Applied Biochemistry and Biotechnology (v.154, #1-3)

Introduction to Session 4: New Biofuels and Biomass Chemicals by Hans Blaschek; Akwasi Boateng (1-2).

Characterization of Fast Pyrolysis Bio-oils Produced from Pretreated Pine Wood by El-barbary M. Hassan; Philip H. Steele; Leonard Ingram (3-13).
The pretreatment of biomass prior to the fast pyrolysis process has been shown to alter the structure and chemical composition of biomass feed stocks leading to a change in the mechanism of biomass thermal decomposition. Pretreatment of feed stocks prior to fast pyrolysis provides an opportunity to produce bio-oils with varied chemical composition and physical properties. This provides the potential to vary bio-oil chemical and physical properties for specific applications. To determine the influence of biomass pretreatments on bio-oil produced during fast pyrolysis, we applied six chemical pretreatments: dilute phosphoric acid, dilute sulfuric acid, sodium hydroxide, calcium hydroxide, ammonium hydroxide, and hydrogen peroxide. Bio-oils were produced from untreated and pretreated 10-year old pine wood feed stocks in an auger reactor at 450 °C. The bio-oils’ physical properties of pH, water content, acid value, density, viscosity, and heating value were measured. Mean molecular weights and polydispersity were determined by gel permeation chromatography. Chemical characteristics of the bio-oils were determined by gas chromatography–mass spectrometry and Fourier transform infrared techniques. Results showed that the physical and chemical characteristics of the bio-oils produced from pretreated pine wood feed stocks were influenced by the biomass pretreatments applied. These physical and chemical changes are compared and discussed in detail in the paper.
Keywords: Pretreatment; Bio-oil; Pine wood; Fast pyrolysis; Chemical characterization

In this paper, the thermodynamic equilibrium models for biomass gasification applicable to various gasifier types have been developed, with and without considering char. The equilibrium models were then modified closely matching the CH4 only or both CH4 and CO compositions from experimental data. It is shown that the modified model presented here based on thermodynamic equilibrium and taking into account local heat and mass considerations can be used to simulate the performance of a downdraft gasifier. The model can also be used to estimate the equilibrium composition of the syngas. Depending on the gasifier type and internal fluid flow, heat and mass transfer characteristics, with proper modification of the equilibrium model, a simple tool to simulate the operation and performance of varying types of biomass gasifier can be developed.
Keywords: Biomass; Gasification; Chemical equilibrium; Modeling; Downdraft gasifier; Syngas

Process Modeling of Comprehensive Integrated Forest Biorefinery—An Integrated Approach by Hua-Jiang Huang; Weilu Lin; Shri Ramaswamy; Ulrike Tschirner (26-37).
The key to expanding the energy supply, increasing energy security, and reducing the dependency on foreign oil is to develop advanced technologies to efficiently transform our renewable bioresources into domestically produced bioenergy and bioproducts. Conventional biorefineries, i.e., forest products industry’s pulp and paper mills with long history of sustainable utilization of lignocellulose (wood), offer a suitable platform for being expanded into future integrated forest biorefineries. Due to the pre-existing infrastructure in current forest products operations, this could present a very cost-effective approach to future biorefineries. In order to better understand the overall process, technical, economic, and environmental impacts, a detailed process modeling of the whole integrated forest biorefinery is presented here. This approach uses a combination of Aspen Plus®, WinGEMS®, and Microsoft Excel® to simulate the entire biorefinery in detail with sophisticated communication interface between the three simulations. Preliminary results for a simple case study of an integrated biorefinery show the feasibility of this approach. Further investigations, including additional details, more process options, and complete integration, are currently underway.
Keywords: Process modeling; Biorefinery modeling; Integrated forest biorefinery; Pulping; Ethanol; Aspen Plus; WinGEMS

Cellulosic Films Obtained from the Treatment of Sugarcane Bagasse Fibers with N-methylmorpholine-N-oxide (NMMO) by Denise S. Ruzene; Daniel P. Silva; António A. Vicente; José. A. Teixeira; Maria T. Pessoa de Amorim; Adilson R. Gonçalves (38-47).
Ethanol/water organosolv pulping was used to obtain sugarcane bagasse pulp that was bleached with sodium chlorite. This bleached pulp was used to obtain cellulosic films that were further evaluated by Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). A good film formation was observed when temperature of 74 °C and baths of distilled water were used, which after FTIR, TGA, and SEM analysis indicated no significant difference between the reaction times. The results showed this to be an interesting and promising process, combining the prerequisites for a more efficient utilization of agro-industrial residues.
Keywords: Agro-industrial residue; Sugarcane bagasse; Cellulose fiber; Cellulose film; NMMO

Five strains of the yeast Phaffia rhodozyma, NRRL Y-17268, NRRL Y-17270, ATCC 96594 (CBS 6938), ATCC 24202 (UCD 67–210), and ATCC 74219 (UBV-AX2) were tested for astaxanthin production using the major sugars derived from corn fiber. The sugars tested included glucose, xylose, and arabinose. All five strains were able to utilize the three sugars for astaxanthin production. Among them, ATCC 74219 was the best astaxanthin producer. Kinetics of sugar utilization of this strain was studied, both with the individual sugars and with their mixtures. Arabinose was found to give the highest astaxanthin yield. It also was observed that glucose at high concentrations suppressed utilization of the other two sugars. Corn fiber hydrolysate obtained by dilute sulfuric acid pretreatment and subsequent enzyme hydrolysis was tested for astaxanthin production by strain ATCC 74219. Dilution of the hydrolysate was necessary to allow growth and astaxanthin production. All the sugars in the hydrolysate diluted with two volumes of water were completely consumed. Astaxanthin yield of 0.82 mg/g total sugars consumed was observed.
Keywords: Phaffia rhodozyma ; Astaxanthin; Fuel ethanol co-products; Lignocellulosic biomass; Corn fiber

Treatment Variable Effects on Supercritical Gasification of High-Diversity Grassland Perennials by Bo Zhang; Zhigang Zhang; Marc von Keitz; Kenneth Valentas (59-66).
Low-input high-diversity (LIHD) mixtures of native grassland perennials were subjected to a supercritical treatment process with the aim of obtaining hydrogen-rich gases. The process was studied based on the following treatment variables: reaction temperature (374 °C to 575 °C, corresponding to a pressure range of 22.1 to 40 MPa), residence time (10 to 30 min), biomass content in the feed, and catalysts (0% to 4% NaOH and solid alkali CaO–ZrO2). The gaseous phase produced from gasification of LIHD primarily consisted of hydrogen (H2), with a mixture of carbon monoxide (CO), methane (CH4), and carbon dioxide (CO2). The statistical significance of treatment variables was evaluated using analysis of variance (ANOVA). It showed that at the level of P < 0.05, temperature, catalysts, and biomass content in the feed significantly affected gas yields, while residence time was not significant.
Keywords: Supercritical gasification; Biodiversity; Hydrogen; Statistical significance

Fast Biodiesel Production with One-Phase Reaction by Ji-Yeon Park; Deog-Keun Kim; Zhong-Ming Wang; Jin-Suk Lee (67-73).
The feasibility of fatty acid methyl ester (FAME) as a co-solvent used to increase the mass transfer between oil and methanol was investigated. FAME, as the co-solvent, does not require additional separation after the reaction because it is the end product of the reaction. To examine intermediate phenomena during the transesterification of soybean oil, the agitation speed was controlled at a slow rate. When the molar ratio of oil to methanol was 1:6 at 0.8wt.% of KOH to oil, oil was at the bottom layer, and methanol and the catalyst were at the top layer. Under the slow agitation process, the contact surface became initially darkened with the production of FAME and glycerol. After a few minutes, the entire top layer became dark. The top layer, containing methanol, KOH, FAME, and glycerol, fell to the bottom layer and then formed the one-phase system. When 0, 5, and 10 wt.% of FAME to oil was initially introduced to the reaction mixture, the FAME content rapidly increased with the FAME concentration level. After forming the one-phase system, the rate of increase of the FAME content was very slow. The time required for the formation of the one-phase system decreased with the amount of FAME and KOH and with temperature.
Keywords: Biodiesel; Transesterification; Co-solvent; Fatty acid methyl ester; One-phase reaction

Biodiesel Synthesis via Esterification of Feedstock with High Content of Free Fatty Acids by Marcella S. Souza; Erika C. G. Aguieiras; Mônica A. P. da Silva; Marta A. P. Langone (74-88).
The objective of this work was to study the synthesis of ethyl esters via esterification of soybean oil deodorizer distillate with ethanol, using solid acid catalysts and commercial immobilized lipases, in a solvent-free system. Three commercially immobilized lipases were used, namely, Lipozyme RM-IM, Lipozyme TL-IM, and Novozym 435, all from Novozymes. We aimed for optimum reaction parameters: temperature, enzyme concentration, initial amount of ethanol, and its feeding technique to the reactor (stepwise ethanolysis). Reaction was faster with Novozym 435. The highest conversion (83.5%) was obtained after 90 min using 3 wt.% of Novozym 435 and two-stage stepwise addition of ethanol at 50°C. Four catalysts were also tested: zeolite CBV-780, SAPO-34, niobia, and niobic acid. The highest conversion (30%) was obtained at 100°C, with 3 wt.% of CBV-780 after 2.5 h. The effects of zeolite CBV 780 concentration were studied, resulting in a conversion of 49% using 9 wt.% of catalyst.
Keywords: Esterification; SODD; Biodiesel; Ethanol; Immobilized lipase; Zeolite

Biomass processing plants have a trade-off between two competing cost factors: as size increases, the economy of scale reduces per unit processing cost, while a longer biomass transportation distance increases the delivered cost of biomass. The competition between these cost factors leads to an optimum size at which the cost of energy produced from biomass is minimized. Four processing options are evaluated: power production via direct combustion and via biomass integrated gasification and combined cycle (BIGCC), ethanol production via fermentation, and syndiesel via Fischer Tropsch. The optimum size is calculated as a function of biomass gross yield (the biomass available to the processing plant from the total surrounding area) and processing cost (capital recovery and operating costs). Higher biomass gross yield and higher processing cost each lead to a higher optimum size. For most cases, a small relaxation in the objective of minimum cost, 3%, leads to a halving of plant size. Direct combustion and BIGCC each produce power, with BIGCC having a higher capital cost and conversion efficiency. When the delivered cost of biomass is high, BIGCC produces power at a lower cost than direct combustion. The crossover point at which this occurs is calculated as a function of the purchase cost of biomass and the biomass gross yield.
Keywords: Biomass availability; Optimum plant size; Biomass processing cost; Economy of scale; BIGCC; Power from biomass; Lignocellulosic ethanol; Fischer Tropsch; Biomass syndiesel

Assessment of Xylanase Activity in Dry Storage as a Potential Method of Reducing Feedstock Cost by William A. Smith; David N. Thompson; Vicki S. Thompson; Corey W. Radtke; Brady Carter (108-122).
Enzymatic preprocessing of lignocellulosic biomass in dry storage systems has the potential to improve feedstock characteristics and lower ethanol production costs. To assess the potential for endoxylanase activity at low water contents, endoxylanase activity was tested using a refined wheat arabinoxylan substrate and three commercial endoxylanases over the water activity range 0.21–1.0, corresponding to water contents of 5% to >60% (dry basis). Homogeneously mixed dry samples were prepared at a fixed enzyme to substrate ratio and incubated in chambers at a variety of fixed water activities. Replicates were sacrificed periodically, and endoxylanase activity was quantified as an increase in reducing sugar relative to desiccant-stored controls. Endoxylanase activity was observed at water activities over 0.91 in all enzyme preparations in less than 4 days and at a water activity of 0.59 in less than 1 week in two preparations. Endoxylanase activity after storage was confirmed for selected desiccant-stored controls by incubation at 100% relative humidity. Water content to water activity relationships were determined for three lignocellulosic substrates, and results indicate that two endoxylanase preparations retained limited activity as low as 7% to 13% water content (dry basis), which is well within the range of water contents representative of dry biomass storage. Future work will examine the effects of endoxylanase activity toward substrates such as corn stover, wheat straw, and switchgrass in low water content environments.
Keywords: Endoxylanase; Water activity; Water content; Biomass; Lignocellulose; Storage; Feedstock; Preprocessing; Stability

Session 6: Advances in Enzyme Science and Technology by Gisella Zanin; Kevin Gray (123-124).