SERIISP-231-3579 UC Category: 244 BE89009472 Aquatic Species Program Annual Report W. S. Bollmeier S. Sprague September 1989 Prepared under Task No. BF911010 FTP No. 656 Solar Energy Research lnstitute A Division of Midwest Research Institute 1617 Cole Boulevard Golden, Colorado 80401-3393 Prepared for the U.S. Department of Energy Contract No. DE-AC02-83CH10093 NOTICE This report was prepared as an account of work sponsored by an agency of the United States government. Neither the United States government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, com- pleteness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not necessarily con- stitute or imply its endorsement, recommendation, or favoring by the United States government or any agency thereof. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States government or any agency thereof. Printed in the United States of America Available from: National Technical lnformation Service U.S. Department of Commerce 5285 Port Royal Road Springfield, VA 22161 Price: Microfiche A01 Printed Copy A08 Codes are used for pricing all publications. The code is determined by the number of pages in the publication. lnformation pertaining to the pricing codes can be found in the current issue of the following publications which are generally available in most libraries: Energy Research Abstracts (ERA); Govern- ment Reports Announcements and Index (GRA and I); Scientificand Technical Abstract Reports (STAR); and publication NTIS-PR-360 available from NTlS at the above address. This report summarizes the progress and research accomplishments of the Aquatic Species Program, field managed by the Solar Energy Research Institute (SERI), through May 1989. This report includes an overview of the entire program and a summary of individual research projects. The program receives its funding through the Biofuels and Municipal Waste Technology Division of the Department of Energy. For further details, contact the SERI Biofuels Program Office Aquatic Species Program Coordinator, Warren Bollmeier, at (303) 231-7669. iii SUMMARY When stimulated by environmental stress, many species of aquatic microalgae produce lipids, or oils, that can be processed into diesel oil or gasoline. These algae have growth rates as high as five times those of most terrestrial plants, and some species flourish in saline or brackish water unsuitable for human. or traditional agricultural use. In addition, microalgae require large quantities of carbon dioxide for growth and lipid production, offering a way to mitigate increases in the concentration of atmospheric carbon dioxide, a major greenhouse gas. The Aquatic Species Program, funded by the Department of Energy and managed by the Solar Energy Research Institute, was designed to take advantage of these attributes. The overall goal of the program is to provide the technology base for the cost effective production of diesel fuel from microalgae by the year 2010. The specific objectives are to identify or develop strains and ,culture (growth) requirements for high biomass and lipid production under fluctuating outdoor conditions, to develop design criteria for large scale culture systems, and to develop efficient harvesting, handling and conversion methods. Moreover, since the first applications of this technology are most likely to occur in the U. S. southwest where there are high levels of solar radiation, a large land base, and an inexpensive water source (saline), the research is driven by the conditions found in this area. Significant progress has been made since the program focused exclusively on the production of fuels from microalgae in 1982. More than 3000 strains collected from desert and saline environments have been studied, resulting in the selection of numerous strains for improvement or development research. The strains selected grow over a wide range of salinities, produce significant quantities of lipid oils and achieve growth rates of nearly three doublings per day. Some of these strains can tolerate freezing temperatures and some as high as 100 degrees (F) or higher. The most promising of these algal strains are maintained in a culture collection at SERI. This is the only collection of algae in the country that is focused on oil products for processing to fuels and serves as a resource for the program. However, many of these strains are distributed to industrial users, including shellfish growers and others interested in exploring microalgae for high- value products. Analyses indicate that economic fuel production will require the microalgae to be grown in intensive culture in large outdoor ponds. The program has established an Outdoor Test Facility (OW) to evaluate pond designs and to translate laboratory successes in strain development to outdoor conditions. The preliminary design consists of a 6-m-deep, raceway-shaped pond (0.25 acre) with a paddle wheel to circulate the water and systems for injecting carbon dioxide and other nutrients into the culture. The OTF became fully operational in 1988 and has two of the 0.25-acre ponds. Recognizing the increased concern with global warming and the fact that large additions of carbon dioxide are required to obtain the production levels required for the technology, SERI has conducted a preliminary analysis coupling microalgae production facilities with power plants in the southwest. The preliminary results are promising and indicate that the carbon dioxide emissions from all the power plants in New Mexico and Arizona could be trapped by microalgae farms covering about 4% of the combined land area of both states. Ongoing analysis will be used to guide and focus the research and to identify opportunities and other potential benefits of the technology. Research is now focused on applying genetic techniques to enhance the lipid production of microalgae. This effort builds on extensive strain characterization research, as well as biochemical studies of the metabolic pathways for lipid synthesis. Using nongenetic methods, scientists have already improved the lipid content of the cell from the 5%-20% found in nature to more than 60% in the laboratory and 40% in outdoor culture. Recent results suggest that microalgae production goals of 70 dry tons per acre per year and lipid content of 60% can be achieved in outdoor culture. Research results suggest that if these goals are reached, oil produced from microalgae could be competitive with petroleum after the year 2010. TABLE OF CONTENTS Page PROGRAM OVERVIEW .................................................... 1 SERI Program Overview Warren Bollmeier; SERI ........................................... 3 TECHNICAL AND ECONOMIC ISSUES ....................................... 7 Microalgal Mass Culture and the Greenhouse Effect: Resources Update Paulanne Chelf, Lewis M. Brown; SERI .............................. 9 C02 Sources for Microalgae-Based Liquid Fuel Production Daniel A. Feinberg, Michael E. Karpuk; TDA, Inc. .................... 17 The Economic Impact of Brine Disposal on Algal Fuel Facilities Pau1Bergeron;SERI .............................................. 21 Design and Operation of an Outdoor Microalgae Test Facility Joseph C. Weissman, David M. Tillett; Microbial Products, Inc. ......... 41 ORGANISM EVALUATION ................................................. 59 Culture Collection Status LewisM. Brown;SERI ............................................ 61 Characterization of Growth and Lipid Yield in Microalgae from the Southwest Using High Salinity Media S. B. Ellingson, P. L. Tyler, M. R. Sommerfield; Arizona State University ....................................................... 71 GENETICS ............................................................... 83 Strategies for Genetic Improvement of Microalgae with Ability to Grow in Outdoor Mass Culture Lewis M. Brown, Terri G. Dunahay, Eric E. Jarvis; SERI ............... 85 The Mitochondria1 Genome of the Exsymbiotic Chlorella-Like Green Alga NlA James A. Waddle, Anne M. Schuster, Kit W. Lee, Russell H. Meints; University of Nebraska-Lincoln .................... 9 1 vii TABLE OF CONTENTS (concluded) Page LIPID BIOCHEMISTRY ..................................................... 1 19 Purification and Characterization of Acetyl-CoA Carboxylase from ..he Diatom Cyctotella cryptica PaulG.Roessler;SERI ............................................ 121 Triglyceride Accumulation and the Cell Cycle in Microalgae K. E. Cooksey, J. B. Guckert, R. Thomas; Montana State University ....................................................... 135 CONVERSION ............................................................ 155 Microalgal Fuel Production Processes: Analysis of Lipid Extraction and Conversion Methods Nick Nagle, Peter Lemke; SERI .................................... 157 viii 1.0 PROGRAM OVERVIEW Researchers in the Aquatics Species Program focus on the use of microalgae as a feedstock for producing renewable, high-energy liquid fuels such as diesel. It is important for the United States to develop alternative renewable oil sources because 42% of the current energy market in the United States is for liquid fuels, and 38% of these fuels are imported.