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Teaching a Microbe to Make Fuel

Genetically modified organism could turn carbon dioxide or waste products into a gasoline-compatible transportation fuel.

David Chandler, MIT
September 03, 2012  |  12 Comments

A humble soil bacterium called Ralstonia eutropha has a natural tendency, whenever it is stressed, to stop growing and put all its energy into making complex carbon compounds. Now scientists at MIT have taught this microbe a new trick: They've tinkered with its genes to persuade it to make fuel — specifically, a kind of alcohol called isobutanol that can be directly substituted for, or blended with, gasoline.

Christopher Brigham, a research scientist in MIT’s biology department who has been working to develop this bioengineered bacterium, is currently trying to get the organism to use a stream of carbon dioxide as its source of carbon, so that it could be used to make fuel out of emissions. Brigham is co-author of a paper on this research published this month in the journal Applied Microbiology and Biotechnology.

Brigham explains that in its natural state, when the microbe’s source of essential nutrients (such as nitrate or phosphate) is restricted, “it will go into carbon-storage mode,” essentially storing away food for later use when it senses that resources are limited.

“What it does is take whatever carbon is available, and stores it in the form of a polymer, which is similar in its properties to a lot of petroleum-based plastics,” Brigham says. By knocking out a few genes, inserting a gene from another organism, and tinkering with the expression of other genes, Brigham and his colleagues were able to redirect the microbe to make fuel instead of plastic.

While the team is focusing on getting the microbe to use CO2 as a carbon source, with slightly different modifications the same microbe could also potentially turn almost any source of carbon, including agricultural waste or municipal waste, into useful fuel. In the laboratory, the microbes have been using fructose, a sugar, as their carbon source.

At this point, the MIT team — which includes chemistry graduate student Jingnan Lu, biology postdoc Claudia Gai, and is led by Anthony Sinskey, professor of biology — have demonstrated success in modifying the microbe’s genes so that it converts carbon into isobutanol in an ongoing process.

“We’ve shown that, in continuous culture, we can get substantial amounts of isobutanol,” Brigham says. Now, the researchers are focusing on finding ways to optimize the system to increase the rate of production and to design bioreactors to scale the process up to industrial levels.

Unlike some bioengineered systems in which microbes produce a wanted chemical within their bodies but have to be destroyed to retrieve the product, R. eutropha naturally expels the isobutanol into the surrounding fluid, where it can be continuously filtered out without stopping the production process, Brigham says. “We didn’t have to add a transport system to get it out of the cell,” he says.

A number of research groups are pursuing isobutanol production through various pathways, including other genetically modified organisms; at least two companies are already gearing up to produce it as a fuel, fuel additive or a feedstock for chemical production. Unlike some proposed biofuels, isobutanol can be used in current engines with little or no modification, and has already been used in some racing cars.

The work is funded by the U.S. Department of Energy’s Advanced Research Projects Agency – Energy (ARPA-E).

12 Comments

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Maxx On
Maxx On
September 20, 2012
I agree with peter-bradshaw, this would have to be combined with photosynthesis for it to be really sustainable. However, I would prefer other technologies that aren't as polluting and as noisy.
Javier Kienzle
Javier Kienzle
September 10, 2012
I appreciate Cliff's academic and honest input on the feasibility and true impact of this example of microbial fuel production. I see too many idealists and greenwashed folks on this site that push ideas with no serious factual consideration.

I'm a firm supporter of Solazyme, although the solutions they are developing may not be ready for several decades.
Peter Bradshaw
Peter Bradshaw
September 6, 2012
If these bacteria can be combined with some photosynthesis action (there are bacteria that do this, also phytoplankton, and of course plants), this could be a worthwhile system to turn CO2 and sunlight into usable fuel. After all, the earth did this a few 100 million years ago, and built up all those fossil fuel resources we are currently burning back to CO2 (and H2O).

If some process like this doesn't work, some future generation of our offspring will be forced to use electric cars, or just bicycles, to get around.
Cliff Claven
Cliff Claven
September 5, 2012
@Joel, @Lauryn: First point is that this microbe process is very similar to the micro-algae and bacterium efforts underway in many labs with the target outputs being either alcohols or lipids. Second point is that we cannot ignore hydrogen and focus only on carbon. Hydrogen is the chief energy carrier. Third point is that, while waste can be a source of both hydrogen and carbon (I mentioned urea in my post above which is generally from organic waste), it is a poor feedstock for energy. Hydrogen and carbon are only energy carriers if they are in chemical bonds that have exothermic energy to release. CO2 does not. H2O does not. Energy must be pumped in from some source to reform CO2 and H2O into CH4 or a lesser fuel such as C4H9OH (butanol), where the hydrogen and carbon are again true energy carriers. Recovering energy from a waste stream is laudable, but it can produce only a tiny fraction of the energy that the generators of that waste stream need. If you get enough to power the waste-handling facility, you are doing well. A cow cannot live off its waste because it has already extracted most of the usable energy from it, and the waste can only be reformed with the help of an entire ecosystem and years of light energy from the sun. Civilization is a living organism and likewise cannot live off of its own waste. The article mentioned the goal of this research being biobutanol to blend with gasoline--that wasn't my leap. We'll be blending each barrel with an eyedropper.
ANONYMOUS
September 5, 2012
What is so new about this??

see http://escholarship.org/uc/item/6wt5z3h9.pdf

Appl Microbiol Biotechnol (2010) 87:1045–1055
DOI 10.1007/s00253-010-2522-6

Received: 28 December 2009 / Revised: 20 February 2010 / Accepted: 23 February 2010 / Published online: 8 April 2010

ABSTRACT:
The production of isobutanol in microorganisms has recently been achieved by harnessing the highly active 2-keto acid pathways. Since these 2-keto acids are precursors of amino acids, we aimed to construct an isobutanol production platform in Corynebacterium glutamicum, a well-known amino-acid-producing microorganism. Analysis of this host's sensitivity to isobutanol toxicity revealed that C. glutamicum shows an increased tolerance to isobutanol relative to Escherichia coli. Overexpression of alsS of Bacillus subtilis, ilvC and ilvD of C. glutamicum, kivd of Lactococcus lactis, and a native alcohol dehydrogenase, adhA, led to the production of 2.6 g/L isobutanol and 0.4 g/L 3-methyl-1-butanol in 48 h. In addition, other higher chain alcohols such as 1-propanol, 2-methyl-1-butanol, 1-butanol, and 2-phenylethanol were also detected as byproducts. Using longer-term batch cultures, isobutanol titers reached 4.0 g/L after 96 h with wild-type C. glutamicum as a host. Upon the inactivation of several genes to direct more carbon through the isobutanol pathway, we increased production by ?25% to 4.9 g/L isobutanol in a ?pyc?ldh background. These results show promise in engineering C. glutamicum for higher chain alcohol production using the 2-keto acid pathways.
Lauryn S
Lauryn S
September 5, 2012
Cliff, it does say that "In the laboratory, the microbes have been using fructose, a sugar, as their carbon source," which implies that this does not have to be the carbon source. Joel is correct in noting that the article states "While the team is focusing on getting the microbe to use CO2 as a carbon source, with slightly different modifications the same microbe could also potentially turn almost any source of carbon, including agricultural waste or municipal waste, into useful fuel."

Every form of energy involves some loss. Ideally, there would at some point be NO fossil fuel energy used to make the sugar powering this reaction, and rather, waste products could be used instead. Obviously this process has not been perfected yet; the article is about what has already been accomplished, and what still needs to be done, IN ORDER for this to be a viable energy source. No one is claiming that the way it's being done in the lab right now is the end goal. If you don't learn to see possibilities, you'll miss them.
Joel Fairstein
Joel Fairstein
September 5, 2012
Cliff, the hydrogen can come from organic waste. You seem to think the research is about sugar conversion, which the article clearly implies is not the end goal.
C. Craig Morris
C. Craig Morris
September 5, 2012
The article doesn't do justice to the concept. And I'm sure MIT scientists don't believe in perpetual motion machines. CO2 is probably of interest for carbon sequestering during transition out of fossil fuel dependence. The vast, exponentially growing, quantity of garbage produced by our planet's inhabitants contains a wealth of renewable, potentially clean, energy. It's easy enough to see where these ideas are going, if you keep an open mind.
Cliff Claven
Cliff Claven
September 5, 2012
No, Joel, you misunderstand the chemistry. CO2 can be a source of carbon, but not a source of energy. CO2 is a combustion product, after all, and the exothermic energy in the chemical bonds has already been released. The energy being input to these microbes is from the sugar and is mostly in its hydrogen, with a smaller contribution from the sugar carbon. Hydrogen is the primary energy carrier in combustion and decomposition chemistry. Look up the stoichiometry of this microbe or algae and trace the hydrogen mass balance. It's being fed in the form of sugar or urea or ammonia. Follow the hydrogen, that will show you where the energy is flowing and help point out attempts at perpetual motion in chemistry--like this one.
Joel Fairstein
Joel Fairstein
September 5, 2012
Cliff, you misunderstood the research. Sugar to isobutanol conversion was only used for proof of concept. The long-range goal is to convert carbon dioxide, not sugar.

Read the article!
Cliff Claven
Cliff Claven
September 5, 2012
Before these scientists go too far down this path, they need to understand and disclose to the reader that there is a large loss of energy in feeding a microbe sugar to get it to make isobutanol. This article presents this process as if this could become a viable source of fuel. It takes more fossil fuel energy to make the sugar, whether it is cultivated or synthesized, than the end-product isobutanol or hydrotreated hydrocarbon will yield back. We need biologists (and reporters) who understand thermodynamics. There actually is a discipline called 'biodynamics' that applies physics to biological systems. See Kooijman, S. A. L. M. Dynamic Energy and Mass Budgets in Biological Systems. Cambridge University Press, 2000. Solazyme is doing the same thing with micro-algae. They feed them sugar instead of using sunlight and photosynthesis. The resulting fuel is $61.33 a gallon.
Robert Pavel Oimeke
Robert Pavel Oimeke
September 5, 2012
This is an exciting and interesting research that promises to hold a major stake in commercial production of biofuels (Isobutal).
The next major challenge is to make this commercially viable!

I wish to seek clarification on "when the microbe's source of essential nutrients (such as nitrate or phosphate) is restricted, "it will go into carbon-storage mode," essentially storing away food for later use when it senses that resources are limited." but when the microbe is "tinkered with" it will produce Isobutal and still not store it for future use despite the essential nutrients lacking. What does the microbe store as food, an alternate, for future survival in such circumstances?

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