By: Shwetha Sivakaminathan and Lauren Harroff

Last week we had the opportunity to attend a workshop at the UTEX Culture Collection of Algae located at the University of Texas in Austin. Algae include a very diverse range of organisms with differing sizes, shapes, colors and structural components. Microalgae fall into two broad categories: Cyanobacteria or Blue-green algae and Eukaryotic algae or true algae.

Outside the Culture Collection

The UTEX Culture Collection contains approximately 3,000 different strains of living algae, representing most major algal taxa. The workshop we attended focused on managing algae cultures, sterile culture techniques, basic biology of algae, and measuring growth—all important subjects for algal biofuels. We were able to tour the impressive collection, listen to lectures, and even try out the techniques ourselves. Below is just a sample of some of the things we learned:

Lauren practices pulling Pasteur pipettes into tiny micropippetes to isolate single cells for creating axenic cultures.

Some of the common terminologies used for microalgae are:

Culture (noun): A population of living microorganisms that is maintained away from its natural habitat under a set of conditions that is (to at least some extent) controlled.

Culture (verb): The process of maintaining and/or managing a culture.

Unialgal culture: A culture that contains one and only one kind of algae. It may contain other non-algal microorganisms.

Axenic culture (pure culture): A culture that contains only a single kind of microorganism.
Anaxenic culture of an alga would not contain any bacterial contaminants.

Species: A population of living microorganisms that is very similar as determined especially by their DNA sequences, but also by other characteristics. (The species we use is Chlorella protothecoides)

Strain: A population of living microorganisms, all of which were isolated from the same location at the same time, and all of which are virtually identical according to their DNA sequences. (The strain we use is Chlorella protothecoides UTEX 256.)

Subculturing: (Passaging or Transferring) A subculture is a new culture made by transferring some fraction of cells from a previous culture to fresh growth medium. Sub culturing is used to establish long term cell cultures and/or expand the number of cells in the culture.

Algae cultures are maintained either on agar slants or as liquid cultures depending on the nature of the strain. The incubator and storage rooms for all are maintained at room temperature of around 20 degrees C.

Most of the fresh water and soil algae that need to be stored for a long period of time go into cryopreservation i.e. storage in liquid nitrogen (below -196C). Cryopreservation minimizes maintenance cost, reduces space requirements, diminishes risk of contamination and prevents genetic changes.




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Thanks to the crew down at Croc’s for partnering with the Clemson Sustainable Biofuels initiative! We wish them the best in their new venture in Tiger Town. Croc’s offers a huge selection of wings and draft beers, as well as live music in downtown Clemson. Check them out this Saturday after homecoming for music from Cravin Melon.

Corey Benson, of the Clemson Sustainable Biofuels creative inquiry program is pictured here with Mathew, kitchen manager and kind giver of the grease. When you stop in be sure to thank them for partnering with Clemson Sustainable Biofuels! Stay tuned for an announcement of a grease appreciation night in early December to thank Croc’s for supporting our program. Once a month we will be highlighting one of our partners. On this day we will gather for food and libations to thank these local businesses for their support. This is an idea we’ve borrowed from our friends at Piedmont Biofuels 😉

Stay Tuned!


Biosystems Engineering seniors Kaitlyn, Steven and Anne scooping grease after our big win over FSU!Well, the build is under way! After a morning collecting used cooking oil from our Death Valley stadium and HUGE win over Florida State, Anne, Kait, Steven and I headed back to home base to unload our grease and load our mobile 5kw generator over to the Folks at Habitat for Humanity who were excited to use our locally produced biodiesel to build the annual Habitat house here on campus.

 From 8am-5pm this little gennie purs running on B100 and powering tablesaws, circular saws, battery charges and anything else requiring power for the build. Over the next week we will also be fueling the light towers and generators supplied by United Rentals for our homecoming float building and celebration. We will be fueling this equipment with a blend of 20% biodiesel and 80% diesel fuel.

Building a Habitat house on B100!


Holly and Dex have redemption. Through our Creative Inquiry course on value added coproducts from glycerol, we finally have some useful soap coming down the line. Our first batch looked like  failed batch of brownies. But alas, we retooled, enhanced our quality control and anlytics, and today Holly cut the soap into bars to cure. This soap is made from glycerol that we refined in our lab to remove free fatty acids, remove methanol, remove water, filter to 30 micron, and adjust to a pH of 7. We then pressed some Chinese Tallow Seed and Sunflower Seeds to make up the rest of the oil for our batch. Some teatree oil was also used for providing fragrance.

The bars are a brownish color and smell fantastic. It will be a few weeks before we can begin using these bars, but in the meantime we will be refining our recipe and making molds to shape our soaps. Tiger Paw soap anyone?

Holly wafting the sweet success of the Teatree aroma!


CU Sustainability Partners!As we roar into summer, we have our first downtown Clemson restaurants ready to donate their grease to our mission for sustainability. Walker Massey of C.U. Economic Development helped us with this design and we love it! This week we’ll be delivering a 250 gallon grease dumpster to DaWaat Indian Cuisine on Keith St. We’re excited to support DaWaat in this partnership and we look forward to supporting them by dining at their fine restaurant. I hope you folks will too! And tell them thank you for choosing to recycle their used cooking oil to our local sustainability project! The C.U. campus will be much cleaner for their contribution.


As the sun prepared to set Friday evening, Holly Garret and I put together a quick video and project summary for the Discovery Channel.

We explained how we use off grid renewable energy to power our process, and we focus on the environmentally favorable non-food feedstock for biodiesel and ethanol production. For those of you who don’t know, our analytics lab is 70% off grid thanks to the Photovoltaic Solar array that Dr. Terry Walker was able to purchase and install for us. We also have a solar hot water panel awaiting installation to provide our heat for vegetable oil settling and moisture removal.

Welcome to the Clemson Sustainbale Biofuels Iniative! - Featuring Environmental Engineering Masters Student holly Garret!

Our Mobile Biodiesel Production Lab is 100% off grid and powered by B100 made by myself with the help of our dedicated students. We have also plumbed the generator to capture waste heat that we then divert to heat our process. Using the generator for both electricity and heat is referred to Co-generation, and co-generation is part of every sustainable energy system involving combustion.

We also discussed our research involving algae cultivation. Specifically our goal of creating a plug flow photobioreactor to mount onto combustion engines. This bioreactor will capture CO2 emissions produced during combustion, and feed it back to the algae. By the time the algae reaches the end of the bioreactor we will be able to harvest it for oil (to make biodiesel!) and biomass pellets for testing as a feedstock for gassification.

Ultimately we’d like to install a gassifier on campus for steam and electricity production and capture emissons for the exhaust to feed our algae cultivators, that will in turn provide us with biodiesel feedstock and feedstock for our gassifier.

One of the issues with gassification is the transportation of the feedstock. It is currently not a favorable energy balance due to the energy involved in processing, pelletizing and transporting forest products to feed gassifiers. Algae has the potential to improve this energy balance because it is a feedstock that can be generated on-site with significantly less acrage than forest products.


Thanks to the amazing Creative Inquiries program at C.U. we were able to fund an undergrad over the summer to help run our 90 gallon per week biodiesel processor here on Campus. Dexter Pearson is a rising Biosystems Engineering  junior and participated in the C.I. program this spring. He was a great student and always stayed late or came in for extra hours because he was genuinely interested in the process. Now after a month of running the plant together, Dexter is autonomous.

I feel our lab is in good hands and I know that the biodiesel produced for the CU Facilities fleet will meet the highest quality specifications with Dexter at the helm.

Recent activities include oilseed crushing for soap production. Dexter crushed about 10 lbs of chinese tallow seed and 15 lbs of sunflower seed to give us some high quality oil for converting our glycerol co-product into soap. We purchased the soapmaking handbook “Journey Into Soapmaking” from theabundancefoundation.org because it specifically details how to use biodiesel glycerin for high quality soap making. With our first successful batch soon coming off the line, we hope to plan for a Fall Creative Inquiry to make different soaps such as detergents, dishsoap, laundry soap and garden scrubs from our glorious co-products.

50% by weight oil and waxes! That's perfect for soap!


By Holly Garret

Now that school is out for the summer, Clemson’s cafeterias aren’t nearly at the oil volume as during the year.  We’ve been doing some outreach to a few places in downtown Clemson which will hopefully prove fruitful and get us back running a few batches a week.  As part of my Master’s research, I’m also doing a feasibility study just to see how much waste oil is produced in Clemson on a given week.  (Of course, this will fluctuate throughout the year, as football season and students come and go.)  But a rough estimate is what I want right now.  I’ve started contacting all the restaurants in the Clemson area and talking with the managers or owners about how much oil they use a week.  Again this is a rough estimate, as most of them are telling me how much they USE, rather than how much they DUMP after its been used (which is the stuff we’d want to use).  I am meeting today with TD’s and he is going to give me both figures, how much they buy and how much is collected by Carolina Byproducts, their waste oil contractor.  This way I can get a rough percentage of how much is lost in the frying process.  2%? 5%?   Another technical kink is that most places receive containers which weigh 35 pounds.  This is not gallons and to figure it out, I’d need to know what type of oil it was and its density.  I may need to go back and ask those places what type of oil it was.   I hope to have this data by the end of the summer, compiled with all the assumptions checked out.


Why not? For the first time in recent history biodiesel and biodiesel blends are actually less expensive than petroleum diesel fuel. A blend of B5 is compatible with any diesel engine type, and most engines are approved for use up to B20. Just a little biodiesel goes a long way to improve emissions by reducing CO2 and unburned hydrocarbons, while also boosting Cetane value and adding lubricity. If you want to see biodiesel in our CAT bus fleet, please write to
Keith Moody [KMoody@cityofclemson.org] and say “Biodiesel in the CAT bus please” in the title.