Impact of Non-Ionic Surfactants on Enzymatic Hydrolysis of ... · PDF fileImpact of Non-Ionic...

International Forest Biorefining Conference (IFBC) May 9-11, 2017 Thunder Bay, Ontario, Canada

Impact of Non-Ionic Surfactants on Enzymatic Hydrolysis of Pulp Alaa Alhammad, Lew Christopher

Biorefining Research Institute, Lakehead University, Thunder Bay, Canada [email protected]

Abstract: Lignocellulose hydrolysis to fermentable sugars for biorefinery applications requires the use of enzymes that are currently still too expensive to allow a cost-effective bioconversion process. The addition of surfactants to enzymatic hydrolysis of lignocellulose has been reported to increase the conversion efficiency of lignocellulosic biomass to soluble sugars. Due to their hydrophobic interaction with lignin, surfactants serve as lignin blockers that reduce the non-specific enzyme adsorption to lignin which in turn increases the productive binding of cellulolytic enzymes to cellulose. This work compares the impact of four non-ionic surfactants: polyethylene glycol (PEG8000), polyethylene glycol 4000 (PEG4000), Triton X-100, and Tween 20) on the enzymatic hydrolysis of poplar pulp with a commercial cellulase preparation applied at a charge of 5 wt%. Each surfactant was used at three loadings (1, 5, and 8 wt% on pulp) and three levels of pulp consistency (5, 10, and 15 wt%). The highest glucose titer of 33.1 g /L was attained in the presence of 1 wt% PEG4000 at 5 wt% pulp consistency and 96 h hydrolysis time. This result represents a significant improvement of 48.8% in the enzymatic conversion of poplar pulp to glucose as compared to the control without surfactant.


2017 IFBC - Abstract Submission Form

Advanced Membrane Technologies for Biorefinery and Bioenergy Production: Challenges and Opportunities

Alnour Bokhary1, Baoqiang Liao1 1Department of Chemical Engineering, Lakehead University, Canada, [email protected]

Abstract: Decreasing competitiveness of the forest products industries and the growing environmental concerns have motivated the research to focus on exploitation of the different wood components in an efficient way. This situation led to the study and development of integrated forest biorefinery (IFBR) which utilizes forest-based biomass rather than fossil fuels to produce value-added chemicals and bioenergy. This new business paradigm offers a broad range of potentially attractive products. However, the biorefinery will only ever be truly economically viable if advanced separation technologies that deliver the hemicellulose and lignin fractions in a useable form are developed. In many instances, an effective separation of lignocellulosic materials can be the single biggest factor affecting the overall success and commercialization of biorefineries. Thus, a number of separation and purification approaches have been proposed. Among the proposed separation methods, membrane technologies have been studied in a wider range of perspectives, and have been shown to play a key role in products recovery and purification in biorefining processes. Compared with the commonly applied separation methods, membrane technologies offer excellent fractionation, short processing steps, less chemical utilization and considerable energy saving. Also, membrane separation processes can be integrated with the existing operating units, not to mention their easy operation and scale-up. This poster highlights the applications and the recent advancement in the membrane technologies for biorefinery and bioenergy production. The paper also discusses the challenges and opportunities of this new business paradigm of forest industries.

mailto:[email protected]



Biochar from Non-Conventional Biomass (Fruit Wastes)

Animesh Dutta, Bharat Regmi 1School of Engineering, University of Guelph, Canada, [email protected]

Abstract: Wastes are resources waiting for opportunity to be utilized. For last decade, most research done on HTC using lignocellulosic biomass as feedstock. However, fruit wastes which contain high amount of sugar content is also a promising feedstock for the process. The characteristics and yield of biochar from fruit waste via HTC has not been investigated yet. Researchers have reported that the fuel characteristics improved with increase in temperature of HTC process but at expense of mass yield through process. Thus the purpose of this study is an attempt to characterized biochar produced by HTC using various fruit wastes feedstocks. In this research, various non-conventional biomasses such as rotten apple, apple chip pomace (AP) and grape pomace (GP) was used to produce biochar. Samples were mixed with deionized water in ratio of 1:10 (w/w) and was then pretreated at various temperature of 190, 225 and 260°C for 15mins using Parr hydrothermal reactor. Characterisation of biochar was done in terms of mass yield, energy density, proximate and ultimate analysis Combustion behaviour of different biochar was done using QWM (Macro-TGA) reactor. Biochar obtained via HTC of different fruit waste in a batch reactor has shown highly improved physicochemical and combustion properties of biomass indicating higher potential to replace coal at power plant without any modification. Unlike other biomass, HTC of fruit waste at higher temperatures resulted in higher mass yield likely due to presence of higher glucose content which undergoes for carbonization above 190°C. The TGA analysis showed that the derivative mass loss increases with increase in HTC temperature indicating the faster rate of combustion. Further maximum weight loss percentage per degree temperature increases with increase in HTC temperature.




Utilization of Paper Mill Primary Sludge for the Production of Sugars

Amit Nair1, Amanda Bakke2 and Sudip K. Rakshit1

1Department of Engineering and the Biorefining Research Institute Lakehead University, Thunder Bay, Canada – ON P7B5E1

2Department of Biochemistry, University of Guelph, Guelph, Canada ON N1G 2W1 [email protected]

Abstract: With its abundance of forests, Canada is a leading paper producing country with products ranging newspapers, specialty papers, market pulp and other wood products. For every ton of paper produced in many of these mills, approximately 30 kg of dry primary sludge is being produced. The dry sludge is then either shipped to landfills or incinerated, generating pollution. The primary sludge consists primarily of cellulose with some hemicellulose and lignin in the secondary sludge. Cellulose can be hydrolyzed through enzymatic processes to produce reducing sugars. Ash content and other components in the primary sludge can affect the yield of reducing sugars during hydrolysis process. Some pretreatment processes are required for efficient hydrolysis and increased yields of sugars. This paper will present some initial work done using primary sludge from a large paper mill in Thunder Bay and it conversion to fermentable sugars.




Biocarbon as a Green Substitute for Mineral Filled Automotive Parts

Ehsan Behazin1,2, Manjusri Misra1,2, Amar K. Mohanty1,2

1Bioproducts Discovery and Development Centre, University of Guelph, Canada, [email protected]

2School of Engineering, University of Guelph, Canada Abstract: Utilization of biorefinery systems such as pyrolysis would help farmers to attain required energy for their greenhouses while managing agricultural waste in a more eco-friendly manner. However, generating a considerable amount of by-product in such systems causes sustainability issues in long runs. In this study, the by-product of pyrolysis (biocarbon) is intended to be used as filler in plastic industry targeting toughened polypropylene based automotive parts. The biocarbon particle size, type, and the amount of compatibilizer were optimized to achieve biocomposites with balanced mechanical properties. Among the investigated factors, the functional polymer type had the greatest impact on the properties. By optimizing the concentration of the polypropylene-based functional polymer the impact toughness of the composites improved by 120 %. Results demonstrated that substituting traditional petroleum based glass fiber and talc filled composites with biocarbon biocomposites would provide the required mechanical properties with added benefits of around 15% lower density, 30% higher bio-based content and cheaper price. These benefits would translate into improved fuel economy of cars and reduction in greenhouse gas production. Considering the high-volume usage of plastic parts especially in automotive industry, this approach would complete the biomass refinery cycle and makes it sustainable.



Extraction and Characterization of Rice Husk Lignin Obtained in a Biorefinery Process

Eliana P. Dagnino1, Mariano J. Sequeira1, Ester R. Chamorro1, María C. Area2

1Centro de Investigación en Química Orgánica Biológica, Instituto de Modelado e Innovación Tecnológica, IMIT (UTN-CONICET-UNNE), Universidad Tecnológica Nacional, Argentina.

[email protected]

2Programa de pulpa y papel, Instituto de Materiales de Misiones, IMAM (UNAM-CONICET), Universidad Nacional de Misiones, Argentina.

Abstract: The biorefinery of lignocellulosic wastes looks for the optimal separation of the main components of these materials (lignin, hemicelluloses, cellulose, extractives, and inorganics) to achieve their best possible use. On the other hand, husk produced in the industrialization of rice constitutes a very abundant lignocellulosic residue in the northeast region of Argentina and its large accumulation makes its disposition difficult, causing potential fire risk and proliferation of vermin, among others. Numerous investigations have been carried out regarding possible pretreatments to achieve effective separation of the different components. The fractionation involving an acid-alkaline sequence achieves the effective separation of hemicelluloses in the first step and lignin in the second. The optimization of the above-mentioned pretreatments was carried out in previous works, whereas lignin separation from the process liquor and its chemical modification as a function of time and temperature are presented in this one. The recovery of lignin from the liquor was effected by dropping the pH from about 10 to 3 and the precipitated solid (recovered lignin) was removed by vacuum filtration. Lignin quantification was performed gravimetrically (mass of recovered lignin/lignin mass in the untreated material) and the chemical characterization was carried out by the determination of the concentration of phenolic oxides by UV spectroscopy, based on the absorption difference of phenolic units in neutral and alkaline solutions at 300 and 360nm. Results show that temperature makes decrease the total OH concentration from 8.91% at 140 ° C to 8.05% at 160 ° C. In contrast, the reaction time showed no significant effect on the percentage of OH. The percentages of OH structures are higher than those found by other authors.


Aminated lignin as a flocculant for effluent of textile industry

Fangong Kong, Shoujuan Wang, Mohan Konduri, Pedram Fatehi Department of Chemical Engineering, Lakehead University, 955 Oliver Road, Thunder Bay,

Ontario, P7B5E1, [email protected]

Abstract: The amination of polymers was regarded as an effective method to improve their performance in various applications. Kraft lignin can be modified to function as a flocculant. In this work, the amination of kraft lignin with glycidyl-trimethylammonium chloride (GTMAC) was assessed in an aqueous solution under altered conditions. The conditions investigated were temperature, time, pH, GTMAC to lignin molar ratio, and lignin concentration. The optimized conditions based on charge density and solubility of cationic lignin were found to be 70 °C, 1 h, 12.5 pH, 2/1 GTMAC/lignin molar ratio, and 1.0 wt.% lignin concentration. The solubility of the resulting aminated lignin reached 90 % in 1 wt.% lignin concentration, and the charge density reached 1.10 meq/g under the optimized conditions. The properties of aminated lignin were analyzed using various tools including NMR, FTIR, and TGA. The performance of aminated lignin was assessed as a flocculant for dye removal from simulated solutions containing Remazol Brilliant Violet, Reactive Black or Direct Yellow.




Algal Heterotrophic Metabolism for Oil Production

Fatimah Alsayahan, Lew Christopher, and Ladislav Malek

Department of Biology and Biorefining Research Institute, Lakehead University, Thunder Bay, Canada, [email protected]

Abstract: The challenges associated with the use of conventional sources of oil increase the demand for alternative fuel sources. Biofuels offer the chance of replacing the depleting fossil fuel. However, the food-versus-fuel conflict that exists with the use of food-based crops as source of biofuels means that non-edible algal biomass can be an attractive source of oil that ensures high oil production without trading off food production. Algal biomass may also ensure year-round production and supply, and can use minimal space if produced in fermentors. The addition of glucose to otherwise autotrophically grown algae has been shown to increase oil accumulation. Optimal algal mixotrophic production of oil is based on the availability of appropriate organism, sugar addition, and other culture conditions. Recently, cellulose was shown to be metabolized by algae. In this work, we studied the heterotrophic algal metabolism of the known oil producing Auxenochlorella pyrenoidosa and several other algae isolated from decaying wood and pulp mill waste water treatment plants. We chose to address the question whether cellulose in the form of cellulosic waste or the model water-soluble substrate carboxymethyl cellulose can substitute for pure glucose in boosting oil synthesis. We believe that the outcomes of this study will add important new information and contribute toward development of a commercially-viable process for mixotrophic production of algal oil.




Mesoporous Carbon Materials Derived from Lignin: Synthesis and Electrochemical Applications

Guosheng Wu, Zhaoyang Zhang, Cassandra K Ostrom, Xin Chang, Aicheng Chen

Department of Chemistry, Lakehead University, Canada, [email protected]

Abstract: Carbon is a very important material that has various applications in many industries, as catalyst supports, absorbents, and electrodes. Carbon based materials may be produced from different resources including wood, oil, natural gas, etc. Recently, lignin has attracted considerable attention as a sustainable resource, in serving as a starting material for the fabrication of carbon based materials. One of the most important examples is lignin-derived carbon fibres that are generated via an electrospinning process. Further, activated carbon and carbon nanoparticles have been fabricated from lignin through various approaches. Here, we report on novel mesoporous carbon materials which are fabricated from kraft lignin through a facile and rapid solution combustion reaction method, followed by an annealing treatment under inert gas flow without acid or alkali activation. In a typical fabrication process, urea was employed as a fuel, and gas bubbles that were generated during the solution combustion reaction acted to produce pore structures within the carbon materials, which were retained following the annealing treatment. In addition, unlike other methods for fabricating mesoporous structures, a template was not required for this process. Parameters such as the quantity of urea, concentration of lignin, and temperature were optimized to achieve the highest surface area and porous structure. Our results revealed that the surface areas of the prepared mesoporous carbon materials were much higher than that of commercial carbon black. These mesoporous carbon materials fabricated from a sustainable resource have various applications as an alternative to carbon black, such as catalyst supports, adsorbents, etc. Our electrochemical studies have shown that the mesoporous carbon and Pt-modified mesoporous carbon exhibit much higher activity than that of carbon black and Pt deposited carbon black in terms of hydrogen adsorption/desorption and methanol oxidation, promising for energy and environmental applications.




Enhanced Production of Lignocellulolytic Enzymes in Three

Bacillus Strains by Using Algae as Both Carbon Source and

Inducer

Haipeng Guo1,2, Yanwen Wu1, Bingsong Zheng3, Dean Jiang2, Wensheng Qin1 1 Department of Biology, Lakehead University, 955 Oliver Road, Thunder Bay, Ontario, P7B

5E1, Canada, [email protected] 2 State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences,

Zhejiang University, Hangzhou 310058, China. 3 State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, Linan,

Hangzhou 311300, China. Abstract: Lignocellulosic biomass consisting of carbohydrate polymers (cellulose and hemicellulose) and an aromatic polymer (lignin) is the most abundant and sustainable raw material on the Earth for the production of biofuels and chemicals. However, the biomass recalcitrance in decomposition has posted a main challenge in biofuel production due to lignin carbohydrate complexes formed by lignin intimately interlaced with carbohydrates through ester and ether bonds to prevent enzymatic hydrolysis to fermentable sugars. Lignocellulolytic enzymes produced by bacteria are able to break the biomass recalcitrance, making cellulose and hemicellulose available for producing biofuels through fermentation. Therefore, high efficient production of these lignocellulolytic enzymes from bacteria is particularly important for cost saving in the industrial process. In this study, the production abilities of lignocellulolytic enzymes in LB medium without algae were used as reference for comparison. We tested three Bacillus strains (A4, K1 and X4) for enzyme activities. In LB medium without algae, the highest activities of exoglucanases (CMCase), endoglucanases (FPase), xylanase, and laccase were 2.74, 0.07, 0.85 U ml-1 and 241.6 U L-1, in Bacillus A4. In Bacillus K1, they were 5.21, 2.30, 56.75 U ml-1, and 87.03 U L-1, respectively. And in Bacillus X4, they were 6.80, 2.39, 39.66 U ml-1, and 88.15 U L-1, respectively. When the three Bacillus strains (A4, K1 and X4) were cultured in mineral salt medium containing 5% (w/v) algae as carbon source and inducer, their highest CMCase activities were 2.97, 5.45 and 5.09 U ml-1, respectively. Their maximum FPase activities were 0.09, 3.58 and 2.42 U ml-1. Their optimal xylanase activities were 0.89, 33.04 and 27.97 U ml-1. And their corresponding highest laccase activities were 1783.34, 308.89 and 205.56 U L-1. The results suggest algae could be both as carbon source and inducer in mineral salt medium for the high production of ligninolytic enzyme cocktails by using Bacillus strains.



Ultrasonic-Assisted Extraction of High-Value Betulinic Acid from Birch Based Chaga Mushroom

Hanin Alhazmi1, Peter Adewale2, Sai Swaroop Dalli 3 and Sudip Kumar Rakshit 1,3 1Department of Biology, Lakehead University, Thunder Bay, ON, Canada P7B 5E1

[email protected] 2Biorefining Research Institute, Lakehead University, Thunder Bay, ON, Canada P7B 5E1 3Department of Chemistry and Material Sciences, Lakehead University, Thunder Bay, ON,

Canada P7B 5E1

Abstract: Chaga mushroom (Inonotus obliquus) is known as a white-rot fungus growing on birch trees. It is found mainly in the forests of Canada, the United States, Russia, Korea and Eastern Europe. It is edible and also regarded as a traditional medicinal mushroom. Betulinic acid has been identified as one of the phytochemicals found in chaga mushroom. The acid is a pentacyclic triterpenoid with several biological properties such as antiparasitic activity, anticancer, etc. This study aims to enhance the extraction of betulinic acid from birch based chaga using ultrasonication. The cavitation effect of ultrasound technique is suitable to disintegrate the substrate and improve adequate contact between the extraction solvent and the desired substance. After some water refluxing pre-treatment, the ultrasonic systems helped to extract the betulinic acid in large quantities using solvent extraction. Following the optimization of the extraction solvents and ultrasound parameters, it was found that the use of ultrasound further enhanced the extraction of betulinic acid from birch based chaga.




Renewable Natural Resources, Biomass Refining and Analytics

Joni Lehto1, Hannu Pakkanen1, Jarmo Louhelainen1, and Raimo Alén1 1Department of Chemistry, University of Jyväskylä, Finland, [email protected]

Abstract: The research carried out by the Laboratory of Applied Chemistry (LAC), Department of Chemistry, University of Jyväskylä has recently concentrated on

• composition of wood and non-wood materials, • characterization of various process streams from pulping, oxygen delignification, and

bleaching, as well as their process chemistry, • paper chemistry (including applied molecular modeling of papermaking chemistry), • development of monitoring methods for the pulp and paper industry, • environmental chemistry, and • production of useful chemicals from lignocellulosic biomass.

Renewable biomass resources can be industrially utilized in a number of ways to produce chemicals, fuels, and other products. Along with the research for improving further the processes applied in the conventional pulp and paper industry or chemical industry, novel process concepts related to biorefining are also being developed. In all these cases, generally a better understanding of process chemistry as well as detailed knowledge on the properties of feedstocks and products is needed. In addition, various environmental aspects on the processes are of great importance. These kinds of research and education activities belong to the core areas of the research program “Renewable Natural Resources and Chemistry” in the Department of Chemistry. It can be concluded that the LAC research team has especially involved in complex separation and identification problems. Research infrastructure of LAC comprises comprehensive facilities including premises (wood/sample handling and storage, cooking and bleaching laboratories, and analytical laboratories) and research equipment (modern analytical spectroscopic and chromatographic instrumentation and analysis methods) generally applied in wood and pulping chemistry. In addition, close cooperation is done with the forest industry production units, especially in terms of acquisition of wood samples, industrial grade cooking liquors, and other materials needed for the research. Cooperation with other research groups within LAC and other laboratories/departments located in the University of Jyväskylä is strengthened by collaboration performed with domestic and foreign research



Techno-Economic Analysis and Supply Chain Design for a Forest Biorefinery to Produce Value-Added Bio-based Products from

Lignin and Forestry Residues

Luana Dessbesell1,2, Zhongshun Yuan2, Mathew Leitch1, Reino Pulkki1, Chunbao (Charles) Xu2

1Faculty of Natural Resources Management, Lakehead University, Canada, [email protected]

2Institute for Chemicals and Fuels from Alternative Resources (ICFAR), Western University, Canada

Abstract: The Canadian forest industry is seeking to diversify its portfolio to remain competitive, as bioenergy alone is not enough to transform the forest industry. Higher-value products such as bio-based chemicals and materials can augment lower-value bioenergy. Besides diversifying the portfolio, we can replace petroleum-based products, and decrease petroleum dependence and footprint. For example, kraft lignin (KL) from pulpmill recovery boilers can be refined into bio-based polyols and phenols. However, recent progress in conversion technologies is not enough to introduce bio-based products to the market. Challenges persist and examples of technical difficulties are bio-based feedstock yield, composition, processing and logistics; likewise, commercial and strategic challenges are subsidy limitations, integration into current value chains, and uncertainty in confronting a new area. To assist with these challenges, our ongoing work aims to generate a techno-economic analysis for a KL biorefinery (BR) to produce bio-based phenols and polyols, and also to develop a BR supply chain with a capacity design model to deal with uncertainties in supply and demand. The process used in the intended BR consists of KL depolymerization and further oxypropylation (proprietary process). The economic indicators calculated are Net Present Value (NPV), Pay Back Period (PBP) and Internal Rate of Return (IRR). The investment in the KL BR was found feasible for the base case scenario (KL cost 700 US$⋅t-1, bio-based polyol price 2300 US$⋅t-1, and bio-based phenol price 1700 US$⋅t-1) generating a positive, attractive NPV, IRR, and PBP. A sensitivity analysis showed that the NPV is highly affected by fluctuations in polyol and phenol selling price, capital investment, and KL cost, respectively. By complementing this study with the supply chain design we expect to assist the forest-based industry to improve the value of forest resources, and also contribute to introducing bio-based value-added products to the market.




Antioxidant, Antimicrobial and Anticancer Activities of Lichen Extracts

Lyndon Letwin1,2,3, Erik Yeash4, Lada Malek2,3, Zacharias Suntres2,3, Kerry Knudsen5, Lew Christopher1,2,4

1Department of Biology, Lakehead University, Thunder Bay, ON, Canada, [email protected] 2Biorefining Research Institute, Lakehead University, Thunder Bay, ON, Canada

3Northern Ontario School of Medicine, Lakehead University, Thunder Bay, ON, Canada 4South Dakota School of Mines and Technology, Rapid City, SD, USA

5University of California-Riverside, Riverside, CA, USA

Abstract: Lichens contain many complex biochemical pathways which can provide a large array of compounds with the potential for pharmaceutical development. In the present study, extracts from three previously undescribed lichen species were examined for antioxidant, antibacterial and anticancer activity. The results from this study demonstrated the following: (1) the acetone extract of Lobothallia alphoplaca inhibited the growth of cultured breast cancer MCF-7 cells at a relatively low concentration; (2) the acetone and ethyl acetate extracts of Xanthoparmelia mexicana inhibited Gram-positive bacteria; and (3) Acarospora socialis ethanol extract exhibited significant DPPH antioxidant scavenging activity. Further work is warranted into the Lobothallia alphoplaca extracts which inhibited neoplastic MCF-7 cells. Further investigation in order to identify specific active compounds and the range of selectivity of this toxic effect against primary cells and a wider variety of cultured cancer cells could lead to new and improved anti-cancer compounds.




Anticancer and Antibacterial Activities of Umbilicaria muhlenbergii Lichen

Lyndon Letwin1,2,3, Lada Malek1,2, Zacharias Suntres2,3, and Lew Christopher1,2 1Department of Biology, Lakehead University, Thunder Bay, ON, Canada, [email protected]

2Biorefining Research Institute, Lakehead University, Thunder Bay, ON, Canada 3Northern Ontario School of Medicine, Lakehead University, Thunder Bay, ON, Canada

Abstract: Lichens contain many complex biochemical pathways which can provide a large array of compounds with the potential for pharmaceutical development. In the present study, extracts from an edible Northern Ontario lichen, Umbilicaria muhlenbergii, were examined for antibacterial and anticancer activity. The results demonstrate that the crude extract of Umbilicaria muhlenbergii exhibits antibacterial activity against gram-positive bacteria, while also exhibiting anti-cancer effects against MCF-7 breast cancer cells. The crude extract was then separated into fractions via silica gel column chromatography, and each fraction was analyzed for anti-cancer activity. From this, two fractions of particular relevance were selected for further investigation. They demonstrate anti-cancer activity with a greater efficacy than the crude extract, warranting further separation. Further work will include the identification of compounds within the fractions.


Lipid and Carotenoid Synthesis by the Oleaginous Yeast, Rhodosporidium dibovatum, Grown on Glucose versus Glycerol

Maryam Mirzaie1, Irene Fakankun1, Nima Nasirian2, and David B. Levin1

1 Department of Biosystems Engineering, University of Manitoba, Winnipeg, MB, Canada, R3T 5V6, [email protected]

2 Department of Agricultural Mechanization, Shoushtar Branch, Islamic Azad University, Shoushtar, Iran

Abstract: Production of microbial single cell oils (triacylglycerides) has been the focus of much research because of their structural similarities to vegetable oils. Carotenoids are also high-value products synthesized by microorganisms, such as microalgae and yeast, and have important applications in animal feed, in human nutrition and nutraceuticals, and in medical research. Rhodosporidium diobovatum is a “red” yeast that can synthesize and accumulate high concentrations of both triacylglycerides (TAGs) and the carotenoid -carotene. Thus, in this study, we assessed the relationships between lipid and carotenoid synthesis by R. dibovatum 08-225 in nitrogen-limiting medium (GMY) containing equal mass (40 g/L) of glucose or glycerol. The cultures were also supplemented with additional substrate at 72 hours (h) post-inoculation (pi) and continuing adding it every 24 hours for more 5 days. After 72 h pi, for 5 days, the glucose and glycerol cultures produced approximately equal amounts of cell mass, but the glycerol cultures produced slightly greater amounts of lipid (38.31 ± 0.1 % dcw) than the glucose cultures (37.48 ± 0.09 % dcw) at 192 h pi. Glycerol cultures produced higher amounts of ß-carotene at 48 h pi (245.80 ± 6.95 µg/g dcw, versus 201.44 ± 0.89 µg/g dcw at 24 h pi in Glucose media, respectively). After 48 h pi, ß-carotene production significantly decreased in both cultures, but after the addition of fresh substrates at 72 h pi, ß-carotene concentrations stayed constant at around 80 µg/g dcw in these cultures, with a sharply increased in lipid concentrations.




Superabsorbent Biodegradable Materials Produced From Wood-Derived Cellulosic Fibers

Md Nur Alam* and Lew P. Christopher

Biorefining Research Institute, Lakehead University, Canada, [email protected]

Abstract: There has been a great interest in producing superabsorbent materials from cellulose over the past few decades. Superabsorbent products already exist on the market; however, in most cases they are produced by incorporating superabsorbing polymeric particles to the fibers, which are either based on synthetic polymers, such as polyacrylates, sulfonated polystyrene, polyvinyl alcohol, etc, or on natural polymers, such as carboxyalkyl cellulose, gum, carboxyalkyl starch, cellulose sulfate, etc. All these products suffer from three major disadvantages: 1) lack of complete biodegradability; 2) physical dislodgement of the particles from the fibers during manufacturing and transportation that results in a diminished efficiency; and 3) limited water absorbency that ranges from 10 to 100 g water/g material. Here we describe a novel, aqueous-based and environmentally-friendly process for production of superabsorbent hydrogels from wood-derived, cross-linked cellulosic fibers. The conditions for hydrogel preparation (cross-linking parameters, swelling time, drying method, rewetting, etc.) were optimized to attain a water absorption capacity of up to 500 times the dry weight of the superabsorbent material. The novel highly-absorbent and fast-swelling cellulose-based hydrogels can find applications in high-value hygiene and biomedical products.

mailto:mnalam

mailto:mnalam



Designing Kraft Lignin Based Dispersant for Clay Suspension

Mohan Konduri, Pedram Fatehi Department of Chemical Engineering, Lakehead University, 955 Oliver Road, Thunder Bay,

Ontario, P7B5E1, [email protected] Abstract: Kraft lignin valorization is a crucial way to convert kraft pulping processes to biorefineries. Kraft lignin can be modified to function as a dispersant for various applications. To generate useful dispersants, it is important to identify the correlation between the properties of suspensions and dispersants. In this study, kraft lignin was modified by oxidation using nitric acid under varied conditions to produce oxidized lignin products with different charge densities and molecular weights. Then, the effect of anionic charge density (1.2-3.6 meq/g) and molecular weight (11,000-88,000 g/mol) of the lignin-based products on their dispersion performance was evaluated using different advanced techniques. It was discovered that the lignin-based product with the charge density of 3.6 meq/g and molecular weight of 30,000 g/mol was the most effective dispersant for clay suspensions. In this work, the adsorption of lignin-based dispersants on clay particles and the impact of adsorption on the particle size, zeta potential, relative turbidity and sedimentation of clay particles in different slurries will be discussed.




Biopolyesters Derived from Biorefinery Resources as Impact Modifiers for Biodegradable Poly (Butylene Succinate) Bioplastic

Oscar Valerio1,2, Manjusri Misra1,2, Amar Mohanty1,2

1School of Engineering, University of Guelph, Canada, [email protected] 2Bioproducts Discovery and Development Centre, University of Guelph, Canada

Abstract: Poly (butylene succinate) (PBS) is a compostable commercially available bioplastic with excellent mechanical properties similar to common petroleum based polymers like polypropylene. The major limitations for the usage of poly (butylene succinate) in commercial applications are its relative high cost when compared with petroleum based polymers, as well as a low impact resistance (~30 J/m on notched Izod impact). Thus, improving the impact resistance of poly (butylene succinate) by the addition of a biobased elastomer produced from cheap and abundant monomers such as glycerol could help in producing a biobased polymeric material suitable for applications where PBS cannot meet impact requirements. For this purpose, biobased poly (glycerol succinate) gel polyesters have been synthesized in a green synthesis procedure in absence of solvents or catalysts. These elastomeric polyesters were employed as impact modifiers for PBS by means of melt blending processing. A key advantage of this technology in terms of sustainability is the employment of glycerol, the main co-product of biodiesel production on biorefineries, as well as succinic acid, produced from fermentation of biomass resources, as monomers for the synthesis of PGS. It was found that after the addition of 30 wt% of PGS to PBS the notched Izod impact of PBS was significantly increased by 244% (from 31.9 to 110 J/m) whereas the elongation at break of the material remained unchanged. PGS particles were distributed uniformly on the PBS matrix with no evident agglomeration as evidenced by scanning electron microscopy imaging. This was explained by the presence of hydroxyl groups on the backbone of PGS which are thought to enable a good interfacial adhesion between the blend components and prevent PGS coalescence during injection molding. The final biobased 70/30 wt% PBS/PGS blend material displayed a 4-fold increased impact resistance while retaining a high elongation at break, which makes it suitable for molding of flexible and tough plastic parts.



Characterization of Biochars from Pyrolysis of Soy Hulls and Coffee Chaff

Peter Quosai, Andrew Anstey, Amar Mohanty, Manjusri Misra

Bioproduct Development and Discovery Centre, Department of Plant Agriculture and School of Engineering, University of Guelph, Canada, [email protected]

Abstract: Biochar is created as a primary product of slow pyrolysis of biomass and a coproduct of biooil production by fast pyrolysis. The product’s physical properties vary widely with the biomass used, temperature, and duration of pyrolysis. Biochar can be used for composite applications such as a bio-based replacement for carbon black, carbon sequestration, soil amendment, greenhouse growth media, filtration, or a precursor to activated carbon; the possible applications depend on the specific properties of the biochar. There remains a great deal unknown about the chemical changes which occur through the pyrolysis process. To better understand how specific conditions and biomass can be used to create specific biochar products, fundamental research is necessary to characterize new biochars from novel carbon sources, such as agricultural wastes. By identifying the conditions that create these properties, feedstocks can be identified for creating biochar optimized for specific applications. In this study, two agricultural products were investigated: soy hulls and coffee chaff. Pyrolysis was conducted in a horizontal tube furnace purged with a continuous flow of nitrogen at both 500 and 900 degrees Celsius, at two unique holding times. Using various physical and chemical characterization techniques this study describes how feedstock characteristics and pyrolysis conditions affect the biochar produced. We have found that for all feedstocks, a higher temperature of pyrolysis imparts the following properties: an increase in carbon content, a reduction in surface functionality, an increase in ash content, an increase in thermal stability, and an increase in electrical conductivity. The biochars created from both materials share similar thermal stabilities and material properties. The goal of this work is to contribute to the understanding of the effect which feedstock has on biochar and to identify higher value applications for biochar by which pyrolysis can represent a sustainable method of disposal for agricultural wastes.

Metabolic Engineering of Pseudomonas putida LS46 for Enhanced Production of Polyhydroxyalkanoate Polymers

Riffat I. Munir, Parveen K. Sharma, David B. Levin

Department of Biosystems Engineering, University of Manitoba, Winnipeg, MB, Canada, R3T 5V6, [email protected]

Abstract: Pseudomonas putida strains have been widely studied for their ability to synthesize medium chain length polyhydroxyalkanoate (PHA) polymers with different monomer compositions from a wide range of carbon sources. P. putida is known to synthesize PHA polymers either via the β-oxidation pathway when fatty acids are used as carbon source or the fatty acid de novo biosynthesis pathway in the presence of glucose or glycerol. Previously, studies have identified pha J1 and pha J4 genes (R-specific enoyl-CoA hydratases) in the β-oxidation pathway, and pha G gene (hydroxyl acyl ACP CoA transacylase) in de novo biosynthesis pathway to play a significant role in the production of PHAs. These genes encode important “monomer offering” enzymes that supply substrate to PHA synthase (encoded by pha C), the enzyme that polymerizes PHA polymers. In addition, PhaG has been shown to directly link the fatty acid de novo biosynthesis with PHA biosynthesis pathway. Genetic manipulation of these genes via gene knockouts and genetic recombination, which may enhance PHA production, are being investigated.




Electrochemical and Photoelectrochemical Oxidation of Lignin Model Compounds

Shuai Chen, Min Tian, Daniel Liba, Aicheng Chen

Department of Chemistry, Lakehead University, Canada, [email protected]

Abstract: Lignin, known as a byproduct of the pulp and paper industry, is a tridimensional amorphous phenolic polymer composed of phenylpropane units linked together by various bonds. Lignin is potentially a major renewable, nonfossil source of aromatic and cyclohexyl compounds. Electrochemical and photoelectrochemical (PEC) approaches have been explored to modify lignin to value-added products. However, the inherent complexity of the lignin polymer makes it difficult to evaluate in terms of reaction kinetics. Consequently, relatively simple molecules that contain isolated functional groups or typical lignin bonding patterns can be utilized to reveal detailed information of mechanistic processes. In this study, we chose the following two lignin model compounds: 3-hydroxy-1-(3,4-dimethoxyphenoxy)-2-(2-methoxyphenoxy)-1,3-propanone (HDM) and 1-(3,4-dimethoxyphenoxy)-2-(2-methoxyphenoxy)-1,3-propanediol (DMP). The structures of the two model compounds are shown in the figure. The aim of this study was to provide some mechanistic insights of the electrochemical and PEC oxidation of lignin. In this study, we have investigated the abrasive stripping voltammetry of the two lignin model compounds and lignin in the room temperature ionic liquids (RTILs) on a gold macrodisk and in 0.1 M H2SO4 and 0.1 M NaOH on a boron-doped diamond (BDD) macroelectrode, with the hope of using the voltammetry to fingerprint the functional groups within the lignin molecule. The PEC oxidation of the lignin model compounds was carried out using titanium dioxide nanotubes. UV-Vis spectroscopy was employed to monitor the time-dependent concentration changes during the PEC process. A new band appeared at ~304 nm during the PEC oxidation of DMP, revealing intermediate formation. The rate of DMP intermediate formation was increased with the increase of the initial concentrations, while it was decreased with increased temperature. On the other hand, despite the similarity in structure between HDM and DMP, there were only small increases in absorbance during the PEC oxidation of HDM, suggesting that HDM is less reactive. Finally, quantum chemical calculation based on the density functional theory was performed and the results were consistent with our PEC study.




Bioactives from the Outer Bark of Betula papyrifera and Their Potential for Use in the Cosmetic Industry

Statton Eade1, Sudip Rakshit2

1Biorefining Research Institution, Lakehead University, Canada, [email protected] Abstract: In an effort to revitalize the wood and pulp/paper industry, the integrated biorefinery concept proposes using all parts and components of trees including the bark and foliage. Though rich in valuable bioactive compounds, the bark of Betula (birch) species trees is generally underexploited and burned as fuel to reduce energy costs. By utilizing these residues for the production of high value compounds for pharmaceuticals, food additives, and cosmetic agents, significantly more value can be injected into the forestry industry as well as the bioeconomy. Historically, birch has been used in folk remedies to produce oils for the treatment of skin diseases (such as psoriasis and eczema) as well as teas for the treatment of digestive tract infections. This presentation will focus on the extraction of high value compounds from the outer bark of white birch (Betula papyrifera) such as triterpenes (betulin, betulinic acid, and lupeol) and investigate their potential as cosmetic/cosmeceutical ingredients. Flavonoid and phenol content of the bark will also be discussed. Many of the compounds present in the outer bark of Betula species exhibit a variety of bioactivities which are considered beneficial to the development of cosmetic products, specifically those pertaining to skin health. Triterpenes, which are present in the outer bark of boreal birch at significantly high levels (20-35%), are known inhibitors of elastase (one of the key enzymes involved in skin aging) and have been shown to exhibit anti-inflammatory and anti-cancer activity coupled with a remarkably low toxicity. Using solvent extraction, crude ethanolic extracts were obtained and a compositional analysis was performed to determine the triterpene, total phenol, and total flavonoid content. The ethanolic extracts will be investigated further for their ability to scavenge reactive species such as nitric oxide (NO) and singlet oxygen (1O2), their capacity to inhibit elastase, as well as their UV-protecting potential. Results will be compared to the commercially available French maritime pine bark extract Oligopin® as a positive control. The overall anti-aging potential of the extracts due to their anti-oxidant, elastase-inhibiting, and UV-protecting properties will be discussed in this presentation.

Development of an Oxygen-limited High Cell Density Cultivation

Process for Pseudomonas putida LS46 to Increase Biopolymer Productivity

Warren Blunt1, Chris Dartiailh1, Richard Sparling2, Daniel Gapes3, David B. Levin1,

Nazim Cicek1 1Department of Biosystems Engineering, University of Manitoba,

Winnipeg, MB, Canada, R3T 5V6, [email protected] 2Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada, R3T 2N2

3Scion Research, Te Papa Tipu Innovation Park, 49 Sala Street, Private Bag 3020, Rotorua, New Zealand 3046

Abstract: During production of medium chain length polyhydroxyalkanoates (mcl-PHA) from fatty acids using Pseudomonas putida LS46, it was recently shown that both the polymer content of the cell and synthesis rate are increased under microaerophilic conditions. However, that work was conducted in batch bioreactors with low overall volumetric productivity. In the present study, we have increased the volumetric productivity using an oxygen-limited fed batch strategy. A feeding method was developed in which the carbon source and nitrogen source were fed in small, accurate pulses in response to off-gas CO2 concentrations using a Titrimetric Off-Gas Analyzer (TOGA) sensor system. Carbon and nitrogen were fed to ensure balanced growth as long as the dissolved oxygen remained above 0% air saturation, and subsequently oxygen limited conditions were allowed to prevail in order to increase polymer content. Using this method with maximal mass transfer capability of the bioreactor (aeration at 2 VVM using compressed air, mixing at 1200 rpm), 27.3 ± 3.7 g/L cell dry mass (CDM) was obtained in 27 hours of cultivation, with an average cellular mcl-PHA content of 61.4 ± 9.8 %. The resulting volumetric productivity was 0.68 ± 0.14 g/L/hour. This was nearly a ten-fold improvement in total biomass and productivity from the results obtained in batch experiments. Future work will investigate further improvement to total biomass and overall volumetric productivity by increasing the driving force for oxygen transfer.




Enzymatic Hydrolysis of Negative-Cost Lignocellulosic Material for

Cellulosic Sugar Production

Yang Yu, Ashraf Albahri, Alaa Alhammad, Lew Christopher

Biorefining Research Institute, Lakehead University, Canada, [email protected] Abstract: Worldwide pulp and paper production is about 300 to 350 million ton annually, of which 4.3% is lost in the form of pulp and paper sludge (PPS) because of the unsuitable short fibers for paper-making. PPS is usually disposed by landfilling or incineration. Landfilling is costly for about USD$30 per wet ton and the heating value of cellulose is low, making both methods less feasible for PPS disposal. Alternatively, PPS can be used as a feedstock for cellulosic sugar production, which both generates revenue for pulp/paper mills by selling sugars and reduces the disposing cost. Compared to the conventional feedstock, PPS contains lignin-limited short cellulose fibers, which have big surface area and are much more amenable to enzymatic hydrolysis. In addition, PPS is a negatively-valued reliable feedstock and the process of sugar production can be easily integrated into the pulp/paper processes. General conditions for enzymatic hydrolysis, such as temperature and pH, have been optimized by the enzyme producers but the feedstock-specific factors have to be determined. In this project, the possibility of using PPS as a feedstock for sugar production was studied by investigating the impact of several factors on enzymatic hydrolysis using the commercial enzyme cocktails Cellic® CTec2 (Novozymes, North America), such as washing, autoclaving, addition of surfactants, enzyme doses and feedstock concentration. It was demonstrated that pH adjustment of PPS was required for enzymatic hydrolysis but the sugar production was not improved by washing and autoclaving. Addition of surfactants improved sugar production and yields except 5% Tween80, and the best yields of 23.6% and 21.4% were observed by adding 1% Triton X100 and 5% PEG 8000 respectively. Sugar production and yields increased and the PPS liquefication time reduced by increasing enzyme doses. When PPS concentration increased, the sugar production increased but the yields decreased. By addition of PEG8000, sugar production and yields increased at lower but inhibited at higher enzyme doses and PPS concentrations possibly by the synergistic effect with glucose. The best sugar yield of 74.7% was achieved with 10% PPS concentration, 3% enzyme doses and 5% PEG8000. In conclusion, washing is not required for enzymatic hydrolysis of PPS but pH adjustment is needed. Both sugar production and yield increased, and the time for PPS liquefaction decreased by increasing enzyme doses. Sugar production increased but sugar yield decreased by increasing PPS concentrations. Surfactants increased sugar production and yields at lower but inhibited at higher enzyme doses and PPS concentrations. The trade-off balance between sugar production and yield can be levered with PPS concentration, enzyme dose, and the hydrolysis time. The mechanism of the effect of PEG8000 on hydrolysis and optimization of sugar production and yields will be studied in the future.

https://plus.google.com/u/1/114233691187432537864?prsrc=4




Development of Lipid Extraction Methods and Comparison of

Extraction Efficiency Using Fresh, Air Dried, and Lyophilized

Algal Biomass

Yanwen Wu1,2, Haipeng Guo1,3, Jinchi Zhang2, Wensheng Qin1* 1 Department of Biology, Lakehead University, Thunder Bay, Ontario, P7B 5E1, Canada,

[email protected] 2 Collaborative Innovation Center of Sustainable Forestry in Southern China of Jiangsu

Province, Nanjing Forestry University, Nanjing 210037, China 3 College of Life Sciences, Zhejiang University, Hangzhou 310058, China

Abstract: Global climate change and growing consumption of fossil fuels force humans to pay more and more attentions to the renewable and sustainable fuels. The biodiesel produced from agricultural crops, animal fat, and microalgae has been hotly studied as an alternative energy source. Microalgae are considered more promising than the other alternative sources for biodiesel production, owing to their high biomass yields, short growth time, and low requirements for land and freshwater, especially microalgae contain high contents of lipids. Lipid extraction has been considered as the technical key for the industrial process of microalgal biodiesel. Before lipid extraction, the wet microalgae biomasses are often dried. Various methods have been developed to dry the microalgae; however, few studies focused on optimization of parameters used in the process of drying and oil recovery rate. In this study, two species of oil-rich microalgae (Neochloris oleoabundans and Chlorococcum pamirum) were dried at 4, 22, 40, and 80 °C and then pretreated by using microwave and osmotic shock to evaluate the effects of different drying temperatures on the oil recovery rate. The fresh algae and non-pretreated samples were designed as the control. The total lipids from the two species of microalgae were extracted using the solvent of chloroform: methanol: water (1: 1: 0.9, v/v/v) and 1-butanol. The oil extraction yields using the same pretreatment method and extraction method from different temperatures for dried algae were compared. The results showed that there was no significant difference when the microalgae were dried at 4, 22, and 40 °C, while the oil extraction yield of microalgae dried at 80 °C exhibited a significant reduction. For instance, N. oleoabundans showed the oil extraction yield from 44.74 - 46.96 % when the drying temperature ranging from 4 - 40 °C, while the oil extraction yield was 36.74 % at 80 °C (osmotic shock & solvent of chloroform: methanol: water (1: 1: 0.9, v/v/v)). This study suggested that when algae were dried below 40 °C, the oil extraction yields were similar and stable, and significantly better than dried in higher temperature. This result can help guide industrial algal lipid extraction.


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