Markus Pauly
Markus Pauly
Biofuels, Carbon Sequestration
Associate Professor, Fred Dickinson Chair of Wood Science and Technology
Energy Biosciences Building 212C
2151 Berkeley Way, Berkeley, California 94720-5230
Phone 510.642.1722
Lab Phone 510.643.1027
Fax 510.642.1490

PhD  Technical University Aachen, Germany
M.S.  Technical University Aachen, Germany, 1993

The most abundant renewable plant resource on this planet are the lignicellulosics present in the plant cell wall. They present a sophisticated, highly complex material consisting mainly of various polysaccharides and polyphenols. The Pauly lab uses a synthetic biology approach whereby all necessary components of the biosynthetic machinery of hemicellulosic and pectinaceous polysaccharides are identified. To achieve this goal we use various genetic approaches including forward, reverse, and chemical genetics with the model organism Arabidopsis, but also maize as a grass species.

Plant Cell Wall feedstocks

Research

Plant Cell Walls as a renewable resource for Biofuels and other commodity Chemicals

With an estimated annual production of 180 billion tons per year plant cell walls (lignocellulosics) present an enormous renewable resource, of which only 2% are utilized by humans. The major bottleneck in utilizing this carbon neutral resource for the sustainable production of second generation biofuels is the recalcitrance of the wall material to degradation. With our various research projects we aim at characterizing the structural features of the wall that lead to this recalcitrance. In the long run such knowledge will aid us in overcoming them.

The structure of the Plant Cell Wall

Higher plant cells are encased in cell walls that define their shape and contribute to the strength and structural integrity not only of individual cells, but also of the entire plant. Despite its necessary rigidity, the cell wall is a highly dynamic entity that is metabolically active. It plays crucial roles in diverse cell activities such as growth, differentiation, cell-to-cell communication and transport, senescence, abscission, and plant-pathogen interactions. Microcrystalline cellulose is embedded in a hydrated matrix consisting of coextensive networks of complex heteropolysaccharides and sometimes glycoproteins and polyphenols, such as lignin.

Our research entails the establishment of an analytical platform for the analysis of wall polysaccharides, a forward genetic approach to identify novel wall structures and reverse genetic approaches to gain insights into wall biosynthesis.

Analytical Platform for the Microanalysis of Wall Polysaccharides

Our lab uses mainly classical carbohydrate chemistry based methods to describe the structure of particular wall polysaccharides. These methods encompass solubilization of various wall polymers using sequential extraction procedures that make use of wall degrading enzymes and chemicals. The resulting fractions are then analysed using techniques such as monosaccharide composition and glycosidic linkage analysis see videos (Lignin analysis: http://www.jove.com/details.php?id=1745; Carbohydrate analysis http://www.jove.com/details.php?id=1837). We also determine the presence of ester substituents (such as O-acetyl-substituents). Since such an analysis is rather labor-intensive and time consuming, an oligosaccharide mass profiling method (OLIMP) using specific polysaccharide hydrolases in combination with mass spectrometry has been developed (Video:http://www.jove.com/details.php?id=2046). The sensitivity of OLIMP allows for the rapid assessment of even minute amount of tissue-materials. A profile can be obtained from preparations of as little as 500 Arabidopsis cells prepared by a laser-dissection catapulting instrument.

Forward Genetic Approach: Identification of Structural Wall Mutants

We identify wall mutants by screening of populations of chemically mutagenised seeds for novel structural wall mutants.

An Arabidopsis mutants population was screened using OLIMP. It has lead to the identification of 36 distinct mutants with altered xyloglucan structures including the abundance of ester-substituents. Map-based cloning of the mutated genes gave valuable insights into biosynthesis, metabolism and function of structural variations of xyloglucan. For example, it became evident that the microheterogeneity of wall polysaccharides is dependent not on its biosynthetic machinery but on apoplastic "trimming" of the sidechains by glycosidases.      

A maize mutant population was screened for altered sugar content. Indeed, one of the mutants, termed "candy-leaf 1" or Cal-1, exhibited a 250% increase in glucan content. Cal-1 is currently tested in the field for enhanced yields of biofuel production. 

Massive parallel sequencing: Cell Wall Biosynthesis

Although information about the structural components of cell walls has increased considerably in recent years, very little is known about the biosynthesis of individual wall components on a molecular level. We employed Ilumina sequencing of plant species that synthesize only a single polysaccharide. This approach lead to the identification of genes in whole biosynthetic pathways. A reverse genetic approach is then employed with the correspondent Arabidopsis genes/mutants to ascertain the function of the gene and its role in polysaccharide biosynthesis. Currently, numerous novel genes involved in the synthesis of nucleotide sugars, the substrates for polysaccharide synthesis, have been identified through bioinformatic means by comparison to gene-sequences of well-characterized bacterial enzymes. In addition, we have identified genes that are responisble for polysaccharide O-acetylation. This substitution is an impediment to enzymatic degradation in a biorefinery and once released presents an inhibitor to many fermenting organisms.

The overall goal of these projects is to be able to assemble and reconstruct whole plant polysaccharide pathways in yeast that would produce the particular hemicellulose. 

Recent Publications

(last 5 years, out of 88 total)

Xiong G, Cheng K, Pauly M, 2013, Reduction in xylan O-acetylation results in increased recalcitrance to saccharification as indicated by the Arabidopsis mutant tbl29, in press in Molecular Plant

Gille S, Sharma V, Baidoo EEK, Keasling JD, Scheller HV, Pauly M, 2013, Arabinosylation of an AGP-like polymer impacts root growth as exemplified by the Arabidopsis glycosyltransferase mutant ray1, in press in Molecular Plant

Cheng K, Sorek H, Zimmermann H, Wemmer DE, Pauly M, 2013, Solution-state 2D NMR spectroscopy of plant cell walls enabled by a DMSO-d6[Emim]OAc solvent, Analytical Chemistry, 85 (6) 3213-3221

Liu Z, Padmanabhan S, Cheng K, Xie H, Schwyter P, Pauly M, Bell AT, Prausnitz JM, 2012, Aqueous-ammonia delignification of miscanthus and enzymatic hydrolysis to sugars, Bioresource Technology, 135 23-29

Handford M, Furlán CR, Marchant L, Segura M, Gómez D, Alvarez-Buylla E, Xiong G, Pauly M, Orellana A, 2012, Arabidopsis plants lacking AtUTr7, a Golgi-localized UDP-glucose/UDP-galactose transporter, exhibit alterations in lateral roots, Molecular Plant, 5 (6) 1263-1280

Park SH, Mei C, Pauly M, Ong RG, Dale BE, Sabzikar R, Fotoh H, Nguyen T, Sticklen M, 2012, Down regulation of maize cinnamoyl-CoA reductase via RNAi technology creates brown midrib and improves AFEX-pretreated conversion into fermentable sugars for biofuels, Crop Science 52 (6) 2687-2701

Chiniquy D, Sharma V, Schultink A, Baidoo EE, Rautengarten C, Cheng K, Carroll A, Ulvskov P, Harholt J, Keasling JD, Pauly M, Scheller HV, Ronald PC, 2012, XAXT1, a grass specific xylan:xylosyltransferase in the glycosyltransferase family 61, Proceedings of the National Academy of the USA 109 (42) 17117-17122

Jensen JK, Schultink A, Keegstra K, Wilkerson CG, Pauly M, 2012, RNA-Seq. of developing nasturtium seeds (Tropaeolum majus): Identification and characterization of an additional glactosyltransferase involved in xyloglucan biosynthesis, Molecular Plant, 5 (5) 984-992

Gille S, Pauly M, 2012, Mechanism of plant cell wall O-acetylation, Frontiers in Plant Physiology 3:12, doi: 10.3389/fpls.2012.00012

Gille S, Souza A, Xiong G, Benz M, Schultink A, Ida-Reca B, Pauly M, 2011, O-acetylation of xyloglucan requires AXY4/AXY4L, proteins with a TBL and DUF231 domain, Plant Cell 23 (11) 4041-4053

Chuck G, Tobias C, Kraemer F, Sun L, Li C, Arora R, Singh S, Dibble D, Vogel J, Simmons B, Pauly M, Hake S, 2011, Overexpression of the maize conrgrass1 microRNA gene prevents flowering, improves digestability and increases starch content of biofuel crop plants, Proceedings of the National Academy of the USA 108 (42) 17550-17555

Gunl M, Neumetzler L, Kraemer F, Souza A, Schultink A, Pena M, York WS, Pauly M, 2011, AXY8 encodes an a-fucosidase, underpinning the importance of apoplastic metabolism on the fine structure of plant cell wall polysaccharides, Plant Cell 23 (11), 4025-4040

Troncoso-Ponce MA, Kilaru A, Cao X, Durrett T, Fan J, Jensen J, Pauly M, Wilkerson C, Ohlrogge J, 2011, Comparative deep transcriptional profiling of four developing oilseeds, Plant Journal 68, 1014-1027

Gille S, Kun C, Skinner ME, Liepman AH, Wilkerson C, Pauly M, 2011, Deep Sequencing of Voodoo Lily (Amorphophallus konjac): An approach to identify relevant genes involved in the synthesis of the hemicellulose glucomannan, Planta, 234, 515-526

Manabe Y, Nafisi M, Verhertbruggen Y, Orfila C, Gille S, Rautengarten C, Cherk C, Marcus SE, Somerville S, Pauly M, Knox JP, Sakuragi Y, Scheller HVS, 2011, Loss-of-function mutation of reduced wall acetylation 2 in Arabidopsis leads to reduced cell wall acetylation and increased resistance to Botrytis cinerea, Plant Physiology, 155 (3), 1068-1078
 
Gunl M, Pauly M, 2011, AXY3 encodes a a-xylosidase that impacts the abundance and accessibility of the hemicellulose xyloglucan in Arabidopsis plant cell walls, Planta, 233 (4), 707-719
 
Gunl M, Kraemer FJ, Pauly M, 2011, Oligosaccharide mass profiling (OLIMP) of cell wall polysaccharides by MALDI-TOF/MS, Methods in Molecular Biology Vol 715: The Plant Cell Wall, 43-54, Editor Popper Z, Humana Press
 
Perrson S, Sorensen I, Moller I, Willats W, Pauly M, 2011, Dissection of the Plant Cell Wall by High Throughput Methods, Annual Plant Reviews Vol 41: Plant Polysaccharides: Biosynthesis and Bioengineering, 43-64, Editor Ulvskov P, Wiley-Blackwell
 
Suen G, Scott  JJ, Aylward FO, Adams SM, Tringe SG, Pinto-Tomas AA, Foster CE, Pauly M, Weimer PJ, Barry KW, Goodwin LA, Bouffard P, Li L, Osterberger J, Harkins TT, Slater SC, Donohue TJ, Currie CR, 2010, An insect herbivore microbiome with high plant biomass degrading capacity, PLoS Genetics, 6 (9), 433-443 
 
Pauly M, 2010, A blue-print of the protoplast’s dwelling, Plant Physiology, Classic collection, 154, 1
 
Kaida R, Serada S, Norioko N, Norioka S, Neumetzler L, Pauly M, Sampedro J, Zarra I, Hayashi T, Kaneko TS, 2010, Potential role for purple acid phosphatase in the dephosphorylation of wall proteins in tobacco cells, Plant Physiology, 153 (2), 603-610
 
Lopes FJF, Pauly M, Lau EY, Diola V, Passos JL, Loureiro ME, Brommonshenkel SH, 2010, The EgMUR3 xyloglucan galactosyltransferase from Eucalyptus grandis complements the mur3 cell wall phenotype in Arabidopsis thaliana, Tree Genetics and Genomes, 6, 745-756
 
Gunl M, Gille S, Pauly M, 2010, OLIigo Mass Profiling (OLIMP) of extracellular polysaccharides, in production in Journal of visualized Experiments Journal of visualized Experiments, http://www.jove.com/index/details.stp?id=2046, doi: 10.3791/2046
 
Pauly M, Keegstra K, 2010, Plant cell walls as precursors for biofuels, Current Opinion in Plant Biology, 13 (3), 305-312
 
Santoro N, Cantu SL, Tornqvist CEI, Falbel TG, Bolivar JL, Patterson SE, Pauly M, Walton JD, 2010, A high throughput platform for screening milligram quantities of plant biomass for lignocellulosic digestability, BioEnergy Research, 3 (1), 93-102    
 
Foster CE, Martin T, Pauly M, 2010, Comprehensive compositional analysis of Plant Cell Walls (lignocellulosic biomass); Part II: carbohydrates, Journal of visualized Experiments, 37, http://www.jove.com/index/details.stp?id=1837, doi: 10.3791/1837
 
Foster CE, Martin T, Pauly M, 2010, Comprehensive compositional analysis of Plant Cell Walls (lignocellulosic biomass); Part I: lignin, Journal of visualized Experiments, 37; http://www.jove.com/index/details.stp?id=1745, doi: 10.3791/1745
 
Li M, Xiong G, Li R, Tan D, Zhang B, Cui J, Pauly M, Cheng Z, Zhou Y, 2009, Rice Cellulose Synthase-Like D4 Is Essential for Normal Cell Wall Biosynthesis and Plant Growth, Plant Journal 60, 1055-1069
 
Gille S, Haensel U, Ziemann M, Pauly M, 2009, Identification of plant cell wall mutants by means of a forward chemical genetic approach using hydrolases, Proc. Nat. Academy Sciences U.S.A. 106 (34), 14699-14704

Obel N, Erben V, Schwarz T, Kuehnel S, Fodor A, Pauly M, 2009, Microanalysis of plant cell wall polysaccharides, Molecular Plant 2 (5), 922-932
 
Abasolo W, Eder M, Yamauchi K, Obel N, Reinecke A, Neumetzler L, Dunlop JWC, Mouille G, Pauly M, Hoefte H, Burgert I, 2009, Pectins may hinder the unfolding of xyloglucan chains during cell elongation – implications of the mechanical performance of Arabidopsis hypocotyls with pectin alterations, Molecular Plant 2 (5), 990-999

Pauly M, Keegstra K, 2008, Tear down this wall, Current Opinion in Plant Biology 11 (3), 233-235

Cavalier DM, Lerouxel O, Neumetzler L, Yamauchi K, Reinecke A, Freshour G, Zabotina O, Hahn MG, Burgert I, Pauly M, Raikhel N, Keegstra K, 2008, Disruption of two Arabidopsis thaliana xylosyltransferase genes results in plants deficient in xyloglucan, a major primary cell wall component, Plant Cell 20:1519-1537

Jensen J, Sorensen S, Harholt J, Geshi N, Sakuragi Y, Moller I, Zandleven J, Bernal AJ, Jensen NB, Sorensen C, Pauly M, Beldman G, Willats WGT, Scheller HV, 2008, Identification of a xylogalactuonan xylosyltransferase involved in pectin biosynthesis in Arabidopsis, Plant Cell 20: 1289-1302

Wen F, Rhodesia MC, Nguyen T, Zeng, W, Keegstra K, Immerzeel P, Pauly M, Hawes MC, 2008, Inducible expression of Pisum sativum xyloglucan fucosyltransferase in the pea root cap meristem, and effects of antisense mRNA expression on root cap cell wall structural integrity, Plant Cell Reports 27:1125-1135

Pauly M, Keegstra K, 2008, Cell wall carbohydrates and their modification as raw materials for biofuels, Plant Journal 54, 559-568

Leboeuf E, Immerzeel P, Gibon, Y, Steup M, Pauly M, 2008, High throughput functional assessment of polysaccharide-active enzymes using MALDI-TOF mass spectrometry as exemplified on plant cell wall polysaccharides, Analytical biochemistry, 373 (1):9-17

 

Recent Teaching

Bio1A - Biology - Molecules of Life/ the cell

MCB 102 - Principles of Biochemistry - Metabolism

199 - Supervised Independent Study

299 - Supervised Independent Study

PMB 122 - Bioenergy