Navigation Links
Enzyme detectives uncover new reactions, products
Date:9/8/2008

UPTON, NY -- If your experiment doesn't go the way you expect, take a closer look -- something even more interesting may have happened. That strategy has led scientists at the U.S. Department of Energy's (DOE) Brookhaven National Laboratory to discover a fundamental shift in an enzyme's function that could help expand the toolbox for engineering biofuels and other plant-based oil products. The results will be published online the week of September 8, 2008, in the Proceedings of the National Academy of Sciences.

The Brookhaven scientists were trying to understand the factors that affect where carbon-carbon double bonds are placed in fatty acids, the building blocks of oils and fats, when they are "desaturated" -- that is, when a desaturase enzyme removes hydrogen from the carbon chain.

"Placing double bonds in different positions allows you to change the structure of the fatty acids to make products with different potential applications," explained Brookhaven biochemist John Shanklin, who led the research. The ultimate goal: engineering designer plant oils to be used as biofuels and/or raw materials to reduce the use of petroleum.

To try to change the position of a double bond, the Brookhaven team modified a desaturase enzyme, changing three of the 363 amino acids in its protein sequence. But when they tested the modified enzyme and looked for the expected product with its altered double-bond position, it wasn't there.

They could have moved on and made different amino acid changes to accomplish the initial goal. But Brookhaven research associate Edward Whittle was determined to figure out what was going on with the unusual result. "The substrate, or starting material, had been used up, so something was being produced -- substrates can't just disappear," Whittle said. "If it wasn't the product we were looking for, what was it?"

Whittle's detective work uncovered a remarkable discovery. Instead of producing a shift in double-bond position, the enzyme modification had yielded three completely new products -- two variations of a hydroxylated product called an allylic alcohol and a fatty acid containing two double bonds. "This was a profound shift in enzyme function," noted Shanklin, who has been working with modified enzymes for 15 years. "Usually you make changes very gradually, getting a few percent of a new product mixed with the original product. This was more like throwing a switch, making the change in function close to complete."

The discovery is also notable because the starting enzyme, like other soluble (membrane-independent) desaturases, can ordinarily perform only its one specified reaction -- desaturation. This is unlike desaturase enzymes that reside within the cell membrane, which appear to be more versatile, performing a range of reactions. The soluble and membrane enzymes, however, do share one key feature: both perform reactions that require the production of a highly reactive form of oxygen.

"Since both classes of enzymes produce activated oxygen, in theory the soluble enzymes, like their membrane counterparts, should be able to perform a variety of reactions as well," Shanklin said. "Our work demonstrates that this is indeed the case. Making small changes to the enzyme's amino acid sequence has unlocked the soluble desaturase's potential to facilitate a wider range of chemistry than has been seen before," Shanklin said.

The challenge is to figure out how these structural changes to the enzyme lead to the observed changes in reaction chemistry. Computer-generated models combining the known structure of the starting enzyme in conjunction with its new substrates are helping the scientists understand how the enzyme works. The next step is to obtain real 3-D crystal structures of enzyme-substrate complexes, using the National Synchrotron Light Source at Brookhaven Lab, to see how they match up with the predictions.

Analyzing the structures of soluble enzymes is much simpler than obtaining structures for membrane enzymes. So, in effect, this work is a fast-track approach for correlating structure with function, which should help scientists gain general mechanistic insights relevant to both classes of enzymes. "Understanding how nature has figured out how to do this very difficult chemistry, and how to control that chemistry," Shanklin said, "would be extremely satisfying from a purely scientific perspective. But applying this knowledge could have benefits for us all."

"Right now, the materials we use -- the plastics, foams, nylons -- have been limited by the structures of petroleum-based chemical feedstocks. But if we understand how to engineer designer desaturase-like plant enzymes, we can tailor-make feedstocks with optimal properties, instead of relying on the properties of preexisting raw materials," said Shanklin. "We'd no longer have to say, 'this is what we have, so this is what we can make.' Instead, we could make the best feedstock for a particular application by designing the raw materials that will yield it."


'/>"/>

Contact: Karen McNulty Walsh
kmcnulty@bnl.gov
631-344-8350
DOE/Brookhaven National Laboratory
Source:Eurekalert

Related biology news :

1. Green tea boosts production of detox enzymes, rendering cancerous chemicals harmless
2. MGH researchers describe new way to identify, evolve novel enzymes
3. Enzyme alerts cells powerful army to repair DNA damage
4. Scientists learn structure of enzyme in unusual virus
5. Scientists spy enzyme that makes us unique
6. Unlocking the function of enzymes
7. A key enzyme helps keep the synapse on track
8. Team of chemists receives $5 million grant to develop enzyme mimics
9. Iowa State researcher studies how enzymes break down cellulose
10. Researchers probe a DNA repair enzyme
11. Birth of an enzyme
Post Your Comments:
*Name:
*Comment:
*Email:
(Date:3/18/2016)... , March 18, 2016 --> ... of Biometrics, ICT, Manned & Unmanned Vehicles, Physical infrastructure and ... security companies in the border security market and the continuing ... and Europe has led visiongain to ... improved success. --> defence & security companies ...
(Date:3/15/2016)... New York , March 15, 2016 ... new market report published by Transparency Market Research "Digital Door ... Trends and Forecast 2015 - 2023," the global digital door ... US$ 731.9 Mn in 2014 and is forecast to grow ... 2023. Growth of micro, small and medium enterprises (MSMEs) across ...
(Date:3/14/2016)... NXTD ) ("NXT-ID" or the "Company"), ... the airing of a new series of commercials on Time ... 21 st .  The commercials will air on Bloomberg TV, ... the Street show. --> NXTD ) ("NXT-ID" or ... market, announces the airing of a new series of commercials ...
Breaking Biology News(10 mins):
(Date:4/28/2016)... NEW YORK , April 28, 2016 /PRNewswire/ ... biotechnology acceleration company reports the Company,s CEO  was ... capital titled Accelerators Enter When VCs Fear To ... Life Science Leader magazine is an ... work for everything from emerging biotechs to Big ...
(Date:4/27/2016)... ... 2016 , ... Cambridge Semantics, the leading provider of Smart Data ... has been named to The Silicon Review’s “20 Fastest Growing Big Data Companies of ... serves the needs of end users facing some of the most complex data challenges ...
(Date:4/27/2016)... RESEARCH TRIANGLE PARK, N.C. , April 27, 2016 ... announced today that Martine Rothblatt , Ph.D., Chairman ... an overview and update on the company,s business at ... Conference. The presentation will take place on ... and can be accessed via a live webcast on ...
(Date:4/27/2016)... ... ... Global Stem Cells Group CEO Benito Novas announced that Duncan Ross, Ph.D. ... Labs in Miami. , In 2004, Ross received his Ph.D. in Immunology from the ... the suppression of graft vs. host disease (GVHD) under UM Professor Robert Levy Ph.D. ...
Breaking Biology Technology: