Clearwater Conference

The program started with an opening speech given by the program Chairman Thomas who gave a tribute to the father of Biofuels, Ray Katzen whom the 32nd Symposium on Biotechnology for fuels and chemicals was dedicated. He said that according to Dr. Katzen, black crude is bad, green biomass is good!; this is the first law of motor fuel according to Ray Katzen. In 1978, the US government was desperate to establish a new road map for biofuels. Ray Katzen conducted a study and sold 30,000 copies; one of the maximum copies sold ever by the US government. Ray is also known as “Doctor who turned on the lights”. He established industries in different parts of the world such as Cuba, and Switzerland to name a few. His proposed Project 20 aims to produce 20 billion gallons of biofuels by 2020. I was highly inspired by the works done by Dr. Katzen and thought that this opening speech was a great way to start the symposium.

Blog #1: Monday, Session 1 @ 1 pm, Increasing ethanol productivity from xylose in recombinant Saccharomyces cerevisiae by protein engineering.

This talk was based on the mechanism of pre-treatment that reduces inhibitors such as furfural, which comes from hemicelluloses. The purpose of their study was to engineer a protein involved in the metabolism in order to maximize yields and to increase rates. Cellulose is broken down into glucose (hexose) and hemicellulose is broken down into pentoses. Their research was focused mainly on the intermediate product, pyruvate. Metabolic evolution of E.coli was done to get the desired E. Coli, mutants that were resistant to furfural from xylose and arabinose and hydroxy methyl furfural (HMF) from glucose, fructose, mannose, galactose, etc. Betaine was used to prevent the evolved E. coli cells from bursting. Furfural resistant strains of E. coli were cross-resistant to HMF.

They did a micro-array analysis of RNA of an E. coli to check for the expression analysis of the E. coli. Proteins that are NADPH-dependent oxido-reductases that reduce furfural and HMF were studied. Expression of both genes is reduced in furfural-resistant E. coli. The researchers then deleted certain genes and looked at the expression of certain genes as well as mutanized genes done by evolution. They detected the expression of these genes through microarrays. They also checked the activity of the promoter and the expression and regulation of genes via real-time PCR. Firefly luciferase promoter activity was used to detect the promoter. This presentation was mostly about changing the pathway that makes pyruvate and looking at the effect on the product formation. Normally, amino acids such as cysteine and thionine are not made in large quantity. Genes that were able to reduce furfural and HMF were determined. Expression of such genes from the determined promoter (yqhD) was aldehyde inducible.

Blog #2: Tuesday, Session 4 @ 8:30-9am, A novel, cost-effective method for producing ethanol from carbon dioxide in hybrid algae.

Scientific basis of direct to ethanol process: This talk was from a person representing Algenol Biofuels Inc., which was founded in 2006 and has collaborations with multiple countries across the world such as Germany, Spain, Switzerland, USA- Baltimore and Florida where the headquarter is located. The speaker based his study from Coleman and Deng (1999) who studied linking the Calvin cycle with exogenous fermentation pathway. According to the speaker, the biggest challenge is in producing ethanol in a commercial scale. Their goal was to produce ethanol from carbon dioxide using hybrid algae that are grown in seawater and contained in enclosed bioreactors. The main approach in doing so was to channel as much pyruvate as possible into the fermentation pathway in algae especially targeting the alcohol dehydrogenase. Their study used large flexible plastic bag into which seawater and algae are placed. Nutrients were then introduced periodically into the culture and during the course of the day; the bioreactor was heated, where ethanol is produced from the algae. Heat transfer across the plastic occurs and condensate forms in the surface when the temperature drops during the day. The condensate that is produced is 2-3 times more concentrated ethanol. The condensate is then collected which is run through a processor, which will boost ethanol yield. In order to maintain the salinity within the tolerance of the algae, fresh water must be returned back into the bioreactor. According to the speaker, this utilizes less gallons of fresh water than ethanol produced which is therefore a water saving process.Their goals to be achieved are as follows: Ethanol produced of 6,000-10,000 ga/acre/yr; Capital costs under 5$/ga capacity; Operating cost less than $1/ga/yr, initially hope to be around 1.25$/ga/yr. Another advantage of their method was that there was no competition with feedstock, and marginal or abandoned lands were used.

Blog #3: Tuesday, Session 5 @ 1:30pm, Molecular factors associated with altered cell wall chemistry in Populus.

In order to better understand how plant cell walls are formed and how the composition and properties of lignocellulosic feedstock materials are controlled, a research strategy was developed. The speakers talked about this study that consisted the use of –omics, spectroscopy, sugar release assay, imaging and bioinformatics approaches to characterize a selected set of Populus samples. Populus trees vary greatly in their cell wall compositions and salient individuals were used in sugar release assays. They also characterized a wild-type Populus plants under tension stress in controlled greenhouse conditions; such usage of wood would help in understanding the wood’s tension capabilities, lignin content, cellulose content and xylem composition. New genes in cellulose biosynthesis pathway were determined via the Illumina-based transcriptome and LC-MS/MS proteome profiling datasets. The speaker then presented preliminary results from functional genomics investigations using transgenic Populus plants.

Blog #4: Wednesday, Session 7 @ 10 am, An integrated analysis tool to guide sustainable biomass production.

Iowa used to be the maid of prairie during 1833-1890. According to the speaker, land use change responds to increase in agricultural commodity prices in ways such as: 1. Extensification, 2. Substitution, and 3. Intensification. Extensification relates to bringing new land into production. CRP acres are present in a lot of quantity in Iowa. Although CRP land is less good land for corn, it is not horrible especially those in Iowa. Hill slopes in Iowa that are more prone to erosion were studied. Soybean is more susceptible to erosion than corn. If crops that are planted in the land are changed, you change the chances of erosion in those lands too. E.g. if soybean is replaced with corn or switch grass, erosion is greatly reduced. Substitution means changing the crops planted in the land, e.g. putting perennial grasses into land. If a landscape is re-shaped into one way, it affects the moisture and rainfall in other way in different landscape. The ethanol plants and the corn-corn plants do not coincide. Soybeans do not take up much nitrogen and so you take corn-corn rotation with distance from ethanol plants. This relates to intensification where the nitrogen in the soil is increased. The speaker also discussed some of the integrated tools to guide land use choices. Soil carbon model was presented as well as the social and economic impacts of land use changes were discussed.

One of the statements said by the speaker drew my attention: “Land use is driven by complex social-economic-technological changes.” It made me realize that it is a vicious circle as to how people use land for different purposes at the same time, different social changes affects people’s attitudes regarding land use changes as well.

Blog #5: Thursday, Session 9 @ 9am, Biomass supply and logistics: A Tennessee perspective.

The talk by Mr. Jackson, representing Genera Energy in Knoxville, Tennessee was about Tennessee and the people’s perspective for biofuels. He collaborated with a company that works with local framers to develop switch grass production, which is a potential biofuel feedstock. In order to provide sufficient feedstock, the state has a statewide commitment of $70.5 million. The speaker then talked about how they would go about in working with the farmers to grow ample switchgrass that can make sufficient biofuels. Their current crop is corn but they are planning to use switchgrass pretty soon. The speaker also mentioned about how the farmers are educated and taught through seminars; the farmers are paid $450/acre/year for costs of production and opportunity costs while the switchgrass seeds and the technical expertise are provided to them for free. The drawbacks of using switchgrass are that their cost is increasing as well as the seeds are hard to find. Moreover, the right herbicide for switchgrass hasn’t been found and so trying to figure this out would increase the cost of the switchgrass since the yield might be low. His talk pointed out that much of the research is needed to make the use of biofuels possible which starts with trying to find the right feedstock and in adequate quantity.

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