[tt] technologyreview: more on e.coli that produce butanol

[Alejandro Dubrovsky]

(
http://www.technologyreview.com/printer_friendly_article.aspx?id=20073
)

Wednesday, January 16, 2008
Better Bugs for Making Butanol
Engineered <em>E. coli</em> proves efficient at churning out the
biofuel.
By Alexandra M. Goho

In a push to find better biofuels to reduce gasoline consumption and
lower greenhouse-gas emissions, scientists have genetically engineered
E. coli that is highly efficient in producing butanol, a promising new
type of biofuel. The new technology could speed up the development of
butanol biofuels into a cost-effective alternative to ethanol.

While ethanol is the main biofuel on the market today, energy firms are
increasingly looking to alternatives such as butanol. "It has many
attractive properties," says Jim McMillan, manager of biorefining
process R&D at the National Renewable Energy Laboratory's National
Bioenergy Center, in Golden, CO. Because butanol packs more energy per
gallon than ethanol does, cars running on butanol get better mileage.
And, unlike ethanol, it doesn't mix with water, so it can be shipped in
existing petroleum pipelines without causing problems.

A number of research groups are engineering microbes that can convert
sugar from various feedstocks into butanol. Most of these groups rely on
the bacterium Clostridium acetobutylicum, which naturally makes a form
of butanol called 1-butanol. "But Clostridium is not easy to deal with,"
says James Liao, a chemical engineer at the University of California,
Los Angeles. "It grows slowly, it's very fastidious, and it's not easy
to genetically manipulate." Despite decades of tinkering by scientists,
the microbe still can't produce enough butanol to make it economically
viable as a transportation fuel, Liao says.

Instead, he and his colleagues turned to E. coli. Although the bacterium
does not produce butanol naturally, it is easy to modify and grows fast.
Instead of tweaking the pathway that the microbes employ for fermenting
sugar into alcohol, Liao reasoned that he could program E. coli to
produce butanol by diverting some of the microorganism's metabolites
into alcohol production. These metabolites, called keto acids, are
involved in the synthesis of amino acids, the building blocks of
proteins.

To make butanol from keto acids, the researchers inserted two different
nonnative genes into E. coli. The first gene came from a microbe
commonly used in the production of cheese. The gene codes for an enzyme
that converts keto acids into aldehydes. The second gene, derived from
yeast, codes for an enzyme that converts aldehydes into butanol.

Initially, when linked together in E. coli, the two genes allowed the
microbe to produce small amounts of butanol. With further genetic
modifications, Liao was able to dramatically increase the efficiency of
the process. For instance, deleting certain genes and boosting the
activity of others increased the amount of keto acids available for
conversion into butanol. With all the combined manipulations, the
engineered microbes achieved an efficiency high enough for industrial
use, says Liao.

Gevo, a biofuels startup based in Pasadena, CA, has acquired an
exclusive license to commercialize Liao's technology. (Liao is on the
company's scientific advisory board.) "It's a real breakthrough," says
Mathew Peters, Gevo's chief scientific officer. Not only did Liao
improve the efficiency of the process, but he also designed his microbes
to produce a particular form of butanol called isobutanol. "We believe
isobutanol is a superior fuel," says Peters. Compared with 1-butanol,
isobutanol has a higher octane number, which reduces knocking in the
vehicle's engine.

What's more, the biochemical pathway Liao designed for making isobutanol
can be transferred to other microbes. In addition to investigating E.
coli, Gevo is looking at different microorganisms that could be modified
in the same way. "We're interested in any organism that will make the
process cheaper," says Peters.

Gevo isn't alone in its pursuit of a better butanol-producing bug. In
June 2006, BP and DuPont joined efforts to develop butanol. .

Last June, BP and DuPont, along with Associated British Foods, announced
their plans to build a biobutanol pilot plant at an existing BP site in
England. The plant, which will use sugar beet as a feedstock, is
expected to begin operations in 2009, with the ultimate goal of
commercializing butanol after 2010.

According to Peters, Gevo plans to make a decision by the end of the
year on whether to go ahead with its own plans to build a butanol plant.
In the meantime, certain technological hurdles still need to be overcome
to make butanol cost competitive, he says. Mainly, the microbes need to
get faster at producing butanol, and their tolerance to isobutanol,
which is toxic to the organisms, must improve. Still, Peters expects
Gevo to resolve these issues in the coming months.
Copyright Technology Review 2008.