Ethanol Yield Research Articles

Adapted feeding strategies in fed-batch fermentation improve sugar delivery and ethanol productivity

ABSTRACTBioethanol is a renewable fuel widely used in road transportation and is generally regarded as a clean energy source. Although fermentation is one of the major processes in bioethanol production, studies on improving its efficiency through operational design are limited, especially compared to other steps (pretreatment and hydrolysis/saccharification). In this study, two adapted feeding strategies, in which feed medium addition (sugar delivery) was adjusted to increase the supply of fermentable sugar, were developed to improve ethanol productivity in 5-L fed-batch fermentation by Saccharomyces cerevisiae. Specifically, a linear adapted feeding strategy was established based on changes in cell biomass, and an exponential adapted feeding strategy was developed based on cell biomass accumulation. By implementing these two feeding strategies, the overall ethanol productivity reached 0.88±0.04 and 0.87±0.06 g/L/h, respectively. This corresponded to ~20% increases in ethanol productivity compared to fixed pulsed feeding operations. Additionally, there was no residual glucose at the end of fermentation, and final ethanol content reached 95±3 g/L under the linear adapted operation and 104±3 g/L under the exponential adapted feeding strategy. No statistical difference was observed in the overall ethanol yield (ethanol-to-sugar ratio) between fixed and adapted feeding strategies (~91%). These results demonstrate that sugar delivery controlled by adapted feeding strategies was more efficient than fixed feeding operations, leading to higher ethanol productivity. Overall, this study provides novel adapted feeding strategies to improve sugar delivery and ethanol productivity. Integration into the current practices of the ethanol industry could improve productivity and reduce production costs of fermentation processes.

Integrating surfactants with low enzyme loading to increase the glucan conversion and ethanol concentration of reed after combined pretreatment

In this study, in order to obtain high concentration bioethanol, reed pretreated with liquid hot water and ammonia oxygen (LHW-NH3·H2O/O2) was used as the substrate for fermentation. Enzymatic hydrolysis becomes difficult by the ineffective lignin adsorption and the decline in cellulase activity. It is worth noting that even under low enzyme loading (9 FPU/g pretreated solid), this study was able to maintain a higher glucan conversion rate (97.60%). In addition, under high solid loading conditions, the combined effect of PEG 6000 and rhamnolipids increased ethanol yield (78.40%) and ethanol concentration (71.5 g/L). The combined LHW-NH3·H2O/O2 pretreatment with fed-batch semi-simultaneous saccharification and fermentation technique may be utilized to industrialize the production of bioethanol from reed since we concentrated on the maximum exploitation of biomass resources and reduction in energy consumption.

Co-Fermentation of Glucose–Xylose–Cellobiose–XOS Mixtures Using a Synthetic Consortium of Recombinant Saccharomyces cerevisiae Strains

The efficient conversion of cellulosic sugars is vital for the economically viable production of biofuels/biochemicals from lignocellulosic biomass hydrolysates. Based on comprehensive screening, Saccharomyces cerevisiae RC212 was chosen as the chassis strain for multiple integrations of heterologous β-glucosidase and β-xylosidase genes in the present study. The resulting recombinant BLN26 and LF1 form a binary synthetic consortium, and this co-culture system achieved partial fermentation of four sugars (glucose, xylose, cellobiose, and xylo-oligosaccharides). Then, we developed a ternary S. cerevisiae consortium consisting of LF1, BSGIBX, and 102SB. Almost all four sugars were efficiently fermented to ethanol within 24 h, and the ethanol yield is 0.482 g g−1 based on the consumed sugar. To our knowledge, this study represents the first exploration of the conversion of mixtures of glucose, xylose, cellobiose, and xylo-oligosaccharides by a synthetic consortium of recombinant S. cerevisiae strains. This synthetic consortium and subsequent improved ones have the potential to be used as microbial platforms to produce a wide array of biochemicals from lignocellulosic hydrolysates.

Evaluation of Sweet Sorghum Hybrids for Juce Yield, Ethanol Yield and its Attributing Traits

An investigation was carried out to assess the extent of heterosis for juice yield and its attributing traits in 45 new hybrids of sweet sorghum developed by crossing 3 male sterile lines with 15 restorer in L x T mating design and were sown in randomized block design (RBD) with three replications during Kharif 2018 at Botany Farm, Post Graduate Institute, M.P.K.V., Rahuri. The significant average heterosis, heterobeltiosis and standard heterosis recorded for total biomass, juice yield and its contributing characters in fourty five crosses. The cross CMS-1409 x RSSV-512 showed the highest significant average heterosis (224.84%) and heterobeltiosis (286.36%) for juice yield. The extent of standard heterosis observed for juice yield over the check, CSH-22SS was upto 59.15 per cent. The highest significant standard heterosis was recorded by the cross CMS-1409 x RSSV-512 (59.15%), followed by cross ICMS-479 x RSSV-355 (43.58%) and CMS-1409 x RSSV-355 (24.30%). Based on per se performance and heterotic performance for yield and its principal components in the F1 hybrids the cross combinations viz; CMS-1409 x RSSV-512, ICMS-479 x RSSV-355 and CMS-1409 x RSSV-499 CMS-1409 x RSSV-498 and CMS-1409 x RSSV-355 were appeared to be promising.

Bio-ethanol Production from Strawberry by Saccharomyces cerevisiae in Repeated Batch Abstract

Bio-ethanol, is considered the cleanest fluid fuel, and can be a dependable substitution to non-renewable energy sources. The maturation of sugar parts of plant materials produces it. Due to their high sugar content and abundance of nutrients, berries are thought to be an excellent source of fermentation products. This examination saw how well the yeast "Saccharomyces cerevisiae" created ethanol from strawberries (Fragaria ananassa) in rehashed bunches. According to the findings, strawberry juice had a total sugar concentration of 41.87 g/l. The final bioethanol percentage, ethanol yield, and maximum productivity were, respectively, 6 g/l/h (g ethanol per liter of strawberry juice per hour), 35 g/l (g ethanol per liter of raspberry juice), and 7 g/l (g ethanol per liter of strawberry juice).

High-Efficient Production of Cellulosic Ethanol from Corn Fiber Based on the Suitable C5/C6 Co-Fermentation Saccharomyces cerevisiae Strain

As a potential alternative to fossil-based fuels, cellulosic ethanol has attracted much attention due to its great benefit to energy sustainability and environmental friendliness. However, at present, the industrial competitiveness of cellulosic ethanol production is still insufficient compared with fossil-based fuels because of the higher costs. Expanding the range of lignocellulosic biomass may be a promising measure to promote the economical production of cellulosic ethanol. Corn fiber, a byproduct from the corn deep-processing, is an attractive feedstock for cellulosic ethanol production because of its rich carbohydrate content (generally exceeding 65% of dry weight), almost no transportation cost, and low lignin content allow it to be easily handled. This study first optimized the hydrolysis conditions, including the pretreatment and enzymolysis process based on dilute sulfuric acid, to achieve a high sugar yield. Then, the corn fiber hydrolysates obtained under different hydrolysis conditions were suitably fermented by different C5/C6 co-fermentation Saccharomyces cerevisiae, indicating that the hydrolysate at high solid loading (20%) needs to detoxification to a certain extent but not low solid loading (10%) to achieve high ethanol yield. Finally, the fermentation of the 20% solid loading hydrolysates with resin detoxification was performed in a 50 L bioreactor, achieving the sugar (glucose and xylose) metabolic rate of 2.24 g L −1 h −1 and ethanol yield of 92% of the theoretical value, which are the highest reported levels to date. This study provided a potential process route for cellulosic ethanol production from corn fiber from the perspective of the suitability between the upstream hydrolysis process and the downstream fermentation strain.

Evaluating performance of CrCl3-catalyzed ethanol pretreatment of poplar on cellulose conversion

As green and effective depolymerization processes, CrCl3 and CrCl3-catalyzed ethanol pretreatment were comprehensively investigated to improve cellulose conversion from pretreated residues in poplar in the present study. The influence of pretreatment temperature on delignification, structural characteristics of lignin, and the yield of glucose and ethanol was comparatively analyzed to emphasize the synergy effect of CrCl3 and ethanol solvent. Particularly, CrCl3-catalyzed ethanol pretreatment at 200 °C resulted in the pretreated substrates with >94% removal of hemicellulose and lignin, consequently reaching 88.88% and 56.11% yields of glucose and ethanol, respectively. Lignin's structural change was observed by Gel permeation chromatography and 2D nuclear magnetic resonance spectroscopy and its relationship with the efficiency of enzymatic hydrolysis and fermentation was determined. Results indicated that the increased pretreatment temperature enhanced the lignin depolymerization with smaller molecular weight, fewer β-O-4 linkages, and higher S/G ratio, especially in CrCl3-catalyzed ethanol pretreatment, which was positively associated with poplar cellulose conversion. In addition to offering a comprehensive illustration of the sugar/ethanol generation during CrCl3-catalyzed organosolv pretreatment, this work also provided theoretical support for biofuel and biochemical production from poplar.

Steam Explosion of Eucalyptus grandis Sawdust for Ethanol Production within a Biorefinery Approach

In this work, Eucalyptus grandis sawdust was subjected to steam explosion as the first step in cellulosic ethanol production within a biorefinery approach. The effect of the moisture content in the eucalypt sawdust (8 and 50%) and pretreatment process variables, such as temperature and residence time, were evaluated along with the influence of the water washing of steam-exploded solids on enzymatic hydrolysis and C6 fermentation yields. All other process streams were characterized to evaluate the recovery yield of valuable co-products. A recovery of nearly 100% glucans in the solid fraction and 60% xylans in the liquid fraction, mainly as partially acetylated oligomers, was obtained. The best enzymatic hydrolysis efficiencies (66–67%) were achieved after pretreatment at 205 °C for 10 min. The washing of pretreated sawdust with water improved the hydrolysis efficiencies and ethanol production yields by 10% compared to the unwashed pretreated solids under the same experimental condition. The highest ethanol yields were achieved after pretreatment of the sawdust with an 8% moisture content at 205 °C for 10 min, enzymatic hydrolysis at 13 wt% total solids with 25 FPU/g glucans, and fermentation with S. cerevisiae PE-2. In this case, 227 L ethanol and 40 kg total xylose (including xylo-oligomers) were obtained per ton of dry eucalypt sawdust.

Reduction of ethanol content in wine with an improved combination of yeast strains and process conditions

One interesting strategy to address the increasing alcohol content of wines, associated with climate change, is to reduce the ethanol yield during fermentation. Within this strategy, the approach that would allow the clearest reduction in alcohol content is the respiration of part of the grape sugars by yeasts. Non-Saccharomyces species can be used for this purpose but suffer from a limited ability to dominate the process and complete fermentation. In turn, Saccharomyces cerevisiae shows a high production of acetic acid under the growth conditions required for respiration. Previously proposed procedures used combinations of non-Saccharomyces and S. cerevisiae starters, or a strain of S. cerevisiae (PR1018), with unique metabolic properties. In both cases, precise management of oxygen availability was required to overcome the acetic acid problem. In this work, we have developed a laboratory scale process to take advantage of the properties of PR1018 and a strain of Metschnikowia pulcherrima. This process is more robust than the previous ones and does not rely on strict control of oxygenation or even the use of this particular strain of S. cerevisiae. Aeration can be interrupted instantly without impairing the volatile acidity. Under the selected conditions, an ethanol reduction of around 3% (v/v) was obtained compared to the standard fermentation control.

High starch duckweed biomass production and its highly-efficient conversion to bioethanol

This paper carried out a new attempt which combined nutrient starvation, uniconazole and elevated CO2 to increase duckweed biomass and starch content. At laboratory level, the highest growth rate and starch content reached up to 9.4 ± 0.2 g m−2 d−1 and 75.9 ± 3.3 %, leading to an estimated starch yield of 30.8 t ha−1 y−1. Furthermore, without any pretreatment, up to 88.4 ± 0.4 % of glucose from high starch duckweed biomass can be released. The enzymatic hydrolyzates can be efficiently converted to bioethanol by yeast and the ethanol yield based on duckweed biomass input (YE/B) reached up to 0.38 ± 0.01 g g−1 applying simultaneous saccharification and fermentation (SSF), which, to our knowledge, is the highest duckweed-to-ethanol yield thus far reported. Based on the data above-mentioned, an annual duckweed-based ethanol yield of 16.0 × 103 L ha−1 can be achieved, about 2.6 times higher than maize-based ethanol yield. This approach has achieved quick high starch accumulation in duckweed and demonstrated that duckweed can serve as a competitive starch feedstock for bioethanol production.

Metabolomics of Escherichia coli for Disclosing Novel Metabolic Engineering Strategies for Enhancing Hydrogen and Ethanol Production.

The biological production of hydrogen is an appealing approach to mitigating the environmental problems caused by the diminishing supply of fossil fuels and the need for greener energy. Escherichia coli is one of the best-characterized microorganisms capable of consuming glycerol-a waste product of the biodiesel industry-and producing H2 and ethanol. However, the natural capacity of E. coli to generate these compounds is insufficient for commercial or industrial purposes. Metabolic engineering allows for the rewiring of the carbon source towards H2 production, although the strategies for achieving this aim are difficult to foresee. In this work, we use metabolomics platforms through GC-MS and FT-IR techniques to detect metabolic bottlenecks in the engineered ΔldhΔgndΔfrdBC::kan (M4) and ΔldhΔgndΔfrdBCΔtdcE::kan (M5) E. coli strains, previously reported as improved H2 and ethanol producers. In the M5 strain, increased intracellular citrate and malate were detected by GC-MS. These metabolites can be redirected towards acetyl-CoA and formate by the overexpression of the citrate lyase (CIT) enzyme and by co-overexpressing the anaplerotic human phosphoenol pyruvate carboxykinase (hPEPCK) or malic (MaeA) enzymes using inducible promoter vectors. These strategies enhanced specific H2 production by up to 1.25- and 1.49-fold, respectively, compared to the reference strains. Other parameters, such as ethanol and H2 yields, were also enhanced. However, these vectors may provoke metabolic burden in anaerobic conditions. Therefore, alternative strategies for a tighter control of protein expression should be addressed in order to avoid undesirable effects in the metabolic network.

Investigating the potential of avocado seeds for bioethanol production: A study on boiled water delignification pretreatment

The increasing need for alternative fuels to replace fossil fuels has made bioethanol a promising option. Although numerous sources of sugar generation and agricultural wastes can be converted into ethanol, Avocado Seeds (AS) are particularly attractive as raw materials due to their abundance, high carbohydrate content, and lack of interactions with the food chain. Therefore, this study investigated the potential of AS for bioethanol production using several steps, including boiled water delignification pretreatment, catalytic hydrolysis, and fermentation with Saccharomyces cerevisiae. The delignification pretreatment of AS involved soaking in 4% (w/v) sodium hydroxide liquor for 24 hours. Then the mixture was heated to 80°C and stirred slowly for 2.5 hours and after that washing with boiled water at 100 oC for 1.5 hours and screening the mixture. Subsequently, catalytic hydrolysis and fermentation were carried out using two different concentrations of Saccharomyces cerevisiae as yeast, namely 10% (w/v) and 15% (w/v). Qualitative sample analysis was conducted using scanning electron microscopy (SEM) to observe the effect of delignification pretreatment, while FTIR analysis using Thermo Scientific Nicolet iS50 was used to test for glucose functional groups. Quantitative analysis was performed using gas chromatography 7890b mass spectrophotometry 5977A, Agilent DBVRX to determine hydrolysate fermentation. The results revealed that the highest ethanol yield was achieved through fermentation with 15% (w/v) yeast and 40% (v/v) catalyst, resulting in an ethanol yield of 83.755% of the theoretical maximum.

Engineering Thermoanaerobacterium aotearoense SCUT27 with the deficiency of a hypothetic protein regulated by ArgR1864 for enhanced ethanol production from lignocellulosic hydrolysates

Arginine repressor (ArgR1864) is a multifunctional regulator in Thermoanaerobacterium aotearoense SCUT27 (SCUT27). For better understanding its function in SCUT27, the transcriptome of SCUT27/ΔargR1864 under xylose was further investigated. 2014 (V518_2014) denoted as a hypothetic protein gene was mined, which played important roles on xylose metabolism and carbon metabolic flux distribution in SCUT27. In 2014 deletion mutant, no lactic acid could be produced along with enhanced ethanol formation. Same changes were also observed in SCUT27/ΔargR1864. ArgR1864 is the direct regulator of 2014 identified by electrophoretic mobility shift assays (EMSA). When 2014 was complemented in SCUT27/ΔargR1864, the fermentation traits of SCUT27/ΔargR1864/2014 were returned to wild-type levels, suggesting that the improved xylose utilization and ethanol formation of SCUT27/ΔargR1864 were attributed to the low expression of 2014. When cultured under sorghum stalk, corn cob and wheat straw hydrolysates, SCUT27/Δ2014 also showed the excellent capability for sugar utilization and ethanol production with the consumption rate of xylose and glucose increased by 20%–37.93 and 64.29%–190.91% separately, the ethanol production improved by 326.61%–547.89% and ethanol yield enhanced by 216.67%–337.50%. Thus, 2014, as a hypothetic protein gene, could be used as the new genetic reference to predict functional genes to engineer the microorganism for biofuel production from lignocellulosic hydrolysates.

Synergetic effect of hydrothermal and deep eutectic solvents (DES) pretreatments on Robinia wood fractionation for the manufacture of bioethanol and cellulose nanocrystals

This study dealt with the use of environmentally friendly processes based on microwave-assisted autohydrolysis (MAA) and deep eutectic solvents (DES) for the selective fractionation of Robinia pseudoacacia wood (RW) within a biorefinery approach. MAA enabled the recovery of 76% of hemicelluloses in the form of oligomers. Afterwards, different conditions were assessed for the optimal delignification of RW with the DES choline chloride combined with lactic acid reaching delignification ratios up to 86%. Two different methods were accomplished to valorize the cellulosic-rich solid fraction after delignification: (i) bioethanol via enzymatic-fermentative pathway (attaining 53.3 g ethanol/L, about 83% of ethanol yield), and (ii) cellulose nanocrystals (length of 27–550 nm, width of 2–12 nm). Hence, this study presents a novel multiproduct biorefinery to selectively separate the main components of RW and valorize its cellulosic fraction using eco-friendly procedures.

An integration of machine learning models and life cycle assessment for lignocellulosic bioethanol platforms

The availability and accuracy of data play a critical role in the development of Life Cycle Assessment (LCA) models, as they directly impact the effectiveness and reliability of the model. Machine learning (ML) approach has the potential to improve the data quality, leading to enhanced performance of LCA models. In this study, ML algorithms (random forest, extreme gradient boosting, and artificial neural network) were applied to predict the ethanol yield and NaOH consumption, which were key life cycle inventories for lignocellulosic bioethanol process. The prediction models exhibited a decent level of accuracy for ethanol yield (accuracy ∼ 0.85) and NaOH consumption (accuracy > 0.8). In addition, ML algorithms (genetic algorithms, particle swarm optimization, and simulate annealing) were applied to identify optimal feedstock characteristics and conditions for maximum ethanol yield. The optimization models showed an 18% improvement compared to the highest yield from the dataset (0.41 g/g). The ML models (prediction and optimization algorithms) were further integrated with the LCA model (GREET1) to enhance the ethanol yield and to reduce greenhouse gas (GHG) emission for target feedstocks. This resulted in a 6% and 19% improvement in ethanol yield and an approximately 8% and 14% reduction of GHG emissions for grass and corn stover, respectively. This study demonstrates the feasibility of utilizing ML to predict LCA metrics for the development of LCA models and to optimize the process parameters to minimize environmental impact.

Comparison of marine and rushton impeller in high solid loading of two-step pretreated oil palm trunk using simultaneous saccharification and fermentation for ethanol production

Substrate loading enhancement in fermenter generates difficulty in mixing that results in low ethanol yield owing to insufficient mass transfer. Thus, this study evaluated the efficiency of a six-blade Rushton turbine and a three-blade marine type propeller in a 3 L stirred tank bioreactor (STBR) by using steam exploded and alkaline pretreated oil palm trunk (OPT) fibers in a unique batch and fed-batch strategy. The fed-batch began with 5% (w/v) solid loading and after 12 h of simultaneous saccharification and fermentation (SSF) reaction, 2.5% (w/v) solid was added in the bioreactor with 6 h interval. The marine and Rushton impellers were compared at 5% (w/v) batch and 20% (w/v) fed-batch SSF, using agitation rate of 100 rpm. At 5% (w/v) solid loading, the highest ethanol concentration and theoretical yield were 15.66 ± 1.71 g/L and 61.69 ± 2.34%, and 12.10 ± 0.98 g/L and 54.01 ± 3.52%, respectively for marine and Rushton impeller. At high solid loading (20% w/v), the marine impeller achieved maximum ethanol concentration of 41.33 ± 2.02 g/L at 72 h of fermentation, with a low power consumption of 54.73 kW. The Rushton impeller resulted in ethanol concentration of 39.08 ± 1.47 g/L at 96 h of fermentation with relatively higher power consumption of 83.62 kW. The marine impeller is therefore efficient with the fed-batch strategy.

Acetic acid, growth rate, and mass transfer govern shifts in CO metabolism of Clostridium autoethanogenum.

Syngas fermentation is a leading microbial process for the conversion of carbon monoxide, carbon dioxide, and hydrogen to valuable biochemicals. Clostridium autoethanogenum stands as a model organism for this process, showcasing its ability to convert syngas into ethanol industrially with simultaneous fixation of carbon and reduction of greenhouse gas emissions. A deep understanding on the metabolism of this microorganism and the influence of operational conditions on fermentation performance is key to advance the technology and enhancement of production yields. In this work, we studied the individual impact of acetic acid concentration, growth rate, and mass transfer rate on metabolic shifts, product titres, and rates in CO fermentation by C. autoethanogenum. Through continuous fermentations performed at a low mass transfer rate, we measured the production of formate in addition to acetate and ethanol. We hypothesise that low mass transfer results in low CO concentrations, leading to reduced activity of the Wood-Ljungdahl pathway and a bottleneck in formate conversion, thereby resulting in the accumulation of formate. The supplementation of the medium with exogenous acetate revealed that undissociated acetic acid concentration increases and governs ethanol yield and production rates, assumedly to counteract the inhibition by undissociated acetic acid. Since acetic acid concentration is determined by growth rate (via dilution rate), mass transfer rate, and working pH, these variables jointly determine ethanol production rates. These findings have significant implications for process optimisation as targeting an optimal undissociated acetic acid concentration can shift metabolism towards ethanol production. KEY POINTS: • Very low CO mass transfer rate leads to leaking of intermediate metabolite formate. • Undissociated acetic acid concentration governs ethanol yield on CO and productivity. • Impact of growth rate, mass transfer rate, and pH were considered jointly.

Engineering All-Round Cellulase for Bioethanol Production.

One strategy to decrease both the consumption of crude oil and environmental damage is through the production of bioethanol from biomass. Cellulolytic enzyme stability and enzymatic hydrolysis play important roles in the bioethanol process. However, the gradually increased ethanol concentration often reduces enzyme activity and leads to inactivation, thereby limiting the final ethanol yield. Herein, we employed an optimized Two-Gene Recombination Process (2GenReP) approach to evolve the exemplary cellulase CBHI for practical bioethanol fermentation. Two all-round CBHI variants (named as R2 and R4) were obtained with simultaneously improved ethanol resistance, organic solvent inhibitor tolerance, and enzymolysis stability in simultaneous saccharification and fermentation (SSF). Notably, CBHI R4 had a 7.0- to 34.5-fold enhanced catalytic efficiency (kcat/KM) in the presence/absence of ethanol. Employing the evolved CBHI R2 and R4 in the 1G bioethanol process resulted in up to 10.27% (6.7 g/L) improved ethanol yield (ethanol concentration) than non-cellulase, which was far more beyond than other optimization strategies. Besides bioenergy fields, this transferable protein engineering routine holds the potential to generate all-round enzymes that meet the requirement in biotransformation and bioenergy fields.

The effect of pretreatment choice on cellulosic ethanol production from sugarcane straw: An insight into environmental impact profile and GHG emissions mitigation potential in Brazil

The exploration of residual lignocellulosic biomass for biofuel production is crucial to achieve significant greenhouse gas (GHG) emissions mitigation within the following decades, translating to diminished agricultural environmental impact and land-use change dynamics. On the other hand, cellulosic ethanol production pathways are typically resource-intensive, in energy and chemical terms, which directly influence its carbon intensity. Pretreatment choice, to this end, is key, since it dictates the overall process performance and yield, and may include crucial flows, under the life-cycle perspective, such as solvents and other chemicals. This work, then, aims to evaluate the effect of pretreatment choice, namely hydrothermal (HT), steam explosion (SE), and alkaline (AK), in the technical and environmental performance of cellulosic ethanol production from sugarcane straw (SCS), extending this analysis to the final GHG emissions mitigation potential for gasoline and fossil-generated electricity substitution, under the Brazilian context in São Paulo. Results show that, while AK provided the highest ethanol yield, this pretreatment option gave the lowest electricity generation surplus, and its sodium hydroxide usage was identified as an important environmental hotspot in most impact categories, which narrowed down its GHG emission mitigation gap for gasoline substitution. HT and SE presented similar ethanol and electricity yields, with SE being the most balanced option in terms of productivity and environmental impact profile. By selecting the SE route, all of the available SCS in São Paulo could be converted into 10% of the Brazilian annual ethanol production, and mitigate 5.4 MtCO2e of gasoline emissions, 15% of the Brazilian Biofuel Policy (RenovaBio) target for 2022.

Evaluation of Generic Fertiliser as an Alternative Inorganic Nitrogen Source for Ethanolic Glucose Fermentation by Saccharomyces cerevisiae

In the studies and production of bioethanol, the preferred fermenting yeast (Saccharomyces cerevisiae) is usually cultured in liquid broth that contains yeast extract and peptone. However, the use of these laboratory and scientific grade chemicals is costly, making them impractical for mass bioethanol production. Therefore, this study was conducted to evaluate the feasibility of glucose ethanolic fermentation by S. cerevisiae using generic fertiliser formulations to provide inorganic nitrogen, phosphorus, potassium and trace elements (NPK-TE). Fermentation media of different generic fertiliser strength at 0.5X, 1.0X and 2.0X Fertiliser Nitrogen Equivalents (FNE), as compared to the conventional Yeast Extract-Peptone (YEP) medium as control, was used as fermentation broth during the ethanolic fermentation of glucose. Based on the results, S. cerevisiae cultured in YEP broth produced the highest cell concentration for both wet (21.93 g/L) and dry cells (3.87 g/L), with rapid increment observed in the first 72 h of fermentation. By the end of the fermentation period, lactic acid (3.14 g/L) and acetic acid (0.96 g/L) levels were recorded to be the lowest in YEP medium while their concentration (lactic acid, 8.08 g/L) and (acetic acid, 2.67 g/L) were highest in 2.0X FNE fertiliser medium. Results indicated that the best theoretical ethanol yield (TEY) among the fertiliser media was achieved when fermentation was performed in the 0.5X FNE fertiliser medium, with a TEY of 86.18%. TEY yields were 78.68% and 51.54% in broth with 1.0X and 2.0X FNE, respectively. In general, all three fertiliser media supported ethanolic fermentation of glucose, with the 0.5X FNE fertiliser broth showing a yield that is significantly close to the conventional YEP medium, as seen in the statistical analysis. Similarities in other fermentation profiles such as acetic acid, lactic acid, and biomass production, as well as glucose utilisation, between the results from the YEP samples and samples from the fertiliser broths (at 0.5X and 1.0X FNE) have also shown that generic fertiliser has the potential to be used as an alternative medium to replace the conventional YEP to produce ethanol at a lower cost.