Sugarcane-Based Biofuels and Bioproducts

Sugarcane has garnered much interest for its potential as a viable renewable energy crop. While the use of sugar juice for ethanol production has been in practice for years, a new focus on using the fibrous co-product known as bagasse for producing renewable fuels and bio-based chemicals is growing in interest. The success of these efforts, and the development of new varieties of energy canes, could greatly increase the use of sugarcane and sugarcane biomass for fuels while enhancing industry sustainability and competitiveness. Sugarcane-Based Biofuels and Bioproducts examines the development of a suite of established and developing biofuels and other renewable products derived from sugarcane and sugarcane-based co-products, such as bagasse. Chapters provide broad-ranging coverage of sugarcane biology, biotechnological advances, and breakthroughs in production and processing techniques. This text brings together essential information regarding the development and utilization of new fuels and bioproducts derived from sugarcane. Authored by experts in the field, Sugarcane-Based Biofuels and Bioproducts is an invaluable resource for researchers studying biofuels, sugarcane, and plant biotechnology as well as sugar and biofuels industry personnel.

Effect of pretreatment on saccharification of sugarcane bagasse by complex and simple enzyme mixtures

Saccharification of sugarcane bagasse pretreated at the pilot-scale with different processes (in combination with steam-explosion) was evaluated. Maximum glucan conversion with Celluclast 1.5 L (15–25 FPU/g glucan) was in the following order: glycerol/HCl > HCl > H2SO4 > NaOH, with the glycerol system achieving ∼100% conversion. Surprisingly, the NaOH substrate achieved optimum saccharification with only 8 FPU/g glucan. Glucan conversions (3.6–6%) obtained with mixtures of endo-1,4-β-glucanase (EG) and β-glucosidase (βG) for the NaOH substrate were 2–6 times that of acid substrates. However, glucan conversions (15–60%) obtained with mixtures of cellobiohydrolase (CBH I) and βG on acidified glycerol substrate were 10–30% higher than those obtained for NaOH and acid substrates. The susceptibility of the substrates to enzymatic saccharification was explained by their physical and chemical attributes. Acidified glycerol pretreatment offers the opportunity to simplify the complexity of enzyme mixtures required for saccharification of lignocellulosics.

Sweet sorghum : opportunities for a new, renewable fuel and food industry in Australia

Sweet sorghum is receiving significant global interest as an agro-industrial crop because of its capacity to co-produce energy, food, and feed products in integrated biorefineries. This report assesses the opportunities to develop a sweet sorghum industry in Australia, reports on research demonstrating the production of energy, food, and feed products, and assesses the potential economic and sustainability benefits of sweet sorghum biorefineries in the Australian context.

Biorefineries – creating economic opportunities from biofuels and bio-products

Biorefineries, producing fuels, green chemicals and bio-products, offer great potential for improving the profitability and sustainability of tropical agricultural industries. Biomass from tropical crops like sugarcane, sweet sorghum, palm and cassava offer great potential because of the high biomass growth potential under favourable climatic conditions. Biorefineries aim to convert waste residues through biochemical and enzymatic processes to low cost fermentable sugars which are a platform for value-adding. Through subsequent fermentation utilising microbial biotechnologies or chemical synthesis, the sugars can be converted to fuels including ethanol and butanol, oils, organic acids such as lactic and levulinic acid and polymer precursors. Other biorefinery products can include food and animal feeds, plastics, fibre products and resins. Pretreatment technologies are a key to unlocking this potential and new technologies are emerging. This paper will address the opportunities available for tropical biorefineries to contribute to the future profitability of tropical agricultural industries. The importance of pretreatment technologies will be discussed.

Assembly of Human Papillomavirus Type-16 Virus-Like Particles: Multifactorial Study of Assembly and Competing Aggregation

Pentameric capsomeres of human papillomavirus capsid protein L1 expressed in Escherichia coli self-assemble into virus-like particles (VLPs) in vitro. A multifactorial experimental design was used to explore a wide range of solution conditions to optimize the assembly process. The degree of assembly was measured using an enzyme-linked immunosorbent assay, and a high-throughput turbidity assay was developed to monitor competing aggregation. The presence of zinc ions in the assembly buffer greatly increased the incidence of aggregation and had to be excluded from the experiment for meaningful analysis. Assembly of VLPs was optimal at a pH of about 6.5, calcium and sodium ions had no measurable effect, and dithiothreitol and glutathione inhibited assembly. Tryptophan fluorescence spectroscopy demonstrated that an increase in urea concentration reduced the rate of VLP formation but had no effect on the final concentration of assembled VLPs. This study demonstrates the use of the hanging-drop vapor-diffusion crystallization method to screen for conditions that promote aggregation and the use of tryptophan fluorescence spectroscopy for real-time monitoring of the assembly process.

Green cane impact on sugar processing : ISSCT process workshop 2008

The ISSCT Process Section workshop held in Réunion 20–23 October 2008 was attended by 51 delegates from 10 countries. The theme was Green cane impact on sugar processing. The workshop provided a valuable and timely opportunity to review and discuss the impact on factory operations and performance from a green cane supply that could include significant levels of trash. It was particularly relevant to those mills that were considering options to boost their biomass intake for increased co-generation capacity. Several of the speakers related their experiences with processing ‘whole of crop’ cane supplies through the factory. Speakers detailed the problems and increased losses that were incurred when processing cane with high trash levels. The consensus of the delegates was that the best scenario would involve a cane-cleaning plant at the factory so that only clean cane would be processed through the factory. The forum recommended that more research was required to address the issues of increased impurities in the process streams associated with high trash levels. Site visits to the two factories and a cane-delivery station were arranged as part of the workshop.

Clarification of cane juice for fermentation

Fermentation feedstocks in the sugar industry are based on cane juice, B molasses or final molasses. Brazil has been producing ethanol by directing sugarcane juice to fermentation directly or using lower quality juice as a diluent with B molasses to prepare the fermentation broth. One issue that has received only limited interest particularly from outside Brazil is the most appropriate conditions for clarification of the juice going to fermentation. Irrespective of whether the juice supply is the total flow from the milling tandem or a diffuser station or a part of the total flow, removal of the insoluble solids is essential. However, the standard defecation process used by sugar factories around the world to clarify juice can introduce unwanted calcium ions and remove other nutrients such as phosphorus and nitrogen that are considered essential for the fermentation process. An investigation was undertaken by SRI to assess the effects on the constituents of cane juice when subjected to the typical clarification process in an Australian factory and what conditions would be needed to provide a clarified juice suitable for fermentation. Typical juices from one factory were clarified in laboratory trials under a range of pH conditions and the resulting clarified juices analysed. The results indicated that pH had a major effect on the residual concentrations of key constituents in the clarified juice and that the selected clarification conditions are determined by the nominated quality criteria of clarified juice feedstock for fermentation. Further trials were conducted in overseas factories to confirm the results obtained in Australia. It became apparent that the preferred specifications for clarified juice going to fermentation varied from country to country. Each supplier of fermentation technology had criteria applying to clarified juice feedstock that would have a major impact on the standard of clarification required to achieve compliance with the criteria.

Understanding mild acid pretreatment of sugarcane bagasse through particle scale modeling

Sugarcane bagasse is an abundant and sustainable resource, generated as a by-product of sugarcane milling. The cellulosic material within bagasse can be broken down into glucose molecules and fermented to produce ethanol, making it a promising feedstock for biofuel production. Mild acid pretreatment hydrolyses the hemicellulosic component of biomass, thus allowing enzymes greater access to the cellulosic substrate during saccharification. A particle-scale mathematical model describing the mild acid pretreatment of sugarcane bagasse has been developed, using a volume averaged framework. Discrete population-balance equations are used to characterise the polymer degradation kinetics, and diffusive effects account for mass transport within the cell wall of the bagasse. As the fibrous material hydrolyses over time, variations in the porosity of the cell wall and the downstream effects on the reaction kinetics are accounted for using conservation of volume arguments. Non-dimensionalization of the model equations reduces the number of parameters in the system to a set of four dimensionless ratios that compare the timescales of different reaction and diffusion events. Theoretical yield curves are compared to macroscopic experimental observations from the literature and inferences are made as to constraints on these “unknown” parameters. These results enable connections to be made between experimental data and the underlying thermodynamics of acid pretreatment. Consequently, the results suggest that data-fitting techniques used to obtain kinetic parameters should be carefully applied, with prudent consideration given to the chemical and physiological processes being modeled.

Modified alumina nanofiber membranes for protein separation

Large-scale purification/separation of bio-substances is a key technology required for rapid production of biological substances in bioengineering. Membrane filtration is a new separation process and has potential to be used for concentration (removal of solvent), desalting (removal of low molecular weight compounds), clarification (removal of particles), and fractionation (protein-protein separation). In this study, we developed an efficient membrane for protein separation based on ceramic nanofibers. Alumina nanofibers were prepared on a porous support and formed large flow passages. The radical changes in membrane structure provided new ceramic membranes with a large porosity (more than 70%) due to the replacement of bulk particles with fine fibers as building components. The pore size had an average of 11 nm and pure water flux was approximately 360 L•h-1•m-2•bar-1. Further surface modification with a self-assembled monolayer of (3-aminopropyl) triethoxysilane enhanced the membrane filtration properties. Characterization with SEM, FTIR, contact angle, and proteins separation tests indicated that the fibril layers uniformly spread on the surface of the porous support. Moreover, the membrane surface was changed from hydrophilic to hydrophobic after silane groups were grafted. It demonstrated that the silane-grafted alumina fiber membrane can reject 100% BSA protein and 92% cellulase protein. It was also able to retain 75% trypsin protein while maintaining a permeation flux of 48 L•h-1•m-2•bar-1.

Revenue diversification opportunities from sugarcane

Sugarcane is a major global agricultural crop that produces significant quantities of sugar and biomass in tropical and sub-tropical regions. Over many centuries, the crop has been grown primarily for its high sugar content which traditionally contributes over 95% of the revenue derived from the crop. While the production of renewable electricity from bagasse and rum from molasses has a long history, in more recent decades significant advances have been made in the production of cogeneration products and fuel ethanol at large scale. Sugarcane biorefineries producing fuels, green chemicals, biopolymers and bio-products offer great potential for improving the profitability of sugarcane production. This paper will address the opportunities available for sugarcane biorefineries to contribute to future profitability and sustainability of the sugarcane industry.

The economic value of new tropical biorefinery industries in Australia

Biorefineries, co-producing fuels, green chemicals and bio-products, offer great potential for enhancing agricultural value, and developing new industries in the bioeconomy. Biomass biorefineries aim to convert agricultural crops and wastes through biochemical and enzymatic processes to low cost fermentable sugars and other products which are platforms for value-adding. Through subsequent fermentation or chemical synthesis, the bio-based platforms can be converted to fuels including ethanol and butanol, oils, organic acids such as lactic and levulinic acid and polymer precursors. Other biorefinery products can include food and animal feeds, plastics, fibre products and resins. In 2014, QUT commissioned a study from Deloitte Access Economics and Correlli Consulting to assess the potential future economic value of tropical biorefineries to Queensland. This paper will report on the outcomes of this study and address the opportunities available for tropical biorefineries to contribute to the future profitability and sustainability of tropical agricultural industries in Queensland and more broadly across northern Australia.

Accumulation of recombinant cellobiohydrolase and endoglucanase in the leaves of mature transgenic sugar cane

A major strategic goal in making ethanol from lignocellulosic biomass a cost-competitive liquid transport fuel is to reduce the cost of production of cellulolytic enzymes that hydrolyse lignocellulosic substrates to fermentable sugars. Current production systems for these enzymes, namely microbes, are not economic. One way to substantially reduce production costs is to express cellulolytic enzymes in plants at levels that are high enough to hydrolyse lignocellulosic biomass. Sugar cane fibre (bagasse) is the most promising lignocellulosic feedstock for conversion to ethanol in the tropics and subtropics. Cellulolytic enzyme production in sugar cane will have a substantial impact on the economics of lignocellulosic ethanol production from bagasse. We therefore generated transgenic sugar cane accumulating three cellulolytic enzymes, fungal cellobiohydrolase I (CBH I), CBH II and bacterial endoglucanase (EG), in leaves using the maize PepC promoter as an alternative to maize Ubi1 for controlling transgene expression. Different subcellular targeting signals were shown to have a substantial impact on the accumulation of these enzymes; the CBHs and EG accumulated to higher levels when fused to a vacuolar-sorting determinant than to an endoplasmic reticulum-retention signal, while EG was produced in the largest amounts when fused to a chloroplast-targeting signal. These results are the first demonstration of the expression and accumulation of recombinant CBH I, CBH II and EG in sugar cane and represent a significant first step towards the optimization of cellulolytic enzyme expression in sugar cane for the economic production of lignocellulosic ethanol.