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Table 4 Enzyme activities for AFEX-SG hydrolysis measured by oxime-NIMS

From: Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling

  G1 (mM) G2 (mM) G3 (mM) G4 (mM) Total (mM) Average (mM) Stdev   
No enzyme 0.2 0.0 0.0 0.0 0.58   
CMX00_3a 4.6 5.8 0.2 0.1 8.54     
  4.5 5.0 0.1 0.0 7.67 10.2 0.8   
XynY 0.0 1.3 1.5 0.2 0.76     
  0.0 1.0 1.0 0.2 0.54 2.6 0.6   
XynA 0.0 0.0 0.0 0.0 0.02     
  0.0 0.0 0.0 0.0 0.00 0.0 0.0   
  1. Activity reported as µmol of reducing sugar released per hour per µmol of purified enzyme determined by oxime-NIMS as described in “Methods” section. Standard deviation indicated for the duplicate samples
  2. AFEX-SG AFEX-treated switchgrass, G1 glucose, G2 cellobiose, G3 cellotriose, G4 cellotetraose, P1 pentose (i.e., likely xylose), P2 pentobiose, P3 pentotriose, P4 pentotetraose, P5 pentopentose, P6 pentohexose
  3. aXyloglucan-derived oligosaccharides identified using abbreviations from [59]; X, Xyl–Glu; XG, Xyl–Glu–Glu; X2–G, Xyl–Xyl–Glc, X3–G, Xyl–Xyl–Xyl–Glc
  P1 (mM) P2 (mM) P3 (mM) P4 (mM) P5 (mM) P6 (mM) Total (mM) Average (mM) Stdev
No enzyme 0.0 0.0 0.0 0.0 0.0 0.0 0.0
CMX00_3a 3.2 14.5 17.3 6.7 1.6 0.3 43.6   
  3.1 15.3 17.0 6.6 1.1 0.2 43.3 43.4 0.2
XynY 94.5 127.9 40.5 10.5 1.1 0.1 274.6   
  93.1 143.3 40.1 12.0 0.9 0.0 289.5 282.0 10.5
XynA 36.7 94.5 0.3 2.2 10.7 1.0 145.3   
  39.3 95.0 0.1 1.5 11.0 0.3 147.3 146.3 1.4
  1. Activity reported as µmol of reducing sugar released per hour per µmol of purified enzyme determined by oxime-NIMS as described in “Methods” section. Standard deviation indicated for the duplicate samples
  2. AFEX-SG AFEX-treated switchgrass, G1 glucose, G2 cellobiose, G3 cellotriose, G4 cellotetraose, P1 pentose (i.e., likely xylose), P2 pentobiose, P3 pentotriose, P4 pentotetraose, P5 pentopentose, P6 pentohexose
  3. aXyloglucan-derived oligosaccharides identified using abbreviations from [59]; X, Xyl–Glu; XG, Xyl–Glu–Glu; X2–G, Xyl–Xyl–Glc, X3–G, Xyl–Xyl–Xyl–Glc
  Xa (mM) XG (mM) X2-G (mM) X3-G (mM) Xyl-Rha-GalA-Xyl (mM) Total (mM) Average (mM) Stdev  
No enzyme 0.0 0.0 0.0 0.0 0.0 0.0  
CMX00_3a 2.9 0.1 1.4 0.2 0.6 5.2    
  1.4 0.1 1.1 0.2 0.6 3.4 4.3 1.3  
XynY 0.4 0.0 0.2 0.1 2.6 3.3    
  0.4 0.0 0.2 0.1 2.5 3.1 3.2 0.1  
XynA 0.4 0.0 0.0 0.0 0.1 0.5    
  0.2 0.0 0.0 0.0 0.1 0.3 0.4 0.1  
  1. Activity reported as µmol of reducing sugar released per hour per µmol of purified enzyme determined by oxime-NIMS as described in “Methods” section. Standard deviation indicated for the duplicate samples
  2. AFEX-SG AFEX-treated switchgrass, G1 glucose, G2 cellobiose, G3 cellotriose, G4 cellotetraose, P1 pentose (i.e., likely xylose), P2 pentobiose, P3 pentotriose, P4 pentotetraose, P5 pentopentose, P6 pentohexose
  3. aXyloglucan-derived oligosaccharides identified using abbreviations from [59]; X, Xyl–Glu; XG, Xyl–Glu–Glu; X2–G, Xyl–Xyl–Glc, X3–G, Xyl–Xyl–Xyl–Glc