Lignocellulosic biomass for bioethanol production: Current perspectives,potential issues and future prospects

During the most recent decades increased interest in fuel from biomass in the United States and worldwide has emerged each time petroleum derived gasoline registered well publicized spikes in price. The willingness of the U.S. government to face the issues of more heavily high-priced foreign oil and climate change has led to more investment on plant-derived sustainable biofuel sources. Biomass derived from corn has become one of the primary feedstocks for bioethanol production for the past several years in the U.S. However, the argument of whether to use food as biofuel has led to a search for alternative non-food sources. Consequently, industrial research efforts have become more focused on low-cost large-scale processes for lignocellulosic feedstocks originating mainly from agricultural and forest residues along with herbaceous materials and municipal wastes. Although cellulosic-derived biofuel is a promising technology, there are some obstacles that interfere with bioconversion processes reaching optimal performance associated with minimal capital investment. This review summarizes current approaches on lignocellulosic-derived biofuel bioconversion and provides an overview on the major steps involved in cellulosic-based bioethanol processes and potential issues challenging these operations. Possible solutions and recoveries that could improve bioprocessing are also addressed. This includes the development of genetically engineered strains and emerging pretreatment technologies that might be more efficient and economically feasible. Future prospects toward achieving better biofuel operational performance via systems approaches such as risk and life cycle assessment modeling are also discussed.     

Fermentation of wheat and triticale hydrolysates: A comparative study

In bioethanol production four wheat varieties were investigated: NS 40S, Renesansa, Rapsodija and Dragana, as well as four triticale varieties: Oganj, Jutro, Odisej and NST 21/06. The samples were grinded and mixed with water under conditions prescribed for bioethanol processing. Liquefaction of wheat samples was conducted at 65 °C and 60 °C for triticale samples. All the investigated samples were prepared with or without the addition of technical enzymes: thermostable α-amylase (Thermamyl SC) and glucoamylase (SAN Super 360 L). After liquefaction and saccharification, the samples were subjected to fermentation with Saccharomyces cerevisiae (dry active instant yeast). After the fermentation step ended, ethanol content was determined, and autoamylolytical quotient was calculated. The following values of autoamylolytical quotient were obtained: 71.79% for variety NS 40S; 72.22% for variety Dragana; 62.15% for variety Rapsodija and 81.46% for variety Renesansa. The obtained results revealed that Renesansa is the most suitable wheat variety for bioethanol production due to the largest amount of its native amylolytical enzymes. The following autoamylolytical quotients were obtained for triticale: 99.30% for variety Oganj; 98.65% for variety Jutro; 99.55% for variety Odisej and 94.24% for variety NST 21/06. The results implied that, in the case of triticale, technical enzymes were not needed for starch degradation. It is possible to conduct preparation of triticale at 60 °C.     

Potential development of bioethanol production in Vojvodina

The Autonomous Province of Vojvodina is an Autonomous Province in Serbia, containing about 27% of its total population according to the 2002 Census. Contribution of renewable energy sources in total energy consumption of Vojvodina contemporary amounts to less than 1%, apropos 280 GWh/year. By combining of methods of introduction of new and renewable sources, systematic application of methods for increasing of energetic efficacy, as well as of introduction of the new technologies, percentage of contribution of the non-conventional energy sources in Vojvodina could be increased to as much as 20%. This paper presents the potential of development of bioethanol production in Vojvodina. Production of bioethanol on small farms can be successfully applied for processing of only 30 kg of corn per day, with obtaining of crude ethanol in the so-called “brandy ladle” and use of lygnocellulosic agricultural wastes as an energy source. In a case of construction of a larger number of such plants, the only possible solution is seen in the principle of construction of the so-called “satellite plants”, which will on small farm produce crude ethanol, with obtaining and consumption of stillage for animal feeding, and consumption of agricultural wastes as energetic fuels. If stillage is to be used as feed in wet feeding, it is estimated that, because of restrictions established by the magnitude of animal farm, the upper limit of capacity of such enterprises that process is at some 10–15 tons of corn per day, and production of 3000–3500 hL of absolute ethanol per day. In such a case, for animal feeding necessary is to have herd with 1300–1700 of milking cows or 5000–25,000 heads of sheep and/or pigs. Technological model of separate grain processing ad bioethanol production from dextrose hydrolysates of starch is interesting for countries possessing plants for bioethanol production from molasses and plants for cereals processing into starch and dextrose hydrolysates of starch.     

Critical analysis of the European Union directive which regulates the use of biofuels: An approach to the Spanish case

For more than a decade we have lived in a period where the so-called “sustainability” is crucial and is motivated primarily by the social awareness of achieving a balance between human development and the conservation of the environment. This philosophy has a direct and inevitable impact on business and politics. Governments have long since been developing standards and encouraging various diverse initiatives whose aim is to defend the environment.In recent times, the global debate on the environment has been centred on CO₂ emissions. This gas is the major cause of the “greenhouse effect” and people are more concerned with the idea that the emissions of this gas should be minimized. As a result of this concern, the Kyoto Protocol was enacted and subscribed to by many countries, setting the maximum gas emissions for them.Fossil fuels are a major source of CO₂ emissions. In 2003 the European Union (EU) directive 2003/30/EC [2003/30/EC Directive of the European Parliament and the Council—8th may 2003. On the promotion of the use of biofuels or other renewable fuels for transport] was developed with the aim of promoting the use of biofuels as a substitute for diesel or petrol among European Union countries as well as to contribute to fulfilling the commitments on climate change, security of supply in environmentally friendly conditions and the promotion of renewable energy sources.In order to achieve these goals, the directive forces all EU members to ensure that at least 5.75% of all petrol and diesel fuels sold for transport purposes are biofuels before December 31 of 2010. European Union countries have social and economic characteristics unique to themselves. The energy dependence from foreign sources, the features of the agricultural sector or the degree of industrialization varies greatly from one country to another. In this context, it is questionable whether the obligation imposed by this directive applies to achieve uniform and/or identical goals in each of the countries involved and whether the actions of the various governments are also aligned with these goals.     

Ethanol-tolerant Saccharomyces cerevisiae strains isolated under selective conditions by over-expression of a proofreading-deficient DNA polymerase δ

Ethanol damages the cell membrane and functional proteins, gradually reducing cell viability, and leading to cell death during fermentation which impairs effective bioethanol production by budding yeast Saccharomyces cerevisiae. To obtain more suitable strains for bioethanol production and to gain a better understanding of ethanol tolerance, ethanol-tolerant mutants were isolated using the novel mutagenesis technique based on the disparity theory of evolution. According to this theory evolution can be accelerated by affecting the lagging-strand synthesis in which DNA polymerase δ is involved. Expression of the pol3-01 gene, a proofreading-deficient of DNA polymerase δ, in S. cerevisiae W303-1A grown under conditions of increasing ethanol concentration resulted in three ethanol-tolerant mutants (YFY1, YFY2 and YFY3), which could grow in medium containing 13% ethanol. Ethanol productivity also increased in YFY strains compared to the wild-type strain in medium containing 25% glucose. Cell morphology of YFY strain cells was normal even in the presence of 8% ethanol, whereas W303-1A cells were expanded by a big vacuole. Furthermore, two of these mutants were also resistant to high-temperature, Calcofluor white and NaCl. Expression levels of TPS1 and TSL1, which are responsible for trehalose biosynthesis, were higher in YFY strains relative to W303-1A, resulting in high levels of intracellular trehalose in YFY strains. This contributed to the multiple-stress tolerance that makes YFY strains suitable for the production of bioethanol.     

Pretreatment and hydrolysis of cellulosic agricultural wastes with a cellulase-producing Streptomyces for bioethanol production

Production of reducing sugar by hydrolysis of corncob material with Streptomyces sp. cellulase and ethanol fermentation of cellulosic hydrolysate was investigated. Cultures of Streptomyces sp. T3-1 improved reducing sugar yields with the production of CMCase, Avicelase and ??-glucosidase activity of 3.8, 3.9 and 3.8 IU/ml, respectively. CMCase, Avicelase, and ??-glucosidase produced by the Streptomyces sp. T3-1 favored the conversion of cellulose to glucose. It was recognized that the synergistic interaction of endoglucanase, exoglucanase and ??-glucosidase resulted in efficient hydrolysis of cellulosic substrate. After 5 d of incubation, the overall reducing sugar yield reached 53.1 g/100 g dried substrate. Further fermentation of cellulosic hydrolysate containing 40.5 g/l glucose was performed using Saccharomyces cerevisiae BCRC 21812, 14.6 g/l biomass and 24.6 g/l ethanol was obtained within 3 d. The results have significant implications and future applications regarding to production of fuel ethanol from agricultural cellulosic waste.     

Homolytic cleavage C-C bond in the electrooxidation of ethanol and bioethanol

Nowadays, the studies are focused on the search of better electrocatalysts that promote the complete oxidation of ethanol/bioethanol to CO₂. To that end, amorphous bi-catalytic catalysts of composition Ni₅₉Nb₄₀Pt_1−xY_x (Y = Cu, Ru, x = 0.4% at.) have been developed, obtained by mechanical alloying, resulting in higher current densities and an improvement in tolerance to adsorbed CO vs. Ni₅₉Nb₄₀Pt₁ catalyst. By using voltammetric techniques, the appearance of three oxidation peaks can be observed. The first peak could be associated with the electrooxidative process of ethanol/bioethanol to acetaldehyde, the second peak could be the oxidation of acetaldehyde to acetic acid, and the last peak might be the final oxidation to CO₂. Chrono-amperometric experiments show qualitative poisoning of catalytic surfaces. However, the in situ Fourier Transformed Infrared Spectroscopy, FTIR, is used for the quasi-quantitative determination with which can be observed the appearance and evolution of different vibrational bands of carbonyl and carboxylic groups of different species, as it moves towards anodic potential in the electrooxidative process.     

Influence of impregnation with lactic acid on sugar yields from steam pretreatment of sugarcane bagasse and spruce, for bioethanol production

Lignocellulosic biomass can be utilized to produce ethanol, a promising alternative energy source produced through fermentation of sugars. However, in order to achieve high sugar and ethanol yields, the lignocellulosic material must be pretreated before the enzymatic hydrolysis and fermentation. Dilute acid pretreatment, using SO₂, is one of the most promising methods of pretreatment for softwood and agricultural residues. However, handling the high acidity of the slurry obtained from pretreatment and difficulty in recycling/degradation of the impregnating agent are some of the drawbacks of the dilute acid processes. In the present study the influence of utilization of a weak organic acid (lactic acid), as impregnating agent, on the sugar yield from pretreatment, with and without addition of SO₂, was investigated. The efficiency of pretreatment was assessed by enzymatic hydrolysis of the slurry obtained by pretreatment, using sugarcane bagasse and spruce, stored for one and two months in the presence of lactic acid separately, as feedstocks. Pretreatment of bagasse after storage with 0.5% lactic acid resulted in an overall glucose yield, i.e. after enzymatic hydrolysis, of 79% of theoretical based on the amount available in the raw material. This was as good as pretreatment using SO₂ as impregnating agent. However, storage of spruce with lactic acid before pretreatment, with and without addition of SO₂, was not efficient and resulted in lower sugar yields than pretreatment using SO₂ only.     

Process optimization for bioethanol production from cassava starch using novel eco-friendly enzymes

Although cassava (Manihot esculenta Crantz) is a potential bioethanol crop, high operational costs resulted in a negative energy balance in the earlier processes. The present study aimed at optimizing the bioethanol production from cassava starch using new enzymes like Spezyme^® Xtra and Stargen™ 001. The liquefying enzyme Spezyme was optimally active at 90 °C and pH 5.5 on a 10% (w/v) starch slurry at levels of 20.0 mg (280 Amylase Activity Units) for 30 min. Stargen levels of 100 mg (45.6 Granular Starch Hydrolyzing Units) were sufficient to almost completely hydrolyze 10% (w/v) starch at room temperature (30 ± 1 °C). Ethanol yield and fermentation efficiency were very high (533 g/kg and 94.0% respectively) in the Stargen + yeast process with 10% (w/v) starch for 48 h. Raising Spezyme and Stargen levels to 560 AAU and 91.2 GSHU respectively for a two step loading [initial 20% (w/v) followed by 20% starch after Spezyme thinning]/initial higher loading of starch (40% w/v) resulted in poor fermentation efficiency. Upscaling experiments using 1.0 kg starch showed that Stargen to starch ratio of 1:100 (w/w) could yield around 558 g ethanol/kg starch, with a high fermentation efficiency of 98.4%. The study showed that Spezyme level beyond 20.0 mg for a 10% (w/v) starch slurry was not critical for optimizing bioethanol yield from cassava starch, although an initial thinning of starch for 30 min by Spezyme facilitated rapid saccharification-fermentation by Stargen + yeast system. The specific advantage of the new process was that the reaction could be completed within 48.5 h at 30 ± 1 °C.