title: Direct and up-close views of plant cell walls show a leading role for lignin-modifying enzymes on ensuing xylanases.
abstract: BACKGROUND
  A key barrier that limits the full potential of biological processes to create new, sustainable materials and fuels from plant fibre is limited enzyme accessibility to polysaccharides and lignin that characterize lignocellulose networks. Moreover, the heterogeneity of lignocellulosic substrates means that different enzyme combinations might be required for efficient transformation of different plant resources. Analytical techniques with high chemical sensitivity and spatial resolution that permit direct characterization of solid samples could help overcome these challenges by allowing direct visualization of enzyme action within plant fibre, thereby identify barriers to enzyme action.
  
  
  RESULTS
  In the current study, the high spatial resolution (about 30 nm) of scanning transmission X-ray microscopy (STXM), and the detection sensitivity (ppm) of time-of-flight secondary ion mass spectrometry (ToF-SIMS), were harnessed for the first time to investigate the progression of laccase, cellulase and xylanase activities through wood samples, and to evaluate complementary action between lignin-modifying and polysaccharide-degrading enzymes. In particular, complementary insights from the STXM and ToF-SIMS analyses revealed the key role of laccase in promoting xylanase activity throughout and between plant cell walls.
  
  
  CONCLUSIONS
  The spatial resolution of STXM clearly revealed time-dependent progression and spatial distribution of laccase and xylanase activities, whereas ToF-SIMS analyses confirmed that laccase promoted protein penetration into fibre samples, leading to an overall increase in polysaccharide degradation. Spectromicroscopic visualizations of plant cell wall chemistry allowed simultaneous tracking of changes to lignin and polysaccharide contents, which provides new possibilities for investigating the complementary roles of lignin-modifying and carbohydrate-active enzymes.
authors.affiliation: Department of Sustainable Bioproducts, Mississippi State University, Starkville, MS 39759 USA.
authors.forename: Dragica
authors.initials: D
authors.lastname: Jeremic
authors.affiliation: Department of Biochemistry, Chemistry and Physics, Niagara University, Lewiston, NY 14109 USA.
authors.forename: Robyn E
authors.initials: RE
authors.lastname: Goacher
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, ON M5S 3E5 Canada.
authors.forename: Ruoyu
authors.initials: R
authors.lastname: Yan
authors.affiliation: Canadian Light Source Inc., 44 Innovation Boulevard, Saskatoon, SK S7N 2V3 Canada.
authors.forename: Chithra
authors.initials: C
authors.lastname: Karunakaran
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, ON M5S 3E5 Canada.
authors.forename: Emma R
authors.initials: ER
authors.lastname: Master
keywords: Cellulase; Laccase; Lignocellulose; Scanning transmission X-ray microscopy; Spectromicroscopy; Time-of-flight secondary ion mass spectrometry; Wood; Xylanase