Research proposal on ethanol production

Ethanol production from agricultural wastes using Sacchromyces

when they grew a specially engineered lab strain on pure xylose and added the supplements, the ethanol yield jumped by more than 50%—a result consistent with the complete consumption of xylose. and to challenge their method, they used an inbred laboratory strain of yeast known to be particularly sensitive to ethanol. experiments confirmed their reinterpretation: just adding potassium and reducing acidity—with no phosphate present—gave the same remarkable boost in ethanol production. in addition, they have promising initial results showing production increases from samples of mixed raw materials actually used in industrial fermentations. by their results, the researchers decided to apply their methods to other issues encountered in biofuels production.

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tests described so far involve the ethanol-sensitive lab strain of yeast. “ethanol tolerance is one of those traits that’s sort of mysterious,” says lam. researchers already have pushed up ethanol productivity from the levels reported here. “butanol, for example, has a higher energy content per liter; and unlike ethanol, it can be used as a direct substitute for gasoline in today’s cars. in other “firsts,” the researchers described the mechanism by which alcohols poison yeast; they defined two genes that control ethanol tolerance; and they modified those genes in lab yeast to make them out-produce the industrial strains—even without the supplements.

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adding either potassium chloride (to increase potassium) or potassium hydroxide (to reduce acidity) pushes up ethanol output significantly from the baseline with no supplements. the top two bars show production from the basic lab strain, without and with the supplements. researchers believe that their findings reveal the mechanism by which ethanol and other alcohols kill yeast at levels relevant to biofuel production. however, when assisted by the supplements, the lab strain produces far more ethanol than the commercial strains do. curves in the top figure at the right show ethanol production from a single culture growing for 72 hours under four conditions: on the standard laboratory medium with no supplement (blue); with added potassium chloride (red); with added potassium hydroxide (light blue); and with both potassium chloride and potassium hydroxide (black).

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Research on Ethanol Production from Sugarcane Wastes

adding potassium and an acidity-reducing compound helps the yeast tolerate higher concentrations of the ethanol they’re making without dying. diagram shows the amount of ethanol produced as a function of the viable population of yeast cells over the course of fermentation. the next pairs of bars show results from cultures involving yeast strains used in bioethanol production in brazil (middle pair) and the united states (bottom pair).” such high-energy alcohols are even more toxic to yeast than ethanol is, but the researchers found that the boost in tolerance extended to such alcohols as well. is already producing ethanol from sugar cane juice, but the use of wastes will result in more effective climate change prevention.

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in cultures grown under unsupplemented conditions, the commercial strains produce more ethanol than the lab strain does. table shows ethanol production from standard lab yeast and from yeast strains used in bioethanol production in brazil and in the united states. the goal of this project is to contribute to mitigating climate change through sustainable fuel production, and extension to other regions such as southeast asia is also planned. how could they make yeast resistant to higher concentrations of ethanol? when all three receive the supplements, the ethanol yields are comparable.

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on ethanol production from sugarcane wastessugarcane wastes: producing sustainable energy from residues. but production from the lab strain with the supplements is significantly greater than the baseline production from the two commercial strains. certain yeast that were somewhat ethanol-tolerant showed high activity in genes related to phosphate utilization. but the largest boost—by far—comes from individual cells being able to withstand higher levels of ethanol. is increasing demand for bioethanol fuel produced from crops such as sugarcane and corn as an effective means for mitigating climate change without increasing the amount of co2 in the atmosphere.

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the impact of the potassium on ethanol tolerance occurred too quickly for cells to be mounting a biological response. indeed, using yeast with genetically augmented pumps but no added supplements pushed ethanol production up by 27%—not as high as the level reached with unaltered yeast plus the supplements, but still higher than the levels reached with the brazil and us commercial strains. given the already high performance of certain industrial yeast strains, making those changes could bring even more dramatic increases in the production of ethanol—and perhaps second-generation biofuels such as butanol—than have been demonstrated in the lab. “the per-cell production rate hardly changes, but the cells stay alive much longer; and the longer they live, the more ethanol they make,” says lam. “so the ability to endure toxicity is a principle determinant of ethanol output—and it’s the primary trait strengthened by our supplements,” says fink.

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but yeast strains used commercially are carefully selected for their genetic predisposition toward ethanol tolerance. “altering a single gene has not prevented diabetes—or made yeast more ethanol-tolerant. of optimum fuel ethanol production system for the use of bagasse and straws. examination showed that the elevated potassium plus reduced acidity results in an increase in cell viability and thus in ethanol productivity. but the boost in ethanol production persisted even when they used genetic methods to disable phosphate metabolism in the yeast.

Ethanol production from agricultural wastes using Sacchromyces

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“the biggest limitation on cost-effective biofuels production is the toxic effect of alcohols such as ethanol on yeast,” says gregory stephanopoulos, the willard henry dow professor of chemical engineering. the data show a direct correlation between total viable population and ethanol production. of adding potassium chloride and potassium hydroxide on ethanol output. to right: gregory stephanopoulos of chemical engineering, gerald fink of biology and the whitehead institute, and felix lam of chemical engineering are developing new insights and techniques that could one day dramatically increase the amount of ethanol, butanol, and other biofuels that yeast can produce from raw materials such as corn and sugar cane. thus, adding the supplements to the growth medium influences ethanol output more than do any genetic differences among the yeast strains.

but there’s a problem: at certain concentrations, the ethanol kills the yeast that make it. each supplement pushed up ethanol output significantly, and adding the two together brought an increase of 80%. engineers and biologists at mit have found a simple way to make yeast produce more ethanol from sugars: spike the mixture they’re growing on with two common chemicals. in addition, assisting the commercial strains with the supplements pushes their ethanol yields up to roughly the same level as yields from the lab strain with supplements. not surprisingly, the baseline ethanol production from those strains is higher than that from the basic lab strain.

” at concentrations achieved in biofuel production, alcohols do not dissolve the yeast’s cell membrane but rather make it porous. challenge is to make fuels that are better suited than ethanol to today’s transportation needs. “everyone knows that ethanol is ultimately a pretty lousy fuel,” says lam. if larger-scale laboratory or pilot-plant tests take place and are successful, lam notes that it should be relatively easy to implement the approach at existing bioethanol plants: just increase potassium levels and control acidity inside the bioreactor. curves show ethanol production from laboratory yeast growing on glucose for 72 hours.

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