Innovative co-production of polyhydroxyalkanoates and methane from broken rice.

Broken rice, a low-cost starchy residue of the rice industry, can be an interesting substrate to reduce the polyhydroxyalkanoates (PHAs) production cost. However, since the most common PHAs-producing strains lack amylases, this waste must be firstly hydrolysed by additional commercial enzymes. In this work, the acidogenesis phase of the anaerobic digestion was exploited as efficient hydrolysis step to convert broken rice into volatile fatty acids (VFAs) to be used as PHAs carbon source by Cupriavidus necator DSM 545, one of the most promising PHAs-producing microbes. Broken rice, both non-hydrolysed and enzymatically hydrolysed, was processed in two continuous stirred tank reactors, at hydraulic retention times (HRT) of 5, 4 and, 3 days, to produce VFAs. The highest VFAs levels were obtained from non-hydrolysed broken rice which was efficiently exploited for PHAs accumulation by C. necator DSM 545. PHAs contents were higher after 96 h of incubation and, noteworthy, reached the highest value of 0.95 g/L in the case of 4 days HRT without any chemicals supplementation, except vitamins. Moreover, in view of a biorefinery approach, the residual solid fraction was used for methane production resulting in promising CH4 levels. Methane yields were very promising again for 4 days HRT. As such, this HRT resulted to be the most suitable to obtain effluents with high promise in terms of both PHAs accumulation and CH4 production. In addition, these results demonstrate that broken rice could be efficiently processed into two valuable products without any costly enzymatic pre-treatment and pave the way for future biorefining approaches where this by-product can be converted in a cluster of added-value compounds. Techno-economical estimations are in progress to assess the feasibility of the entire process, in view of supporting the low-cost conversion of organic waste into valuable products.

Enhanced fermentative lactic acid production from source-sorted organic household waste: Focusing on low-pH microbial adaptation and bio-augmentation strategy.

Lactic acid (LA) production at low pH could significantly reduce the need for neutralizing agents, leading to reduction of operational costs. In the present study, LA production at acidic conditions was investigated using source-sorted organic household waste (SSOHW). Controlling the pH at low value (i.e. 5.0) and bio-augmenting with Pediococcus acidilactici led to a concentration of 39.3 ± 0.5 g-LA/L with a yield of 0.75 ± 0.02 g-LA/g-sugar. In contrast, secondary fermentation at higher pH level (i.e. 5.5 and 6.0) resulted in complete LA degradation. Subsequently, consecutive batch fermentations were conducted to adapt P. acidilactici to SSOHW and improve the LA production. Results showed that P. acidilactici could successively adapt in the SSOHW reaching a relative abundance above 2.8% at adaptation process. The added P. acidilactici ensured a high concentration of LA at three consecutive generations, achieving an increment above 18% compared to control test (abiotic augmentation). Moreover, adaptation processes (i.e. maintaining pH at 4.0 or stepwise decreasing the pH from 5.0 to 4.0) significantly improved LA concentration and productivity at the pH of 4.0. Overall, the results provide a promising method to reduce the LA production costs using residual resources.

Valorization of municipal organic waste into purified lactic acid.

Municipal organic waste (biowaste) consists of food derived starch, protein and sugars, and lignocellulose derived cellulose, hemicellulose, lignin and pectin. Proper management enables nutrient recycling and sustainable production of platform chemicals such as lactic acid (LA). This review gathers the most important information regarding use of biowaste for LA fermentation covering pre-treatment, enzymatic hydrolysis, fermentation and downstream processing to achieve high purity LA. The optimal approach was found to treat the two biowaste fractions separately due to different pre-treatment and enzyme needs for achieving enzymatic hydrolysis and to do continues fermentation to achieve high cell density and high LA productivity up to 12 g/L/h for production of both L and D isomers. The specific productivity was 0.4 to 0.5 h-1 but with recalcitrant biomass, the enzymatic hydrolysis was rate limiting. Novel purification approaches included reactive distillation and emulsion liquid membrane separation yielding purities sufficient for polylactic acid production.

Multicomponent nanoparticles as means to improve anaerobic digestion performance.

Sufficient quantity of trace metals is essential for a well performing anaerobic digestion (AD) process. Among the essential trace elements in active sites of multiple important enzymes for AD are iron and nickel ions. In the present study, iron and nickel in the form of Fe2O3 and NiO were coated on TiO2 nanoparticles to be used in batch and continuous operation mode. The effect of TiO2, Fe2O3-TiO2, and NiO-TiO2 nanoparticles on each step of AD process was assessed utilizing simple substrates (i.e. cellulose, glucose, acetic acid, and mixture of H2-CO2) as well as complex ones (i.e. municipal biopulp). The hydrolysis rate of cellulose substrate increased with higher dosages of the coated TiO2 with both metals. For instance, the hydrolysis rate was increased up to 54% at Fe2O3-TiO2 and at a concentration of 23.5 mg/L for NiO-TiO2 it was increased up to 58%, while higher dosage suppressed the hydrolytic activity. Experimental results revealed that low dosages of NiO-TiO2 increased the accumulated methane production up to 24% probably by increasing the enzymatic activity of acetoclastic methanogenesis. NiO-TiO2 showed positive effect on batch and continuous AD of biopulp and improved methane yield up to 8%.

Impact of storage duration and micro-aerobic conditions on lactic acid production from food waste.

Food waste (FW) is an abundant resource with great potential for lactic acid (LA) production. In the present study, the effect of storage time on FW characteristics and its potential for LA production was investigated. The largest part of sugars was consumed during 7 to 15 days of FW storage and the sugar consumption reached 68.0% after 15 days. To enhance the LA production, micro-aerobic conditions (13 mL air/g VS) and addition of ß-glucosidase were applied to improve polysaccharides hydrolysis, resulting to increase of monosaccharides content to 76.6%. Regarding fermentative LA production, the highest LA titer and yield of hydrolyzed FW was 32.1 ± 0.5 g/L and 0.76 ± 0.01 g/g-sugar, respectively. Furthermore, L-LA isomer was higher than 70% when FW was stored for up to 7 days. However, attention should be paid on controlling the FW storage to approximately one week.

Potassium inhibition during sludge and biopulp co-digestion; experimental and model-based approaches.

Process instability with consecutive low methane production are common challenges of the anaerobic digestion (AD) of municipal wastes. In the present study, the co-digestion of sewage sludge and municipal biopulp was investigated at batch and continuously fed digesters. At batch tests, the highest methane yield for co-digestion (467 ± 17 mLCH4/gVS) was achieved when biopulp contributed to 80% of organic matter content and sludge the remaining 20%. At continuous mode operation, co-digestion achieved 0.91 ± 0.11 L/(L·d) methane productivity, while mono-digestion of sludge achieved 0.62 ± 0.05 L/(L·d). Potassium inhibition was investigated at the most efficient co-digestion scenario and was found that the half maximal inhibitory concentration (IC50) occurred at 8 g-K+/L. Subsequently, the effect of K+ was investigated at different scenarios at continuous operation. Simulations based on BioModel described the inhibitory effect of K+ by introducing non-competitive inhibition of methanogens. Simulation results confirmed the strongly inhibitory effect of potassium to the AD process.

Anti-algal activity of Fe2O3-TiO2 photocatalyst on Chlorella vulgaris species under visible light irradiation.

Many industries located in coastal areas use a large amount of seawater. Algal biofouling can be a major problem that hinders the efficiency of these industrial facilities. In most cases, seawater requires algal removal pre-treatment to avoid or mitigate biofilm formation. To remediate green microalgae, Fe2O3-TiO2 nanoparticles with 2.5% w/w Fe2O3 were applied as a visible light driven photocatalyst. The anti-algal activity of the photocatalytic pre-treatment using green microalgae, Chlorella vulgaris was tested. The experiments were carried out in freshwater, artificial seawater, and real seawater. Effect of photocatalyst dosage, visible light intensity, and water salinity on the removal of microalgae was investigated. The highest inactivation efficiency of Chlorella vulgaris was achieved under 55 W/m2 visible light irradiation when 0.25 g/L of Fe2O3-TiO2 photocatalyst was used. The photocatalytic removal kinetics of Chlorella vulgaris followed the pseudo first order Langmuir-Hinshelwood model. The results revealed that the efficiency of photocatalytic removal of algae decreased with increasing of seawater salinity. The anti-algal activity of Fe2O3-TiO2 nanoparticles was attributed to the generation of reactive oxygen species (ROS) through the photocatalytic process. H+ radical was shown to be the most important ROS that nanoparticles produced in the aqueous media. Using Fe2O3-TiO2 nanoparticles in photocatalytic pre-treatment could be an efficient environmental-friendly method for micro-algal remediation in seawater under visible light.

Carbon dioxide anion radical as a tool to enhance lignin valorization.

Lignin is particularly recalcitrant for valorization via the existing pretreatment methods due to its complex cross-linking polymeric network. The aim of this study is to develop a novel integrated pretreatment strategy to exploit the potential of lignocellulosic biomass as resource for production of biofuels and aromatic chemicals. In this regard, a novel UV/TiO2/HCOOH reaction was proposed to systematically generate hydroxyl radical (OH) and carbon dioxide radical anion (CO2-) to depolymerize lignin. Usage of 2,3-dihydrobenzofuran as a simple probe molecule showed cleavage ß-O-4 linkage occurred via H abstraction mechanism. The addition of methyl viologen as CO2- scavengers proved the presence of CO2- in this UV/TiO2/HCOOH reaction. Lignin and wheat straw were used to investigate the effect of different parameters, including formic acid concentration and TiO2 dosage, on the efficiency of the reaction. At optimized conditions, the highest phenolic concentrations attained were 173.431 and 66.802 mg/g lignin and wheat straw, respectively. A cycle test was designed with the aim to favor the complete consumption of formic acid through more pretreatment cycles for producing the highest possible Total Phenolic Compounds (TPC) in the liquid phase. After the third consecutive cycle, 103.651 ±â€¯5.964 mg-TPC/g, was obtained. Meanwhile it was found the remaining wheat straw solid fibers used for biogas production, showed 11.0% increase biogas production and increased degradation rate compared to the untreated wheat straw.

Application of nano-structured materials in anaerobic digestion: Current status and perspectives.

Nanotechnology is gaining more attention in biotechnological applications as a research area with a huge potential. Nanoparticles (NPs) can influence the rate of anaerobic digestion (AD) as the nano-sized structures, with specific physicochemical properties, interact with substrate and microorganisms. The present work has classified the various types of additives used to improve the AD processes. Nanomaterials as new additives in AD process are classified into four categories: Zero-valent metallic NPs, Metal oxide NPs, Carbon based nanomaterials, and Multi-compound NPs. In the following, application of nanomaterials in AD process is reviewed and negative and positive effects of these materials on the AD process and subsequently biogas production rate are discussed. This study confirms that design and development of new nano-sized compounds can improve the performances of the AD processes.

Graphene based ZnO nanoparticles to depolymerize lignin-rich residues via UV/iodide process.

In this work, potassium iodide (KI) and graphene oxide (GO) were utilized to promote the selectivity of photocatalytic process for alkali lignin oxidation over ZnO. Different concertation of GO was added during the microwave synthesis procedure of ZnO, and the characterization results revealed that graphene can shift the conduction band to more reducing potential, resulting to higher production of superoxide anion radicals (O2-) compared to OH. Response Surface Methodology revealed the most suitable interaction among loading of GO, KI and irradiation time on lignin and total phenolic compound (TPC) degradation. Specifically, the optimal conditions (i.e. maximum lignin (52%) and minimum TPC (55%) degradation) were at [KI] = 0.64 mM; GO content into ZnO 1.2 mg/mL; 240 min of irradiation time. The results showed that higher addition of graphene into structure of ZnO could preserve more phenolics from degradation due to less production of OH. Furthermore, the addition of KI at optimized conditions could enhance the selectivity of degradation of lignin and phenolics via producing I radicals and quenching the excess amount of generated OH, respectively. The lower generation of OH at optimized conditions was quantitatively confirmed by a photoluminescence simplified technique. In addition, the effect of the photocatalytic process on substrate's anaerobic degradability was examined in order to evaluate the suitability of the pretreated solution for energy recovery. Indeed, besides the higher TPC concentration, the biogas production of treated straw at optimized conditions was increased by 35% compared to the untreated sample.

Miscanthus straw as substrate for biosuccinic acid production: Focusing on pretreatment and downstream processing.

The main aim of this study was to optimize pretreatment strategies of Miscanthus × giganteus for biosuccinic acid production. A successful pretreatment with organosolv method (80% w/w of glycerol, 1.25% of H2SO4), prevented sugars conversion to furfurals and organic acids, and thereby resulted in high sugar recovery (glucan > 98%, xylan > 91%) and biomass delignification (60%). Pretreated biomass was subjected to hydrolysis with various cellulolytic enzyme cocktails (Viscozyme® L, Carezyme 1000L®, ß-Glucanase, Cellic® CTec2, Cellic® HTec2). The most effective enzymes mixture composed of Cellic® CTec2 (10% w/w), ß-Glucanase (5% w/w) and Cellic® HTec2 (1% w/w) resulted in high glucose (93.1%) and xylose (69.2%) yields after glycerol-based pretreatment. Succinic acid yield of 75-82% was obtained after hydrolysates fermentation, using Actinobacillus succinogenes 130Z. Finally a successful downstream concept for succinic acid purification was proposed. The succinic acid recovery with high purity (>98%) was developed.

Co-digestion of Laminaria digitata with cattle manure: A unimodel simulation study of both batch and continuous experiments.

This work investigated the thermophilic (55 °C) co-digestion performance both in batch and continuous mode operation. The biochemical methane potentials of L. digitata and cattle manure were 308 ±â€¯24 and 203 ±â€¯33 mL CH4/g VS, respectively. The optimum co-digestion feedstock ratio was found to be 80% macroalgae: 20% manure on a volatile solids basis, which produced 290 ±â€¯19 mL CH4/g VS under long-term and stable continuous operation at an organic loading rate of 2 g VS/L/d and hydraulic retention time of 15 days. Simulations of the batch and continuous experiments were, for the first time, carried out using an integrated anaerobic bioconversion model without structural modifications. Close fits between measured and simulated data provided mutual confirmation of experimental reliability and model robustness, and provided new perspectives for the use of the software tool.

Nickel spiking to improve the methane yield of sewage sludge.

The presence of micro-nutrients can be stimulatory for the anaerobic digestion (AD) of hardly degradable wastes and thus, improve process performance. Among the essential trace elements, nickel is involved in multiple important enzymes necessary for efficient AD. The present study investigates the effect of nickel spiked sewage sludge on batch and continuous mode operation. Metal spiking was conducted in the form of nanoparticles (Ni-NPs) and salt (NiCl2·6H2O). Results from batch assays showed that 5 mgNi-Salt/kgVS in the presence of Nitrilotriacetic acid (NTA) enhanced the methane yield by ∼10% compared to the untreated sample. The impact of Ni-NPs in the AD process was also positive, but slightly lower compared to the effect of NiCl2·6H2O. The stimulatory impact of Ni was also revealed in continuously fed digester boosting the methane yield by ∼8%. Overall, the improved methane production indicated that methanogenic archaea were favoured by the simultaneous supplementation of Ni and NTA.

Photocatalytic inactivation of Vibrio fischeri using Fe2O3-TiO2-based nanoparticles.

Biofouling is a major problem in water membrane processes, especially in seawater reverse osmosis plants. Inactivation of Vibrio fischeri (a well-known marine bacterium forming biofilm) through photocatalysis via visible light was investigated in this work using active Fe2O3-TiO2 nanoparticles. Five Fe2O3-TiO2 photocatalysts with different weight percentage of Fe2O3 (0-5 wt%) were synthesized using an ultrasonic-assisted co-precipitation method. The photocatalysts were characterized by powder X-ray diffraction (XRD), BET surface area, transmission electron Æ (TEM) plus selected area electron diffraction (SAED) patterns, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX) and diffuse-reflectance spectroscopy (DRS). Based on the design of experiments, the synthesized photocatalysts were tested for inactivation of V. fischeri under visible light irradiation at different temperatures (25-35 °C) and different photocatalyst dosage (0.1-2 g/L). The photocatalytic microbial inactivation experiments were performed in artificial seawater appropriate for growth of the marine bacterium. The results revealed that the highest inactivation efficiency of V. fischeri was achieved when 1 g/L of 2.5 wt% Fe2O3-TiO2 were used, at 35 °C. Photocatalytic inactivation of microorganisms using visible light-driven Fe2O3-TiO2 photocatalysts, could introduce an innovative green method in pretreatment units of reverse osmosis plants to control the membrane biofouling.

Integrated production of cellulosic bioethanol and succinic acid from rapeseed straw after dilute-acid pretreatment.

The aim of this study was to develop an integrated biofuel (cellulosic bioethanol) and biochemical (succinic acid) production process from rapeseed straw after dilute-acid pretreatment. Rapeseed straw pretreatment at 20% (w/v) solid loading and subsequent hydrolysis with Cellic® CTec2 resulted in high glucose yield (80%) and ethanol output (122-125 kg of EtOH/Mg of rapeseed straw). Supplementation the enzymatic process with 10% dosage of endoxylanases (Cellic® HTec2) reduced the hydrolysis time required to achieve the maximum glucan conversion by 44-46% and increased the xylose yield by 10% compared to the process with Cellic® CTec2. Significantly higher amounts of succinic acid were produced after fermentation of pretreatment liquor (48 kg/Mg of rapeseed straw, succinic acid yield: 60%) compared to fermentation of xylose-rich residue after ethanol production (35-37 kg/Mg of rapeseed straw, succinic yield: 68-71%). Results obtained in this study clearly proved the biorefinery potential of rapeseed straw.

Electricity generation and microbial communities in microbial fuel cell powered by macroalgal biomass.

The potential of macroalgae Laminaria digitata as substrate for bioelectricity production was examined in a microbial fuel cell (MFC). A maximum voltage of 0.5 V was achieved without any lag time due to the high concentration of glucose and mannitol in the hydrolysate. Total chemical oxygen demand removal efficiency reached over 95% at the end of batch run. Glucose and mannitol were degraded through isobutryrate as intermediate. The 16S rRNA gene high throughout sequencing analysis of anodic biofilm revealed complex microbial composition dominated by Bacteroidetes (39.4%), Firmicutes (20.1%), Proteobacteria (11.5%), Euryarchaeota (3.1%), Deferribacteres (1.3%), Spirochaetes (1.0%), Chloroflexi (0.7%), Actinobacteria (0.5%), and others (22.4%). The predominance of Bacteroidetes, Firmicutes and Proteobacteria demonstrated their importance for substrate degradation and simultaneous power generation. These results demonstrate that macroalgae hydrolysate can be used as a renewable carbon source of microbial electrochemical systems for various environmental applications.

Process performance and modelling of anaerobic digestion using source-sorted organic household waste.

Three distinctive start-up strategies of biogas reactors fed with source-sorted organic fraction of municipal solid waste were investigated to reveal the most reliable procedure for rapid process stabilization. Moreover, the experimental results were compared with mathematical modeling outputs. The initial inoculations to start-up the reactors were 10, 50 and 100% of the final working volume. While a constant feeding rate of 7.8gVS/d was considered for the control reactor, the organic loading rate for fed-batch reactors with 10 and 50% inoculation was progressively increased during a period of 60 and 13days, respectively. The results clearly demonstrated that an exponentially feeding strategy, considering 50% inoculation relative to final volume, can significantly decrease the alternatively prolonged period to reach steady conditions, as observed by high biogas and methane production rates. The combination of both experimental and modelling/simulation succeeded in optimizing the start-up process for anaerobic digestion of biopulp under mesophilic conditions.

Amino acids production focusing on fermentation technologies - A review.

Amino acids are attractive and promising biochemicals with market capacity requirements constantly increasing. Their applicability ranges from animal feed additives, flavour enhancers and ingredients in cosmetic to specialty nutrients in pharmaceutical and medical fields. This review gives an overview of the processes applied for amino acids production and points out the main advantages and disadvantages of each. Due to the advances made in the genetic engineering techniques, the biotechnological processes, and in particular the fermentation with the aid of strains such as Corynebacterium glutamicum or Escherichia coli, play a significant role in the industrial production of amino acids. Despite the numerous advantages of the fermentative amino acids production, the process still needs significant improvements leading to increased productivity and reduction of the production costs. Although the production processes of amino acids have been extensively investigated in previous studies, a comprehensive overview of the developments in bioprocess technology has not been reported yet. This review states the importance of the fermentation process for industrial amino acids production, underlining the strengths and the weaknesses of the process. Moreover, the potential of innovative approaches utilizing macro and microalgae or bacteria are presented.

In-situ biogas upgrading process: Modeling and simulations aspects.

Biogas upgrading processes by in-situ hydrogen (H2) injection are still challenging and could benefit from a mathematical model to predict system performance. Therefore, a previous model on anaerobic digestion was updated and expanded to include the effect of H2 injection into the liquid phase of a fermenter with the aim of modeling and simulating these processes. This was done by including hydrogenotrophic methanogen kinetics for H2 consumption and inhibition effect on the acetogenic steps. Special attention was paid to gas to liquid transfer of H2. The final model was successfully validated considering a set of Case Studies. Biogas composition and H2 utilization were correctly predicted, with overall deviation below 10% compared to experimental measurements. Parameter sensitivity analysis revealed that the model is highly sensitive to the H2 injection rate and mass transfer coefficient. The model developed is an effective tool for predicting process performance in scenarios with biogas upgrading.

A systematic methodology to extend the applicability of a bioconversion model for the simulation of various co-digestion scenarios.

Detailed simulation of anaerobic digestion (AD) requires complex mathematical models and the optimization of numerous model parameters. By performing a systematic methodology and identifying parameters with the highest impact on process variables in a well-established AD model, its applicability was extended to various co-digestion scenarios. More specifically, the application of the step-by-step methodology led to the estimation of a general and reduced set of parameters, for the simulation of scenarios where either manure or wastewater were co-digested with different organic substrates. Validation of the general parameter set involved the simulation of laboratory-scale data from three continuous co-digestion experiments, treating mixtures of different organic residues either at thermophilic or mesophilic conditions. Evaluation of the results showed that simulations using the general parameter set fitted experimental data quite well, indicating that it offers a reliable reference point for future simulations of anaerobic co-digestion scenarios.