The combination of plant-expressed cellobiohydrolase and low dosages of cellulases for the hydrolysis of sugar cane bagasse

Background The expression of biomass-degrading enzymes (such as cellobiohydrolases) in transgenic plants has the potential to reduce the costs of biomass saccharification by providing a source of enzymes to supplement commercial cellulase mixtures. Cellobiohydrolases are the main enzymes in commercial cellulase mixtures. In the present study, a cellobiohydrolase was expressed in transgenic corn stover leaf and assessed as an additive for two commercial cellulase mixtures for the saccharification of pretreated sugar cane bagasse obtained by different processes. Results Recombinant cellobiohydrolase in the senescent leaves of transgenic corn was extracted using a simple buffer with no concentration step. The extract significantly enhanced the performance of Celluclast 1.5 L (a commercial cellulase mixture) by up to fourfold on sugar cane bagasse pretreated at the pilot scale using a dilute sulfuric acid steam explosion process compared to the commercial cellulase mixture on its own. Also, the extracts were able to enhance the performance of Cellic CTec2 (a commercial cellulase mixture) up to fourfold on a range of residues from sugar cane bagasse pretreated at the laboratory (using acidified ethylene carbonate/ethylene glycol, 1-butyl-3-methylimidazolium chloride, and ball-milling) and pilot (dilute sodium hydroxide and glycerol/hydrochloric acid steam explosion) scales. We have demonstrated using tap water as a solvent (under conditions that mimic an industrial process) extraction of about 90% recombinant cellobiohydrolase from senescent, transgenic corn stover leaf that had minimal tissue disruption. Conclusions The accumulation of recombinant cellobiohydrolase in senescent, transgenic corn stover leaf is a viable strategy to reduce the saccharification cost associated with the production of fermentable sugars from pretreated biomass. We envisage an industrial-scale process in which transgenic plants provide both fibre and biomass-degrading enzymes for pretreatment and enzymatic hydrolysis, respectively.

SOTrain: A new program for training supervisors and operators in Australian sugar factories

The provision of effective training of supervisors and operators is essential if sugar factories are to operate profitably and in an environmentally sustainable and safe manner. The benefits of having supervisor and operator staff with a high level of operational skills are reduced stoppages, increased recovery, improved sugar quality, reduced damage to equipment, and reduced OH&S and environmental impacts. Training of new operators and supervisors in factories has traditionally relied on on-the-job training of the new or inexperienced staff by experienced supervisors and operators, supplemented by courses conducted by contractors such as Sugar Research Institute (SRI). However there is clearly a need for staff to be able to undertake training at any time, drawing on the content of online courses as required. An improved methodology for the training of factory supervisors and operators has been developed by QUT on behalf of a syndicate of mills. The new methodology provides ‘at factory’ learning via self-paced modules. Importantly, the training resources for each module are designed to support the training programs within sugar factories, thereby establishing a benchmark for training across the sugar industry. The modules include notes, training guides and session plans, guidelines for walkthrough tours of the stations, learning activities, resources such as videos, animations, job aids and competency assessments. The materials are available on the web for registered users in Australian Mills and many activities are best undertaken online. Apart from a few interactive online resources, the materials for each module can also be downloaded. The acronym SOTrain (Supervisor and Operator Training) has been applied to the new training program.

Calcium phosphate flocs and the clarification of sugar cane juice from whole of crop harvesting

Sugar cane biomass is one of the most viable feedstocks for the production of renewable fuels and chemicals. Therefore, processing the whole of crop (WC) (i.e., stalk and trash, instead of stalk only) will increase the amount of available biomass for this purpose. However, effective clarification of juice expressed from WC for raw sugar manufacture is a major challenge because of the amounts and types of non-sucrose impurities (e.g., polysaccharides, inorganics, proteins, etc.) present. Calcium phosphate flocs are important during sugar cane juice clarification because they are responsible for the removal of impurities. Therefore, to gain a better understanding of the role of calcium phosphate flocs during the juice clarification process,the effects of impurities on the physicochemical properties of calcium phosphate flocs were examined using small-angle laser light scattering technique, attenuated total reflectance Fourier transformed infrared spectroscopy, and X-ray powder diffraction. Results on synthetic sugar juice solutions showed that the presence of SiO2 and Na+ ions affected floc size and floc structure. Starch and phosphate ions did not affect the floc structure; however, the former reduced the floc size, whereas the latter increased the floc size. The study revealed that high levels of Na+ ions would negatively affect the clarification process the most, as they would reduce the amount of suspended particles trapped by the flocs. A complementary study on prepared WC juice using cold and cold/intermediate liming techniques was conducted. The study demonstrated that, in comparison to the one-stage (i.e., conventional) clarification process, a two-stage clarification process using cold liming removed more polysaccharides (≤19%),proteins (≤82%), phosphorus (≤53%), and SiO2 (≤23%) in WC juice but increased Ca2+ (≤136%) and sulfur (≤200%)

Soluble mediators in platelet concentrates modulate dendritic cell inflammatory responses in an experimental model of transfusion

The transfusion of platelet concentrates (PCs) is widely used to treat thrombocytopenia and severe trauma. Ex vivo storage of PCs is associated with a storage lesion characterized by partial platelet activation and the release of soluble mediators, such as soluble CD40 ligand (sCD40L), RANTES, and interleukin (IL)-8. An in vitro whole blood culture transfusion model was employed to assess whether mediators present in PC supernatants (PC-SNs) modulated dendritic cell (DC)-specific inflammatory responses (intracellular staining) and the overall inflammatory response (cytometric bead array). Lipopolysaccharide (LPS) was included in parallel cultures to model the impact of PC-SNs on cell responses following toll-like receptor-mediated pathogen recognition. The impact of both the PC dose (10%, 25%) and ex vivo storage period was investigated [day 2 (D2), day 5 (D5), day 7 (D7)]. PC-SNs alone had minimal impact on DC-specific inflammatory responses and the overall inflammatory response. However, in the presence of LPS, exposure to PC-SNs resulted in a significant dose associated suppression of the production of DC IL-12, IL-6, IL-1a, tumor necrosis factor-a (TNF-a), and macrophage inflammatory protein (MIP)-1b and storage-associated suppression of the production of DC IL-10, TNF-a, and IL-8. For the overall inflammatory response, IL-6, TNF-a, MIP-1a, MIP-1b, and inflammatory protein (IP)-10 were significantly suppressed and IL-8, IL-10, and IL-1b significantly increased following exposure to PC-SNs in the presence of LPS. These data suggest that soluble mediators present in PCs significantly suppress DC function and modulate the overall inflammatory response, particularly in the presence of an infectious stimulus. Given the central role of DCs in the initiation and regulation of the immune response, these results suggest that modulation of the DC inflammatory profile is a probable mechanism contributing to transfusion-related complications.

Serum levels of soluble receptor for advanced glycation end products and of S100 proteins are associated with inflammatory, autoantibody, and classical risk markers of joint and vascular damage in rheumatoid arthritis

Introduction: The receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin superfamily of cell surface receptor molecules. High concentrations of three of its putative proinflammatory ligands, S100A8/A9 complex (calprotectin), S100A8, and S100A12, are found in rheumatoid arthritis (RA) serum and synovial fluid. In contrast, soluble RAGE (sRAGE) may prevent proinflammatory effects by acting as a decoy. This study evaluated the serum levels of S100A9, S100A8, S100A12 and sRAGE in RA patients, to determine their relationship to inflammation and joint and vascular damage. Methods: Serum sRAGE, S100A9, S100A8 and S100A12 levels from 138 patients with established RA and 44 healthy controls were measured by ELISA and compared by unpaired t test. In RA patients, associations with disease activity and severity variables were analyzed by simple and multiple linear regressions. Results: Serum S100A9, S100A8 and S100A12 levels were correlated in RA patients. S100A9 levels were associated with body mass index (BMI), and with serum levels of S100A8 and S100A12. S100A8 levels were associated with serum levels of S100A9, presence of anti-citrullinated peptide antibodies (ACPA), and rheumatoid factor (RF). S100A12 levels were associated with presence of ACPA, history of diabetes, and serum S100A9 levels. sRAGE levels were negatively associated with serum levels of C-reactive protein (CRP) and high-density lipoprotein (HDL), history of vasculitis, and the presence of the RAGE 82Ser polymorphism. Conclusions: sRAGE and S100 proteins were associated not just with RA inflammation and autoantibody production, but also with classical vascular risk factors for end-organ damage. Consistent with its role as a RAGE decoy molecule, sRAGE had the opposite effects to S100 proteins in that S100 proteins were associated with autoantibodies and vascular risk, whereas sRAGE was associated with protection against joint and vascular damage. These data suggest that RAGE activity influences co-development of joint and vascular disease in rheumatoid arthritis patients.

Review and future options for computer modelling in the sugar industry

Computer modelling has been used extensively in some processes in the sugar industry to achieve significant gains. This paper reviews the investigations carried out over approximately the last twenty five years, including the successes but also areas where problems and delays have been encountered. In that time the capability of both hardware and software have increased dramatically. For some processes such as cane cleaning, cane billet preparation, and sugar drying, the application of computer modelling towards improved equipment design and operation has been quite limited. A particular problem has been the large number of particles and particle interactions in these…

Practical determination of cane wagon derailment forces for the Australian sugar industry

Derailments are a significant cost to the Australian sugar industry with damage to rail infrastructure and rolling stock in excess of $2 M per annum. Many factors can contribute to cane rail derailments. The more prevalent factors are discussed. Derailment statistics on likely causes for cane rail derailments are presented with the case of empty wagons on the main line being the highest contributor to business cost. Historically, the lateral to vertical wheel load ratio, termed the derailment ratio, has been used to indicate the derailment probability of rolling stock. When the derailment ratio reaches the Nadal limit of 0.81 for cane rail operations, there is a high probability that a derailment will occur. Contributing factors for derailments include the operating forces, the geometric variables of the rolling stock and the geometric deviations of the railway track. These combined, have the capacity to affect the risk of derailment for a cane rail transport operating system. The derailment type that is responsible for creating the most damage to assets and creating mill stops is the flange climb derailment, as these derailments usually occur at speed with a full rake of empty wagons. The typical forces that contribute to the flange climb derailment case for cane rail operations are analysed and a practical derailment model is developed to enable operators to better appreciate the most significant contributing factors to this type of derailment. The paper aims to: (a) improve awareness of the significance of physical operating parameters so that these principles can be included in locomotive driver training and (b) improve awareness of track and wagon variables related to the risk of derailment so that maintainers of the rail system can allocate funds for maintenance more effectively.

Improving factory performance using an integrated sugar factory model

A whole of factory model of a raw sugar factory was developed in SysCAD software to assess and improve factory operations. The integrated sugar factory model ‘Sugar-SysCAD’ includes individual models for milling, heating and clarification, evaporation, crystallisation, steam cycle, sugar dryer and process and injection water circuits. These individual unit operation models can be either used as standalone models to optimise the unit operation or in the integrated mode to provide more accurate prediction of the effects of changes in any part of the process on the outputs of the whole factory process. Using the integrated sugar factory model, the effect of specific process operations can be understood and practical solutions can be determined to address process problems. The paper presents two factory scenarios to show the capabilities of the whole of factory model.

Development of molecular tools for the improvement of transgene expression in sugar cane

Biotechnology has the potential to improve sugar cane, one of the world's major crops for food and fuel. This research describes the detailed characterisation of introns and their potential for enhancing transgene expression in sugar cane via intron-mediated enhancement (IME). IME is a phenomenon whereby an intron enhances gene expression from a promoter. Current knowledge on the mechanism of IME or its potential for enhancing gene expression in sugar cane is limited. A better understanding of the factors responsible for IME will help develop new molecular tools that facilitate high levels of constitutive and tissue-specific gene expression in this crop.

The role of added sugar in a healthy diet and implications for health inequalities

The role of added sugar in a healthy diet and implications for health inequalities Sugars provide a readily available, inexpensive source of energy, can increase palatability and help preserve some foods. However added sugars also dilute the nutrient density of the diet. Further, consumption of sugar-sweetened beverages is associated with increased risk of weight gain and reduced bone strength, and high or frequent consumption of added sugars is associated with increased risk of dental caries, particularly in infants and young children. The products of the 2013 NHMRC Dietary Guidelines work program at www.eatforhealth.gov.au include the comprehensive evidence base about food, diet and health relationships and the dietary modeling used to inform recommendations. This presentation will detail the scientific evidence underpinning the revised dietary recommendations on consumption of foods and drinks containing added sugar and compare recommendations with the most recently available relevant Australian dietary intake and trend data. Differences in intakes of relevant food and drinks across quintiles of social disadvantage and in particular between Aboriginal and Torres Strait Islander groups and non-Indigenous Australians will also be explored.

The influence of impurities on calcium phosphate floc structure and size in sugar solutions

Settling, dewatering and filtration of flocs are important steps in industry to remove solids and improve subsequent processing. The influence of non-sucrose impurities (Ca2+, Mg2+, phosphate and aconitic acid) on calcium phosphate floc structure (scattering exponent, Sf), size and shape were examined in synthetic and authentic sugar juices using X-ray diffraction techniques. In synthetic juices, Sf decreases with increasing phosphate concentration to values where loosely bound and branched flocs are formed for effective trapping and removal of impurities. Although, Sf did not change with increasing aconitic acid concentration, the floc size significantly decreased reducing the ability of the flocs to remove impurities. In authentic juices, the flocs structures were marginally affected by increasing proportions of non-sucrose impurities. However, optical microscopy indicated the formation of well-formed macro-floc network structures in sugar cane juices containing lower proportions of non-sucrose impurities. These structures are better placed to remove suspended colloidal solids.