Abstract

Hydrocarbons are readily oxidized by bacteria utilizing the AlkB gene and thus the actions of AlkB represent one of nature's best defenses against the environmental effects of oil spills. The mechanism by why AlkB works is not well understood. Nor is it well understood why AlkB, which is thought to be structurally similar to enzymes that catalyze the conversion of single C-C bonds to double C=C bonds, catalyzes the addition of an OH group to an alkane instead. The goal of this project is to determine the three-dimensional structure of AlkB, using X-ray crystallography, to develop a model of the relationship between structure and function in this important metalloenzyme. To accomplish this goal, mechanistic work using diagnostic substrates, and spectroscopic characterizations, will also be carried out.

Public Health Relevance

This project focuses on understanding how the enzyme that catalyzes the first step in the transformation of oil to carbon dioxide works. It will also explore how this enzyme, which is structurally similar to enzymes important in human lipid metabolism, functions in comparison with that important class of enzymes as well.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
2R15GM072506-02
Application #
7714890
Study Section
Special Emphasis Panel (ZRG1-BCMB-B (02))
Program Officer
Anderson, Vernon
Project Start
2005-03-01
Project End
2013-08-31
Budget Start
2009-09-01
Budget End
2013-08-31
Support Year
2
Fiscal Year
2009
Total Cost
$210,000
Indirect Cost

  Institution

Name
Bates College
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
058951401
City
Lewiston
State
ME
Country
United States
Zip Code
04240

  Publications

Hsieh, Chun H; Huang, Xiongyi; Amaya, José A et al. (2017) The Enigmatic P450 Decarboxylase OleT Is Capable of, but Evolved To Frustrate, Oxygen Rebound Chemistry. Biochemistry 56:3347-3357
Austin, Rachel Narehood; Saito, Mak A (2014) Metals in marine biochemistry. Metallomics 6:1105-6
Austin, Rachel Narehood; Kenney, Grace E; Rosenzweig, Amy C (2014) Perspective: what is known, and not known, about the connections between alkane oxidation and metal uptake in alkanotrophs in the marine environment. Metallomics 6:1121-5
Naing, Swe-Htet; Parvez, Saba; Pender-Cudlip, Marilla et al. (2013) Substrate specificity and reaction mechanism of purified alkane hydroxylase from the hydrocarbonoclastic bacterium Alcanivorax borkumensis (AbAlkB). J Inorg Biochem 121:46-52
Cooper, Harriet L R; Mishra, Girish; Huang, Xiongyi et al. (2012) Parallel and competitive pathways for substrate desaturation, hydroxylation, and radical rearrangement by the non-heme diiron hydroxylase AlkB. J Am Chem Soc 134:20365-75
Austin, Rachel N; Luddy, Kate; Erickson, Karla et al. (2008) Cage escape competes with geminate recombination during alkane hydroxylation by the diiron oxygenase AlkB. Angew Chem Int Ed Engl 47:5232-4
Chakrabarty, Sarmistha; Austin, Rachel N; Deng, Dayi et al. (2007) Radical intermediates in monooxygenase reactions of rieske dioxygenases. J Am Chem Soc 129:3514-5
Rozhkova-Novosad, Elena A; Chae, Jong-Chan; Zylstra, Gerben J et al. (2007) Profiling mechanisms of alkane hydroxylase activity in vivo using the diagnostic substrate norcarane. Chem Biol 14:165-72
Austin, Rachel N; Deng, Dayi; Jiang, Yongying et al. (2006) The diagnostic substrate bicyclohexane reveals a radical mechanism for bacterial cytochrome P450 in whole cells. Angew Chem Int Ed Engl 45:8192-4
Bertrand, Erin; Sakai, Ryo; Rozhkova-Novosad, Elena et al. (2005) Reaction mechanisms of non-heme diiron hydroxylases characterized in whole cells. J Inorg Biochem 99:1998-2006