Increase in Fermentable Sugars of Olive Tree Pruning Biomass for Bioethanol Production: Application of an Experimental Design for Optimization of Alkaline Pretreatment

Olive Tree Pruning (OTP) biomass can be considered a suitable source of fermentable sugars for the production of second-generation bioethanol. The present study proposes a remarkable alternative for the valorization of olive tree pruning residues. OTP biomass was processed using a sequential calcium hydroxide pretreatment/enzymatic hydrolysis. A 24–1 half fractional factorial design was adopted for the screening of process variables and a central composite design was used for the optimization stage. Temperature and lime loading resulted statistically significant. The following optimal conditions were obtained: 0.01 g of Ca(OH)2/g of dry material, 20 g of H2O/g of dry material at 160 °C for 2 h. The mathematical model that governs this alkaline pretreatment was obtained with a 76% adjusted determination coefficient, which means that it is a good representation of the process. Under optimal operating conditions, 13% of the cellulose and 88% of the hemicellulose was solubilized. Moreover, the fermentable sugar content increased 1800% compared with the initial conditions, obtaining 240 g of glucose per kg of OTP residue. The fermentable sugars obtained after the calcium hydroxide pretreatment and enzymatic hydrolysis of OTP biomass yielded 2.8 g of ethanol/100 g of raw material.

Gracilaria gracilis (Gracilariales, Rhodophyta) from Dakhla (Southern Moroccan Atlantic Coast) as Source of Agar: Content, Chemical Characteristics, and Gelling Properties

Agar is a sulfated polysaccharide extracted from certain marine red algae, and its gel properties depend on the seaweed source and extraction conditions. In the present study, the seaweed Gracilaria gracilis (Gracilariales, Rhodophyta) from Dakhla (Moroccan Atlantic Coast) was investigated for its agar content, structure, and gel properties. The agar yields of G. gracilis were 20.5% and 15.6% from alkaline pretreatment and native extraction, respectively. Agar with alkaline pretreatment showed a better gelling property supported by higher gel strength (377 g·cm−2), gelling (35.4 °C), and melting (82.1 °C) temperatures with a notable increase in 3,6-anhydro-galactose (11.85%) and decrease in sulphate (0.32%) contents. The sulfate falling subsequent to alkaline pretreatment was verified through FT-IR spectroscopy. The 13C NMR spectroscopy showed that alkaline-pretreated agar has a typical unsubstituted agar pattern. However, native agar had a partially methylated agarose structure. Overall, this study suggested the possibility of the exploitation of G. gracilis to produce a fine-quality agar. Yet, further investigation may need to determine the seasonal variability of this biopolymer according to the life cycle of G. gracilis.

Techno-economic analysis of recombinant Endo-β-1,4-Glucanase production from Escherichia coli Eg-RK2 culture using oil palm empty fruit bunch

A techno-economic analysis of recombinant cellulase production from E. coli Eg-RK2 was conducted to support the fulfilling of Indonesia’s energy roadmap for ethanol production. The plant utilizes OPEFB as a primary substrate in cellulase production, with an expected lifetime of 12 years. The plant is assumed to be built in Indonesia and it will fulfill 1% of the total market demand. The effect of different pretreatment processes (alkaline, steam explosion, and sequential acid-alkaline) on the profitability parameter was also studied. A simulation using SuperPro Designer was used to calculate the mass and energy balance based on the kinetic parameters of E. coli EgRK2. A technology evaluation showed that alkaline pretreatment provides the highest yield with no known inhibitors formed. The steam explosion pretreatment offers the lowest rate of lignin and hemicellulose removal, and it is understood to form known fermentation inhibitors. The NPVs of the alkaline, steam explosion and sequential acid-alkaline pretreatments are USD 32,121,000, USD -36,841,000, and USD 384,000, respectively, which means the alkaline pretreatment is economically very feasible for the production of cellulase.

Investigation of the effects of pretreatment on the elemental composition of ash derived from selected Nigerian lignocellulosic biomass

Lignocellulosic biomass is an important source of renewable energy and a potential replacement for fossil fuels. In this work, the X-ray fluorescence (XRF) method was used to analyze the elemental composition of raw and pretreated lignocellulosic biomass of cassava peels, corn cobs, rice husks, sugarcane bagasse, yam peels, and mixtures of cassava peels and yam peels, corn cobs and rice husks and all five biomass samples combined. The influence of particle size on elemental properties was investigated by screening the selected biomass into two size fractions, of an average of 300 and 435 µm, respectively. The total concentration of Mg, Al, Si, P, S, Cl, Ca, Ti, Cr, Mn, Fe, Co, Cu, Zn, Sn, Ni, Br, Mo, Ba, Hg, and Pb were determined for each of the biomass samples before and after the different pretreatments adopted in this study. From the results of the analysis, there was a significant reduction in the concentration of calcium in all the analyzed biomass after the alkaline pretreatment with rice husks biomass having the lowest concentration of 66 ppm after the alkaline pretreatment. The sulfur content of the acid pretreated biomass increased considerably which is likely due to the sulfuric acid used for the acid pretreatment. The fact that a mixture of biomass feedstock affects the properties of the biomass after pretreatment was validated in the mixed biomass of cassava peels and yam peels biomass as an example. The concentration of Mg in the mixed biomass was 1441 ppm but was 200 ppm and 353 ppm in individual cassava peels and yam peels respectively. The results of this study demonstrated that pretreated mixtures of biomass have varied elemental compositions, which could be an important factor affecting downstream processes, especially if a hybrid feedstock is used in a large-scale application.

Polyhydroxyalkanoate production from rice straw hydrolysate obtained by alkaline pretreatment and enzymatic hydrolysis using Bacillus strains isolated from decomposing straw

Rice straw is an important low-cost feedstock for bio-based economy. This report presents a study in which rice straw was used both as a source for isolation of bacteria producing the biodegradable polyester polyhydroxyalkanoate (PHA), as well as the carbon source for the production of the polymer by the isolated bacteria. Of the 100 bacterial isolates, seven were found to be positive for PHA production by Nile blue staining and were identified as Bacillus species by 16S rRNA gene sequence analysis. Three isolates showed 100% sequence identity to B. cereus, one to B. paranthracis, two with 99 and 100% identity to B. anthracis, while one was closely similar to B. thuringiensis. For use in PHA production, rice straw was subjected to mild alkaline pretreatment followed by enzymatic hydrolysis. Comparison of pretreatment by 2% sodium hydroxide, 2% calcium hydroxide and 20% aqueous ammonia, respectively, at different temperatures showed maximum weight loss with NaOH at 80 °C for 5 h, but ammonia for 15 h at 80 °C led to highest lignin removal of 63%. The ammonia-pretreated rice straw also led to highest release of total reducing sugar up to 92% on hydrolysis by a cocktail of cellulases and hemicellulases at 50 °C. Cultivation of the Bacillus isolates on the pretreated rice straw revealed highest PHA content of 59.3 and 46.4%, and PHA concentration of 2.96 and 2.51 g/L by Bacillus cereus VK92 and VK98, respectively.