Category Archives: Biotechnology

Background:
Since its inception, the carbohydrate-active enzymes database (CAZy; http://www.cazy.org) has described the families of enzymes that cleave or build complex carbohydrates, namely the glycoside hydrolases (GH), the polysaccharide lyases (PL), the carbohydrate esterases (CE), the glycosyltransferases (GT) and their appended non-catalytic carbohydrate-binding modules (CBM). The recent discovery that members of families CBM33 and family GH61 are in fact lytic polysaccharide monooxygenases (LPMO), demands a reclassification of these families into a suitable category.
Results:
Because lignin is invariably found together with polysaccharides in the plant cell wall and because lignin fragments are likely to act in concert with (LPMO), we have decided to join the families of lignin degradation enzymes to the LPMO families and launch a new CAZy class that we name "Auxiliary Activities" in order to accommodate a range of enzyme mechanisms and substrates related to lignocellulose conversion. Comparative analyses of these auxiliary activities in 41 fungal genomes reveal a pertinent division of several fungal groups and subgroups combining their phylogenetic origin and their nutritional mode (white vs. brown rot).
Conclusions:
The new class introduced in the CAZy database extends the traditional CAZy families, and provides a better coverage of the full extent of the lignocellulose breakdown machinery.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/41

Background:
Pretreatment is a key step to decrease the recalcitrance of lignocelluloses and then increase the digestibility of cellulose in second-generation bioethanol production. In this study, wood chips from triploid poplar were biopretreated with white rot fungus Trametes velutina D10149. The effects of incubation duration on delignification efficiency and structural modification of cellulose were comparably studied, as well as the digestibility of cellulose by simultaneous saccharification and fermentation (SSF).
Results:
Although microbial pretreatments did not significantly introduce lignin degradation, the data from SSF exhibited higher cellulose conversion (21-75% for biopretreated samples for 4--16 weeks) as compared to the untreated poplar (18%). In spite of the essential maintain of crystallinity, the modification of lignin structure during fungal treatment undoubtedly played a key role in improving cellulose bioconversion rates. Finally, the ethanol concentration of 5.16 g/L was detected in the fermentation broth from the cellulosic sample biodegraded for 16 weeks after 24 h SSF, achieving 34.8% cellulose utilization in poplar.
Conclusion:
The potential fungal pretreatment with Trametes velutina D10149 was firstly explored in this study. It is found that the biopretreatment process had a significant effect on the digestibility of substrate probably due to the removal and unit variation of lignin, since the crystallinities of substrates were rarely changed. Additional investigation is still required especially to improve the selectivity for lignin degradation and optimize the digestibility of cellulose.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/42

Background:
It is widely believed that reducing the lignocellulosic biomass particle size would improve the biomass digestibility by increasing the total surface area and eliminating mass and heat transfer limitation during hydrolysis reactions. However, past studies demonstrate that particle size influences biomass digestibility to a limited extent. Thus, this paper studies the effect of particle size (milled: 2 mm, 5 mm, cut: 2 cm and 5 cm) on rice straw conversion. Two different Ammonia Fiber Expansion (AFEX) pretreament conditions, AFEX C1 (low severity) and AFEX C2 (high severity) are used to pretreat the rice straw (named as AC1RS and AC2RS substrates respectively) at different particle size.
Results:
Hydrolysis of AC1RS substrates showed declining sugar conversion trends as the size of milled and cut substrates increased. Hydrolysis of AC2RS substrates demonstrated opposite conversion trends between milled and cut substrates. Increasing the glucan loading to 6% during hydrolysis reduced the sugar conversions significantly in most of AC1RS and AC2RS except for AC1RS-2 mm and AC2RS-5 cm. Both AC1RS-2 mm and AC2RS-5 cm indicated gradual decreasing trends in sugar conversion at high glucan loading. Analysis of SEM imaging for URS and AFEX pretreated rice straw also indicated qualitative agreement with the experimental data of hydrolysis. The largest particle size, AC2RS-5 cm produced the highest sugar yield of 486.12 g/kg of rice straw during hydrolysis at 6% glucan loading equivalent to 76.0% of total theoretical maximum sugar yield, with an average conversion of 85.9% from total glucan and xylan. In contrast, AC1RS-5 cm gave the lowest sugar yield with only 107.6 g/kg of rice straw, about 16.8% of total theoretical maximum sugar yield, and equivalent to one-quarter of AC2RS-5 cm sugar yield.
Conclusions:
The larger cut rice straw particles (5 cm) significantly demonstrated higher sugar conversion when compared to small particles during enzymatic hydrolysis when treated using high severity AFEX conditions. Analysis of SEM imaging positively supported the interpretation of the experimental hydrolysis trend and kinetic data.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/40

Background:
The use of Ionic liquids (ILs) as biomass solvents is considered to be an attractive alternative for the pretreatment of lignocellulosic biomass. Acid catalysts have been used previously to hydrolyze polysaccharides into fermentable sugars during IL pretreatment. This could potentially provide a means of liberating fermentable sugars from biomass without the use of costly enzymes. However, the separation of the sugars from the aqueous IL and recovery of IL is challenging and imperative to make this process viable.
Results:
Aqueous alkaline solutions are used to induce the formation of a biphasic system to recover sugars produced from the acid catalyzed hydrolysis of switchgrass in imidazolium-based ILs. The amount of sugar produced from this process was proportional to the extent of biomass solubilized. Pretreatment at high temperatures (e.g., 160[degree sign]C, 1.5 h) was more effective in producing glucose. Sugar extraction into the alkali phase was dependent on both the amount of sugar produced by acidolysis and the alkali concentration in the aqueous extractant phase. Maximum yields of 53% glucose and 88% xylose are recovered in the alkali phase, based on the amounts present in the initial biomass. The partition coefficients of glucose and xylose between the IL and alkali phases can be accurately predicted using molecular dynamics simulations.
Conclusions:
This biphasic system may enable the facile recycling of IL and rapid recovery of the sugars, and provides an alternative route to the production of monomeric sugars from biomass that eliminates the need for enzymatic saccharification and also reduces the amount of water required.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/39

Background:
Understanding the effects of pretreatment on anaerobic digestion of sludge waste from wastewater treatment plants is becoming increasingly important, as impetus moves towards the utilization of sludge for renewable energy production. Although the field of sludge pretreatment has progressed significantly over the past decade, critical questions concerning the underlying microbial interactions remain unanswered. In this study, a metagenomic approach was adopted to investigate the microbial composition and gene content contributing to enhanced biogas production from sludge subjected to a novel pretreatment method (maintaining pH at 10 for 8 days) compared to other documented methods (ultrasonic, thermal and thermal-alkaline).
Results:
Our results showed that pretreated sludge attained a maximum methane yield approximately 4-fold higher than that of the blank un-pretreated sludge set-up at day 17. Both the microbial and metabolic consortium shifted extensively towards enhanced biodegradation subsequent to pretreatment, providing insight for the enhanced methane yield. The prevalence of Methanosaeta thermophila and Methanothermobacter thermautotrophicus, together with the functional affiliation of enzymes-encoding genes suggested an acetoclastic and hydrogenotrophic methanogenesis pathway. Additionally, an alternative enzymology in Methanosaeta was observed.
Conclusions:
This study is the first to provide a microbiological understanding of improved biogas production subsequent to a novel waste sludge pretreatment method. The knowledge garnered will assist the design of more efficient pretreatment methods for biogas production in the future.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/38

Background:
Microalgal biomass contains a high level of carbohydrates which can be biochemically converted to biofuels using state-of-the-art strategies that are almost always needed to employ a robust pretreatment on the biomass for enhanced energy production. In this study, we used an ultrasonic pretreatment to convert microalgal biomass (Scenedesmus obliquus YSW15) into feasible feedstock for microbial fermentation to produce ethanol and hydrogen. The effect of sonication condition was quantitatively evaluated with emphases on the characterization of carbohydrate components in microalgal suspension and on subsequent production of fermentative bioenergy.MethodScenedesmus obliquus YSW15 was isolated from the effluent of a municipal wastewater treatment plant. The sonication durations of 0, 10, 15, and 60 min were examined under different temperatures at a fixed frequency and acoustic power resulted in morphologically different states of microalgal biomass lysis. Fermentation was performed to evaluate the bioenergy production from the non-sonicated and sonicated algal biomasses after pretreatment stage under both mesophilic (35[degree sign]C) and thermophilic (55[degree sign]C) conditions.
Results:
A 15 min sonication treatment significantly increased the concentration of dissolved carbohydrates (0.12 g g-1), which resulted in an increase of hydrogen/ethanol production through microbial fermentation. The bioconvertibility of microalgal biomass sonicated for 15 min or longer was comparable to starch as a control, indicating a high feasibility of using microalgae for fermentative bioenergy production. Increasing the sonication duration resulted in increases in both algal surface hydrophilicity and electrostatic repulsion among algal debris dispersed in aqueous solution. Scanning electron microscope images supported that ruptured algal cell allowed fermentative bacteria to access the inner space of the cell, evidencing an enhanced bioaccessibility. Sonication for 15 min was the best for fermentative bioenergy (hydrogen/ethanol) production from microalga, and the productivity was relatively higher for thermophilic (55[degree sign]C) than mesophilic (35[degree sign]C) condition.
Conclusion:
These results demonstrate that more bioavailable carbohydrate components are produced through the ultrasonic degradation of microalgal biomass, and thus the process can provide a high quality source for fermentative bioenergy production.Source:
http://www.biotechnologyforbiofuels.com/content/6/1/37