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Going to Extremes

Mead has been lucky before. While working as a staff scientist in a UW-Madison chemistry lab, he helped develop a technology known as TA cloning that became a multi-million dollar product for Invitrogen, the California biotech company that licensed the approach. This success, as well as subsequent research experiences, inspired him to start Lucigen in his basement. “But you know what they say about lightning striking twice,” he laughs.

Until you come up with a product to sell, (the discovery of an enzyme) doesn’t really matter.

In 2000, Mead hired his first employee, Schoenfeld, who proposed the idea of bioprospecting among Yellowstone’s hot springs for viral enzymes. “When I first started thinking about this type of research,” says Schoenfeld, “I pulled out some review papers that said people were detecting (viruses in) ocean water and lake water, but nobody had even thought about hot spring water.” Before joining Lucigen, he’d already proposed this idea at two other Madison-area biotech companies and been shot down. Scooping up a super-bug containing a super-enzyme not only relies on luck, but some tricky lab work. Before it can be identified, the gene that encodes that super-enzyme must survive the sample preparation process, which often involves chopping all of the sample’s genetic material into more manageably sized pieces. Finally, should the enzyme be found, there remains the monumental task of developing it into a useful product that people will buy.

“Until you come up with a product to sell, (the discovery of an enzyme) doesn’t really matter,” says Schoenfeld. “And you need to make it user-friendly so the customer can just open up a kit and make it work.”

While Mead was undaunted by the challenge, he says it was tough finding funding for Lucigen’s field expeditions. Even when they finally succeeded, winning a Small Business Innovation Research grant from the federal government, the reviewers didn’t hesitate to make their reservations known. “Reviewers don’t usually fund fishing expeditions,” says Schoenfeld. “But in this case they said, ‘Sure, it’s a fishing expedition, but if it works, it would be worth it.'”

Schoenfeld applied for a research permit from Yellowstone, which approves between 30 and 50 such permits for microbial research each year. Early on, they met with some luck. Just as their first grant was running out, they discovered a new type of DNA polymerase-an enzyme similar to the famous Taq polymerase, but with some promising differences-in one of the springs along the park’s Firehole River. They spent five years developing it into a basic PCR kit for DNA analysis. Now, capitalizing on this enzyme’s unique properties, Schoenfeld is in the process of developing a 30-minute diagnostic test that can be used to detect a number of viral and microbial infections, including HIV and tuberculosis. It would require no equipment, and if he can get it to work, Schoenfeld is optimistic that he can make it precise enough to recognize one flu strain from another.

But in many ways, bioenergy is an even bigger gamble. Mead and Lucigen scientist Phil Brumm began building a library of enzymes for the industry several years ago, but they did so recognizing that the industry they hope to sell to does not yet exist. To date, no industrial process for the conversion of plant material to biofuels has proved cost-effective, and research on new methods of bioconversion remains in its infancy. Even as Mead fills his bottles with potentially promising bacteria, he does so with the knowledge that it may take years of lab work before he can say what he has-and whether it can play a meaningful role in making plant-based ethanol a commercial reality.

And that’s one of the hard realities of bioprospecting. Although the possibilities are enticing, the work involves a level of delayed gratification. On their visits to Yellowstone, Mead and Schoenfeld stick to their daily routines, performing the repeated tasks of setting up equipment, sampling and packing up with cold efficiency. The only inspirational moments come from the setting-the vast western sky, the steaming landscape, the glimpses of eagles and elk. Otherwise, the hours are filled with hiking and waiting-and hoping that the next bottle will pull up the microbial Moby Dick.

The trips usually span three or four days, but they can still feel incessantly long. The hours getting into and out of the park. The hikes laden with 40-pound packs of equipment. The nights spent at the kitchen sink, re-filtering samples to separate bacteria from viruses. Then an early bedtime so they can rise and repeat the whole thing the next day.

By the end of their seventh trip to Yellowstone, Schoenfeld will have collected five samples of concentrated viruses, and Mead will have filled more than 20 plastic bottles with bacteria-laced spring water, covering nearly 10 miles of trail in the process. From this catch, they’ll continue the search for Lucigen’s first million-dollar enzyme, a goal that-as long as it’s still ahead of them-will lead them to hot springs sites year after year. They don’t plan to stop until they find what they are looking for. It’s business, of course, but also something more.

“It’s like hunting or fishing in a way,” explains Schoenfeld. “The same brain chemical that makes people fish makes us go back for more (enzymes). You want to get the big one.”

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