A mathematical model to design a lignocellulosic biofuel supply chain system with a case study based on a region in Central Texas

This study formulates a model to maximize the profit of a lignocellulosic biofuel supply chain ranging from feedstock suppliers to biofuel customers. The model deals with a time-staged, multi-commodity, production/distribution system, prescribing facility locations and capacities, technologies, and material flows. A case study based on a region in Central Texas demonstrates application of the proposed model to design the most profitable biofuel supply chain under each of several scenarios. A sensitivity analysis identifies that ethanol (ETOH) price is the most significant factor in the economic viability of a lignocellulosic biofuel supply chain.     

Label free quantitative proteomic analysis of secretome by Thermobifida fusca on different lignocellulosic biomass

Solid state fermentation of lignocellulosic biomass by filamentous microorganisms to induced enzyme production has been recognized as an attractive and cost effective technology. The secretion profile of lignocellulolytic enzymes by thermostable filamentous Thermobifida fusca (T. fusca) in solid state fermentation of different lignocellulosic biomasses, such as corn stover, hay; saw dust; sugarcane bagasse; wood chips; and un-dried green plant were explored using label-free exponentially modified protein abundance index (emPAI) based quantitative proteomics. Comparative analyses of T. fusca secretion profiles between cellulose and the various lignocellulosic biomasses showed induced expression of cellulolytic proteins by cellulose, and expression of hemicellulose, pectin and lignin degrading enzymes were induced by lignocellulosic biomasses. The solid state fermentation by T. fusca on lignocellulosic biomasses also revealed increased expressions of various transport proteins and hypothetical proteins. The Bray-Curtis similarity indices, clustering, and multidimensional scaling plot explicated differential protein expressions by T. fusca on different lignocellulosic biomasses, indicating that protein secretion by T. fusca is reliant on substrate complexity.     

Ethanol from lignocellulosic materials by a simultaneous saccharification and fermentation process (SFS) with Kluyveromyces marxianus CECT 10875

Simultaneous saccharification and fermentation (SSF) process for ethanol production from various lignocellulosic woody (poplar and eucalyptus) and herbaceous (Sorghum sp. bagasse, wheat straw and Brassica carinata residue) materials has been assayed using the thermotolerant yeast strain Kluyveromyces marxianus CECT 10875. Biomass samples were previously treated in a steam explosion pilot plant to provide pretreated biomass with increased cellulose content relative to untreated materials and to enhance cellulase accessibility. SSF experiments were performed in laboratory conditions at 42 °C, 10% (w/v) substrate concentration and 15 FPU/g substrate of commercial cellulase. The results indicate that it is possible to reach SSF yields in the range of 50–72% of the maximum theoretical SSF yield, based on the glucose available in pretreated materials, in 72–82 h. Maximum ethanol contents from 16 to 19 g/l were obtained in fermentation media, depending on the material tested.     

Supercritical carbon dioxide pretreatment of corn stover and switchgrass for lignocellulosic ethanol production

Supercritical CO₂ (SC-CO₂), a green solvent suitable for a mobile lignocellulosic biomass processor, was used to pretreat corn stover and switchgrass at various temperatures and pressures. The CO₂ pressure was released as quickly as possible by opening a quick release valve during the pretreatment. The biomass was hydrolyzed after pretreatment using cellulase combined with β-glucosidase. The hydrolysate was analyzed for the amount of glucose released. Glucose yields from corn stover samples pretreated with SC-CO₂ were higher than the untreated sample’s 12% glucose yield (12 g/100 g dry biomass) and the highest glucose yield of 30% was achieved with SC-CO₂ pretreatment at 3500 psi and 150 °C for 60 min. The pretreatment method showed very limited improvement (14% vs. 12%) in glucose yield for switchgrass. X-ray diffraction results indicated no change in crystallinity of the SC-CO₂ treated corn stover when compared to the untreated, while SEM images showed an increase in surface area.     

2,3-Butanediol production using Klebsiella oxytoca ATCC 8724: Evaluation of biomass derived sugars and fed-batch fermentation process

For this study, 2,3-butanediol (BD) fermentation from pure and biomass-derived sugar were optimized in shake-flask and 5-L bioreactor levels using Klebsiella oxytoca ATCC 8724. The results showed that 70 g/L of single sugar (glucose or xylose) and 90 g/L of mixed-sugar (glucose:xylose = 2:1) were optimum concentrations for efficient 2,3-BD fermentation. At optimum sugar concentrations, 2,3-BD productivities were 1.03, 0.64 and 0.50 gL⁻¹ h⁻¹, and yields were 0.43, 0.36 and 0.35 g/g in glucose, xylose and mixed-sugar medium, respectively. The lack of simultaneous utilization of glucose and xylose led to the lowest productivity in the mixed-sugar medium. Detoxification of biomass hydrolyzates was necessary for efficient 2,3-BD fermentation when sugar concentrations in the medium was 90 g/L or higher, but not with sugar concentrations of 30 g/L or less. A fed-batch fermentation using glucose medium led to an increase 2,3-BD titer to 79.4 g/L and yields 0.47 g/g, while productivity decreased to 0.79 gL⁻¹ h⁻¹. However, the fed-batch process was inefficient using mixed-sugar and biomass hydrolyzates because of poor xylose utilization. These results indicated that appropriate biomass processing technologies must be developed to generate separate glucose and xylose streams to produce high 2,3-BD titer from biomass-derived sugar using a fed-batch process.     

The proteomes of feedstocks used for the production of second‐generation ethanol: a lacuna in the biofuel era

Maize, sorghum, sugarcane, switchgrass and miscanthus are the main crops suggested as potential sources of lignocellulosic biomass for the production of second‐generation ethanol. The attention these crops have received has been concentrated in the field of genomics, and very little research has been performed in the field of proteomics, particularly in the cell wall proteomic, despite the importance of these crops in biofuel production. New mass spectrometry‐based proteomic methods allow the identification and quantification of thousands of proteins in complex mixtures, as well as the detection of post‐translational changes in complex proteomes, providing important insight into the downstream consequences of gene expression. Together with other ‘omic’ approaches, proteomic might be decisive to bring new information in the study of cell wall formation. Here, we briefly highlight proteomic techniques and review the research that has been completed on the proteomes of these five crops.     

木质纤维素生物转化产氢技术现状与发展趋势

氢能是一种清洁能源,利用木质纤维素类生物质生产氢气,在生产可再生绿色能源的同时,避免了木质纤维素资源未被充分利用而造成的环境污染和资源浪费,它的开发与应用对人类未来能源与经济发展具有十分重要意义。以下综述了木质纤维素生物转化产氢技术的研究现状,提出了木质纤维素生物转化产氢的总体构想与对产业发展方向的建议。     

Strategies for the production of cell wall‐deconstructing enzymes in lignocellulosic biomass and their utilization for biofuel production

Microbial cell wall‐deconstructing enzymes are widely used in the food, wine, pulp and paper, textile, and detergent industries and will be heavily utilized by cellulosic biorefineries in the production of fuels and chemicals. Due to their ability to use freely available solar energy, genetically engineered bioenergy crops provide an attractive alternative to microbial bioreactors for the production of cell wall‐deconstructing enzymes. This review article summarizes the efforts made within the last decade on the production of cell wall‐deconstructing enzymes in planta for use in the deconstruction of lignocellulosic biomass. A number of strategies have been employed to increase enzyme yields and limit negative impacts on plant growth and development including targeting heterologous enzymes into specific subcellular compartments using signal peptides, using tissue‐specific or inducible promoters to limit the expression of enzymes to certain portions of the plant or certain times, and fusion of amplification sequences upstream of the coding region to enhance expression. We also summarize methods that have been used to access and maintain activity of plant‐generated enzymes when used in conjunction with thermochemical pretreatments for the production of lignocellulosic biofuels.     

Characteristics of hydrogen and methane production from cornstalks by an augmented two- or three-stage anaerobic fermentation process

This paper presents the co-production of hydrogen and methane from cornstalks by a two- or three-stage anaerobic fermentation process augmented with effective artificial microbial community. Two-stage fermentation by using the anaerobic sludge and DGGE analysis showed that effective and stable strains should be introduced into the system. We introduced Enterobacter aerogens or Clostridium paraputrificum into the hydrogen stage, and C. paraputrificum was proven to be more effective. In the three-stage process consisting of the improved hydrolysis, hydrogen and methane production stages, the highest soluble sugars (0.482 kg/kg cornstalks) were obtained after the introduction of Clostridium thermocellum in the hydrolysis stage, under the thermophilic (55 °C) and acidic (pH 5.0) conditions. Hydrolysates from 1 kg of cornstalks could produce 2.61 mol (63.7 l) hydrogen by augmentation with C. paraputrificum and 4.69 mol (114.6 l) methane by anaerobic granular sludge, corresponding to 54.1% energy recovery.     

Single cell oil production from low-cost substrates: The possibility and potential of its industrialization

Currently, single cell oils (SCO) attract much attention because of their bi-function as a supplier of functional oils and feedstock for biodiesel production. However, high fermentation costs prevent their further application, and the possibility and potential of their industrialization is suspected. Therefore, various low-cost, hydrophilic and hydrophobic substrates were utilized for SCO production. Of these substrates, lignocellulosic biomass, which is the most available and renewable source in nature, might be an ideal raw material for SCO production. Although many reviews on SCO have been published, few have focused on SCO production from low-cost substrates or evaluated the possibility and potential of its industrialization. Therefore, this review mainly presents information on SCO and its production using low-cost substrates and mostly focuses on lignocellulosic biomass. Finally, the possibility and potential of SCO industrialization is evaluated.     

Lignocellulolytic characterization and comparative secretome analysis of a Trichoderma erinaceum strain isolated from decaying sugarcane straw

The fungus Trichoderma reesei is employed in the production of most enzyme cocktails used by the lignocellulosic biofuels industry today. Despite significant improvements, the cost of the required enzyme preparations remains high, representing a major obstacle for the industrial production of these alternative fuels. In this study, a new Trichoderma erinaceum strain was isolated from decaying sugarcane straw. The enzyme cocktail secreted by the new isolate during growth in pretreated sugarcane straw-containing medium presented higher specific activities of β-glucosidase, endoxylanase, β-xylosidase and α-galactosidase than the cocktail of a wild T. reesei strain and yielded more glucose in the hydrolysis of pretreated sugarcane straw. A proteomic analysis of the two strains' secretomes identified a total of 86 proteins, of which 48 were exclusive to T. erinaceum, 35 were exclusive to T. reesei and only 3 were common to both strains. The secretome of T. erinaceum also displayed a higher number of carbohydrate-active enzymes than that of T. reesei (37 and 27 enzymes, respectively). Altogether, these results reveal the significant potential of the T. erinaceum species for the production of lignocellulases, both as a possible source of enzymes for the supplementation of industrial cocktails and as a candidate chassis for enzyme production.     

Assessment of a dry and a wet route for the production of biofuels from microalgae: energy balance analysis

In this study, the energy balance of two microalgae-to-biofuel concepts, one via a so called “dry route” (oil extraction from dried algae) and one via a “wet route” (oil extraction in the water phase), are assessed. Both routes are intended to convert the chemical energy contained in the microalgae into high-value biofuels with minimal fossil energy consumption. The analysis shows that the drying process in the dry route and the oil extraction process in the wet route consume a significant amount of energy. By coupling waste heat from a nearby power plant to the process, the energy balance can be improved and a potential fossil energy ratio (FER) up to 2.38 and 1.82 can be reached for the dry and wet route, respectively. The results indicate that based on current available technologies, the dry route has higher FER and the wet route has more potential in producing high valuable biofuels.     

Biofuel production from palm oil with supercritical alcohols: effects of the alcohol to oil molar ratios on the biofuel chemical composition and properties

Biofuel production from palm oil with supercritical methanol (SCM) and supercritical ethanol (SCE) at 400 °C and 15 MPa were evaluated. At the optimal alcohol to oil molar ratios of 12:1 and 18:1 for the SCM and SCE processes, respectively, the biofuel samples were synthesized in a 1.2-L reactor and the resulting biofuel was analyzed for the key properties including those for the diesel and biodiesel standard specifications. Biofuel samples derived from both the SCM and SCE processes could be used as an alternative fuel after slight improvement in their acid value and free glycerol content. The remarkable advantages of this novel process were: the additional fuel yield of approximately of 5% and 10% for SCM and SCE, respectively; the lower energy consumption for alcohol preheating, pumping and recovering than the biodiesel production with supercritical alcohols that use a high alcohol to oil molar ratio of 42:1.     

Syngas fermentation of Clostridium carboxidivoran P7 in a hollow fiber membrane biofilm reactor: Evaluating the mass transfer coefficient and ethanol production performance

Gasification followed by syngas fermentation is a unique hybrid process for converting lignocellulosic biomass into fuels and chemicals. Current syngas fermentation faces several challenges with low gas–liquid mass transfer being one of the major bottlenecks. The aim of this work is to evaluate the performance of hollow fiber membrane biofilm reactor (HFM-BR) as a reactor configuration for syngas fermentation. The volumetric mass transfer coefficient (K_La) of the HFM-BR was determined at abiotic conditions within a wide range of gas velocity/flowrate passing through the hollow fiber lumen and liquid velocity/flowrate passing through the membrane module shell. The K_La values of the HFM-BR were higher than most reactor configurations such as stir tank reactors and bubble columns. A continuous syngas fermentation of Clostridium carboxidivorans P7 was implemented in the HFM-BR system at different operational conditions, including the syngas flow rate, liquid recirculation between the module and reservoir, and the dilution rate. It was found that the syngas fermentation performance such as syngas utilization efficiency, ethanol concentration and productivity, and ratio of ethanol to acetic acid depended not only on the mass transfer efficiency but also the characteristics of biofilm attached on the membrane module (biofouling or abrading of the biofilm). The HFM-BR results in a highest ethanol concentration of 23.93 g/L with an ethanol to acetic acid ratio of 4.79. Collectively, the research shows the HFM-BR is an efficient reactor system for syngas fermentation with high mass transfer.     

生物航煤生产技术的发展现状

由于温室气体的大量排放和对化石燃料的高度依赖,航空业的可持续发展得到了全世界的关注。生物航煤被认为是一种有前景的传统航空燃料替代品。本文概述了制备生物航煤的代表性工艺技术路线、发展现状以及生物航煤产业发展所面临的机遇和挑战。迄今为止,已经有多种生物航煤制备工艺得到美国材料实验协会(American Society for Testing and Materials, ASTM)认证。其中,酯和脂肪酸加氢是目前最为成熟、可以实现完全商业化的路径。考虑到技术经济性和成熟度,短期内,费托合成是比较有发展前景的工艺。     

Compositional analysis of lignocellulosic biomass: conventional methodologies and future outlook

The composition and structural properties of lignocellulosic biomass have significant effects on its downstream conversion to fuels, biomaterials, and building-block chemicals. Specifically, the recalcitrance to modification and compositional variability of lignocellulose make it challenging to optimize and control the conditions under which the conversion takes place. Various characterization protocols have been developed over the past 150?years to elucidate the structural properties and compositional patterns that affect the processing of lignocellulose. Early characterization techniques were developed to estimate the relative digestibility and nutritional value of plant material after ingestion by ruminants and humans alike (e.g. dietary fiber). Over the years, these empirical techniques have evolved into statistical approaches that give a broader and more informative analysis of lignocellulose for conversion processes, to the point where an entire compositional and structural analysis of lignocellulosic biomass can be completed in minutes, rather than weeks. The use of modern spectroscopy and chemometric techniques has shown promise as a rapid and cost effective alternative to traditional empirical techniques. This review serves as an overview of the compositional analysis techniques that have been developed for lignocellulosic biomass in an effort to highlight the motivation and migration towards rapid, accurate, and cost-effective data-driven chemometric methods. These rapid analysis techniques can potentially be used to optimize future biorefinery unit operations, where large quantities of lignocellulose are continually processed into products of high value.     

Robust factor selection in early cell culture process development for the production of a biosimilar monoclonal antibody

This work presents a multivariate methodology combining principal component analysis, the Mahalanobis distance and decision trees for the selection of process factors and their levels in early process development of generic molecules. It is applied to a high throughput study testing more than 200 conditions for the production of a biosimilar monoclonal antibody at microliter scale. The methodology provides the most important selection criteria for the process design in order to improve product quality towards the quality attributes of the originator molecule. Robustness of the selections is ensured by cross‐validation of each analysis step. The concluded selections are then successfully validated with an external data set. Finally, the results are compared to those obtained with a widely used software revealing similarities and clear advantages of the presented methodology. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:181–191, 2017     

Straw utilization for biofuel production: A consequential assessment of greenhouse gas emissions from bioethanol and biomethane provision with a focus on the time dependency of emissions

The shift from straw incorporation to biofuel production entails emissions from production, changes in soil organic carbon (SOC) and through the provision of (co‐)products and entailed displacement effects. This paper analyses changes in greenhouse gas (GHG) emissions arising from the shift from straw incorporation to biomethane and bioethanol production. The biomethane concept comprises comminution, anaerobic digestion and amine washing. It additionally provides an organic fertilizer. Bioethanol production comprises energetic use of lignin, steam explosion, enzymatic hydrolysis and co‐fermentation. Additionally, feed is provided. A detailed consequential GHG balance with in‐depth focus on the time dependency of emissions is conducted: (a) the change in the atmospheric load of emissions arising from the change in the temporal occurrence of emissions comparing two steady states (before the shift and once a new steady state has established); and (b) the annual change in overall emissions over time starting from the shift are assessed. The shift from straw incorporation to biomethane production results in net changes in GHG emissions of (a) ?979 (?436 to ?1,654) and (b) ?955 (?220 to ?1,623) kg CO₂‐eq. per t_{dry matter} straw converted to biomethane (minimum and maximum). The shift to bioethanol production results in net changes of (a) ?409 (?107 to ?610) and (b) ?361 (57 to ?603) kg CO₂‐eq. per t_{dry matter} straw converted to bioethanol. If the atmospheric load of emissions arising from different timing of emissions is neglected in case (a), the change in GHG emissions differs by up to 54%. Case (b) reveals carbon payback times of 0 (0–49) and 19 (1–100) years in case of biomethane and bioethanol production, respectively. These results demonstrate that the detailed inclusion of temporal aspects into GHG balances is required to get a comprehensive understanding of changes in GHG emissions induced by the introduction of advanced biofuels from agricultural residues.     

Observability analysis of biochemical process models as a valuable tool for the development of mechanistic soft sensors

By enabling the estimation of difficult‐to‐measure target variables using available indirect measurements, mechanistic soft sensors have become important tools for various bioprocess monitoring and control scenarios. Despite promising higher process efficiencies and increased process understanding, widespread application of soft sensors has been stalled by uncertainty about the feasibility and reliability of their estimations given present process analytical constraints. Observability analysis can provide an indication of the possibility and reliability of soft sensor estimations by analyzing the structural properties of first‐principle (mechanistic) models. In addition, it can provide a criteria for selection of suitable measurement methods with respect to their information content; thereby leading to successful implementation of soft sensors in bioprocess development and manufacturing environments. We demonstrate the utility of observability analysis for two classes of upstream bioprocesses: the processes involving growth and ethanol formation by Saccharomyces cerevisiae and the process of penicillin production by Penicillium chrysogenum. Results obtained from laboratory‐scale cultivations in addition to in‐silico experiments enable a comparison of theoretical aspects of observability analysis and the real‐life performance of soft sensors. By taking the expected error of measurements provided to the soft sensor into account, an innovative scaling approach facilitates a higher degree of comparability of observability results among various measurement configurations and process conditions. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1703–1715, 2015     

Sources of variability in greenhouse gas and energy balances for biofuel production: a systematic review

Across the energy sector, alternatives to fossil fuels are being developed, in response to the dual drivers of climate change and energy security. For transport, biofuels have the greatest potential to replace fossil fuels in the short‐to medium term. However, the ecological benefits of biofuels and the role that their deployment can play in mitigating climate change are being called into question. Life Cycle Assessment (LCA) is a widely used approach that enables the energy and greenhouse gas (GHG) balance of biofuel production to be calculated. Concerns have nevertheless been raised that published data show widely varying and sometimes contradictory results. This review describes a systematic review of GHG emissions and energy balance data from 44 LCA studies of first‐ and second‐generation biofuels. The information collated was used to identify the dominant sources of GHG emissions and energy requirements in biofuel production and the key sources of variability in published LCA data. Our analysis revealed three distinct sources of variation: (1) ‘real’ variability in parameters e.g. cultivation; (2) ‘methodological’ variability due to the implementation of the LCA method; and (3) ‘uncertainty’ due to parameters rarely included and poorly quantified. There is global interest in developing a sustainability assessment protocol for biofuels. Confidence in the results of such an assessment can only be assured if these areas of uncertainty and variability are addressed. A more defined methodology is necessary in order to allow effective and accurate comparison of results. It is also essential that areas of uncertainty such as impacts on soil carbon stocks and fluxes are included in LCA assessments, and that further research is conducted to enable a robust calculation of impacts under different land‐use change scenarios. Without the inclusion of these parameters, we cannot be certain that biofuels are really delivering GHG savings compared with fossil fuels.