Biochemical Pathway for Ethanol Production

Ethanol from lignocellulosic biomass is produced mainly via biochemical routes. The three major steps involved are pretreatment, enzymatic hydrolysis, and fermentation. 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 ?-glucosidase (?-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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Salman Zafar is the Founder of Cleantech Solutions, and an international consultant, advisor and trainer with expertise in waste management, biomass energy, waste-to-energy, environment protection and resource conservation. His geographical areas of focus include Asia, Africa and the Middle East. Salman has successfully accomplished a wide range of projects in the areas of biogas technology, biomass energy, waste-to-energy, recycling and waste management. Salman has participated in numerous national and international conferences all over the world. He is a prolific environmental journalist, and has authored more than 300 articles in reputed journals, magazines and websites. In addition, he is proactively engaged in creating mass awareness on renewable energy, waste management and environmental sustainability through his blogs and portals. Salman can be reached at or

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