the Ch + Cdb treatment representing a 1.27-fold increase in methane content relative to the control. Carbon dioxide concentration also increased from 11.41 ± 6.24% in the control to 15.01 ± 5.54% in the treated sample, corresponding to a 1.32-fold increase. Total VFA concentration was higher in Ch + Cdb at 596.43 mg L⁻ 1 than in the control (562.24 mg L⁻ 1 ), with acetic acid as the dominant component. Increased concentrations of acetic acid, propionic acid, and butyric acid in the treated system did not inhibit methanogenesis, suggesting improved intermediate conversion and process stability. Biological pretreatment enhanced hydrolysis, accelerated methanogenesis, and improved biogas yield with a stable reactor performance, and highlight the whole-cell biocatalyst potential of cellulose-degrading bacteria for optimum anaerobic digestion of lignocellulosic biomass.