Fermentation Reactions Research Articles

Synergistic sodium alginate- and biochar-immobilized cells for enhancing fermentative hydrogen production from food waste.

An immobilized hydrogen-producing consortium investigated biohydrogen production from food waste using a combination of sodium alginate and cassava rhizome biochar. We investigated the effect of varying the biochar concentration from 0 to 3% and the size of immobilized cells from 1 to 7mm. Immobilized cells were prepared using 50% (v/v) enriched hydrogen-producing consortium, 2% (w/v) sodium alginate, and 0 to 3% (w/v) cassava rhizome biochar. The optimal conditions for achieving the highest hydrogen production in the batch fermentation reactor were identified as a biochar concentration of 2% (w/v) and an immobilized cell size of 2mm. The highest hydrogen yield, maximum hydrogen production rate, and lag time recorded were 0.69mmol H2/g-COD, 0.02mmol H2/g-COD.h, and 41.51h, respectively. This research highlights the potential of cassava biochar technology for efficient biohydrogen production from food waste, contributing to renewable energy generation and sustainable waste management.

Preparation and flavor characteristics of plant-based meat flavoring by mixed fermentation of multiple strains

In this study, plant-based meat flavoring was prepared by mixed fermentation and thermal reaction. Based on the changes of amino acid nitrogen, reducing sugar, and total acid during the fermentation process and combined with sensory evaluation, the optimal fermentation conditions were determined as 4 % total inoculation amount of Monascus purpureus, Actinomucor elegans, and Bacillus subtilis with an inoculation ratio of 1:1:1, and 36 h of fermentation time. The odor characterization of meat flavoring was identified by solid-phase microextraction–gas chromatography–mass spectrometry (SPME–GC–MS), aroma extract dilution analysis (AEDA), quantitative measurements, odor activity value (OAV) and aroma recombination and omission experiments. 64 volatile compounds and 28 odorants were identified. The compounds with FD ≥ 27 were precisely quantified by five-point standard calibration curve and OAVs were calculated, and those with OAV ≥ 1 were subjected to aroma recombination and omission experiments. Finally, furfuryl mercaptan, difurfuryl disulfide, 4-methyl-5-ethylthiazole, 1-octanal, and 2-acetylthiophene were determined to be the key aroma-active compounds. These compounds have been recognized as key aroma-active compounds or detected in other meats or meat flavorings. In this study, we prepared plant-based meat flavoring through mixed fermentation, and provide new ideas for the development of plant-based thermal reaction meat flavoring process.

Modeling, simulations, and Simulink developments in the analysis of optimal control of temperature and pH in a batch ethanol fermentation process

Transfer of laboratory-scale experiments to production-scale ethanol fermentation is time-consuming and involves expensive prototype systems from complex experimental designs that determine optimal operating conditions for minimal substrate and product inhibitions. The study developed and validated a Simulink-based model for optimal pH and temperature control using fuzzy logic and PID controllers respectively and taking advantage of 2D and 1D substrate and product inhibition models from which suitable ethanol fermentation reaction rates models were selected. Temperature and pH levels and substrate, product, and biomass concentrations were measured. Selected inhibition models were linear-product, linear substrate-sudden stop product, and linear substrate for cassava, maize, and sorghum, respectively. Fuzzy logic controller ascertained optimal flow rate of acid and base as 0.000196 ml/s and 0.000204 ml/s, respectively, and pH error and rate of pH error as 0.00334 and 0.00368, respectively. F-test two-sample for variances showed no significant difference between model and experimental curves (cassava: F critical = 0.9704, F calculated = 0.1905; maize: F critical 0.9704, F calculated = 0.2149; sorghum: F critical = 0.9704, F calculated = 0.2488). PID logic controller showed model curves and experimental curves with good fit. F-test two-sample for variances showed no significant difference between model and experimental curves (cassava: F critical = 0.9704, F calculated = 0.1288; maize: F critical = 0.9704, F calculated = 0.2083; sorghum: F critical = 0.9704, F calculated = 0.2016). The study provided an improved approach as solution for optimal pH and temperature conditions in order to mitigate substrate and product inhibitions during ethanol fermentation. It illustrated that the application of artificial intelligence-based controllers provides satisfactory outcomes that are desirable for implementation in the industrial space.Graphical

Digital model of biochemical reactions in lactic acid bacterial fermentation of simple glucose and biowaste substrates

As concerns about the environmental impacts of biowaste disposal increase, lactic acid bacterial fermentation is becoming increasingly popular. Current academic research is aimed at the process optimization by developing digital bioreactors. The primary focus is to develop a digital model mimicking the biochemical reactions. In the light of this, this paper intended to build a digital model of biochemical reactions during the fermentation process of both glucose and biowaste substrates, including white pasta and organic municipal waste. For this purpose, near-infrared (NIR) and mid-infrared (MIR) spectroscopy techniques were used to collect spectral information during the fermentation process. Next, the samples were analyzed by High Pressure Liquid Chromatography (HPLC) to measure their glucose, fructose, arabinose, xylose, disaccharide, lactic acid, and acetic acid contents. The results showed that learning algorithms trained on MIR spectra accurately estimated the biochemical reactions for both glucose and biowaste substrates. For the glucose substrate, the results showed R-squared of 0.97 and RMSE of 4.69 g/L for glucose, and R-squared of 0.98 and RMSE of 2.74 g/L for lactic acid. In the case of biowaste substrate, estimations included glucose (R-squared = 0.97, RMSE = 4.69 g/L), fructose (R-squared = 0.88, RMSE = 1.47 g/L), arabinose (R-squared = 0.98, RMSE = 0.55 g/L), xylose (R-squared = 0.93, RMSE = 1.11 g/L), disaccharide (R-squared = 0.90, RMSE = 0.55 g/L), total sugar (R-squared = 0.98, RMSE = 3.79 g/L), lactic acid (R-squared = 0.98, RMSE = 2.74 g/L), and acetic acid (R-squared = 0.97, RMSE = 0.36 g/L). Regarding NIR spectral data, the predictive models were accurate when the substrate was glucose, however, they failed to accurately estimate the chemical reactions in the case of biowaste substrate. The findings of this study can be used to fulfill the requirements for a continuous fermentation process with the objective of maximizing lactic acid production.

Multiscale modeling and experimental study of molecular weight distribution and monomeric ratio in PHA production

While plastics have brought considerable convenience to our daily lives, most of them are still made up of petroleum-based materials. The plastic production industry accounts for a substantial portion of greenhouse gas emissions, and these products cause serious environmental pollution due to their poor degradability. Hence, bio-derived polymers have gained much research interest to substitute the existing plastic materials. Among them, polyhydroxyalkanoates (PHAs) are regarded as the most promising candidate owing to their biocompatible and biodegradable nature. In particular, poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB) is now in the limelight for its unique mechanical properties. However, understanding the sophisticated mechanism and structure is quite challenging, posing obstacles to its successful commercialization. In response to this difficulty, we devised a multiscale model using the kinetic Monte Carlo (kMC) algorithm to enable capturing the extensive characteristics of PHA, including molecular weight distribution and monomeric ratio. Considering the different time and length scales of the overall process, the model is developed with two layers for the fermentation and polymerization reactions, and they are successfully integrated. The results not only validate the experimental results but also showcase a wide range of PHA information that cannot be measured through instrumental analyses.

Comparative genomic analysis of an emerging Pseudomonadaceae member, Thiopseudomonas alkaliphila.

Thiopseudomonas alkaliphila, an organism recently classified within the Pseudomonadaceae family, has been detected in diverse sources such as human tissues, animal guts, industrial fermenters, and decomposition environments, suggesting a diverse ecological role. However, a large knowledge gap exists in how T. alkaliphila functions. In this comparative genomic analysis, adaptations indicative of habitat specificity among strains and genomic similarity to known opportunistic pathogens are revealed. Genomic investigation reveals a core metabolic utilization of multiple oxidative and non-oxidative catabolic pathways, suggesting adaptability to varied environments and carbon sources. The genomic repertoire of T. alkaliphila includes secondary metabolites, such as antimicrobials and siderophores, indicative of its involvement in microbial competition and resource acquisition. Additionally, the presence of transposases, prophages, plasmids, and Clustered Regularly Interspaced Short Palindromic Repeats-Cas systems in T. alkaliphila genomes suggests mechanisms for horizontal gene transfer and defense against viral predation. This comprehensive genomic analysis expands our understanding on the ecological functions, community interactions, and potential virulence of T. alkaliphila, while emphasizing its adaptability and diverse capabilities across environmental and host-associated ecosystems.IMPORTANCEAs the microbial world continues to be explored, new organisms will emerge with beneficial and/or pathogenetic impact. Thiopseudomonas alkaliphila is a species originally isolated from clinical human tissue and fluid samples but has not been attributed to disease. Since its classification, T. alkaliphila has been found in animal guts, animal waste, decomposing remains, and biogas fermentation reactors. This is the first study to provide an in-depth view of the metabolic potential of publicly available genomes belonging to this species through a comparative genomics and draft pangenome calculation approach. It was found that T. alkaliphila is metabolically versatile and likely adapts to diverse energy sources and environments, which may make it useful for bioremediation and in industrial settings. A range of virulence factors and antibiotic resistances were also detected, suggesting T. alkaliphila may operate as an undescribed opportunistic pathogen.

Pulsed light processing of sugarcane juice: quality evaluation and microbial load assessment.

Freshly extracted sugarcane juice is an ideal substrate for microbial fermentation and browning reactions. The present study is the first report on the potential of pulsed light (PL) processing in improving microbial stability with the retention of major bioactive. PL processing at different levels of voltage (2.1-2.7 kV) and number of pulses (100-200) was explored. The present study aimed to investigate the impact of PL processing on the quality of sugarcane juice, bioactive composition and microbial load. The microbial load, such as aerobic mesophiles, yeast and mold, and total coliform, was reduced to below 1 log colony-forming units mL-1 in juice samples subjected to intense PL treatment at 2.7 kV. The maximum value of the total color difference of the sugarcane juice was below 4.0, even at extreme levels of PL process parameters. In comparison with the unprocessed juice, the reduction in total phenols (Folin ciocalteu reagent assay) and the total antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl free radical scavenging assay) was limited to 6% and 16.7%, respectively, when treated at 2.7 kV/200 pulses. The pH and total soluble solids of the juice remained unaffected in all the processed samples. Among the process parameters considered, the treatment voltage was found to significantly affect the quality parameters and microbial load. PL processing at 2.1 kV/170 pulses gave an optimally processed juice with a microbial load below the permissible limit and desirability value of 0.77. The results suggest that the PL treatment is effective for enhancing the microbial stability and maintaining the bioactive components of the sugarcane juice. Furthermore, the outcomes from the present study are expected to pave the way for further in-depth investigation of the effect of PL treatment on the critical quality attributes and shelf life of sugarcane juice. The technology will be useful for adoption by different stakeholders, including manufacturers and retailers in the food processing sector. © 2024 Society of Chemical Industry.

Produção de aguardente de abacaxi da Amazônia

Pineapple brandy represents an exotic fusion between the traditional Brazilian drink and the tropical flavor of pineapple. Its production involves a process of fermentation and distillation. This study explored the spirit production process on a laboratory and pilot scale, in which it is worth highlighting that no amounts of sugar were used to favor the fermentation reaction, as the brix content of the juice was 14° brix. To obtain the spirit, on a laboratory scale, a blender was used to obtain the juice, precision scales, and beakers for the fermentation process. In the simple distillation process, a distillation flask, heating blanket and a condenser were used. At the pilot scale, a mill, a reactor where fermentation took place and a copper still for distillation were used. The results of the production processes generated a drink with the characteristic appearance of traditional spirits, with an alcohol content of 45% vol/vol and an average density of 0.980969 g/ml. Therefore, its potential as a differentiated product can be seen, with possible application in the beverage market in Brazil.

Electrochemical Characteristics of Microbial Fuel Cells Operating with Various Food Industry Wastewaters

In the present work, four different wastewaters from the food industry were used in parallel, in four identical dual-chamber MFCs, with graphite granules as anodic electrodes. Specifically, a mixture of hydrogenogenic reactor effluents (effluents from a dark fermentation reactor fed with cheese whey (CW), for hydrogen production), CW, and a mixture of expired fruit juices and wastewater from the confectionery industry were simultaneously used in MFCs to evaluate the effect of the type of effluent/wastewater on their efficiency. An electrochemical characterization was performed using electrochemical impedance spectroscopy measurements under open- (OCP) and closed-circuit conditions, at the beginning and end of the operating cycle, and the internal resistances were determined and compared. The results showed that the highest OCP value, as well as the highest power density (Pmax) and Coulombic efficiency (εcb) at the beginning of the operating cycle, was exhibited by the MFC, using a sugar-rich wastewater from the confectionery industry as substrate (sugar accounts for almost 92% of the organic content). This can be correlated with the low internal resistance extracted from the Nyquist plot at OCP. In contrast, the use of CW resulted in a lower performance in terms of OCP, εcb and Pmax, which could be correlated to the high internal resistance and the composition of CW, a substrate rich in lactose (disaccharide), and which also contains other substances (sugars account for almost 72% of its organic content, while the remaining 28% is made up of other soluble compounds).

Analysis of the contribution of different electron transfer pathways for hydrogen production in a bioelectrochemically assisted dark fermentation system

This research analyzed the contribution of different electron transfer pathways for H2 production in a bioelectrochemicallly assisted dark fermentation system (BEDF), and compared to an electric field-driven dark fermentation (EFDF) reactor. The results showed that intrinsic extracellular electron transfer (iEET) via dark fermentation accounted for 76.3% of the H2 production in the BEDF reactor. In comparison, the extracellular electron transfer (bEET) pathway via the bioelectrode enhanced by the bioelectrochemical system and the electric field-driven extracellular electron transfer (eEET) in the bulk solution accounted for 5% and 18.7%, respectively. This suggests that the main electron transfer pathway for H2 production comes from the bulk solution rather than that on the bioelectrodes. Under the condition of electric field, extracellular polymeric substances (EPS) were secreted in large quantities in the bulk solution, and electroactive microorganisms such as Proteobacteria were significantly enriched, which further promote the H2 production through the eEET pathway.

Sewage sludge-derived biocarbons as catalysts of bioanodes in a dual-chamber microbial fuel cell using nejayote as substrate

In this work, nejayote (wastewater from tortilla industry) was evaluated as substrate of a dual-chamber microbial fuel cell (MFC). For this purpose, novel bioanodes were built using Bacillus subtilis (B. subtilis) as the electrochemically active microorganism (EAM), and sewage sludge-derived biocarbon (SSB) and methanol-functionalized SSB (FSSB) as catalysts. The relationship between the physicochemical properties of SSB and FSSB, biocompatibility, electrochemical performance and the percentage of COD removed was analyzed. Up to 70% of chemical oxygen demand (COD) from nejayote was successfully removed using the SSB + B. subtilis bioanode, with the advantage of energy generation by the MFC (Pcell= 1.44 mW m-2 and jcell= 41 mA m-2). An adverse effect was observed when functionalizing SSB to produce FSSB. Despite a fair biocompatibility, the FSSB + B. subtilis bioanode promoted fermentation reactions that increased the COD and charge transfer resistance. The improved performance of SSB + B. subtilis was ascribed to a reasonable biocompatibility, low charge transfer resistance and less limitation to the diffusion of proton species between the biofilm and the substrate. Therefore, the treatment of nejayote in a MFC with the SSB + B. subtilis bioanode was identified as a promising alternative for decreasing COD and generating small amounts of energy, instead of using energy for its remedy.

Application of Recycled Filling to Improve the Purification Performance of Confectionery Wastewater in a Vertical Anaerobic Labyrinth Flow Bioreactor

Anaerobic wastewater treatment is, in many cases, a justified alternative to typical activated sludge processes, from a technological, economic, and ecological point of view. The optimisation of fermentation reactors is primarily concerned with increasing the biodegradation of organic compounds and biogas production, as well as improving efficiency in the removal of nitrogen and phosphorus compounds. The aim of the research was to determine the impact of using low-cost recycled filling on the efficiency of treating real confectionery wastewater in a vertical anaerobic labyrinth flow bioreactor. The experiments focused on selecting the organic loading rate that would allow for the effective biodegradation and removal of pollutants, as well as the efficient production of biomethane. It was found that the tested reactor can operate efficiently at a maximum organic loading rate (OLR) of 7.0–8.0 g of chemical oxygen demand (COD)/L·d. In this OLR range, high efficiency was guaranteed for both wastewater treatment and biogas production. However, increasing the OLR value to 8.0 g COD/L·d had a significant negative effect on the methane (CH4) content in the biogas. The most efficient variants achieved a biodegradation efficiency of around 90% of the organic compounds, a CH4 content of over 70% in the biogas, and a biogas yield of over 400 L/kg of COD removed. A significant influence of the applied OLR on the ratio of free organic acids (FOS) to total alkaline capacity (TAC) and pH was observed, as well as a strong correlation of these indicators with the specific biogas yield and CH4 content. The application of a solution based on the use of a hybrid system of anaerobic granulated sludge and an anaerobic filter resulted in an efficient treatment process and an almost complete elimination of suspensions from the wastewater.

Phase separation as a strategy to prevent sulfide-related drawbacks in methanogenesis: performance and energetic aspects.

The establishment of sulfate (SO42-) reduction during methanogenesis may considerably hinder the efficient energetic exploitation of methane, once removing sulfide from biogas is obligate and can be costly. In addition, sulfide generation can negatively impact the performance of methanogens by triggering substrate competition and sulfide inhibition. This study investigated the impacts of removing SO42- during fermentation on the performance of a second-stage methanogenic continuous reactor (R2), comparing the results with those obtained in a single-stage system (R1) fed with SO42--rich wastewater (SO42- of up to 400mg L-1, COD/SO42- of 3.12-12.50). The organic load (OL) was progressively increased to 5.0g COD d-1 in both reactors, showing completely discrepant performances. Sulfate-reducing bacteria outperformed methanogens in the consumption for organic matter during the start-up phase (OL = 2.5g COD d-1) in R1, directing up to 73% of the electron flow to SO42- reduction. An efficient methanogenic activity was established in R1 only after decreasing the OL to 0.625g COD d-1, after which methanogenesis prevailed by consuming ca. 90% of the removed COD. Nevertheless, high sulfide proportions (up to 3.1%) were measured in biogas. Conversely, methanogenesis was promptly established in R2, resulting in a methane-rich (> 80%) and sulfide-free biogas regardless of the operating condition. From an economic perspective, processing the biogas evolved from R2 would be cheaper, although the techno-economic impacts of managing the sulfur pollution in the fermentative reactor still need to be understood.

Specific Organic Loading Rate Control for Improving Fermentative Hydrogen Production

Inhibiting homoacetogens is one of the main challenges in fermentative hydrogen production because these hydrogen consumers have similar growth features to hydrogen producers. Homoacetogens have been related to the excessive accumulation of biomass in fermentative reactors. Therefore, a suitable food/microorganism ratio has the potential to minimize the homoacetogenic activity. In this work, the specific organic loading rate (SOLR) was controlled in two fermentative fixed-bed up-flow reactors through scheduled biomass discharges. Reactors were differentiated by the bed arrangement, namely, packed and structured conformation. The SOLR decay along the time in both reactors was previously simulated according to the literature data. The volume and volatile suspended solids (VSS) concentration of discharges was estimated from the first discharge, and then additional discharges were planned. Biomass discharges removed 21% of the total biomass produced in the reactors, maintaining SOLR values of 3.0 ± 0.4 and 3.9 ± 0.5 g sucrose g−1 VSS d−1 in the packed-bed and structured-bed reactors, respectively. Such a control of the SOLR enabled continuous and stable hydrogen production at 2.2 ± 0.2 L H2 L−1 d−1 in the packed-bed reactor and 1.0 ± 0.3 L H2 L−1 d−1 in the structured-bed one. Controlling biomass was demonstrated to be a suitable strategy for keeping the continuous hydrogen production, although the fermentative activity was impaired in the structured-bed reactor. The homoacetogenic was partially inhibited, accounting for no more than 30% of the total acetic acid produced in the reactor. Overall, the high amount of attached biomass in the packed-bed reactor provided more robustness to the system, offsetting the periodic suspended biomass losses via the planned discharges. Better characterizing both the VSS composition (aiming to differentiate cells from polymeric substances) and the bed hydrodynamics could be useful to optimize the online SOLR control.

Enhanced Succinic Acid Production and Electronic Utilization Efficiency by Actinobacillus succinogenes 130Z in an ORP-Controlled Microbial Electrolysis Cell System

Microbial electrochemical systems have shown great value as a means of enhancing the efficiency of fermentation reactions, but at present, there is no reliable means to balance the extracellular electron supply and corresponding intracellular demands in these systems. The current work describes the unique use of an oxidation–reduction-potential (ORP)-level-controlled microbial electrolysis cell (MEC) system to successfully balance the extracellular electron supply and succinic acid fermentation via A. succinogenes (130Z). The ORP-controlled MEC system with neutral red (NR) yielded a significant increase in succinic acid production (17.21%). The utilization of NR in this MEC system improved the ORP regulatory sensitivity. The optimal approach to the ORP level control was the use of a −400 mV high-voltage electric pulse-based strategy, which increased the yield of succinic acid by 13.08% compared to the control group, and reduced the energy consumption to 52.29% compared to the potentiostatic method. When compared to the −1 V constant potential MEC system, the high-voltage electric pulse-based ORP strategy for the MEC system control provided sufficient electrons to this system while using less electricity (11.96%) and producing 12.48% (74.43 g/L) more succinic acid during fed-batch fermentation. The electronic utilization efficiency of the ORP-controlled MEC system was 192.02%, which was 15.19 times that of the potentiostatic system. The electronic utilization efficiency is significantly increased in the ORP-controlled MEC system. Succinic acid production is ensured by a high-voltage electric pulse-based method, while the influence on cell growth and power consumption are minimized. Fed-batch fermentation with the high-voltage electric pulse-based ORP strategy for MEC system control is noted to be ideal to achieve a further increase in succinic acid concentration and electronic utilization efficiency.

Polyurethane as an alternative support media to low-density polyethylene in fermentative-sulfidogenic reactors: Beneficial or indifferent?

Sporadically addressed in previous studies, the removal of sulfate during fermentation has been a matter of great interest in new applications of fixed-film two-stage biodigestion as the most efficient strategy to boost methane production from a low-sulfate fermented substrate. This study assessed the use of polyurethane (PU) as the support material in a thermophilic (55 °C) fermentative/sulfidogenic fixed-film reactor subjected to different operating conditions based on the variation of the sulfate loading rate (SLR = 8.0–4.0 kg-SO42− m−3 day−1), HRT (12.0–16.0 h) and liquid upflow velocity (3.42–7.90 m h−1). A maximum sulfate removal efficiency of only 46.8 % was achieved when applying SLR and HRT of 4.0 kg-SO42− m−3 day−1 and 16.0 h, respectively, reaching a total dissolved sulfide concentration of 275 mg L−1. This poor performance resulted from two factors: the interspaced arrangement of PU pieces and the use of ethanol as the only carbon source. While the first triggered excess biomass accumulation, the latter favored the establishment of a very restricted microbial community. Only some Clostridia-like bacterial classes, Proteobacteria, and other non-SRB bacteria belonging to phyla Synergistetes and Thermotogae were identified. Hence, under the studied conditions, using PU did not enhance sulfidogenesis in the fermentative reactor.

Changes in quality and bacterial profiles of Tualang and Kelulut honeys preserved by post‐harvest maturation

SummaryPost‐harvest maturation of two Malaysian honeys, the Tualang and Kelulut was studied by measuring changes in physicochemical and antioxidant properties, hydroxymethylfurfural (HMF) contents and bacterial profiles at room temperature of 23–26 °C. After maturation at the recommended period of 26 weeks, water activity of both honeys increased between 0.89% and 2.34% while free acidity increased between 2.05% and 2.24%. Results suggested the prominence of fermentation reactions in honey during post‐harvest maturation as fructose concentration reduced by 10.6% and 1.05% for the Tualang and Kelulut honey, respectively, while HMF concentrations were kept at a safe limit of 48.00 and 61.23 mg/kg honey. The total phenolic content of Tualang and Kelulut honey increased significantly by 12.61% and 54.66% respectively. The highlight of this post‐harvest maturation process for Kelulut honey was the improvement found in antioxidant properties of DPPH radical scavenging activity by 10.01%–54.74% and also the probiotic‐like potential in terms significant increase in relative abundance of the Bacillus genera to 2.6% and Lactobacillus to 6.25% at 26 weeks. The prolonged maturation process up to 1 year however revealed continuous accumulation of HMF to values above 80 mg/kg honey, surpassing the limits by the Codex Alimentarius Commission despite improvements of antioxidant properties and bacterial profiles.

Towards biotechnological production of bio-based low molecular weight esters: a patent review.

Low molecular weight (LMW) esters, like ethyl acetate, methyl formate or butyl acetate, are widespread bulk chemicals in many industries. Each of them is currently produced in huge amounts (millions of tons per year scale) starting from fossil-based feedstock and they are used mainly because of their low toxicity and complete biodegradability. Energy transition is just half of the story on the path of fighting climate change: 45% of the global greenhouse gas emissions are caused by the production and use of all the products, materials and food necessary for modern human life. If the world is to reach its climate goals, there is the need to leave underground a significant proportion of the fossil feedstock and minimize environmental impacts of chemical manufacturing. This is the reason why a lot of efforts have been made to find novel routes for LMW esters production starting from renewable raw materials (e.g. biomasses or off-gases) and exploiting low-impact manufacturing, such as microbial fermentation or enzymatic reactions. This review reports the most significant patents, in the field of white biotechnology, that will hopefully lead to the commercialization of bio-based LMW esters as well as novel strategies, current problems to be solved, newer technologies, and some patent applications aiming at possible future developments.

Process parameters optimization for biosynthesis of gibberellic acid using orange waste as an alternative substrate

Gibberellic acid (GA3) is the most important gibberellin that affects various plant developmental processes. As such, it has an international market value of $106 million dollars. However, it is costly in the market because of high downstream processing cost that is linked to its current mode of production via submerged fermentation. Solid-state fermentation, in contrast, minimizes this limitation of cost when optimal process parameters are used. Hence, this study utilized orange waste as a cheap substrate in the bio-production of gibberellic acid through the optimization of three process parameters (incubation temperature, pH and substrate concentration). The study also investigated the interactive effects of these parameters on the yield of gibberellic acid. Using Design Expert® software, a 19-run experiment, varied at five levels, was generated. Accordingly, each fermentation reaction that had been supplemented with 0.03% FeSO4.7H2O and 0.01% (NH4)2SO4 was inoculated with a constant fungal inoculum. The highest yield of gibberellic acid was with a combination of 19.5 g of substrate set at 30 °C incubation temperature with a pH of 5.5. The interaction between the three factors was a linear relationship. The bio-production of gibberellic acid using orange waste as an alternative substrate suggests the possibility of a further reduction in cost of production of high-end value metabolites when proper optimization is carried out.

Optimization of methane production from co-digestion of citrus agro-industrial waste in a two-stage system: Investigation of the effects of organic matter concentration and alkalinization

The essential oils in citrus waste poses challenges for the anaerobic digestion application to energy recovery from these wastes due to the potential to inhibit the bioprocess. The two-stage system is a practical alternative to overcome these limitations of anaerobic co-digestion of citrus peel waste and wastewater. This study proposes to overcome drawbacks associated with the use of these substrates in anaerobic digestion by separating the phases, performing parameters optimization of the methanogenic stage. To obtain optimized conditions for the methanogenic stage, the Rotational Central Composite Design (RCCD) and Response Surface Methodology (RSM) were applied, evaluating the effects of diluting the Fermentation Reactor Content (FRC) of the first stage (35.7%–100%) and alkalinization (up to 0.60 g sodium bicarbonate.gCOD−1), in methane production and organic matter removal. Values from 2100.4 to 6320.6 mLCH4.L−1 were obtained in the experimental design assays, with no inhibition identified by the high substrate concentration, nor by the limonene content (1.5–5.0 mg.L−1). Lower dilution of the fermentation reactor content (90%–100%) resulted in higher accumulated methane production, while the variation in the alkalinization did not imply significant effects on this response. The optimized condition for maximum production potential was obtained with 99.3% of fermentation reactor content and 0.58 gNaCHO3.gCOD−1, reaching a production of 6549.7 mLCH4.L−1, with organic matter removal (rCOD) of 87.1%, and 58.6% of reduction in solid waste content. For this condition, acetogenic metabolism predominance was observed, in the conversion of butyric acid and propionic acid into acetic acid. In the analysis of the microbial community of the optimized condition assay, a high relative abundance of hydrolytic and acidogenic bacteria (AUTHM297, Lachnospiraceae, and Hungateiclostridiaceae) was observed, as well as methanogenic archaea related to acetoclastic, hydrogenotrophic, and methylotrophic methanogenesis (Methanoregula, Methanolinea, Methanomassiliicoccus, Candidatus Methanomethylicus, and Methanofolis).