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 2nd generation feedstock.
