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Cold water algae for biofuels – a new source of alternative energy


Leader Yoshihiro Shiraiwa, University of Tsukuba
Researcher Tony Haymet, University of California San Diego Peter Wilson, University of Tasmania
Term April 2014 - March 2015
Research Outline

World-class skills are available at UCSD, the University of Tasmania (UTAS) and the University of Tsukuba (UT) to determine the value of further research into the use of cold water algae as a source of biofuels for the planet. Biofuels research is currently not keeping pace with expected future energy production requirements. Attention has turned to the promising alternative of single-celled algae which grow quickly, need few added nutrients, can be grown on a very large scale and take C02 from the air as part of their growth process. Compared with crops normally used to produce oil such as soybeans or palm, algae can produce over 30 times the amount of oil per acre. Society needs to immediately begin the development of molecular and genetic tools that will allow algae to become an economically viable biofuel source.
• The San Diego Center for Algae Biotechnology (and in particular the Mayfield group at the UCSD) is of prime importance in this area
• There is a large algae library/culture bank associated with both UTAS and nearby CSIRO Marine Research in Hobart
• The University of Tsukuba (the M. Watanabe group, focusing more fresh water microalgae, and the Shiraiwa group, focusing more marine microalgae, in the Faculty of Life and Environmental Sciences) are arguably the world leaders in the science surrounding microalgae and engineering such algae for industrial use.

Each of these groups has a long history of successful study of microalgae and biomass production, but has different and complimentary foci in their research. Their respective research activities are currently outside-funded by a variety of agencies (MEXT, CREST/JST, NSF and DOE) but neither group is currently looking at cold water algae in any detail. Cold water strains are very important to optimize algal biofuel production in well-developed countries which need renewable energy source and where locate under cold climate. Recent algal blooms at high latitudes have suggested that climate change is causing more favorable conditions for large scale growth of algae in clean, cold waters. Both groups however currently have monocultures of various polar algae species and it is only through research cooperation, such as an intensive workshop, where rapid progress will be made. The Shiraiwa group at UT is currently analyzing the ice binding properties of various algae and diatoms and liaising with several overseas agencies (such as the Alfred Wegener Institute in Germany and the Ben Group at the University of Ottawa). Even after Dr. Peter Wilson moved to Australia, we are keeping close scientific collaboration on this project and therefore are continuing to include him as an important member. In order for Japan, Australia and the USA to be together at the forefront of biofuels science and production it is essential to work more closely and quickly determine the best algal species and culture conditions for large scale growth.


Leader Yoshihiro Shiraiwa, University of Tsukuba
Researcher Tony Haymet, University of California San Diego Peter Wilson, University of Tasmania
Term April 2014 - March 2015
Achievements Outline

Professor Wilson’s presentation focused on the effects of freeze-thaw cycles on cold water algae, claiming that biofuel research is currently not keeping pace with the expected future. Recent algal blooms at high latitude have suggested that climate change is causing more favorable conditions for large scale growth of algae in clean, cold water. Furthermore, extracellular macromolecules associated with both Arctic and Antarctic sea ice diatoms have been shown to have ice-binding activities. It has also been found that in many cases viability of species is higher when cold environments are present. However laboratory tests have shown that reproducibility can be decreased by 10% after a single freeze/thaw cycle and this has been attributed to sample degradation and not to instrument variability. The question, then, is whether or not ice growing on surface of potential growth ponds might affect reproducibility.

Professor Wilson outlined an experiment where he tested the nucleation temperature for a given sample of water, with and without algae present. Those algae included both polar and temperate species. He concluded that more than 30 freeze thaw cycles did not change the efficiency of the nucleating site. This was evidenced because the survival curve of the data plotted was still the same shape and asymptotic at each end.

Activity Contents

The following conclusions were drawn from the experiment:
- The presence of diatoms in water increased the temperature for ice formation up to 6 degrees Celsius and up to 10 degrees Celsius in salt water
- Active polar diatoms do not cope well with freezing and multiple freeze/thaw cycles

Policy Paper

Leader Yoshihiro Shiraiwa, University of Tsukuba
Researcher Tony Haymet, University of California San Diego Peter Wilson, University of Tasmania
Term April 2014 - March 2015
Title Algae for biofuels - cold water, inland in ponds or in the lab?

Cold water algae for biofuels are increasingly becoming a new source of alternative energy.  Due to global warming, increases in atmospheric CO2 concentrations cause ocean acidification.  In Florida’s case, for example, a vast amount of coral reefs are being broken up by acidification.  This especially affects high latitude areas, notably Arctic regions.  As a result, Arctic sea ice size is becoming smaller year by year and the thickness of ice in Arctic regions is presently no higher than 5 meters.  The goal, therefore, is to stop the increase of CO2 concentrations.

An observable algae bloom has been occurring in the world’s oceans over the past few years, leading to an explosion of algae research for biofuel production. Accordingly, microalgae photosynthesis is very effective in reducing CO2 levels in the atmosphere.  One such type of microalgae, cocclithophorids, have calcium carbonate crystals on the cell surface.  As such, the inside of cell can became petroleum, which is a source of crude oil.

With regard to determining if cold water algae are viable for use as biofuel stock, firstly we must determine their behavior in ice laden waters.  Much of the period to date has been spent determining who, around the world, has stocks and cultures of cold water microalgae which might be used to assay for antifreeze and ice binding proteins before we look at lipid content and biofuels possibilities.  These algae must withstand multiple freeze-thaw cycles to be able to be used as a source of biofuels in cold surface waters at high latitudes.

  1. The Alfred Wegener Institute in Bremerhaven, Germany studies antifreeze and salt stress tolerant proteins from sea ice diatoms. They note that “some polar diatoms (e.g. Fragilariopsis species)” are able to grow and to divide below freezing temperature and above average sea water salinity. Their work includes a patent application in which there are three nucleic acid sequences coding for salt and cold stress and their respectively proteins.  Dr M. Bayer-Giraldi, who works on this at AWI, and together with Professor Peter Wilson, is working on this and other projects and have they recently written a book chapter together.
  2. In Australia CSIRO houses the Australian National Algae Culture Collection (ANACC) at the Marine and Atmospheric Research laboratories located in Hobart, Tasmania. ANACC maintains over 950 strains, and is a major culture collection in the Australasian region. It is housed in a world-class algal culture facility, the collection is a research resource for investigations into the growth and physiology, taxonomy, biochemistry and molecular genetics of microalgae.  It is managed by Dr Susan Blackburn and both Professor Peter Wilson and Professor Tony Haymet have been in contact with Susan recently about collaborating and using the cultures.
  3. The University of Tasmania is studying the microbial communities inhabiting sea ice ecosystems which currently contribute 10–50% of the annual primary production of polar seas. Brine algae collected from McMurdo Sound (Antarctica) sea ice has been incubated in situ under various carbonate chemistry conditions. They find that projected increases in seawater pCO2, will not adversely impact brine algal communities. Professor Wilson now works at the same institution as Professor Andrew McMinn, the PI on this work.  Professor Haymet, from UCSD Scripps Institution of Oceanography, and Professor Wilson continue to collaborate closely on this project and meet every few months to discuss.
  4. The Japan Science and Technology Agency is currently running a 5-year project to create new basic technologies for bioenergy production using algae and other microorganisms. There are two types of algae: microalgae, which are made up of a single cell, mostly made of cellulose, and cannot produce high amounts of hydrogen, but have more potential to produce biofuels; and macroalgae, which are large algae that can produce hydrocarbons. Microalgae can produce drop-in fuel, which is useful for driving cars by mixing it with gasoline (if carbon chain is liquid, then it can directly be used as fuel).  Professor Shiraiwa has stated that plant oil produces tri-acyl glycerol (TAG) which contains fatty acids. In order to use TAG and produce biodiesel, large amounts of methanol are needed.

The focus of Professor Shiraiwa’s research has been the Emiliania huxleyi bloom in the Bering Sea near Alaska, which has proven to be a useful organism for biofuel production.  Emiliania huxleyi is the best-suited candidate, as alkenes and alkenones are dominant, which enables us to get more crude oil components.  At the University of Tsukuba experiments underway have tried to produce more of this compound using genetic engineering. It is different from other microalgae, as it produces the least amount of lipids (straight chain) and contains no neutral polysaccharides. It is also unique in that it has two pathways: a plant type and animal type of metabolism for production of DHA.  Researchers have collected hundreds of strains of culture collection and have found that the content of these lipid oils (alkenes) is different among strains; as such, they are able to select the best strain and pick the best organism from that.  For example, strains that produce C9-18 alkanes can be used for jet fuel.  The Mirai cruise in 2010 to the Arctic Ocean succeeded in collecting high-latitude strains of E. huxleyi, which is able to survive in cold water and produce lipids as storage compound.

  1. Scripps Institute of Oceanography at the University of California San Diego have carried out experiments which were conducted on Mauna Loa in Hawaii, where CO2 measurements have increased over 40% since 1960, now at over 400 ppm (parts per million). Carbon that’s been buried in the earth has been depleted due to carbon 13 and carbon 14, and fluctuation in CO2 levels is in part due to change in vegetation and the burning of fossil fuels.  It is clear that on Earth we want to be able to burn liquid fuel, but don’t have sufficiently high energy density to fly planes, etc. solely on CO2; as a result, we need to limit the use of liquid fuels. Algae, therefore, is likely  to be the answer to this problem. Algae takes sunlight and CO2 and turns it into fat and protein. Therefore, growing algae on the surface of the earth could be used to reduce CO2 levels. Earth, however, is 71% water, and with one side almost all ocean, there is a lot of potential of growing algae on large bodies of water. Algae, unfortunately, are the least studied of plants, but can be used to solve our liquid fuel problem. Professor Haymet has pointed out that the US Department of Energy is planning to save microalgae biofuels with oil at $55 per barrel, giving the following suggestions:
  • A need for high-value secondary products
  • Scripps researchers are making progress with animal and fish vaccines, which are required at high volumes, and hence perfect for mass production by algae
  • Algae are adept at making things in vast volumes
  • While making fuel simultaneously, competition is nonexistent

The high productivity of microalgae can be used for biotechnological applications, namely biofuels. Diatoms are excellent lipid accumulators and were the featured organism in early US biofuels efforts. AT UCSD the laboratory of Dr Mark Hildebrand has pioneered the application of genetic manipulation approaches to improve lipid productivity to make the technology cost-efficient. HVSP is a diatom-based vaccine that is useful in that:

  • Killed pathogenic vaccines are expensive
  • Live modified or recombinant bacteria or virus vectored vaccines may revert to virulence
  • Target: respiratory disease in cattle, H. somni is a causative agent of the disease, most important cause of economic loss in cattle

Diatom-expressed antigens will be inexpensive to produce regionally on a large scale, can be produced in various locations

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