Simultaneous Saccharification And Fermentation Research Articles

Harnessing Bacillus subtilis QY5 PP784163 for Bioethanol Production from Potato Peel Waste and Nutrient Recovery for Animal Feed: Maximizing Resource Efficiency

The present work focuses on the utilization of potato peel waste for the production of bioethanol. In the present study, extensive screening was undertaken to isolate amylolytic and cellulolytic microbes using starchy biomass. After confirming the chemical composition of potato peel waste (PPW), several trials were performed to enhance the amylase and cellulase production from Bacillus subtilis to hydrolyze the PPW in submerged fermentation. Optimization of physical parameters was performed using both commercial and indigenous media from enzymatically hydrolyzed PPW. Different routes of various combinations were designed to enhance bioethanol production. The maximum ethanol titer of 0.50% and 0.41% was recorded in Route B and A, i.e., separate saccharification and ethanol fermentation and consolidated fermentation. Simultaneous saccharification and fermentation (SSF) also measured a good ethanol yield of 0.46%. The fermented residual cake was checked for nutritional components and showed a high content of protein and amino acids because of the addition of unicellular yeasts. This cake can be utilized as an animal feed supplement.

Stirring the hydrogen and butanol production from Enset fiber via simultaneous saccharification and fermentation (SSF) process

Enset fiber is a promising feedstock for biofuel production with the potential to reduce carbon emissions and improve the sustainability of the energy system. This study aimed to maximize hydrogen and butanol production from Enset fiber through simultaneous saccharification and fermentation (SSF) process in bottles as well as in bioreactor. The SSF process in bottles resulted in a higher butanol concentration of 11.36 g/L with a yield of 0.23 g/g and a productivity of 0.16 g/(L h) at the optimal process parameters of 5% (w/v) substrate loading, 16 FPU/g cellulase loading, and 100 rpm agitation speed from pretreated Enset fiber. Moreover, a comparable result to the bottle experiment was observed in the bioreactor with pH-uncontrolled SSF process, although with a decreased in butanol productivity to 0.095 g/(L h). However, using the pre-hydrolysis simultaneous saccharification and fermentation (PSSF) process in the bioreactor with a 7% (w/v) substrate loading led to the highest butanol concentration of 12.84 g/L with a productivity of 0.104 g/(L h). Furthermore, optimizing the SSF process parameters to favor hydrogen resulted in an increased hydrogen yield of 198.27 mL/g-Enset fiber at atmospheric pressure, an initial pH of 8.0, and 37 °C. In general, stirring the SSF process to shift the product ratio to either hydrogen or butanol was possible by adjusting temperature and pressure. At 37 °C and atmospheric pressure, the process resulted in an e-mol yield of 12% for hydrogen and 38% for butanol. Alternatively, at 30 °C and 0.55 bar overpressure, the process achieved a yield of 6% e-mol of hydrogen and 48% e-mol of butanol. This is the first study to produce hydrogen and butanol from Enset fiber using the SSF process and contributes to the development of a circular bioeconomy.Graphical

Altered sterol composition mediates multiple tolerance of Kluyveromyces marxianus for xylitol production

BackgroundCurrently, the synthesis of compounds based on microbial cell factories is rapidly advancing, yet it encounters several challenges. During the production process, engineered strains frequently encounter disturbances in the cultivation environment or the impact of their metabolites, such as high temperature, acid-base imbalances, hypertonicity, organic solvents, toxic byproducts, and mechanical damage. These stress factors can constrain the efficiency of microbial fermentation, resulting in slow cell growth, decreased production, significantly increased energy consumption, and other issues that severely limit the application of microbial cell factories.ResultsThis study demonstrated that sterol engineering in Kluyveromyces marxianus, achieved by overexpressing or deleting the coding genes for the last five steps of ergosterol synthase (Erg2-Erg6), altered the composition and ratio of sterols in its cell membrane, and affected its multiple tolerance. The results suggest that the knockout of the Erg5 can enhance the thermotolerance of K. marxianus, while the overexpression of the Erg4 can improve its acid tolerance. Additionally, engineering strain overexpressed Erg6 improved its tolerance to elevated temperature, hypertonic, and acid. YZB453, obtained by overexpressing Erg6 in an engineering strain with high efficiency in synthesizing xylitol, produced 101.22 g/L xylitol at 45oC and 75.11 g/L xylitol at 46oC. Using corncob hydrolysate for simultaneous saccharification and fermentation (SSF) at 46oC that xylose released from corncob hydrolysate by saccharification with hemicellulase, YZB453 can produce 45.98 g/L of xylitol, saving 53.72% of the cost of hemicellulase compared to 42oC.ConclusionsThis study elucidates the mechanism by which K. marxianus acquires resistance to various antifungal drugs, high temperatures, high osmolarity, acidity, and other stressors, through alterations in the composition and ratio of membrane sterols. By employing sterol engineering, the fermentation temperature of this unconventional thermotolerant K. marxianus was further elevated, ultimately providing an efficient platform for synthesizing high-value-added xylitol from biomass via the SSF process at temperatures exceeding 45 °C.

Integrating use of sugarcane bagasse with sugarcane juice for efficient l-lactic acid production through fed-batch simultaneous saccharification and fermentation: Process configurations and kinetic analysis

The present study investigated the utilization of sugarcane juice (SJ) and sugarcane bagasse (SB) for L-lactic acid (L-LA) production by a newly isolated Enterococcus gallinarum TSKKU D-6. SJ was first investigated as a single substrate, yielding 43.9 ± 2.9 g-LA/L, with a fermentation efficiency of 72.0 ± 0.1 %, based on sucrose content in SJ. On the other hand, simultaneous saccharification and fermentation (SSF) of SB as a single substrate yielded 68.2 ± 2.7 g-LA/L, with a fermentation efficiency of 74.1 ± 0.0 %, based on cellulose content in SB. SSF of SJ and SB as co-substrates was subsequently carried out to increase the acid production, where SJ was first fermented, followed by the addition of SB to enhance LA titer. Effects of mixing and SB addition strategy on the kinetics and economic feasibility of L-LA production were investigated using a bioreactor equipped with a single and dual impellers, as well as the use of dual impellers under fed-batch SSF. The use of single and dual impellers gave similar L-LA titers of 95.9 ± 1.4 and 102.6 ± 0.6 g/L, respectively. Nevertheless, the use of dual impellers shortened the processing time, enhancing LA productivity from 0.37 to 1.12 g/(L·h). As a result, the economic yield and productivity were improved by 7.1 % and 187.5 %, respectively. Better still, performing fed-batch SSF with the use of dual impellers further improved the economic productivity by 8.7 %. This study demonstrated that integrating use of SB as a low-cost substrate considerably enhanced the acid production and lessened the use of SJ. It was also demonstrated that mixing, and process configuration are critical factors determining the process performance.

Investigation of the effect of specific gravity and pH on the fermentation period of a single and combined algae species as a potential bioethanol feedstock

Algae has garnered significant attention as a promising biomass source for bioethanol production and as a solution to sustainability issues. Nevertheless, the disruption of complex carbohydrates required for yeast fermentation occurs during algae cultivation. The fermentation process, which is critical for producing bioethanol as a cost-effective and environmentally friendly alternative fuel, may be improved by meticulously managing the parameters that influence fermentation, such as specific gravity and pH. Bioethanol was produced from algae species (Navicula sp., Oscillatoria sp. and diatoms) using simultaneous saccharification and fermentation (SSF) processes. A control, single and combined algal species were prepared in the following formation: Oscillatoria sp. and Navicula sp. (sample A), Oscillatoria sp. (sample B), diatom and Oscillatoria sp. (sample C), Navicula sp. (sample D), and diatoms, Navicula sp. and Oscillatoria sp (sample E). The daily bioethanol concentrations in the fermentation medium were measured using a digital optical refractometer, and a Two-way ANOVA analysis (p < 0.05) was conducted to determine the differences between the period of fermentation and the bioethanol concentration. Additionally, the pH and specific gravity were assessed. The Two-way ANOVA analysis conducted indicated that the F-values surpassed the F-critical values and the p-values were all notably below 0.05, suggesting a statistically significant distinction between the samples in their fermentation periods and bioethanol concentration. The highest concentration was observed in sample E at 39.53 g/L, followed by samples C and D at 37.36 g/L, samples A and B at 35.10 g/L, and the control at 4.78 g/L. The findings showed that mixed cultures could potentially improve ethanol yield. The optimal fermentation period was determined to be 72 h. The pH levels in the control and algae samples varied between 6.5 and 6.7 and 7.06 to 8.42, respectively. The specific gravity in the control and algae samples ranged from 1.003 to 1.004 and 1.023 to 1.037, respectively. During algae fermentation, specific gravity initially increased, then stabilized before decreasing. pH levels correlated with fermentation duration. Understanding these factors is crucial for maximizing bioethanol production. This discovery could lead to exploring how different algae species interact to improve cultivation methods.

Biovalorizing felled oil palm trunk as a sole feedstock for lactic acid production through efficient simultaneous saccharification and fermentation

This study aimed to develop practical biorefinery process for bio-valorizing felled oil palm trunk (OPT) as a sole feedstock for lactic acid (LA) production. OPT was separated into oil palm sap (OPS), vascular bundle (OPT-VB), and parenchyma (OPT-PA) fractions. OPS containing high amounts of sugars (38.69 g/L) and nitrogen (0.63 g/L), could be used directly for LA production by Lactobacillus acidophilus and also as base medium for simultaneous saccharification and fermentation (SSF) of OTP-VB and OPT-PA. The repeated SSF efficiently utilized cellulosic OPT-VB and produced LA up to 50–71 g/L during five cycles. Due to high water-absorbing and swelling properties of OPT-PA, it could not be initially added at high loadings. It was then intermittently added through fed-batch SSF, which effectively produced LA of 72.85 ± 1.61 g/L. These strategies have shown the efficient biorefinery process for biovalorization of OPT and may also be applicable to other similar agricultural wastes.

Bioethanol derived from rice straw agricultural waste as alternative energy towards a pollution-free era

Abstract Rice production occupies the first position as raw material for bioethanol production, with a total output throughout Indonesia of 56.63 million tons in 2023 (BPS, 2023). The purpose of this innovation is to create bioethanol derived from new raw materials, namely rice straw, by applying the simultaneous saccharification and fermentation (SSF) method for the conversion of lignocellulose into alcohol, which will produce bioethanol with a purity level of about 99% with a more efficient process. This method goes through several processes, including breaking the size of the rice straw to 0.1 mm and mixing it with several bacteria for fermentation and hydrolysis. The saccharification process is also carried out until the distillation process to make high levels of bioethanol. Bioethanol from the simultaneous saccharification and fermentation (SSF) method is tested using an Octane Number Analyzer to determine the octane number content. The test included mixing bioethanol with Pertamax 92 to prove the effect of adding bioethanol to Pertamax, and the result was 93.1. Therefore, the innovation of making bioethanol from rice straw with the SSF method is expected to be a solution for Indonesia towards a pollution-free era.

The enhancement of ethanol production from NaOH-catalyzed organosolv pretreated poplar with compound additives

In this study, different additives (such as Bovine serum albumin (BSA), PEG 8000, calcium lignosulfonate, tea saponin, and Tween 80) were introduced to enhance the efficiency of simultaneous saccharification and fermentation (SSF) of NaOH-catalyzed organosolv pretreated poplar. Then the influence of two-factor additive compound on SSF was determined, only 75 mg/g BSA had a positive interaction with 150 mg/g tea saponin, which generated the maximal ethanol yield of 92.83 % with ethanol concentration of 18.82 g/L after 24 h, ascribing to the higher relative enzyme activity, free enzyme, and Zeta potential. Furthermore, with the addition of combined additives at an enzyme dosage of 8 FPU/g substrate, the NaOH-catalyzed organosolv pretreated poplar liberated higher ethanol than that without additive at an enzyme dosage of 15 FPU/g substrate. This phenomenon indicated that the use of combined additive can improve fermentation efficiency, while reducing almost 1/2 enzyme dosage. In general, this study provided a facile and efficient method for ethanol production with compound additives.

A Short Review of Second-Generation Isobutanol Production by SHF and SSF

As isobutanol exhibits higher energy density and lower hygroscopicity than ethanol, it is considered a better candidate biofuel. The sustainable supply of inedible biomass and lack of competition with the food supply have stimulated significant worldwide interest in the production of isobutanol from this resource. Both separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) have been applied to isobutanol production to effectively utilize inedible biomass as a feedstock. However, both processes have various challenges, including low isobutanol yield and high production costs. This review summarizes the potential of isobutanol as a biofuel, methods for conferring isobutanol productivity, recent experimental studies, and developments in both SHF and SSF with the isobutanol-producing strains. Challenges to increasing the isobutanol yield and various suggestions for improvements to enable commercial production are also discussed.

Techno-economical valorization of sugarcane bagasse for efficiently producing optically pure D-(−)-lactate approaching the theoretical maximum yield in low-cost salt medium by metabolically engineered Klebsiella oxytoca

Sugarcane bagasse (SCB) was utilized for efficiently producing optically pure D-(−)-lactate by Klebsiella oxytoca KIS004-91T strain. Cellulase (15 U/g NaOH-treated SCB) sufficiently liberated high sugars with saccharifications of 79.8 % cellulose and 52.5 % hemicellulose. For separated hydrolysis and fermentation, D-(−)-lactate was produced at 53.5 ± 2.1 g/L (0.98 ± 0.01 g/g sugar utilized or 0.71 ± 0.01 g/g total sugars) while D-(−)-lactate at 47.2 ± 1.8 g/L (0.78 ± 0.03 g/g sugar used or 0.69 ± 0.01 g/g total sugars) was obtained under simultaneous saccharification and fermentation (SSF). D-(−)-lactate at 99.9 ± 0.9 g/L (0.97 ± 0.01 g/g sugar utilized or 0.78 ± 0.01 g/g total sugars) was improved via fed-batch SSF. Based on mass balance, raw SCB of 7 kg is required to produce 1 kg D-(−)-lactate. Unlike others, D-(−)-lactate production was performed in low-cost salt medium without requirements of rich nutrients. Costs regarding medium, purification, and waste disposal may be reduced. This unlocks economic capability of SCB bioconversion or agricultural and agro-industrial wastes into high valuable D-(−)-lactate.

Efficient Biovalorization of Oil Palm Trunk Waste as a Low-Cost Nutrient Source for Bioethanol Production

This study aimed to efficiently utilize felled oil palm trunk (OPT) for bioethanol and lactic acid production. OPT was separated into two fractions: oil palm sap (OPS) and OPT fiber. OPS contained substantial amounts of sugars (38–40 g/L) and nitrogen (0.60–0.70 g/L), which can serve as a base medium for bioethanol production. As bioethanol production requires high sugar concentrations, OPS was concentrated, supplemented with OPT fiber, and used for bioethanol production through simultaneous saccharification and fermentation (SSF) by Saccharomyces cerevisiae. Repeated-batch SSF for five cycles efficiently utilized OPT fiber and achieved an average ethanol production of 35–42 g/L in each cycle. To increase the accessibility of the enzyme, OPT fiber was acid-pretreated prior to the SSF process. The combined use of acid-pretreated OPT slurry and concentrated OPS provided the maximum ethanol production of 49.63 ± 1.05 g/L. The fermented broth after ethanol recovery, containing mainly xylose, was used to produce lactic acid at a concentration of 18.85 ± 0.55 g/L. These strategies can greatly contribute to the zero-waste biorefinery of OPT and may also be applicable for the efficient biovalorization of other similar agricultural wastes.

Inhibitor Tolerance Capacity of Pichia kudriavzevii NBRC1279 and NBRC1664

The thermotolerant yeast Pichia kudriavzevii (previously known as Issatchenkia orientalis), can produce ethanol from a variety of carbon sources and grows at around 45 °C. Thus, this yeast is considered a useful biocatalyst for producing ethanol from lignocellulose through simultaneous saccharification and fermentation (SSF). SSF has several advantages, such as a simplified manufacturing process, ease of operation and reduced energy input. Using P. kudriavzevii NBRC1279 and NBRC1664, we previously succeeded in producing ethanol through SSF; however, the extent to which inhibitors by-produced from lignocellulose hydrolysis affect the growth and ethanol productivity of the two strains remains to be investigated. In this study, to better understand the inhibitor tolerance capacity of the two strains, spot assay, growth experiment, real-time quantitative PCR (RT-qPCR) analysis and multiple sequence alignment analysis were carried out. When P. kudriavzevii NBRC1279 and NBRC1664, as well as Saccharomyces cerevisiae BY4742 as a control, were cultured on SCD plates containing 17% ethanol, 42 mM furfural, 56 mM 5-hydroxymethylfurfural (HMF) or 10 mM vanillin, only P. kudriavzevii NBRC1664 was able to grow under all conditions. Moreover, the inhibitor tolerance capacity of P. kudriavzevii NBRC1664 was greater than those of other strains using SCD medium containing the same concentrations of various inhibitors. When an RT-qPCR analysis of seven gene sequences from aldehyde dehydrogenase and the aldehyde dehydrogenase family protein (ADHF) was performed using P. kudriavzevii NBRC1664 cultivated in the presence of 56 mM HMF, ADHF1 and ADHF2 were up-regulated in the early logarithmic growth phase. Moreover, a multiple sequence alignment of the amino acid sequences of ADHF1, ADHF2 and the known ADH suggested that ADHF1 and ADHF2 may catalyze the reversible NAD+-dependent oxidation of HMF. Our data may be useful for future studies on the metabolic engineering of more useful strains for ethanol production from lignocellulose.

Bio-2,3-butanediol production from banana waste: Preliminary techno-economic evaluation of processing strategies

This study evaluates different fermentation strategies to produce 2,3-butanediol (2,3-BD) from banana industry waste, such as whole bananas (fruit + peels) and banana peels, selecting the most favorable from a technical and economic point of view. Both residues have enough free sugars (17.8 %–35.8 %), glucan (11.0 %–14.2 %) and hemicellulose (2.8 %–6.3 %), to be promising substrates for 2,3-BD fermentation. Saccharification was studied by comparing enzymatic hydrolysis, hydrothermal pretreatment, and hydrothermal pretreatment followed by enzymatic hydrolysis. Different fermentation scenarios were also compared regarding the 2,3-BD yield and productivity: Separate Hydrolysis and Fermentation (SHF), Simultaneous Saccharification and Fermentation (SSF), and direct fermentation without prior saccharification using Paenibacillus polymyxa DSM-365 as the fermenting microorganism. The results showed that the pretreatment step was not necessary to improve the release of fermentable sugars. Enzymatic hydrolysis was the most effective alternative for maximizing sugar recovery, reaching sugar concentrations of 18.1 g/L (recovery: 92.5 %) for banana peels and 33.3 g/L (recovery: ∼100 %) for whole bananas. The SSF strategy led to higher 2,3-BD concentrations of 15.0 g/L and 26.6 g/L for banana peels and whole bananas, respectively. The preliminary economic analysis indicated that SSF and direct fermentation could be the more cost-effective process alternatives for banana peels and whole bananas, respectively. Thus, it was demonstrated that banana waste is an interesting resource for the production of 2,3-BD. The bioprocess can be competitive when using a low-cost raw material and reducing the number of process steps compared to traditional technologies.

Simultaneous Saccharification and Fermentation (SSF) of Blended Organic Fertilizer from Yam and Sweet Potato Peels

This study shows the production of organic fertilizer by utilization of yam and sweet potatoes peels through Simultaneous Saccharification and Fermentation (SSF) Method. Samples are categorized into sun-dried, sundried and autoclaved, Heat-dried, heat-dried and autoclaved, the fermentation process is carried out using alpha amylase and saccharomyces cerevisiae. Investigation of mineral elements (N, Cd, P, Zn, Pb, As, K, Hg.), in the eight varieties of Organic Fertilizer produced shows concentration of Cadmium (Cd) ranges from 0.0015 mg/kg to 0.0033 mg/kg), Nitrogen (N) in Percentage ranges from 2.3450% to 3.8550%; Phosphorus (P) ranges from 2.345 mg/kg to 3.607 mg/kg; Zinc (Zn) ranges from 2.215 mg/kg to 6.335 mg/kg. Lead (Pb) ranges from 0.00 mg/kg to 0.0015 mg/kg, potassium (K) ranges from 3.952 mg/kg to 6.213 mg/kg; Mercury and Arsenic nil. The study shows that Sweet Potato Peel (SDSPP) contains high Cadmium and potassium compare to other organic fertilizer produced. Sundried and Autoclaved Yam Peel (SDAYP) contains high percentage of Nitrogen (N), Heat Dried and Autoclaved Sweet Potato Peel (HDASPP) contains low Nitrogen, Sundried and Autoclaved Sweet Potato Peel (SDASPP) contains low phosphorus, Sundried Yam Peel (SDYP), Sundried and Autoclaved Yam Peels (SDAYP), Heat Dried Sweet Potato Peel HDSPP and Heat Dried Sweet Potato Peel (HDSPP) has zero trace of Lead (Pb) making it free of food intoxication from heavy metals, SDYP contains high phosphorus. These indicate that the samples will make quality organic fertilizer because it contains high Nitrogen, Phosphorus and potassium (NPK) which are essential elements required in plant structures.

Simultaneous Saccharification and Fermentation for Isobutanol Production from Banana Peel

Each year, near 40 million tons of banana peels are discarded around the world. This plant biomass could potentially be utilized for energy production. Simultaneous saccharification and fermentation (SSF) is an effective method for producing biofuels from plant biomasses. Since SSF with enzymatic hydrolysis and fermentation are performed simultaneously in the same reactor, the production process is simpler than most existing methods. Here, we describe isobutanol production using SSF with hydrothermally treated banana peel samples and an Escherichia coli strain able to utilize glucose and xylose to produce isobutanol. To enhance the glucose and xylose concentrations, the reaction conditions for the enzymatic hydrolysis of plant biomass using two kinds of saccharification enzymes were optimized, including the enzyme unit ratio, reaction temperature and sample gram. When the optimized conditions for enzymatic hydrolysis were applied to SSF, the glucose and xylose produced from the hydrothermally treated samples were consumed, producing isobutanol. Moreover, the isobutanol concentration increased with an increasing initial culture pH, reaching 1.27 g/L at pH 6.5, which was consistent with the optimal initial culture pH for isobutanol production by this E. coli strain. Taken together, these results indicate that the established method is potentially useful for industrial isobutanol production.

Biorefinery-based sustainability assessment of macroalgae waste valorization to polylactic acid: Exergy and exergoeconomic perspectives

This study evaluated the exergetic and exergoeconomic performance of three L-lactic acid (L-LA) production scenarios. All three scenarios produced L-LA from red macroalgae waste, Eucheuma cottonii residue (ECR) with the same species of enzymes, and lactic acid bacteria (LAB). Scenario 1 is the biorefinery process for the L-LA. Scenario 2 is the biorefinery process for the L-LA with fertilizer production and wastewater treatment. Scenario 3 is the polylactic acid (PLA) biorefinery process with fertilizer production and wastewater treatment. From the study, the total production rate for L-LA was 5194 kg/hr, with a total consumption rate of 12079 kg/hr for ECR. In addition, the production rates of organic fertilizer and reused water were 1609 and 1233 kg/hr, respectively, for scenario 2. Then, the production rate of the PLA was 4142 kg/hr for scenario 3. From the comparison, the energy consumption was increased due to the increased demand for process and equipment (from 28272.53 kW to 30296.15 kW). From the exergetic analysis aspect, the highest contributor to the exergy loss was the simultaneous saccharification and fermentation (SSF) unit, totaling 18875.62 kW. For the exergoeconomic aspect, the SSF unit also contributed the highest exergy destruction cost rate (average 70%). The study results provide theoretical guidance for exergetic and economic performance improvement in future L-LA and PLA production.

Comparison of simultaneous saccharification and fermentation with LPMO-supported hybrid hydrolysis and fermentation.

Enzymatic saccharification is used to convert polysaccharides in lignocellulosic biomass to sugars which are then converted to ethanol or other bio-based fermentation products. The efficacy of commercial cellulase preparations can potentially increase if lytic polysaccharide monooxygenase (LPMO) is included. However, as LPMO requires both a reductant and an oxidant, such as molecular oxygen, a reevaluation of process configurations and conditions is warranted. Saccharification and fermentation of pretreated softwood was investigated in demonstration-scale experiments with 10m3 bioreactors using an LPMO-containing cellulase preparation, a xylose-utilizing yeast, and either simultaneous saccharification and fermentation (SSF) or hybrid hydrolysis and fermentation (HHF) with a 24-hour or 48-hour initial phase and with 0.15 vvm aeration before addition of the yeast. The conditions used for HHF, especially with 48h initial phase, resulted in better glucan conversion, but in poorer ethanol productivity and in poorer initial ethanol yield on consumed sugars than the SSF. In the SSF, hexose sugars such as glucose and mannose were consumed faster than xylose, but, in the end of the fermentation >90% of the xylose had been consumed. Chemical analysis of inhibitory pretreatment by-products indicated that the concentrations of heteroaromatic aldehydes (such as furfural), aromatic aldehydes, and an aromatic ketone decreased as a consequence of the aeration. This was attributed mainly to evaporation caused by the gas flow. The results indicate that further research is needed to fully exploit the advantages of LPMO without compromising fermentation conditions.

A Comparison of ethanol production utilizing separated hydrolysis fermentation and simultaneous saccharification and fermentation methods applied to Tanduk banana peel waste

In 2021, Indonesia’s banana production is expected to reach 8.74 million tons. This substantial volume of bananas will generate a notable quantity of banana peel waste, constituting approximately one-third of the total banana fruit mass. Remarkably, banana peel waste retains 18.5% carbohydrates, which can be effectively converted into glucose, serving as a valuable feedstock for ethanol production. This research is focused on the ethanol production derived from Tanduk banana peel waste, employing a comparative analysis of the Separated Hydrolysis Fermentation (SHF) and Simultaneous Saccharification and Fermentation (SSF) techniques. Additionally, it explored varying raw material concentrations, specifically 200, 250, and 300 g/L. The SSF method integrated the use of cellulase enzymes, while the SHF method relied on HCl for the hydrolysis process. Microbial fermentation was facilitated by Saccharomyces cerevisiae, with a fermentation duration of 96 hours and sample collection at 12-hour intervals. The results demonstrated that higher concentrations of banana peel waste yield increased glucose production during the hydrolysis process, consequently leading to greater ethanol produced. The concentration of 300 g/L banana peel waste is optimal for producing ethanol in both procedures. However, the SHF approach demonstrates a superior ethanol concentration of 51%, surpassing the 33.7% achieved by the SSF method.

Co-valorization of discarded wood pinchips and sludge from the pulp and paper industry for production of advanced biofuels

Several lignocellulosic wastes are generated in the pulp and paper industry (PPI), such as small wood chips (pinchips) and paper sludge, presenting a high cellulose content suitable to be converted into biofuels or bio-products in a forest biorefinery scheme. In this work, two schemes of biorefinery were proposed for their valorization, processing small eucalyptus wood pinchips in two different strategies: (i) autohydrolysis at 230ºC, and (ii) autohydrolysis at 195ºC followed by organosolv process (47.7% ethanol-water, 198ºC for 60 min). More than 95% of cellulose was recovered in both schemes. In the combined process, 76% of delignification was achieved and 78% of xylan was solubilized as xylooligosaccharides. To reduce operational cost of lignocellulosic biomass-to-ethanol fermentation, the mixture of the treated eucalyptus pinchips from two processes with sludge was also proposed to increase the initial glucan content and to supply a rich source of nitrogen (present in the sludge). For that, two experimental designs were carried out for ethanol production by simultaneous saccharification and fermentation (SSF) process. Ethanol from SSF assays using sludge as co-substrate at 0.6 g of sludge/g of treated wood pinchips and 16 FPU/g of pretreated solids allowed to obtain 59 g/L (90% of conversion) and 46 g/L (96% of conversion) when blended with the wood from autohydrolysis and with the wood from autohydrolysis followed by organosolv, respectively. Overall, this study shows an alternative process valorization of biomasses derived from PPI for production of advanced biofuels and bio-products (such as xylooligosaccharides and lignin) contributing to achieving a circular economy.

Influence of Inhibitors from Microwave Pretreatment of Oil Palm Frond Pulping (OPFP) on Bioethanol Production

This research aims to analyze the influence of inhibitors from microwave pretreatment of oil palm frond pulping (OPFP) on the efficiency of bioethanol fermentation by S.cerevisiae in the simultaneous saccharification and fermentation (SSF) processes. OPFPs were achieved at different ages: 3-4, 4-7, 7-10, 10-20, and 20-25 years old. OPFP was pretreated with a microwave and sulfuric acid (MW/SF), microwave and hydrogen sulfide (MW/HP), and microwave and water (MW/W). The results showed that the main inhibitors formed during the pretreatment process of OPFP were acetic acid, furfural, 5-hydroxymethylfurfural (HMF), furfural, formic acid, and phenol. The pretreatment of OPFP with MW/W had the lowest concentrations of inhibitors compared to the other pretreatment methods. The highest bioethanol yields at all ages of OPFP were in the range of 0.41-0.42 g-bioethanol/g-glucose, corresponding to more than 80% fermentation efficiency. At these conditions, the concentrations of the acetic acid were 0.09-0.19 g/l, HMF =0, furfural =0, formic acid 0.05-0.28 g/l, and phenol 0.22-0.47 g/l. The MW/W was the suitable pretreatment of OPFP for bioethanol production due to the lowest to generate the inhibitor and high ethanol production yield.