C. Sambusiti

Do furanic and phenolic compounds of lignocellulosic and algae biomass hydrolyzate inhibit anaerobic mixed cultures? A comprehensive review

Nowadays there is a growing interest on the use of both lignocellulosic and algae biomass to produce biofuels (i.e. biohydrogen, ethanol and methane), as future alternatives to fossil fuels. In this purpose, thermal and thermo-chemical pretreatments have been widely investigated to overcome the natural physico-chemical barriers of such biomass and to enhance biofuel production from lignocellulosic residues and, more recently, marine biomass (i.e. macro and microalgae). However, the pretreatment technologies lead not only to the conversion of carbohydrate polymers (ie cellulose, hemicelluloses, starch, agar) to soluble monomeric sugar (ie glucose, xylose, arabinose, galactose), but also the generation of various by-products (i.e. furfural and 5-HMF). In the case of lignocellulosic residues, part of the lignin can also be degraded in lignin derived by-products, mainly composed of phenolic compounds. Although the negative impact of such by-products on ethanol production has been widely described in literature, studies on their impact on biohydrogen and methane production operated with mixed cultures are still very limited. This review aims to summarise and discuss literature data on the impact of pre-treatment by-products on H2-producing dark fermentation and anaerobic digestion processes when using mixed cultures as inoculum. As a summary, furanic (5-HMF, furfural) and phenolic compounds were found to be stronger inhibitors of the microbial dark fermentation than the full anaerobic digestion process. Such observations can be explained by differences in process parameters: anaerobic digestion is performed with more complex mixed cultures, lower substrate/inoculum and by-products/inoculum ratios and longer batch incubation times than dark fermentation. Finally, it has been reported that, during dark fermentation process, the presence of by-products could lead to a metabolic shift from H2-producing pathways (i.e. acetate and butyrate) to non-H2-producing pathways (i.e. lactate, ethanol and propionate) and whatever the metabolic route, metabolites can be all further converted into methane, but at different rates.

Predictive models of biohydrogen and biomethane production based on the compositional and structural features of lignocellulosic materials.

In an integrated biorefinery concept, biological hydrogen and methane production from lignocellulosic substrates appears to be one of the most promising alternatives to produce energy from renewable sources. However, lignocellulosic substrates present compositional and structural features that can limit their conversion into biohydrogen and methane. In this study, biohydrogen and methane potentials of 20 lignocellulosic residues were evaluated. Compositional (lignin, cellulose, hemicelluloses, total uronic acids, proteins, and soluble sugars) as well as structural features (crystallinity) were determined for each substrate. Two predictive partial least square (PLS) models were built to determine which compositional and structural parameters affected biohydrogen or methane production from lignocellulosic substrates, among proteins, total uronic acids, soluble sugars, crystalline cellulose, amorphous holocelluloses, and lignin. Only soluble sugars had a significant positive effect on biohydrogen production. Besides, methane potentials correlated negatively to the lignin contents and, to a lower extent, crystalline cellulose showed also a negative impact, whereas soluble sugars, proteins, and amorphous hemicelluloses showed a positive impact. These findings will help to develop further pretreatment strategies for enhancing both biohydrogen and methane production.

New opportunities for agricultural digestate valorization: current situation and perspectives

In the agricultural sector of many European countries, biogas production through anaerobic digestion (AD) is becoming a very fast-growing market. AD is a simple and robust process that biologically converts an organic matrix into biogas and digestate, the latter corresponding to the anaerobically non-degraded fraction. So far, digestate has been mostly used at farm-scales for improving soils. However, its ever-increasing production induces problems related to transport costs, greenhouse-gas emissions during storage and high nitrogen content that constrains its use to land application only. Consequently, research on alternative valorisation routes to reduce its environmental impact and to improve the economical profitability of AD plants should draw increasing interest in the future. This review therefore focuses on the different alternatives of digestate valorisation, apart from land applications: (I) the use of the digestate liquor for replacing freshwater and nutrients in algae cultivation; (II) the use of solid digestate for energy production through biological (i.e. AD, bioethanol) or thermal processes (i.e. combustion, hydrothermal carbonization and pyrolysis); (III) the conversion of solid digestate into added-value products (char or activated carbons) through a pyrolysis process.

Benefit of sodium hydroxide pretreatment of ensiled sorghum forage on the anaerobic reactor stability and methane production.

The assessment of the pretreatment effect on the anaerobic digestion process is generally based on the results of batch tests, which may fail in truly predicting full-scale anaerobic reactors performance. Therefore, in this study, the effect of alkaline pretreatment on the anaerobic digestion of ensiled sorghum forage was evaluated by comparing the results of two semi-continuous CSTR (Continuously Stirred Tank Reactor) anaerobic reactors. Results showed that an alkaline pretreatment step, prior to the anaerobic digestion of ensiled sorghum forage, can have a beneficial effect both in enhancing methane production (an increase of 25% on methane production was observed, if compared to that of untreated sorghum) and in giving more stability to the anaerobic digestion process.

Enzymatic and metabolic activities of four anaerobic sludges and their impact on methane production from ensiled sorghum forage

Biochemical methane potential (BMP) tests were run on ensiled sorghum forage using four inocula (urban, agricultural, mixture of agricultural and urban, granular) and differences on their metabolic and enzymatic activities were also discussed. Results indicate that no significant differences were observed in terms of BMP values (258±14NmLCH4g(-1)VS) with a slightly higher value when agricultural sludge was used as inoculum. Significant differences can be observed among different inocula, in terms of methane production rate. In particular the fastest biomethanization occurred when using the urban sludge (hydrolytic kinetic constant kh=0.146d(-1)) while the slowest one was obtained from the agricultural sludge (kh=0.049d(-1)). Interestingly, positive correlations between the overall enzymatic activities and methane production rates were observed for all sludges, showing that a high enzymatic activity may favour the hydrolysis of complex substrate and accelerate the methanization process of sorghum.

Sodium hydroxide pretreatment of ensiled sorghum forage and wheat straw to increase methane production.

The aim of this study was to determine the effect of sodium hydroxide pretreatment on the chemical composition and the methane production of ensiled sorghum forage and wheat straw. NaOH pretreatment was conducted in closed bottles, at 40 °C for 24 h. Samples were soaked in a NaOH solution at different dosages (expressed in terms of total solids (TS) content) of 1 and 10% gNaOH/gTS, with a TS concentration of 160 gTS/L. At the highest NaOH dosage the reduction of cellulose, hemicelluloses and lignin was 31, 66 and 44%, and 13, 45 and 3% for sorghum and wheat straw, respectively. The concentration of soluble chemical oxygen demand (CODs) in the liquid phase after the pretreatment was also improved both for wheat straw and sorghum (up to 24 and 33%, respectively). Total sugars content increased up to five times at 10% gNaOH/gTS with respect to control samples, suggesting that NaOH pretreatment improves the hydrolysis of cellulose and hemicelluloses. The Biochemical Methane Potential (BMP) tests showed that the NaOH pretreatment favoured the anaerobic degradability of both substrates. At 1 and 10% NaOH dosages, the methane production increased from 14 to 31% for ensiled sorghum forage and from 17 to 47% for wheat straw. The first order kinetic constant increased up to 65% for sorghum and up to 163% for wheat straw.

Pyrochars from bioenergy residue as novel bio-adsorbents for lignocellulosic hydrolysate detoxification

The robust supramolecular structure of biomass often requires severe pretreatments conditions to produce soluble sugars. Nonetheless, these processes generate some inhibitory compounds (i.e. furans compounds and aliphatic acids) deriving mainly from sugars degradation. To avoid the inhibition of the biological process and to obtain satisfactory sugars conversion level into biofuels, a detoxification step is required. This study investigates the use of two pyrochars derived from solid anaerobic digestates for the detoxification of lignocellulosic hydrolysates. At a pyrochar concentration of 40gL(-1), more than 94% of 5-HMF and 99% of furfural were removed in the synthetic medium after 24h of contact time, whereas sugars concentration remained unchanged. Furfural was adsorbed faster than 5-HMF by both pyrochars and totally removed after 3h of contact. Finally, the two pyrochars were found efficient in the detoxification of corn stalks and Douglas fir wood chips hydrolysates without affecting the soluble sugars concentrations.

One-Pot dry chemo-mechanical deconstruction for bioethanol production from sugarcane bagasse

The aim of this study was the application of an innovative dry chemo-mechanical pretreatment using different mechanical stresses to produce bioethanol from sugarcane bagasse (SB). The effect of different milling methods on physicochemical composition, enzymatic hydrolysis, bioethanol production and energy efficiency was also evaluated. SB was pretreated with NaOH and H3PO4 at high materials concentration (5 kg/L). Results indicate that vibratory milling (VBM) was more effective in the reduction of particles size and cellulose crystallinity compared to centrifugal (CM) and ball (BM) milling. NaOH pretreatment coupling to BM and VBM was preferred to enhance glucose yields and bioethanol production, while CM consumed less energy compared to BM and VBM. Moreover, the highest energy efficiency (η=0.116 kg glucose/kWh) was obtained with NaOH-CM. Therefore, the combination of dry NaOH and CM appears the most suitable and interesting pretreatment for the production of bioethanol from SB.

Harmonization of the quantitative determination of volatile fatty acids profile in aqueous matrix samples by direct injection using gas chromatography and high-performance liquid chromatography techniques: Multi-laboratory validation study

The performance parameters of volatile fatty acids (VFAs) measurements were assessed for the first time by a multi-laboratory validation study among 13 laboratories. Two chromatographic techniques (GC and HPLC) and two quantification methods such as external and internal standard (ESTD/ISTD) were combined in three different methodologies GC/ESTD, HPLC/ESTD and GC/ISTD. Linearity evaluation of the calibration functions in a wide concentration range (10-1000mg/L) was carried out using different statistical parameters for the goodness of fit. Both chromatographic techniques were considered similarly accurate. The use of GC/ISTD, despite showing similar analytical performance to the other methodologies, can be considered useful for the harmonization of VFAs analytical methodology taking into account the normalization of slope values used for the calculation of VFAs concentrations. Acceptance criteria for VFAs performance parameters of the multi-laboratory validation study should be established as follows: (1) instrument precision (RSDINST≤1.5%); (2) linearity (R(2)≥0.998; RSDSENSITIVITY≤4%; REMAX≤8%; REAVER≤ 3%); (3) precision (RSD≤1.5%); (4) trueness (recovery of 97-103%); (5) LOD (≤3mg/L); and (6) LOQ (10mg/L).

Bioethanol fermentation as alternative valorization route of agricultural digestate according to a biorefinery approach.

This study investigates the feasibility of producing bioethanol from solid digestate after a mechanical fractionation (i.e. centrifugal milling), in order to improve the energy recovery from agricultural wastes and the sustainability of anaerobic digestion plants. A bioethanol yield of 37gkg(-1)TS was evaluated for the solid digestate fraction. Mass and energetic balances were performed and compared between two scenarios: (A) one-stage bioethanol fermentation and (B) two-stage anaerobic digestion-bioethanol fermentation, in order to evaluate the feasibility and the advantages of the two-stage process. Results revealed that, compared to the one-stage process, the dual anaerobic digestion-bioethanol process permitted: (i) to diversify biofuels production; (ii) to provide the thermal energy sufficient for drying digestate (13,351kWhthday(-1)), for the subsequent milling step; (iii) to reduce the electric energy requirement for the milling step (from 23,880 to 3580kWhelday(-1)); (iv) to produce extra electrical energy of 8483kWhelday(-1); (v) to improve the reduction of waste streams generated (from 13% to 54% of organic matter removal).