NAABB_2nd Year-End Report (April-2012)

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Jos Olivares and Dick Sayre at Los Alamos National Laboratory

April 15, 2012, by David Schwartz

AlgaeIndustryMagazine.com

s the National Alliance for Advanced Biofuels and Bioproducts, NAABB, comesto the end of the second year in its three-year lifecycle, its looking more and more likeAquatic Species Program: The Next Generation. Armed with a vast array of scientificresources and brainpower, this broadly based, government funded, research program intothe functional future of algae, shows great potential to deliver the goods that the ASPdetermined were possible, or at least not impossible.

The nearly 40-member consortium, connecting national laboratories with universityresearch departments with private industry, targets major objectives in Algal Biology,Cultivation, Harvesting/Extraction, and Conversion to fuels. Co-product development isincluded in the mix, as well as Economic/energy-balance modeling and resourcemanagement components to establish the technologies viability for overall sustainability,as well as profitability and flexibility during the evolution of the biofuels markets.

NAABB members are developing co-products in large scale marketable livestock andmariculture feeds, through the production of lipid-extracted micro and macro-algae (LEA).The industrial Co-products program also involves the development of synthetic natural gas
http://www.algaeindustrymagazine.com/http://www.algaeindustrymagazine.com/


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production, thermal energy from LEA, bioplastics from proteins and lipids and feedstocksfor the nitrogen chemical industry.

Conducting this orchestra of discovery is Los Alamos National Laboratories (LANL)director of biofuels research, Dr. Jos Olivares, who has shepherded this project since the

proposal stage. Working closely with him to coordinate the gargantuan effort is theesteemed biologist, Dr. Richard Sayre, who recently relocated to LANL from his lead roleat the Donald Danforth Plant Science Center, in St. Louis, MO.

We spoke recently with Jos to get his first hand account of the progress so far, and what

to expect going into the third and final year of this landmark project.

We are approaching the end of the second year of the NAABB, the largest

government-funded algal research consortium to date. How would you assess the

progress so far?

Weve had a lot of progress. In the last two years weve met a number of milestones anddeliverables in all of the different areas of research we have going. For example, we hit amajor milestone in delivering a report to the DOE in which we isolated and screened over500 algal isolates from nature. And by February of this year we had actually isolated about1300 strains of algae from prospecting of natural sources around the U.S., and screenedabout 900 of those for their productivity as far as growth and lipid production.

Of those 900, we went into a second tier screening with 60 of those isolates. And of those60, we identified a number of them that look extremely promising, with productivities thatare much higher than our gold standard,Nannochloropsis salina. These are new isolates ofother organisms, three of which we are taking to outdoor cultivation at our Pecos site, at

Texas A&M. We are hoping they are as productive outside as they are in the lab.


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Test candidate strains for growth in different media Brooklyn College

Are you ready to reveal what those superstrains are at this point?

No, Im not. They are being very closely protected, as you can imagine. We will makethem available once Brooklyn College, the owner of that intellectual property, allows us to

make them public. These are natural strains, so the IP is in knowhow, and we want to makesure that our partners get a jump-start on them.

Eventually what we hope for is that we can take them on to some engineering development,but that I suspect is going to be outside of the current NAABB funded project. We onlyhave one more year to go, so were focusing primarily on developing transgenic tools forother organisms that are more popular, such asNannochloropsis salina and Chlorellaprotothecoides.

Another set of milestones in the algal biology side dealt with developing transgenic toolsfor three species: Chlamydomonas reinhardtii, which is our model species,Botryococcus

braunii and Chlorella protothecoides. And we delivered a report to DOE earlier this yearthat showed that we were developing a nice systems biology pipeline towardsunderstanding the genome and transcriptome of these organisms.

We have developed a number of tools that will help us transform these organisms intomore productive, more robust, candidates. In the process of doing that weve also identifieda number of other organisms that were contaminants to these original ones, and thatpresented us some nice new sources of information, for example,Nannochloris and the


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Tetraselmis species of algae, both of which have very different and very particularbiological characteristics that were trying to exploit for the program.

We are developing a strategy to transform Chlorella andNannochloropsis species towardsmore productive and more robust organisms. Its been hard going but weve worked out

several pathways by primarily trying to understand how to engineer the chloroplast of theorganism and by understanding the metabolic function of a number of enzymes, and thiswill help us design new genetic tools into these organisms. This part of the project ishappening here at Los Alamos National Laboratory, as well as at Texas A&M, Universityof Arizona, Phycal and Targeted Growth. There are also minor partners working on newmolecular developments in Chlamydomonas reinhardtii from UCLA, WashingtonUniversity St. Louis, and the Danforth Plant Science Center.

In the non-transgenic area, were developing a couple of methods with our partners at theUniversity of Arizona to adapt current organisms to become better producing and morerobust.

In the area of adaptive evolution weve been working with Chlorella protothecoidesprimarily. The University of Arizona has developed methods to have it grow faster underlimited phosphate conditions. And, as we know, phosphate is going to be a limiting nutrientfor any kind of agriculture. This work has been a series of adaptive evolutions under achemostat selection series that reduced the phosphate availability of the organism andselected for the best growing isolates of the culture, regrowing them with even morelimited availability of phosphate, while keeping their growth rate and productivity veryhigh and effective.

At LANL we are using flow sorting and selecting subpopulations of algae that have

different lipid properties; we can choose either end of a bell curve where we can selectalgae with very high or very low concentrations of lipid. When we select the top part ofthat curve and regrow it we end up with an isolate of algae that has a 50% increase in lipidcontent compared to wild type. The culture stays healthy and can be maintained for manyrounds beyond that.

We had very good lipid productivity through these procedures, so we tried similar methodslooking atNannochloris sp. and selecting for fast growing species, and weve actually beenable to almost triple the productivity of this organism. This process weve developed lookslike a very nice new technology for selection and adapting of species under naturalconditions, which would not need to include any transgenic modification of algae.


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Mini-raceways at Pacific Northwest National Laboratory

What sorts of developments have you had on the cultivation side?

In cultivation we had a major deliverable in showing we could use photobioreactors toachieve growth rates of 5 grams dry weight per liter, with a minimum of 50% lipid content,and in some cases close to 70%. The work was primarily done at New Mexico State

University and at Solix Biofuels, two of our partners. They demonstrated that these levelsof productivity are achievable under appropriate conditions. The tests were performedcontinuously over a year and a half period of time.

Probably the last major deliverable that we had earlier this year was a down selection ofour harvesting and extraction technologies. Weve had seven different technologies forharvesting and extraction in development throughout our partnership five harvesting andtwo extraction. We went through a very quantitative and rigorous method for comparingthese against each other, against benchmarks such as centrifuge techniques and wetextraction techniques that are more popular in the outside world right now. We thendownselected to four harvesting and extraction technologies that would continue on to

Phase II (scale up).

Of these four technologies, three of them are harvesting technologies, including anelectrolytic flocculation technology out of Texas A&M, membrane harvesting technologyfrom Pacific Northwest National Laboratory, and the acoustic harvesting technology at LosAlamos National Laboratory. And then we have a very special extraction technology usingmesoporous nanoparticles, out of Iowa State University, targeting very high valuechemicals within the oils of the algae, especially polyunsaturated fatty acids and special


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compounds such as cotopherol (Vitamin E) which is produced by some algae in fairlyhigh quantities.

So far those are NAABBs major deliverables and accomplishments. There is a new set ofdeliverables being worked on right now for month 24, which is this month (April), and

those are primarily addressing areas of conversion technologies and some animal feedresearch and sustainability issues.

How much of this information has been published since the beginning of the project?

In the past two years we have had over 50 technical publications accepted, in submission,or in the preparation process; and 16 intellectual property disclosures for licensing by ourpartners. And out of those 16, one technology has spun out into a company, PhenometricsLLC, which makes a small, lab-based photobioreactor technology.

Will the original partners stay fixed, or do you bring in new partners over the course

of the project?

Weve added a number of partners in the last year, including Phycal, SRS Energy, IdahoNational Laboratories and Reliance Industries, from India. Weve also entered into anumber of memoranda of understanding for collaborative research between NAABBpartners and a number of other organizations including Argonne National Laboratory,primarily in using their GREET model system. Weve entered into an agreement withAdvanced Biomass R&D Center, in South Korea, and another research collaborativeagreement with Industrial Technologies Research Institute, in Taiwan. Another new partneron the industry side is Pan Pacific, out of Australia, and weve just added the NarionalRenewable Energy Laboratory (NREL) as a partner to the consortium. So, quite a bit of

activity in the partnership arena, and its still growing.


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Solix Biofuels photobioreactor beds at New Mexico State University

If you could extrapolate to the end of the project, what do you think will develop over

the next year?

Our main goal is essentially to understand the techno-economic viability of algae as afeedstock for the biofuels industry. I think at the end of the third year we will havedeveloped a complete analysis based on currently accepted technologies, and then added tothose well have a number of new technologies and innovations that weve been able todevelop over the course of this project. And I believe well show a much more favorablecondition for algae as a feedstock for biofuels.

So, will it be at $2.00 a gallon that everybody would love to see? I dont think so. But Ithink it will be much more favorable than weve been seeing in the last couple of years. Wethink well be able to show that we can trim major cost and major risk out of algae as abiofuel.

Does this seem like a once only type of research project, or do you see this as

something that will be more common from here on?

I think what were seeing is that more and more of these funding opportunities are callingout for consortia type arrangements, and overall the effect has been a very positiveapproach to developing and managing these kinds of efforts. I think well see consortia as amajor way of doing research in the future.


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My vision is that we will see an NAABB-type consortium continue, and it will look a bitdifferent beyond Year Three. I hope that the focus for the future will be more towardsaddressing the objectives of our industrial partners rather than the needs of the fundingfederal agencies. The funding federal agencies are a good way to get things started and toget some momentum going. But now that weve accomplished that, we need to shift our

efforts to understanding the industry better, and the issues the industry is facing, and try todevelop programs that can go after those things the industry feels are the most important towork on those that a consortium like ours can take on.

Dr. Richard Sayre, your co-director of the consortium has recently relocated to your

home base, LANL. What is he working on and how has this affected the NAABB

operations?

Starting with the second question, it has helped tremendously by having him next door tobounce things off on a daily basis, which was not really possible before because of ourphysical locations. From an operations side its helped us gel a bit more. His focus has been

primarily in the area of algal biology, so his team has been focusing on understandingcontaminants, specifically bacterial contaminants in algal cultures and developing methodsfor controlling them. Just today he let me know that they have found a number of peptidesthat will control the bacterial content of algal cultures in a very positive way. I think weregoing to see some new approaches to algal cultivation coming out of this group.

From a LANL perspective, its been a great kick. He brings a stature and an energy withhim thats wonderful. Along with the New Mexico consortium universities, weredeveloping a laboratory around the concept of biology for energy science, and he will bethe first person moving into that laboratory. So he represents our starship for the biofuelsdevelopment effort here at Los Alamos National Laboratory, and were very excited to

have him here.

You have an upcoming conference, the 2nd International Conference on Algal

Biomass, Biofuels and Bioproducts scheduled for June 10-13, in San Diego. What do

you see happening at that conference?

Our primary focus is understanding the latest and greatest in technical and scientificachievements throughout the algal research community internationally.

What well see, first of all, is that its going to be huge. Weve received over 640 abstractsubmissions both for oral and poster presentations for this conference, which overwhelmed

us. Last year the conference attracted 300 people, and it will probably double or triple thisyear.

Our scientific committee has worked very hard on developing the program, and we willhave about 90 oral presentations primarily in two parallel sessions, and a little over fourhundred posters. Its going to be a very high quality scientific and technical conference andSan Diego is going to be a great setting.


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Well see a lot more participation internationally this year than we had last year. Thesubmissions for presentations that came in were almost equal between the U.S. andinternational.

Final thoughts on NAABB, Year Two?

All in all, this has been a great year, and I hope were starting to see an impact from thework being done by the NAABB partners into the algal biofuels community.

Documents

NAABB_2nd Year-End Report (April-2012)