A Hydrotrope Pretreatment for Stabilized Lignin Extraction and High Titer Ethanol Production

The biomass pretreatment strategies using organic acids facilitate lignin removal and enhance the enzymatic digestion of cellulose. However, lignin always suffers a severe and irreversible condensation. The newly generated C-C bonds dramatically affect its further upgrading. In this study, we used a recyclable hydrotrope (p-Toluenessulfonic acid, p-TsOH) to dissolve lignin under mild condition and stabilized lignin with a quenching agent (formaldehyde, FA) during extraction, achieving both value-added lignin extraction and efficient enzymatic saccharification of cellulose. Approximately 63.7% of lignin was dissolved by 80% (wt. %) p-TsOH with 1.5% FA addition at 80 o C, 30 min. The obtained lignin was characterized by FTIR spectroscopy, TGA, 2D HSQC NMR spectroscopy, and GPC. The results indicated that the extracted lignin exhibited excellent properties, such as light color, a low molecular weight (Mw, 5371 g/mol), and a narrow polydispersity (Mw/Mn, 1.63). The pretreated substrate was converted to ethanol via a quasi-simultaneous saccharification and fermentation process (Q-SSF). After fermentation of 60 h, the ethanol concentration reached 38.7±3.3 g/L which was equivalent to a theoretical ethanol yield of 82.9±2.2% based on the glucan content, while the residual glucose concentration was only 4.69±1.4 g/L. In short, this pretreatment strategy protected lignin to form new C-C linkages and improved the enzymatic saccharification of glucan for high-titer ethanol production.

The Performance Of Clostridium Phytofermentans For Biofuels Production From Lignocellulosic Biomass

Ethanol produced from lignocellulosic biomass is an alternative transportation fuel with the potential to lower greenhouse gas emissions and increase energy security. Source-separated organic waste (SSO) from the city of Toronto was investigated for feasibility as a lignocellulosic ethanol feedstock. Clostridium phytofermentans is a mesophilic, cellulolytic and ethanologenic species with potential application for ethanol production from lignocullulosic biomass. C. phytofermentans was considered for biofuels production through experiments on a variety of substrates including soluble sugars and pure celluloses. Results from this study found that C. phytofermentans produced 73% of the theoretical ethanol yield on cellobiose but grew poorly on glucose and xylose. In addition, C. phytofermentans grew marginally on microcrystalline and ball-milled cellulose, but with supplemental enzymes produced 55% of the theoretical ethanol yield.

The Performance Of Clostridium Phytofermentans For Biofuels Production From Lignocellulosic Biomass

Ethanol produced from lignocellulosic biomass is an alternative transportation fuel with the potential to lower greenhouse gas emissions and increase energy security. Source-separated organic waste (SSO) from the city of Toronto was investigated for feasibility as a lignocellulosic ethanol feedstock. Clostridium phytofermentans is a mesophilic, cellulolytic and ethanologenic species with potential application for ethanol production from lignocullulosic biomass. C. phytofermentans was considered for biofuels production through experiments on a variety of substrates including soluble sugars and pure celluloses. Results from this study found that C. phytofermentans produced 73% of the theoretical ethanol yield on cellobiose but grew poorly on glucose and xylose. In addition, C. phytofermentans grew marginally on microcrystalline and ball-milled cellulose, but with supplemental enzymes produced 55% of the theoretical ethanol yield.

Assessing Ethanol Yield from Fermented Juice of Local and Exotic Sweet Sorghum Varieties Grown in Sudan

An experiment was conducted in 2017 at Alwaha Project, Khartoum State to study the potential of ethanol yield from fermented juice of local and exotic sweet sorghums. 40 genotypes were arranged in RCB design. Juice yield, ethanol yield as percentage of fermented juice, actual and theoretical ethanol yields were studied. The Baker’s yeast (Saccharomyces cerevisiae) was used in the fermentation process. ANOVA tests, correlation and regression analysis of actual vs theoretical ethanol yield were performed. Significant differences were detected among genotypes for ethanol yields. High ethanol yields as percentage from fermented juice were obtained, but ethanol yield per hectare was relatively low due to low juice yields. The genotypes BlueRibbon and KensasCollies from exotic materials; 5AbjSG51 and S.154 Ab70 from local materials were identified as promising for ethanol production. Correlation between actual ethanol yield and Juice yield was positive and highly significant. Regression analysis indicated that theoretical ethanol yield was little bit overestimated. It was concluded that juice yield is more crucial than ethanol concentration in maximizing ethanol yield of sweet sorghum. More efforts are needed to develop high juice-yielding genotypes. Juice maximization can also be achieved by using efficient milling devices and optimizing cultural practices.

Utilization of Biomass Derived from Cyanobacteria-Based Agro-Industrial Wastewater Treatment and Raisin Residue Extract for Bioethanol Production

Biofuels produced from photosynthetic microorganisms such as microalgae and cyanobacteria could potentially replace fossil fuels as they offer several advantages over fuels produced from lignocellulosic biomass. In this study, energy production potential in the form of bioethanol was examined using different biomasses derived from the growth of a cyanobacteria-based microbial consortium on a chemical medium and on agro-industrial wastewaters (i.e., dairy wastewater, winery wastewater and mixed winery–raisin effluent) supplemented with a raisin residue extract. The possibility of recovering fermentable sugars from a microbial biomass dominated by the filamentous cyanobacterium Leptolynbgya sp. was demonstrated. Of the different acid hydrolysis conditions tested, the best results were obtained with sulfuric acid 2.5 N for 120 min using dried biomass from dairy wastewater and mixed winery–raisin wastewaters. After optimizing sugar release from the microbial biomass by applying acid hydrolysis, alcoholic fermentation was performed using the yeast Saccharomyces cerevisiae. Raisin residue extract was added to the treated biomass broth in all experiments to enhance ethanol production. Results showed that up to 85.9% of the theoretical ethanol yield was achieved, indicating the potential use of cyanobacteria-based biomass in combination with a raisin residue extract as feedstock for bioethanol production.

Potential of Candida glabrata from ragi as a bioethanol producer using selected carbohydrate substrates

Vincent M, Johnny Q, Adeni DSA, Suhaili N. 2020. Potential of Candida glabrata from ragi as a bioethanol producer using selected carbohydrate substrates. Nusantara Bioscience 12: 1-10. The flexibility and efficiency of fermenting microorganisms to convert substrates to ethanol are important factors in achieving high bioethanol yields during ethanolic fermentation. In this study, Candida glabrata, a common yeast found in fermented food, was evaluated in terms of its capability to produce ethanol using different types of carbohydrates, which included simple saccharides (glucose, maltose, sucrose), polysaccharides (starch and cellulose) and complex carbohydrates (total sago effluent, TSE). Our results indicated that C. glabrata was able to efficiently produce ethanol from glucose at 79.84% TEY (Theoretical Ethanol Yield). The ethanol production from sucrose was low, which was only 6.44% TEY, while no ethanol was produced from maltose. Meanwhile, for complex carbohydrate substrates such as starch and cellulose, ethanol was produced only when supplementary enzymes were introduced. Simultaneous Saccharification and Fermentation (SSF) of starch dosed with amylases resulted in an ethanol yield of 55.08% TEY, whilst SSF of cellulose dosed with cellulases yielded a TEY of 31.41%. When SSF was performed on TSE dosed with amylases and cellulases, the highest ethanol production was recorded within 24 h, with a yield of 23.36% TEY. Lactic acid and acetic acid were found to be at minimal levels throughout the fermentation period, indicating an efficient ethanol conversion. A notable increase in C. glabrata biomass was observed in cultures fed with glucose, starch (with supplementary amylases), and TSE (with supplementary amylases and cellulases). The current study indicates that C. glabrata can be used for bioethanol production from glucose, polysaccharides, and complex starchy lignocellulosic substrates such as TSE via SSF.

Bio-Fuel Potential of Some Sweet Sorghum Genotypes [Sorghum bicolor (L.) Moench ssp. saccharatum]

The fossil energy sources used in the world are gradually decreasing and limited. Fossil fuels cause environmental pollution, and the unit price is constantly increasing. For this reason, demand for cheaper and renewable energy sources that do not pollute the environment is increasing day by day. The sweet sorghum plant has attracted attention in recent years with its high biomass yield, sugar content and bioethanol yield. In this study, it was aimed to determine the bio-fuel potential of some sweet sorghum genotypes in semi-arid climatic conditions. The experiment was set up in randomized complete block design with 4 replicates. Research was carried out in 2015 under Harran Plain (36o 42’ N and 38o 58’ E) second crop conditions, Sanliurfa, Turkey. In the study 49 genotypes of sweet sorghum were used. Stalk yield, juice yield, syrup yield, brix, sugar yield and theoretical ethanol yield were determined in the study. Significant differences were found between the genotypes for tested characteristics (P≤0.01). Stalk yield ranged from 7110.0 kg da-1 to 24262.5 da-1, juice yield from 2550.0 L da-1 to 12187.5 L da-1, syrup yield from 291.4 L da-1 to 2242.5 L da-1. Also, brix value varied between %7.0 and %18.87, sugar yield between 247.7 da-1 and 1906.1 da-1, Theoretical ethanol yield between 131.9 L da-1 and 1014.8 L da-1. Considering to stalk yield, juice and syrup yield, brix, sugar yield and theoretical ethanol yield; Nebraska sugar, Topper 76, Smith, M81E and Corina genotypes were found as the best. As a result of research, 21 genotypes with better performance than the others were selected for further studies.

Characterization of semi-arid Chadian sweet sorghum accessions as potential sources for sugar and ethanol production

Sweet sorghum (Sorghum bicolor (L.) Moench) is an important crop in Chad that plays an economic role in the countryside were stalks are produced mainly for human consumption without any processing. Unfortunately, very little information exists on its genetic diversity and brix content. Studies performed in 2014 and 2015 showed that there were significant variations (p < 0.001) for all assessed quantitative traits. Potential grain yield (0.12–1.67 t ha−1), days to 50% flowering (68.3–126.3 days), and plant height (128.9–298.3 cm) were among traits that exhibited broader variability. Brix content range from 5.5 to 16.7% across accessions, was positively correlated to stalk diameter and plant height, but negatively correlated to moisture content in fresh stalk and potential grain yield. Fresh stalk yield range from 16.8 to 115.7 Mg ha−1, with a mean value of 58.3 Mg ha−1 across accession. Moisture content in fresh stalk range from 33.7 to 74.4% but was negatively correlated to fresh stalk yield. Potential sugar yield range from 0.5 to 5.3 Mg ha−1 across accession with an average of 2.2 Mg ha−1. Theoretical ethanol yield range from 279.5 to 3,101.2 L ha−1 across accession with an average of 1,266.3 L ha−1 which is significantly higher than values reported under similar semiarid conditions. Overall, grain yields were comparatively low. However, two accessions had grain yield of more than 1.5 t ha−1; which is greater than the average 1.0 t ha−1 for local grain sorghum varieties in Chad. These could have multi-purpose uses; grains, sugar and bioenergy production.

Evaluating and engineering Saccharomyces cerevisiae promoters for increased amylase expression and bioethanol production from raw starch

ABSTRACT Bioethanol production from starchy biomass via consolidated bioprocessing (CBP) will benefit from amylolytic Saccharomyces cerevisiae strains that produce high levels of recombinant amylases. This could be achieved by using strong promoters and modification thereof to improve gene expression under industrial conditions. This study evaluated eight endogenous S. cerevisiae promoters for the expression of a starch-hydrolysing α-amylase gene. A total of six of the native promoters were modified to contain a promoter-proximal intron directly downstream of the full-length promoter. Varying results were obtained; four native promoters outperformed the ENO1P benchmark under aerobic conditions and two promoters showed better expression under simulated CBP conditions. The addition of the RPS25A intron significantly improved the expression from most promoters, displaying increased transcript levels, protein concentrations and amylase activities. Raw starch-utilising strains were constructed through co-expression of selected α-amylase cassettes and a glucoamylase gene. The amylolytic strains displayed improved fermentation vigour on raw corn starch and broken rice, reaching 97% of the theoretical ethanol yield and converting 100% of the available carbon to products within 120 h in small-scale CBP fermentations on broken rice. This study showed that enhanced amylolytic strains for the conversion of raw starch to ethanol can be achieved through turnkey promoter selection and/or engineering.

Biotransformation of Carboxylic Acids to Alcohols: Characterization of Thermoanaerobacter Strain AK152 and 1-Propanol Production via Propionate Reduction

Thermoanaerobacter strains have recently gained interest because of their ability to convert short chain fatty acids to alcohols using actively growing cells. Thermoanaerobacter thermohydrosulfuricus strain AK152 was physiologically investigated for its ethanol and other alcohol formation. The temperature and pH optimum of the strain was 70 °C and pH 7.0 and the strain degraded a variety of compounds present in lignocellulosic biomass like monosaccharides, disaccharides, and starch. The strain is highly ethanologenic, producing up to 86% of the theoretical ethanol yield form hexoses. Strain AK152 was inhibited by relatively low initial substrate (30 mM) concentration, leading to inefficient degradation of glucose and levelling up of all end-product formation. The present study shows that the strain produces alcohols from most of the tested carboxylic acids, with the highest yields for propionate conversion to propanol (40.7%) with kinetic studies demonstrating that the maximum conversion happens within the first 48 h of fermentation. Various physiological tests were performed to maximize the acid conversion to the alcohol which reveals that the optimum pH for propionate conversion is pH 6.7 which affords a 57.3% conversion. Kinetic studies reveal that propionate conversion is rapid, achieving a maximum conversion within the first 48 h of fermentation. Finally, by using 13C NMR, it was shown that the addition of propionate indeed converted to propanol.