The Wackett laboratory investigates enzyme transformations for biotechnological applications. The applications focus on biodegradation for environmental purposes and biocatalysis for producing specialty chemicals or detection kits.  The biodegradation research is now directed toward the treatment of waters generated during the process of hydraulic fracturing to obtain oil and gas from shale resources. We also study the biodegradation of s-triazine compounds such as the herbicide atrazine and pool water chemical cyanuric acid. The biocatalysis research is heavily focused on better understanding the enzymatic basis of bacterial hydrocarbon biosynthesis. Renewable hydrocarbons are currently of interest as fuels or feedstocks. The Wackett lab also studies enzymes that degrade food adulterants to use them for developing detection systems. For example, we had previously worked with Bioo Scientific to help develop the MaxSignal Melamine kit for detection melamine in milk and other food products. 

Research Overview

The Wackett laboratory studies microbial enzymes and pathways for biocatalysis and biodegradation and helped build the Biocatalysis/Bioegradation Database.

Cyanuric acid hydrolase X-ray structure showing bound inhibitor in center

Biodegradation research
The Wackett laboratory investigates microbial biodegradative metabolism and enzymes.  In one example, bacteria initiate metabolism of atrazine via the enzyme atrazine chlorohydrolase for which we first reported the structure (Seffernick, et al, 2010). In collaboration with Hideki Aihara’s group, we have defined the novel cyanuric acid hydrolase protein family (Seffernick, et al, 2012) and, more recently, the X-ray structure The Wackett laboratory is also studying the biodegradation of acrylamide and polycyclic aromatic hydrocarbons.

Bioremediation
Our studies on biodegradation provide opportunities for bioremediation of chemical contaminants in water. The chemicals treated are atrazine, cyanuric acid, acrylamide, polycyclic aromatic hydrocarbons, benzene and substituted aromatic compounds. Biodegrading microbes are being deployed in silica microspheres. The silica encapsulation stabilizes the in vivo enzyme activities and makes a formulation that can be stored and used as needed. We are developing specialized silica gels that enhance biotransformation rates and thus have important industrial applications. We have helped found the bioremediation company Minnepura Technologies, Inc.

Bacterial hydrocarbon biosynthesis
Industry is interested in renewable hydrocarbons as specialty chemical products. Our research investigates the fundamental mechanistic and structural issues underlying biological hydrocarbon synthesis. Studies are focused on the biosynthesis of long-chain olefins and diesel-length alkanes.

Bioinformatics
The Wackett laboratory is involved in a multi-investigator project to design the RAPID algorithm. RAPID is defined as Reactive Activity Product Identification and is a bioinformatics project designed to help predict the reactions catalyzed by broad-specificity enzymes that have significance in biosynthesis and biodegradation.

Dodge, A.G., K. Carrasquillo, L. Rivera, L. Xu, L.P. Wackett and M.J. Sadowsky (2015) A rapid detection method for L-abrine in food as a marker for the toxic protein abrin using two microbial enzymes. Appl. Environ. Microbiol. 81: 1610-1615. PDF

Mutlu, B.R., K. Hirschey, L.P. Wackett, and A. Aksan (2015) Optimization of aging, drying and storage conditions for biocatalytic silica gel. J.Sol-Gel Sci. Technol. 74: 823-833. DOI 1007/s10971-015-3690-8. PDF

Vail, A.W., P. Wang, H. Uefuji, D.A. Samac, C.P. Vance, L.P. Wackett, and M.J. Sadowsky (2015) Biodegradation of atrazine by three transgenic grasses and alfalfa expressing a modified bacterial atrazine chlorohydrolase gene. Transgenic Res. 24:475-488. PDF

Schürner, H., J.L. Seffernick, A. Grzybkowska, A. Dybala-Defratyka, L.P. Wackett, and M. Elsner (2015) Characteristic isotope fractionation patterns in s-triazine degradation have their origin in multiple protonation sites in the s-triazine hydrolase TrzN. Environ. Sci. Technol. 49: 3490-3498. PDF

Fenner, K. S. Canonica, L.P. Wackett and M. Elsner (2013) Evaluating pesticide biodegradation in the environment: Blind spots and emerging opportunities. Science 341: 752-758. PDF

Strong, L.C., T. Gould, L. Kasinkas, A. Aksan, and L.P. Wackett (2013) Biodegradation in produced waters: Chemistry, microbiology and engineering. J. Environ. Eng. 140 (5): B4013001-1 to B4013001-8. PDF

Mutlu, B.R., S. Yeom, H.-W. Tong, L.P. Wackett and A. Aksan (2013) Silicon-alkoxide cross-linked nanoparticle gels for encapsulation of bacterial biocatalysts. J. Mater. Chem. A 1: 11051-11060. PDF

Aukema, K, T. Makris, S. Stoian, J. Richman, E Munck, J.D. Lipscomb, and L.P. Wackett (2013) Cyanobacterial aldehyde deformylase oxygenation of aldehydes yields n-1 aldehydes and alcohols in addition to alkanes. ACS Catalysis 3: 2228-2238. PDF

Tong, H.-W., B.R. Mutlu, L.P. Wackett and A. Aksan. (2013) Silica/PVA biocatalytic nanofibers. Materials Lett. 111:234-237. PDF

Dodge, A.G., C.S. Preiner, and L.P. Wackett (2013) Expanding the cyanuric acid hydrolase protein family to the fungal kingdom. J. Bacteriol. 195: 5233-5241. PDF

Tong, H.-W., B.R. Mutlu, L.P. Wackett, and A. Aksan (2014) Manufacturing of bioreactive nanofibers for bioremediation. Biotech. Bioeng. 111(8): 1483-1493. PDF

Aukema, K.A., L. Kasinkas, A. Aksan, L.P. Wackett (2014) Biodegradation of novel hydrocarbon ring structures found in hydraulic fracturing waters using silica-encapsulated Pseudomonas sp. NCIB9816-4. Appl. Environ. Microbiol. 80: 4968-4976. PDF

Cho, S.,  K. Shi, J.L. Seffernick, A.G. Dodge, L.P. Wackett, and H. Aihara (2014) Cyanuric acid hydrolase from Azorhizobium caulinodans ORS 571: Crystal structure and insights into a new class of ser-lys dyad proteins. PLOS ONE 2014 Jun 10;9(6):e99349. PDF

Mutlu, B.R., S. Yeom, L.P. Wackett, and A. Aksan (2015) Modelling and optimization of a bioremediation system utilizing silica gel encapsulated whole-cell biocatalyst. Chem. Eng. J. 259: 574-580. PDF