Highland Heights, OH, United States
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Carrino-Kyker S.R.,Case Western Reserve University | Swanson A.K.,Phycal, Llc | Burke D.J.,Case Western Reserve University
Aquatic Microbial Ecology | Year: 2011

Urban-rural gradients are important ecological settings to investigate how human land use impacts habitats and, indirectly, inherent organisms. Seasonally flooded vernal pools are common throughout temperate forest landscapes and may represent significant sites of forest nutrient cycling. The effects of urbanization on vernal pool microbial communities, important drivers of nutrient cycling, are largely unknown; thus studies to improve our understanding of microbes and their functional roles in these habitats are needed. Eukaryotic microbial communities sampled from 30 vernal pools of the Cuyahoga River watershed (USA), located along a gradient of urban land use, were profiled with denaturing gradient gel electrophoresis, and were compared between pools using nonmetric multidimensional scaling (NMS). Microbial diversity, and, specifically, the richness and diversity of the fungal operational taxonomic units (OTUs), increased with urbanization. Vernal pool eukaryotic microorganisms formed 2 NMS clusters that differed significantly in sub-watershed urban area. However, the significance of urbanization disappeared when fungal and algal communities were analyzed separately. Water conductivity was consistently correlated with different microbial communities (e.g. eukaryotic, fungal, and algal). Fungal communities also appeared related to the carbon content of the substrate, indicating that vegetation at a local scale may be important for community structure. Almost half of the OTUs matched fungal species, which provides taxonomic evidence that the eukaryotic microbial communities of vernal pools are dominated by fungal species. Overall, our data suggest that the eukaryotic microbes of vernal pools are influenced by a variety of factors of the surrounding landscape, including urbanization, water chemistry, and vegetation type. © Inter-Research 2011.


Grant
Agency: Department of Energy | Branch: | Program: STTR | Phase: Phase I | Award Amount: 147.94K | Year: 2010

The cost of dewatering particles is inversely proportional to particle size. Costs dramatically increase as particle size approaches 10


Leyva-Guerrero E.,Phycal, Llc | Narayanan N.N.,Phycal, Llc | Ihemere U.,Donald Danforth Plant Science Center | Sayre R.T.,Los Alamos National Laboratory
Current Opinion in Biotechnology | Year: 2012

Over two hundred and fifty million Africans rely on the starchy root crop cassava (Manihot esculenta) as their primary source of calories. Cassava roots, however, have the lowest protein:energy ratio of all the world's major staple crops. Furthermore, a typical cassava-based diet provides less than 10-20% of the required amounts of iron, zinc, vitamin A and vitamin E. The BioCassava Plus program employed modern biotechnologies to improve the health of Africans through development and delivery of novel cassava germplasm with increased nutrient levels. Here we describe the development of molecular strategies and their outcomes to meet minimum daily allowances for protein and iron in cassava based diets. We demonstrate that cyanogens play a central role in cassava nitrogen metabolism and that strategies employed to increase root protein levels result in reduced cyanogen levels in roots. We also demonstrate that enhancing root iron uptake has an impact on the expression of genes that regulate iron homeostasis in multiple tissues. These observations demonstrate the complex metabolic interactions involved in enhancing targeted nutrient levels in plants and identify potential new strategies for further enhancing nutrient levels in cassava. © 2011.


Tominaga H.,Mie University | Coury D.A.,Mie University | Coury D.A.,Phycal, Llc | Amano H.,Mie University | Kakinuma M.,Mie University
Ecotoxicology | Year: 2010

Synthesis and accumulation of molecular chaperones are universal responses found in all cellular organisms when exposed to a variety of unfavorable conditions. Heat shock protein 70 (Hsp70), which is one of the major classes of molecular chaperones, plays a particularly important role in cellular stress responses, and the Hsp70 system is the most intensely studied in higher plants and algae. Therefore, we isolated and characterized a cDNA clone encoding Hsp70 from a sterile strain of Ulva pertusa (Ulvales, Chlorophyta). The sterile U. pertusa Hsp70 (UpHsp70) cDNA consisted of 2,272 nucleotides and had an open reading frame encoding a polypeptide of 663 amino acid (AA) residues with a molecular mass of 71.7 kDa. Amino acid alignment and phylogenetic analysis of Hsp70s from other organisms showed that UpHsp70 was more similar to cytoplasmic Hsp70s from green algae and higher plants (≥75%) than to those from other algae and microorganisms. Southern blot analysis indicated that the sterile U. pertusa genome had at least four cytoplasmic Hsp70-encoding genes. UpHsp70 mRNA levels were significantly affected by diurnal changes, rapidly increased by high-temperature stress, and gradually increased by exposure to copper, cadmium, and lead. These results suggest that UpHsp70 plays particularly important roles in adaptation to high-temperature conditions and diurnal changes, and is potentially involved in tolerance to heavy metal toxicity. © 2010 Springer Science+Business Media, LLC.


Grant
Agency: NSF | Branch: Standard Grant | Program: | Phase: | Award Amount: 50.00K | Year: 2010

This Small Business Innovation Research Phase I project develops a system for the biosecure production of algal oils in open raceways while utilizing genetically modified algae. Through production of transgenic algae containing specific gene regulatory sequences, this research develops algal strains that conditionally express an essential chloroplast-encoded gene only in the presence of the specific inducer compound. Thus, transgenic algal strains would survive only in the presence of the inducer compound (which will be added to the ponds in our production facility). On escape from the production facility, these algae will stop production of the essential target protein and be unable to reproduce in the environment. This biosecure production system will allow the use of genetically modified algae in open pond production systems without fear of impacting the surrounding systems.

The broader impact of the proposed research will be to enable the use of low cost, open-pond culturing systems for the production of biofuels using microalgae genetically manipulated for maximum oil and biomass production. This will be accomplished in a biosecure manner through the development of molecular biological strategies that prevent the genetically manipulated algal strains from reproducing once they are removed from the production ponds. The use of molecular genetics to both increase the production of oil and ensure biosecure use of the transgenic algae (unable to replicate in the surrounding environment) could enable cost effective production of biofuels using algal systems.


Grant
Agency: NSF | Branch: Standard Grant | Program: | Phase: | Award Amount: 495.76K | Year: 2012

This Small Business Innovation Research Phase II project develops novel technologies for separation and concentration of intrinsically magnetically susceptible algae for production of biofuels and biochemicals. Phase II builds on the feasibility demonstrated in Phase I using a model alga. During Phase II, an algal strain used for production of renewable biofuel feedstock will be utilized. Novel transformation vectors and tools developed for a production strain, Auxenochlorella protothecoides, will be used to make the algae magnetically susceptible. These traits provided an advantage vs. wild-type strains in growth in low iron medium for the model alga. Phase II will test modified algal strains at lab- and subpilot-scale to determine their performance in growth, and competition with wild-type and weedy algal strains. Additionally, strains will be tested for their ability to be separated or harvested magnetically. This separation will be modeled to determine cost efficacy for primary or secondary dewatering. The specificity of this separation will also be evaluated in relation to downstream use in a heterotrophic bioreactor. The OSU collaboration allows use of these strains in novel rare earth magnetic separators. The endpoint will be novel technologies to improve the overall cost structure for the production of algae-derived biofuels and biochemicals.

The broader impact/commercial potential of this Phase 2 research project will be to provide improvements in the economics of producing renewable biofuels using algae as the production system. It directly addresses one of the major issues with algal biofuels, cost effective dewatering. It also provides a potential selective advantage of the modified strains by improving its ability to compete for iron in an open environment (such as open raceways or photobioreactors). The nation has a critical need to improve its energy security and reduce its dependence on fossil fuels. This research will help address both of these needs. The overall purpose of this research project is business related and focused on commercialization of this technology through integration in a biofuel production process. This research project focuses on a high cost portion of the production process, dewatering, as well as a critical unit process, the heterotrophic bioreactor. The collaboration with OSU and the Cleveland Clinic will result in training of students in this area. The company plans to publish the results of this project once proper control of the intellectual property generated is accomplished.


Grant
Agency: Department of Energy | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 100.00K | Year: 2010

Extraction and distillation of oil from algae can be as much as 40% of the total production cost of algal oil potentially rendering it uneconomical as a feedstock for biofuels. Significant contributors to these high cost and energy requirements of extraction and distillation are the very dilute nature of algal cultures, small algal cell sizes, and tough algal cell walls. For extraction, this usually mandates some type of pre-concentration (a.k.a. dewatering or harvesting), aggressive lysing methods, and high ratios of aggressive solvents. This, in turn, requires expensive equipment and a large amount of energy and distillation. Altogether, traditional extraction/distillation methods are not cost effective necessitating the implementation of innovative approaches to produce algal oil at mainland market competitive prices. This proposal develops a method for economical algal oil extraction based on a solid phase lipid extraction using oil absorbing glass. This glass matrix could be used to either directly absorb oil from lysed cells or differentially absorb the oil from emulsions formed between solvent / water / lipid post liquid / liquid extraction. Direct absorption from algal cells could eliminate or mitigate solvent use in the extraction method. The differential extraction of algal lipids from emulsions could eliminate or mitigate separation major problem in liquid/liquid extractions where emulsions of algal oil with solvent and water often form. The objective at the end of Phase II is the development of optimized extraction methods and associated equipment based on the use of an optimized absorbent glass matrix for algal oil extraction in a prototype deployed in our integrated algal biorefinery pilot plant. Commercial Applications & Other Benefits: If successful, this research project will be developed into a turn-key extraction device for use by other algae companies, not just Phycal, to stimulate the entire algal biofuel industry. Economical production of algal oil has major benefits for U.S. energy, environmental, and job security. If this approach could reduce the overall cost associated with extraction and potentially distillation of algal oils it would have a major impact on the economics of algal oil as a feedstock for energy products.


Grant
Agency: NSF | Branch: Standard Grant | Program: | Phase: | Award Amount: 180.00K | Year: 2010

This SBIR Phase I project will develop a system for magnetic separation from growth media and cell disruption of algae to improve the efficiency of harvesting algae for bio-fuels production. The use of magnetic technologies has the potential to improve the energy efficiency of removing algae from growth media. The use of a magnetic separation technology introduces an energy efficient means by which algae can be dewatered and converted into bio-fuels in an integrated bio-refinery. Using algal strains modified to overproduce ferritin (a magnetically susceptible bio-particle) enables the removal of algae from contaminating biomass as well.

The broader/commercial impact of the project will be magnetic harvesting and dewatering of algal product which could lead to higher efficiencies of oil production. This will enable the use of low cost open pond culturing systems for the production of biofuels using microalgae. The development of technologies using magnetically responsive algae (either
direct, or through association with magnetically responsive particles) to either separate them from growth media and contaminants or to induce cell lysis and aide in oil extraction. The project targets the two highest cost factors in algal oil production, namely dewatering and extraction. As such, advances in these two areas can drive algal based biofuels technology to be
cost competitive with petroleum sources.


Grant
Agency: Department of Energy | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 1.00M | Year: 2011

Extraction is a major cost driver in the production of algal oil. Reducing extraction cost is critical for the economic viability of algal biofuels. This project uses solid phase, hydrophobized silica resin to improve extraction of algal lipids. Process streams containing lipid and other hydrophobic compounds are treated to recover lipids and Corp) for extraction of the algal lipid produced after the Heteroboost process. Hydrophobized resins were evaluated with model triacylglycerides (TAGs) and fatty acids (FFA) to efficacy and selectivity of adsorption of TAGs and FFAs. Resins were then tested in complex mixtures of algal lipids, with or without algal cell debris or biomass. The inability to separate silica from cell debris cleanly ruled out use in these complex mixtures. However, the resins effectively removed algal TAGs and FFAs from clarified aqueous process liquids generated in Phycals aqueous extraction process. Additionally, this efficacy was maintained for 10 adsorption/desorption cycles. Phase II will focus on recovery of hydrophobic compounds from two process streams generated by Phycals aqueous extraction process. The ability to capture algal oil that is currently lost directly improves production cost. Recovering polar lipids that are not currently captured in aqueous extraction will provide additional cost savings. The project will focus on validation of the adsorption/desorption process at the subpilot, documentation of any higher value hydrophobic compounds recovered from this process, scale up of the process to the pilot plant, and develop a cost analysis/model to allow transition to the commercial plants to be constructed in Hawaii. The deliverables will be an optimized reactor design for adsorption/desorption on hydrophobized resins, optimized silica resin, optimized process at subpilot-scale, and field testing of the optimized reactor/methods at pilot-scale. Commercial Applications & amp; Other Benefits: Phycal is constructing a pilot-scale algal oil production facility on the Island of Oahu, Hawaii. This facility is designed to incorporate our existing production methods, but also to be available for testing improvements to these methods that will lower the cost of the algal biofuels that will be produced. This project could have a significant and positive impact on the cost of production of the algal oil by reducing the loss of lipid in process wastestreams and capturing of other hydrophobic compounds that have value either as an additional fuel feedstock or higher value product.


Grant
Agency: National Science Foundation | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 100.00K | Year: 2009

This Small Business Innovation Research Phase I project develops a system for the biosecure production of algal oils in open raceways while utilizing genetically modified algae. Through production of transgenic algae containing specific gene regulatory sequences, this research develops algal strains that conditionally express an essential chloroplast-encoded gene only in the presence of the specific inducer compound. Thus, transgenic algal strains would survive only in the presence of the inducer compound (which will be added to the ponds in our production facility). On escape from the production facility, these algae will stop production of the essential target protein and be unable to reproduce in the environment. This biosecure production system will allow the use of genetically modified algae in open pond production systems without fear of impacting the surrounding systems. The broader impact of the proposed research will be to enable the use of low cost, open-pond culturing systems for the production of biofuels using microalgae genetically manipulated for maximum oil and biomass production. This will be accomplished in a biosecure manner through the development of molecular biological strategies that prevent the genetically manipulated algal strains from reproducing once they are removed from the production ponds. The use of molecular genetics to both increase the production of oil and ensure biosecure use of the transgenic algae (unable to replicate in the surrounding environment) could enable cost effective production of biofuels using algal systems. This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).

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