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An interesting finding from the cold water strain collection project was that many species that predominate after the enrichment procedure were the same as the warm water species selected in previous collection efforts. The genera and species that were commonly found in both the cold water and warm water screening projects were C. muelleri, Amphora coffeiformis, Cyclotella, Navicula, and Nitzschia. However, some ochromonids and green coccoid algae were also isolated from the cold water...
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iiiBg efficiency. The author claimed that this was 50 to 100 better than achieved with conventional culture techniques, though the basis for such extrapolations or comparisons was unclear. III.B.2.d. Fourth Year of the Hawaii ARPS Project, 1983-1984 During this year a number of experiments were carried out to determine productivity of the cultures as a function of several variables, this time studied independently Laws 1984b,c see also Laws et al. 1985 . One task was to screen candidate species...
Laboratory Studies Aet
The objectives of this subcontract were to collect microalgal strains from a variety of locations iC the'desert regions' of an to screen them for their ability to grow under conditions in a commercial microalgal biodiesel facility. Studies were also conducted to optimize a fluorometric procedure for CstBecttiig caps p co entwi toApii thi9 85etllll d Ju gregl8a .mew attehg samps containing microalgae were collected from 125 sites in Arizona, California, Nevada, New Mexico, Texas, and Utah. Some...
Technical Highlights of the Program
A unique collection of oil-producing microalgae. The ASP studied a fairly specific aspect of algae their ability to produce natural oils. Researchers not only concerned themselves with finding algae that produced a lot of oil, but also with algae that grow under severe conditions extremes of temperature, pH and salinity. At the outset of the program, no collections existed that either emphasized or characterized algae in terms of these constraints. Early on, researchers set out to build such a...
IIA Collection Screening and Characterization of Microiigde IIA Collection
Optimal Medium Type dependent on temperature and conductivity used V.A. SERI NREL DOE REPORTS AND PUBLICATIONS V.B. ADDITIONAL REFERENCES V.C. PATENTS II.A. Collection, Screening, and Characterization of Microalgaf II.A.1. Collection, Screening, and Characterization of Microalgae by SERIIn-House Researchers Optimal Medium Type dependent on temperature and conductivity used
Future Directions
Put less emphasis on outdoor field demonstrations and more on basic biology Much work remains to be done on a fundamental level to maximize the overall productivity of algae mass culture systems. The bulk of this work is probably best done in the laboratory. The results of this program's demonstration activities have proven the concept of outdoor open pond production of algae. While it is important to continue a certain amount of field work, small scale studies and research on the A Look Back...
Subcontractors
Included in this section are summaries of the research conducted by various subcontractors within the ASP who contributed to the collection, screening, and characterization of microalgal strains for potential use in biofuel production facilities. Initially, a variety of strain isolation and screening procedures were carried out by the various research groups, as there was no established protocol. This lack of uniformity in the screening protocols made comparing the results from one laboratory...
IIIB Introduction
The long-term objective of the ASP was to develop microalgae liquid fuel production processes. Since its inception, the ASP supported laboratory R amp D projects Section II and algal mass culture projects. However, for the most part, the laboratory and outdoor projects were not integrated into a strongly unified program. This reflects in large part the difficulty of such integration. Also, during the early stages of the ASP, too close an integration would have been restrictive, as it was not...
Laboratory Studies Qzz
Zzyzx medium before algal isolation see Thomas et al. 1986 for media compositions . artificial media that were designed to mimic the water from which the strains were originally obtained. The pH of these various media formulations was typically high because of the presence of high levels of carbonate and bicarbonate, and the total dissolved solids ranged from approximately 1.5 g- L to over 260 g- L . The growth of the cultures was visually scored, and nine of the fastest growing strains For...
IIID NREL Studies of Flue Gas CO Utilization by Microalgae
The last major engineering-related activity carried out by the ASP was a PETC-funded study, both laboratory and process design, for microalgae biodiesel production using power plant flue gases. The general arguments for use of microalgae in CO2 mitigation, from high productivity to relatively low water use were reviewed in several reports Brown et al. 1991 Chelf et al. 1991 . The authors concluded that the SOx and NOx impurities in flue gas would likely not have a major effect on algal...
IVA Microalgal Strain Improvement IVA Conclusions
From the early 1980s through the mid-1990s, there was a major effort by ASP researchers and contractors to identify or develop microalgal strains that demonstrated properties conducive to cost-effective biomass and lipid production. The characteristics deemed desirable in these strains include high productivity, high lipid content, competitiveness in outdoor culture, and tolerance to fluctuations in temperature and salinity. Although a number of strains were identified as possible candidates,...
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promotes the accumulation of lipids in certain species. Green algae are the evolutionary progenitors of higher plants, and, as such, have received more attention than other groups of algae. A member of this group, Chlamydomonas reinhardtii and closely related species has been studied very extensively, in part because of its ability to control sexual reproducaiOhh- raU n Ajpwi This geOlipl o f ai gh ,ticamlna yi efe rreIh 4ae s Cfteys yjhym hass wwams ilathe tofirdtatote wo...
IIID Design and Analysis of Microalgae Open Pond Systems
This report Weissman and Goebel 1987 originated from a competition held by the ASP for the development of a pilot plant test facility for microalgae production. As mentioned in Section III.A.4., two companies were selected to develop competing processes Aquasearch, Inc., of San Diego, California, and Microbial Products, Inc., which had carried out the ASP pond project in California. The objectives were to arrive at cost projections for such a test facility, and also for scale-up costs of a...
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disproportionate decrease in the rate of lipid synthesis versus carbohydrate synthesis, and Fra amp ionatiAsbfwlh Stivp dsi cpro funcedl iptdrcst s crattdgifll aahantS 1 lipepsivation resulted in an increase in the proportion of total lipid as neutral lipids, primarily TAGs, from 43 to 63 after only 4 hours of Si deficiency. Analysis of the fatty acid composition of the accumulated lipids also showed changes induced by Si starvation. In Si-deficient cells, there was an increase in the...
producers and for genetic manipulation of microalgae by classical breeding or
Historically, microalgae have been classified based on morphological similarities. enetic engineering. revious studies by Dr. Gallagher and others see Gallagher 1986 suggested significant physiological variability between isolates of a single species. In these studies, various isolates of a species grown under identical conditions to control for environmentally induced changes in gene expression often showed significant differences in characteristics such as nutrient uptake, growth rates, and...
IIIB The Israeli Microalgae Biodiesel Production Project
In the mid-1980s an algal mass culture project for biodiesel production was supported by the ASP in Israel Arad 1984, 1985, 1986 , as a cooperative project amo1ng the M eMnce gnogpsphic and Limnological Research Institute, with Dr. Ben-Amotz, who investigated lipid production at the laboratory and micropond scale 2 Ben-Gurion University of the Negev, with Professor A. Richmond as Principal Investigator, investigating algal mass cultures with outdoor ponds, essentially of the HRP design. 3...
SERIInHouse Researchers
Ben-Amotz, A. Tornabene, T.G. 1983 Chemical profile of algae with emphasis on lipids of microalgae. Aquatic Species Program Review Proceedings of the March 1983 Principal Investigators' Meeting, Solar Energy Research Institute, Golden, -6Gp homaSPW .H. 1985 Chemical profiles of selected species of microalgae with emphasis on lipids. J. Phycol. 21 72-81. Tornabene, T.G. Holzer, G. Peterson, S.L. 1980 Lipid profile of the halophilic alga, Dunaliella salina. Biochem. Biophys. Res. Comm. 96...
IIIA An Integrated System for the Conversion of Solar Energy withSewageGrown
During FY1976-1977, the project described in Section III.A.2. continued with the same overall objective to determine what pond operating factors could allow control over algal species, and thus permit cultivation of algal types that allow low-cost harvesting by microstrainers. The biomass recycling process described earlier continued to be studied, using the 12-m rectangular ponds and various pond AherllbteIfsS0eaae igi 0fxirprtspnga s rWgtghtidDliliimesahi lnllaa smcarc 1gp Chdaiowith dgsy...
Laboratory Studies Tkd
The goal of this research was to understand the biochemistry of lipid accumulation in microalgae, in particular, to provide information on the biochemical triggers that induce lipid synthesis. The Nile Red fluorescence technique developed by Dr. Cooksey's laboratory and at SERI described above in Sections II.A. 1 .f. and III.B. 1 .e. was used to study lipid accumulation in microalgae in response to N or Si depletion. Nile Red fluorescence was used to monitor the lipid levels in batch cultures...
Table IIIA Microalgae harvesting by bioflocculation Small and large pond results
Comparison of performance between two 1,000-m and two 12-m ponds, operated under otherwise similar conditions except that one pond was screened to remove zooplankton and the other one was not so treated. Data averaged for July 1978. Note agreement in terms of nrodnctivitv grams VSS m d. essentially some 90 AFDW
Fertilizer
This 2-year project Benemann et al. 1979 Eisenberg et al. 1980 extended from the end of 1977 to Winter 1979. Initially the approach to establish microstrainable cultures using the 12-m ponds, described in Section III.A.3., continued to be investigated. Essentially the same results as before were obtained detention time was found to be the key environmental variable determining algal colony size but not necessarily species composition and a negative correlation was found between numbers of algal...
Laboratory Studies Uvj
The purpose of this project was to determine if flow cytometry could be used to select for subpopulations of high lipid-producing algae within an algal culture. Flow cytometry is a method that measures the light scattered or emitted by particles as they pass through a laser beam. Scattered light is believed to reflect the size, shape, and refractive properties of cells. Dr. Solomon initially used exponentially growing and nutrient-stressed cells of the chrysophycean alga Boekelovia to...
IIA Collection Screening and Characterization of Microaiga IIA Collection
Transient Expression of Luciferase in Chlorella ellipsoidea The first step in transformating any organism is getting the foreign DNA inside the cell. For organisms with a cell wall, methods must be devised to either remove or permeabilize the wall, or to get DNA into the cell through the intact wall. Bacterial cell walls do not seem to represent a significant barrier to DNA uptake, and can be induced to take up foreign DNA simply by being washed in low osmotic medium and glycerol, followed by a...
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Figure II.B. 11. Western blot, showing the presence of the 30-kDa NPTII protein in transformed cells of C. cryptica T13L and N. saprophila. In this example, C. cryptica and N. saprophila were transformed with pACCNPT10 and pACCNPT5.1, respectively. Crude cell extracts were separated on SDS-polyacrylamide gels, blotted onto a nitrocellulose filter, and NPTII protein was detected using anti-NPTII primary antibodies and alkaline phosphatase-conjugated goat anti-rabbit IgG secondary antibodies. The...
Collection Screening and Characterization of Microalgae
Collection, Screening, and Characterization of Microalgae by SERIIn-House Researchers SERI researchers also started to evaluate various strains by the rapid screening procedure. Initial work focused on 25 partially characterized strains. These strains were analyzed for growth and Nile Red fluorescence in exponentially growing cultures and in cultures grown under N-deficient conditions for 4 days. The results of the SERI and Sommerfeld laboratories cannot be compared directly, because Nile Red...