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Breeanna Urbanowicz
Assistant Professor

Structure and function of plant carbohydrate active enzymes

Telephone: 706-542-4479
Fax: 706-542-4412

Short Biography
Research Interests

Short Biography:
Dr. Urbanowicz received her B.S. in Biology in 2001 from Purdue University and her Ph.D. in 2008 from Cornell University. Prior to her junior faculty position at the Complex Carbohydrate Research Center, Dr. Urbanowicz was a Postdoctoral Fellow (2008-2013) in the Department of Biochemistry and Molecular Biology at the University of Georgia.

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Research Interests:
Biological molecules, including proteins, polysaccharides, and nucleic acids, are assembled to create complex structures with biochemical and biomechanical properties that are greater than the sum of their parts. My current research focuses on understanding the structure and function of plant carbohydrate active enzymes involved in polysaccharide biosynthesis and modification. Plant cell walls are complex macrostructures comprised of cellulose, hemicellulose, and pectins, together with lesser amounts of protein and phenolic molecules. These components assemble and interact with one another to produce dynamic structures with many capabilities, including providing mechanical support to plant structures and determining the size and shape of plant cells.

The research of my group focuses on understanding the integral steps in the molecular pathways used by plants to synthesize complex polysaccharides. A key area of interest is development of methods to express and analyze recombinant plant enzymes, which has allowed us to investigate plant biochemical pathways in vitro. We have now generated a large collection of recombinant enzymes that are able to catalyze highly specific, covalent modifications of polysaccharides, which we utilize as nature-inspired molecular tools for targeted functionalization and labeling of glycopolymers that greatly expand our toolkit for producing glycopolymer-based products with valuable properties.

High throughput expression of plant biocatalysts. In collaboration with Kelley Moremen at the CCRC, we have designed and evaluated over 75 constructs for heterologous expression in HEK293 cells encoding plant glycosyltransferases (GTs) from diverse families in addition to both polysaccharide O-methyltransferases (O-MTs) and O-acetyltransferases (O-AcTs). Assessment of these constructs for both protein expression levels and retention of in vitro activity determined that this is a robust heterologous expression system for plant derived enzymes. We have applied this system to the in vitro synthesis of decorated hemicellulosic polymers, resulting in the first proof-of-concept generation of enzymatically synthesized, high degree of polymerization (DP), substituted plant polysaccharides in vitro (Urbanowicz et al., 2014).

Polysaccharide Methyltransferases. We have shown that Arabidopsis GXMT1 encodes a glucuronoxylan (GX)-specific 4-O-methyltransferase responsible for methylating 75% of the GlcA residues in GX isolated from mature Arabidopsis inflorescence stems. Reduced methylation of GX in gxmt1-1 plants is correlated with altered lignin composition and increased release of GX by mild hydrothermal pretreatment (Urbanowicz et al., 2012). The ability to selectively manipulate polysaccharide O-methylation may provide new opportunities to modulate biopolymer interactions in the plant cell wall. We are currently investigating the role of other members in this family on polysaccharide methyletherfication.

Investigating the mechanism of polysaccharide O-acetylation. Despite the high degree of O-acetyl substituents found in plant glycopolymers, the biochemical and mechanisms of polysaccharide O-acetylation employed by plants are still lacking. An unsolved question is the source of the acetyl group used by acetyltransferases. We have developed robust methods to biochemically analyze O-acetyltransferases and are applying these techniques to investigate the molecular details of polysaccharide methylation.

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Publications: Author's Last Name: Urbanowicz

Journal Articles
Book Chapters are listed at the bottom of this page.

MJ Soto, K Prabahkar, D Gopaldas, C Ruprecht, I Black, M Pena, F Pfrengle, K Moremen, B Urbanowicz, MG Hahn. 2021. AtFUT4 and AtFUT6: Biochemically Redundant Arabinogalactan Protein-Specific Fucosyltransferases . Frontiers in Plant Science-Plant Glycobiology : -.

V Bharadwaj, MF Crowley, MJ Pena, B Urbanowicz, M O'Neill. 2020. Mechanism and Reaction Energy Landscape for Apiose Cross-linking by Boric Acid in Rhamnogalacturonan II. The Journal of Physical Chemistry Nov 12;124 (45): 10117-10125. PMID:33112619

PJ Smith, EO Soto, C Roth, WJ Barnes, BR Urbanowicz, F Pfrengle. 2020. Enzymatic Synthesis of Artificial Polysaccharides for Materials Applications. ACS Sustainable Chemistry & Engineering July 15: DOI:10.1021/-acssuschemeng.0c.

C Ruprecht, MP Bartezko, D Senf, A Lakhina, PJ Smith, M Soto, H Oh, DV Silva, MH Clausen, KW Moremen, MG Hahn, BR Urbanowicz, F Pfrengle. 2020. A Glycan Array-Based Assay for the Identification and Characterization of Plant Glycosyltransferases. Angew Chem Int Ed Engl Jul 20;59(30): 12493-12498. PMID:32396713

PK Prabhakar, HT Wang, PJ Smith, JY Yang, WJ Barnes, MJ Pena, KW Moremen, BR Urbanowicz. 2020. Heterologous expression of plant glycosyltransferases for biochemistry and structural biology. Methods in Cell Biology. Methods Cell Biol 160: 145-165. PMID:32896313

PS Smith, MJ Pena, WS York, BR Urbanowicz. 2020. Analytical Techniques for Determining the Role of Domain of Unknown Function 579 Proteins in the Synthesis of O-Methylated Plant Polysaccharides. Slas Technol Aug;25(4): 345-355. PMID:32204644

V Lunin, HT Wang, VS Bharadwaj, P Alahuhta, MJ Pena, JY Yang, SA Archer-Hartmann, PA Azadi, ME Himmel, KW Moremen, WS York, YJ Bomble, BR Urbanowicz. 2020. Molecular mechanism of polysaccharide acetylation by the AtXOAT1 xylan O-acetyltransferase. Plant Cell Jul;32(7): 2367-2382. PMID:32354790

M O'Neill, I Black, B Urbanowicz, V Bharadwaj, M Crowley, S Koj, M Pena. 2020. Locating Methyl-Etherified and Methyl-Esterified Uronic Acids in the Plant Cell Wall Pectic Polysaccharide Rhamnogalacturonan II. SLAS Technology Aug;25(4): 329-344. PMID:32468908

J Yang, G Bak, T Burgin, WJ Barnes, H Mayes, MJ Pena, B Urbanowicz, E Nielsen. 2020. Biochemical and Genetic Analysis Identify CSLD3 as a Novel β-1,4-glucan Synthase that Functions during Plant Cell Wall Synthesis . Plant Cell May;32(5): 1749-1767. PMID:32169960

V Lunin, HT Wang, V Bharadwaj, PM Alahuhta, MJ Pena, JY Yang, SA Archer-Hartmann, P Azadi, M Himmel, KW Moremen, WS York, YJ Bomble, B Urbanowicz. 2020. Molecular mechanism of polysaccharide acetylation by the Arabidopsis xylan O-acetyltransferase XOAT1. Plant Cell 32(7): 2367-2382. PMID:32354790

MLA De, DL Duewer, A Fung, L.H. Liu, HK Yau, O Potter, GO Staples, K Furuki, R Frenkel, Y Hu, Z Sosic, P. Zhang, F. Altmann, C Gru, C Shao, J. Zaia, W Evers, S Pengelley, D Suckau, A Wiechmann, A Resemann, W Jabs, A Beck, JW Froehlich, YC Huang, Y. Liu, S Sun, Y. Wang, Y Seo, HJ An, NC Reichardt, JE Ruiz, S Archer-Hartmann, P. Azadi, B.L. Bell, Z Lakos, Y An, JF Cipollo, M Pucic-Bakovic, J Stambuk, G Lauc, X. Li, PG Wang, A Bock, R Hennig, E. Rapp, M Creskey, TD Cyr, M Nakano, T Sugiyama, PA Leung, P Link-Lenczowski, J Jaworek, S.-J. Yang, H. Zhang, T.M. Kelly, S Klapoetke, R Cao, JY Kim, HK Lee, JY Lee, JS Yoo, SR Kim, SK Suh, N de, D Falck, GSM Lagveen-Kammeijer, M Wuhrer, RJ Emery, RP Kozak, L LieuLPRoyle, PA Urbanowicz, NH Packer, X. Song, A Everest-Dass, E. Lattova, S Cajic, K Alagesan, D. Kolarich, T Kasali, V Lindo, Y. Chen, K Goswami, B.C. Gau, R Amunugama, R Jones, CJM Stroop, K. Kato, H Yagi, S Kondo, CT Yuen, A Harazono, X Shi, PE Magnelli, BT Kasper, LK Mahal, DJ Harvey, R O'Flaherty, PM Rudd, R Saldova, ES Hecht, DC Muddiman, J. Kang, P Bhoskar, D Menard, A Saati, C Mrele, S.W. Mast, S Tep, J Truong, T Nishikaze, S Sekiya, A Shafer, S Funaoka, M Toyoda, LPS de, C Caron, P Pradhan, NC Tan, Y. Mechref, S Patil, JS Rohrer, R Chakrabarti, D Dadke, M Lahori, C Zou, JPC Cairo, B Reiz, RM Whittal, CB Lebrilla, S.L. Wu, A Guttman, M Szigeti, BG Kremkow, KH Lee, C Sihlbom, B Adamczyk, C Jin, NG Karlsson, J Ornros, G Larson, J Nilsson, B. Meyer, A Wiegandt, E Komatsu, H. Perreault, ED Bodnar, N Said, YN Francois, E Leize-Wagner, LS Maier, A Zeck, AJR Heck, Y. Yang, R Haselberg, YQ Yu, W Alley, JW Leone, H Yuan, SE Stein . 2020. NIST Interlaboratory Study on Glycosylation Analysis of Monoclonal Antibodies: Comparison of Results from Diverse Analytical Methods. Mol Cell Proteomics 19(1): 11-30. PMID:31591262

MJ Soto, BR Urbanowicz, MG Hahn. 2019. Plant Fucosyltransferases and the Emerging Biological Importance of Fucosylated Plant Structures. Critical Reviews in Plant Sciences 38(4): 327-388.

P Magistrado-Coxen, Y Aqeel, A Lopez, JR Haserick, BR Urbanowicz, CE Costello, J Samuelson. 2019. The most abundant cyst wall proteins of Acanthamoeba castellanii are lectins that bind cellulose and localize to distinct structures in developing and mature cyst walls. PLoS May 16;13(5): e0007352-doi: 10.1371. PMID:31095564

JK Jensen, M Busse-Wicher, CP Poulsen, JU Fangel, PJ Smith, JY Yang, M-J Pena, MH Dinesen, HJ Martens, M Melkonian, GK Wong, KW Moremen, CG Wilkerson, HV Scheller, P Dupree, P Ulvskov, BR Urbanowicz, J Harholt. 2018. Identification of an algal xylan synthase KfXYS1 proves that there is functional orthology between algal and plant cell wall biosynthesis. New Phytologist 218: 1049-1060. PMID:29460505

RA Amos, S. Pattathil, JY Yang, MA Atmodjo, BR Urbanowicz, KW Moremen, D. Mohnen. 2018. A two-phase model for the non-processive biosynthesis of homoglacturonan polysaccharides by the GAUT1:GAUT7 complex. J. Biol. Chem. 293: 19047-19063. PMID:30327429

W Muchero, KL Sondreli, JG Chen, BR Urbanowicz, J Zhang, V Singan, Y Yang, RS Brueggerman, J Franco-Coronado, N Abraham, JY Yang, KW Moremen, AJ Weisberg, JH Chang, E Lindquist, K Berry, P Ranjan, SR Jawdy, J Schmutz, GA Tuskan, JM LeBoldus. 2018. Association mapping, transcriptomics, and transient expression identify candidate genes mediating plant-pathogen interactions in a tree. Proc. Natl. Acad. Sci. USA 115: 11573-11578. PMID:30337484

RA Amos, S Pattathil, JY Yang, MA Atmodjo, BR Urbanowicz, KW Moremen, D Mohnen. 2018. A two-phase model for the non-processive biosynthesis of homogalacturonan polysaccharides by the GAUT1:GAUT7 complex. J Biol Chem 293(49): 19047-19063. PMID:30327429

B.R. Urbanowicz, V.S. Bharadwaj, M. Alahuhta, M.J. Pena, V.V. Lunin, Y.J. Bomble, S. Wang, J.Y. Yang, S.T. Tuomivaara, M.E. Himmel, K.W. Moremen, W.S. York, M.F. Crowley. 2017. Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. Plant J. 91: 931-949. PMID:28670741

D. Ndeh, A. Rogowski, A. Cartmell, A.S. Luis, A. Basle, J. Gray, I. Venditto, J. Briggs, X. Zhang, A. Labourel, N. Terrapon, F. Buffetto, S. Nepogodiev, Y. Xiao, R.A. Field, Y. Zhu, M.A. O'Neill, B.R. Urbanowicz, W.S. York, G.J. Davies, D.W. Abbott, M.C. Ralet, E.C. Martens, B. Henrissat, H.J. Gilbert. 2017. Complex pectin metabolism by gut bacteria reveals novel catalytic functions. Nature 544: 65-70. PMID:28329766

P.J. Smith, H.T. Wang, W.S. York, M.J. Pena, B.R. Urbanowicz. 2017. Designer biomass for next-generation biorefineries: Leveraging recent insights into xylan structure and biosynthesis. Biotechnol. Biofuels 10: 286-286. PMID:29213325

P. Dallabernardina, C. Ruprecht, P.J. Smith, M.G. Hahn, B.R. Urbanowicz, J. Harholt. 2017. Automated glycan assembly of plant cell wall oligosaccharides enables partial characterization of antibodies recognizing galactosylated xyloglucan epitopes. Org. Biomol. Chem. 15: 9996-10000. PMID:29177276

A.C. Srivastava, F. Chen, T. Ray, S. Pattathil, M.J. Pena, U. Avci, H. Li, D.V. Huhman, J. Backe, B. Urbanowicz, J.S. Miller, M. Bedair, C.E. Wyman, L.W. Sumner, W.S. York, M.G. Hahn, R.A. Dixon, E.B. Blancaflor, Y. Tang. 2015. Loss of function of folylpolyglutamate synthetase 1 reduces lignin content and improves cell wall digestibility in Arabidopsis. Biotechnol. Biofuels 8: 224-. PMID:26697113

B.R. Urbanowicz, M.J. Pena, H.A. Moniz, K.W. Moremen, W.S. York. 2014. Two Arabidopsis proteins synthesize acetylated xylan in vitro. Plant J. 80: 197-206. PMID:25141999

A. Kulkarni, M.J. Pena, U. Avci, K. Mazumder, B. Urbanowicz, S. Pattathil, Y. Yin, M.A. ONeill, A.W. Roberts, M.G. Hahn, Y. Xu, A.G. Darvill, W.A. York. 2012. The ability of land plants to synthesize glucoronoxylans predates the evolution of tracheophytes. Glycobiology 22: 439-451. PMID:22048859

M.J. Pena, S.T. Tuomivaara, B.R. Urbanowicz, M.A. ONeill, W.S. York. 2012. Methods for structural characterization of the products of cellulose- and xyloglucan-hydrolyzing enzymes. Methods Enzymol. 510: 121-139. PMID:22608724

B.R. Urbanowicz, M.J. Pena, S. Ratnaparkhe, U. Avci, J. Backe, H.F. Steet, M. Foston, H. Li, M.A. O'Neill, A.J. Ragauskas, A.G. Darvill, C. Wyman, H.J. Gilbert, W.S. York. 2012. 4-O-Methylation of glucuronic acid in Arabidopsis glucuronoxylan is catalyzed by a domain of unknown function family 579 protein. Proc. Natl. Acad. Sci. USA 109: 14253-14258. PMID:22893684

D.M. Harris, K. Corbin, H.T. Wang, R. Gutierrez, A.L. Bertolo, C. Petti, D.M. Smilgies, J.M. Estevez, D. Bonetta, B.R. Urbanowicz, D.W. Ehrhardt, C.R. Somerville, J.K.C. Rose, M. Hong, S. DeBolt. 2012. Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1A903V and CESA3T9421 of cellulose synthase. Proc. Natl. Acad. Sci. USA 109: 4098-4103. PMID:22375033

Book Chapters

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