Ethanol Production via Biochemical Route

Ethanol from lignocellulosic biomass is produced mainly via biochemical routes. The three major steps involved are pretreatment, enzymatic hydrolysis, and fermentation as shown in Figure. Biomass is pretreated to improve the accessibility of enzymes. After pretreatment, biomass undergoes enzymatic hydrolysis for conversion of polysaccharides into monomer sugars, such as glucose and xylose. Subsequently, sugars are fermented to ethanol by the use of different microorganisms.

Pretreated biomass can directly be converted to ethanol by using the process called simultaneous saccharification and cofermentation (SSCF). Pretreatment is a critical step which enhances the enzymatic hydrolysis of biomass. Basically, it alters the physical and chemical properties of biomass and improves the enzyme access and effectiveness which may also lead to a change in crystallinity and degree of polymerization of cellulose. The internal surface area and pore volume of pretreated biomass are increased which facilitates substantial improvement in accessibility of enzymes. The process also helps in enhancing the rate and yield of monomeric sugars during enzymatic hydrolysis steps.

Pretreatment methods can be broadly classified into four groups – physical, chemical, physio-chemical and biological. Physical pretreatment processes employ the mechanical comminution or irradiation processes to change only the physical characteristics of biomass. The physio-chemical process utilizes steam or steam and gases, like SO2 and CO2. The chemical processes employs acids (H2SO4, HCl, organic acids etc) or alkalis (NaOH, Na2CO3, Ca(OH)2, NH3 etc). The acid treatment typically shows the selectivity towards hydrolyzing the hemicelluloses components, whereas alkalis have better selectivity for the lignin. The fractionation of biomass components after such processes help in improving the enzymes accessibility which is also important to the efficient utilization of enzymes.

The pretreated biomass is subjected to enzymatic hydrolysis using cellulase enzymes to convert the cellulose to fermentable sugars. Cellulase refers to a class of enzymes produced chiefly by fungi and bacteria which catalyzes the hydrolysis of cellulose by attacking the glycosidic linkages. Cellulase is mixture of mainly three different functional protein groups: exo-glucanase (Exo-G), endo-glucanase(Endo-G) and β-glucosidase (β-G). The functional proteins work synergistically in hydrolyzing the cellulose into the glucose. These sugars are further fermented using microorganism and are converted to ethanol. The microorganisms are selected based on their efficiency for ethanol productivity and higher product and inhibitors tolerance. Yeast Saccharomyces cerevisiae is used commercially to produce the ethanol from starch and sucrose.

Escherichia coli strain has also been developed recently for ethanol production by the first successful application of metabolic engineering. E. coli can consume variety of sugars and does not require the complex growth media but has very narrow operable range of pH. E. coli has higher optimal temperature than other known strains of bacteria.

The major cost components in bioethanol production from lignocellulosic biomass are the pretreatment and the enzymatic hydrolysis steps. In fact, these two process are someway interrelated too where an efficient pretreatment strategy can save substantial enzyme consumption. Pretreatment step can also affect the cost of other operations such as size reduction prior to pretreatment. Therefore, optimization of these two important steps, which collectively contributes about 70% of the total processing cost, are the major challenges in the commercialization of bioethanol from 2nd generation feedstock.

Enzyme cost is the prime concern in full scale commercialization. The trend in enzyme cost is encouraging because of enormous research focus in this area and the cost is expected to go downward in future, which will make bioethanol an attractive option considering the benefits derived its lower greenhouse gas emissions and the empowerment of rural economy.

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What is Lignocellulosic Biomass?

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First-generation biofuels (produced primarily from food crops such as grains, sugar beet and oil seeds) are limited in their ability to achieve targets for oil-product substitution, climate change mitigation, and economic growth. Their sustainable production is under scanner, as is the possibility of creating undue competition for land and water used for food and fibre production.

The cumulative impacts of these concerns have increased the interest in developing biofuels produced from non-food biomass. Feedstocks from ligno-cellulosic materials include cereal straw, bagasse, forest residues, and purpose-grown energy crops such as vegetative grasses and short rotation forests. These second-generation biofuels could avoid many of the concerns facing first-generation biofuels and potentially offer greater cost reduction potential in the longer term.

The largest potential feedstock for ethanol is lignocellulosic biomass, which includes materials such as agricultural residues (corn stover, crop straws and bagasse), herbaceous crops (alfalfa, switchgrass), short rotation woody crops, forestry residues, waste paper and other wastes (municipal and industrial). Bioethanol production from these feedstocks could be an attractive alternative for disposal of these residues. Importantly, lignocellulosic feedstocks do not interfere with food security. Moreover, bioethanol is very important for both rural and urban areas in terms of energy security reason, environmental concern, employment opportunities, agricultural development, foreign exchange saving, socioeconomic issues etc.

Economically, lignocellulosic biomass has an advantage over other agriculturally important biofuels feedstocks such as corn starch, soybeans, and sugar cane, because it can be produced quickly and at significantly lower cost than food crops. Lignocellulosic biomass is an important component of the major food crops; it is the non-edible portion of the plant, which is currently underutilized, but could be used for biofuel production. In short, lignocellulosic biomass holds the key to supplying society’s basic needs for sustainable production of liquid transportation fuels without impacting the nation’s food supply.

Production of Cellulosic Ethanol

The production of biofuels from lignocellulosic feedstocks can be achieved through two very different processing routes. They are:

• Biochemical – in which enzymes and other micro-organisms are used to convert cellulose and hemicellulose components of the feedstocks to sugars prior to their fermentation to produce ethanol;
• Thermo-chemical – where pyrolysis/gasification technologies produce a synthesis gas (CO + H₂) from which a wide range of long carbon chain biofuels, such as synthetic diesel or aviation fuel, can be reformed.

Lignocellulosic biomass consists mainly of lignin and the polysaccharides cellulose and hemicellulose. Compared with the production of ethanol from first-generation feedstocks, the use of lignocellulosic biomass is more complicated because the polysaccharides are more stable and the pentose sugars are not readily fermentable by Saccharomyces cerevisiae. In order to convert lignocellulosic biomass to biofuels the polysaccharides must first be hydrolysed, or broken down, into simple sugars using either acid or enzymes. Several biotechnology-based approaches are being used to overcome such problems, including the development of strains of Saccharomyces cerevisiae that can ferment pentose sugars, the use of alternative yeast species that naturally ferment pentose sugars, and the engineering of enzymes that are able to break down cellulose and hemicellulose into simple sugars.

Lignocellulosic processing pilot plants have been established in the EU, in Denmark, Spain and Sweden. The world’s largest demonstration facility of lignocellulose ethanol (from wheat, barley straw and corn stover), with a capacity of 2.5 Ml, was first established by Iogen Corporation in Ottawa, Canada. Many other processing facilities are now in operation or planning throughout the world.

Ethanol from lignocellulosic biomass is produced mainly via biochemical routes. The three major steps involved are pretreatment, enzymatic hydrolysis, and fermentation as shown in Figure. Biomass is pretreated to improve the accessibility of enzymes. After pretreatment, biomass undergoes enzymatic hydrolysis for conversion of polysaccharides into monomer sugars, such as glucose and xylose. Subsequently, sugars are fermented to ethanol by the use of different microorganisms.

Pretreated biomass can directly be converted to ethanol by using the process called simultaneous saccharification and cofermentation (SSCF).  Pretreatment is a critical step which enhances the enzymatic hydrolysis of biomass. Basically, it alters the physical and chemical properties of biomass and improves the enzyme access and effectiveness which may also lead to a change in crystallinity and degree of polymerization of cellulose. The internal surface area and pore volume of pretreated biomass are increased which facilitates substantial improvement in accessibility of enzymes. The process also helps in enhancing the rate and yield of monomeric sugars during enzymatic hydrolysis steps.

Pretreatment methods can be broadly classified into four groups – physical, chemical, physio-chemical and biological. Physical pretreatment processes employ the mechanical comminution or irradiation processes to change only the physical characteristics of biomass. The physio-chemical process utilizes steam or steam and gases, like SO₂ and CO₂. The chemical processes employs acids (H₂SO₄, HCl, organic acids etc) or alkalis (NaOH, Na₂CO₃, Ca(OH)₂, NH₃ etc). The acid treatment typically shows the selectivity towards hydrolyzing the hemicelluloses components, whereas alkalis have better selectivity for the lignin. The fractionation of biomass components after such processes help in improving the enzymes accessibility which is also important to the efficient utilization of enzymes.

Presently, a ton of dry biomass typically yields 60-70 gallons of bioethanol. The major cost components in bioethanol production from lignocellulosic biomass are the pretreatment and the enzymatic hydrolysis steps. In fact, these two process are someway interrelated too where an efficient pretreatment strategy can save substantial enzyme consumption. Pretreatment step can also affect the cost of other operations such as size reduction prior to pretreatment. Therefore, optimization of these two important steps, which collectively contributes about 70% of the total processing cost, are the major challenges in the commercialization of bioethanol from 2^nd generation feedstock.

Lignocellulosic Biomass

Image via Wikipedia

First-generation biofuels (produced primarily from food crops such as grains, sugar beet and oil seeds) are limited in their ability to achieve targets for oil-product substitution, climate change mitigation, and economic growth. Their sustainable production is under scanner, as is the possibility of creating undue competition for land and water used for food and fibre production.

The cumulative impacts of these concerns have increased the interest in developing biofuels produced from non-food biomass. Feedstocks from ligno-cellulosic materials include cereal straw, bagasse, forest residues, and purpose-grown energy crops such as vegetative grasses and short rotation forests. These second-generation biofuels could avoid many of the concerns facing first-generation biofuels and potentially offer greater cost reduction potential in the longer term.

The largest potential feedstock for ethanol is lignocellulosic biomass, which includes materials such as agricultural residues (corn stover, crop straws and bagasse), herbaceous crops (alfalfa, switchgrass), short rotation woody crops, forestry residues, waste paper and other wastes (municipal and industrial). Bioethanol production from these feedstocks could be an attractive alternative for disposal of these residues. Importantly, lignocellulosic feedstocks do not interfere with food security. Moreover, bioethanol is very important for both rural and urban areas in terms of energy security reason, environmental concern, employment opportunities, agricultural development, foreign exchange saving, socioeconomic issues etc.

Economically, lignocellulosic biomass has an advantage over other agriculturally important biofuels feedstocks such as corn starch, soybeans, and sugar cane, because it can be produced quickly and at significantly lower cost than food crops. Lignocellulosic biomass is an important component of the major food crops; it is the non-edible portion of the plant, which is currently underutilized, but could be used for biofuel production. In short, lignocellulosic biomass holds the key to supplying society’s basic needs for sustainable production of liquid transportation fuels without impacting the nation’s food supply.