Research Articles

We have developed a yeast Pichia pastoris system for the high-level expression of recombinant Rhizopus oryzae lipase (ROL), which is a potentially effective catalyst in the solvent-free production of biodiesel fuel. In the glycerol fed-batch phase, the combination of the dissolved-oxygen-stat and gradient-control glycerol feeding strategies resulted in a higher cell biomass in the P. pastoris culture, with shorter feed times. In the methanol fed-batch phase, a constant methanol concentration of 0.3-0.5% (v/v) was found to be optimal for high ROL activity, maximum protein concentration, and a maximum biomass. Cells were grown at 30 °C and induced by methanol at 22 °C and a pH of 6.0 with the addition of 0.5% (w/v) casein, which reduced the proteolytic degradation of ROL. Using a combined glycerol step-increasing, on-line methanol feeding strategy and these reducing proteolytic degradation methods, a maximum biomass of 96 g DCW/l, a maximum concentration of 2.0 g/L, and an activity of 1302.2 U/mL ROL were attained. The simple culture process is the highest level of ROL expression reported in P. pastoris. Our results strongly demonstrate that the efficiency of the recombinant ROL expression is directly dependent on feed strategies, lower induction temperature, maintaining the proper pH, and the addition of casein.

Use of alkali-pretreated rice straw and untreated rice straw as substrates for enzyme production under solid-state cultivation was investigated. Cellulase produced from untreated rice straw showed higher activity of FPase, CMCase, β-glucosidase, and xylanase at 6.25 U/g substrate, 111.31 U/g substrate, 173.71 U/g substrate, and 433.75 U/g substrate respectively, as compared to 1.72 U/g substrate, 23.01 U/g substrate, 2.18 U/g substrate, and 45.46 U/g substrate for FPase, CMCase, β-glucosidase, and xylanase, respectively, when alkali-pretreated substrate was used. The results of the X-ray diffractogram analysis showed an increase in relative crystallinity of cellulose in alkali-pretreated rice straw (62.41%) compared to 50.81% in untreated rice straw. However, the crystalline structure of cellulose was partially disrupted after alkali pretreatment, resulting in a decrease in absolute crystallinity of cellulose. The higher the crystallinity of cellulose, the more cellulase production was induced. The structural changes of rice straw before and after alkali pretreatment were compared by using Scanning Electron Microscopy. Fungal mycelial growth was also observed for both untreated and alkali-pretreated substrates. The results of this study indicated that untreated rice straw is a better substrate for cellulase and xylanase production under solid-state fermentation with low environmental impact.

Acetic acid (HOAc) was selected as a bio-oil model compound for the steam reforming of bio-oil for hydrogen production. The influence of temperature and steam-to-carbon ratio (S/C) on the steam reforming of HOAc over hydrogen-type Zeolite Socony Mobil-5 (HZSM-5) and the catalyst with added Pd (Pd/HZSM-5) have been investigated in a fixed-bed reactor. Brunauer–Emmett–Teller surface area measurements, scanning electron microscopy, and transmission electron microscopy were performed to characterize the texture and structure of the catalysts. Upon addition of Pd to HZSM-5, the selectivity of the products was modified and the H2 yield was greatly increased. The hydrogen yield and potential hydrogen yield from the steam reforming of HOAc were as high as 60.2% and 87.5%, respectively, under optimized reaction conditions. Both the conversion of HOAc and the H2 yield over Pd/HZSM-5 were significantly enhanced with increasing S/C ratio and reaction temperature below 600 °C, whereas the H2 yield did not significantly increase at temperatures above 600 °C. The mechanism of HOAc decomposition on the Pd(111) surface was calculated via density functional theory. The optimal decomposition route was found to be CH3COOH* → CH3CO* → CH3* + CO*.

Corn cob (CC) was used as a filler in chitosan (CS) biopolymer films. The effect of glutaraldehyde (GLA) as a crosslinking agent was studied in an effort to improve the properties of CS/CC biocomposite films prepared via solvent casting. The tensile strength and elongation at break values decreased, but the modulus of elasticity increased with CC content. However, the tensile properties of CS/CC biocomposite films improved when modified with GLA. The Fourier transform infrared (FTIR) results indicated the presence of imine bonds (C=N) and ethylenic groups due to the cross-linking reaction between CS and GLA. The thermal stability of CS/CC biocomposite films reduced with increasing CC content. The modification of CS/CC with GLA enhanced the thermal stability of the biocomposite films. Moreover, the wettability and adhesion of the CC-CS system were enhanced by modification with GLA, as demonstrated by a morphological study. The crosslinking agent glutaraldehyde positively affected the tensile strength, modulus of elasticity, and thermal stability of the biocomposite films.

A unique approach was used in which polyoxometalate-based ionic liquids (POM-IL) were synthesized and employed for the one-pot dissolution and conversion of biomass. A library of four functional POM-IL compounds was synthesized using two cationic organic groups (1-butyl-3-methylimidazolim and 1-ethyl-3-methylimidazolium) and two anionic POMs (phosphotungstate and tungstosilicate). The POM-IL compound serves a dual-purpose. First, it dissolves approximately 30 wt% of cellulosic biomass in 2 h at 200 °C. Second, analysis using HPLC confirmed that the POM-ILs catalyze conversion of biomass into commodity monosaccharides such as glucose and xylose. All of the prepared POM-IL compounds demonstrated dynamic thermal stabilities exceeding 300 °C and were characterized using IR and thermogravimetric analysis.

The Raffia textilis fiber has a specific strength of 660 MPaŸcm3/g and scales and hollows on its surface. Thus, this fiber is a potential composite reinforcement. The objective of this study was to evaluate the effect of alkaline treatment at room temperature on its microstructure, structure, composition, thermal behavior, mechanical properties, and color. To this end, slack raw fibers were soaked in three NaOH solutions (2.5%, 5%, and 10% by weight) for 12 hours. SEM observations revealed that fibers got more and more clean and smooth when the solution concentration was increased. In comparison with the raw fiber, it was found that fiber treated with 5% NaOH solution exhibited enhanced tensile strength (129%) and strain to failure (175%), in addition to increased yellowness, redness, and thermal stability. Contrariwise, the Young modulus and lightness slightly decreased with the treatment. The Fourier transform infrared spectra and the XRD patterns suggested an incipient allotropic transformation of cellulose for 10% NaOH-treated fibers. These changes could be explained by the gradual dissolution of non-cellulosic components as revealed by the Fourier transform infrared attenuated total reflection spectra and thermal analysis.

Activated carbons (ACs) prepared from apple pulp and apple peel with phosphoric acid as an activation agent under microwave radiation were investigated. The effects of microwave radiation power and time on the adsorption capacities of the ACs were studied. The optimum AC preparation condition was identified by comparing the MB adsorption capacities of the produced ACs. The obtained results show that the microwave radiation power and time had strong effects on the adsorption capacities. Relative to conventional heating methods, microwave-prepared ACs showed higher BET surface areas and mesopore volumes after a shorter activation time due to differences in the type of heat transfer between these two methods. The N2 adsorption isotherms at −196°C and SEM and FTIR results were used to characterize the properties of the prepared ACs. The N2 adsorption results revealed BET surface areas of 1552 m2/g and 1103 m2/g for apple-peel and apple-pulp-based AC, respectively.

The kraft pulps produced from heartwood and sapwood of Eucalyptus globulus at 130 ºC, 150 ºC, and 170 ºC were characterized by wet chemistry (total lignin as sum of Klason and soluble lignin fractions) and pyrolysis (total lignin denoted as py-lignin). The total lignin content obtained with both methods was similar. In the course of delignification, the py-lignin values were higher (by 2 to 5%) compared to Klason values, which is in line with the importance of soluble lignin for total lignin determination. Pyrolysis analysis presents advantages over wet chemical procedures, and it can be applied to wood and pulps to determine lignin contents at different stages of the delignification process. The py-lignin values were used for kinetic modelling of delignification, with very high predictive value and results similar to those of modelling using wet chemical determinations.

Bio-oil and hydrochar were produced through the hydrothermal liquefaction (HTL) of Salix psammophila (SP) branch residues with recycled processing water, in order to address the lack of water in deserts or sandy lands and the difficulty of water treatment in a batch reactor. The results indicated that the recycling of the HTL processing water could significantly improve the yield of bio-oil from 30.3% to 46.9%. The gas chromatography and mass spectrometry analyses of the obtained bio-oil confirmed the presence of value-added chemicals, such as phenolics, acetic acid, and furans. The acetic acid in the processing water played a key role in the HTL. The heavy oil had a high content (maximum of 42.7 wt%) of the low boiling point fraction (<300 °C), indicating its potential for further applications. The higher heating value of the hydrochar was about 27 MJ/kg, equivalent to the heating value of medium-rank and high-rank coals. These results show that HTL using recycled processing water has great potential for utilization of desert biomass wastes.

Empty fruit bunch oil palm (EFBOP) fibres were surface modified by four different methods, propionylation, vinyltrimethoxy silanization, PPgMA dissolution modification, and PPgMA blending, and integrated into a polypropylene (PP) matrix. The designed biocomposites were subjected to an absorption process at different temperatures. Their water uptake behaviour was compared with the unmodified fibre biocomposites. An increased fibre content and temperature resulted in increased water uptake for all of the biocomposites. The biocomposites containing modified fibres showed a reduction in water uptake, rate of diffusion, sorption, and permeation in comparison with unmodified fibre composites. Comparing the 20 wt% fibre composites at ambient temperature, the performance in water absorption followed the sequence silanization < propionylation < PPgMA dissolution modification < PPgMA blending < no modification. Furthermore, the lowest water absorption was obtained from the silanized fibre/PP composite with 40% fibre content at ambient temperature. Dissolution or blending of PPgMA gave similar water uptake results. The reduction of diffusion, sorption, and permeation confirmed that the modification of fibres was potentially effective at resisting water penetration into the composites.