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Table 3 Kinetics of GOOX-VN mutant enzymes on cello- and xylo-oligosaccharides

From: Xylo- and cello-oligosaccharide oxidation by gluco-oligosaccharide oxidase from Sarocladium strictumand variants with reduced substrate inhibition

Substrate Parameter GOOX-VN Y72F Y72A E247A W351A Q353N Q353A Q384N Q384A
Glucose kcat (min-1) 361 ± 5 - - 472 ± 6 (1,280 ± 70)* - - (713 ± 14) -
  Km (mM) 15 ± 1 - - 62 ± 2 (890 ± 60) - - (325 ± 10) -
  kcat/Km (mM-1 min-1) 24 ± 2 - - 7.7 ± 0.3 (1.4 ± 0.1) - - (2.2 ± 0.1) -
Cellobiose kcat (min-1) 372 ± 14 497 ± 7 644 ± 8 480 ± 14 1,040 ± 30 117 ± 2 16 ± 1 916 ± 13 210 ± 4
  Km (mM) 0.05 ± 0.01 1.3 ± 0.1 2.5 ± 0.1 0.17 ± 0.02 13 ± 1 11.3 ± 0.4 1.1 ± 0.2 0.63 ± 0.03 8.8 ± 0.3
  kcat/Km (mM-1 min-1) 7,000 ± 1,000 370 ± 20 250 ± 12 2,800 ± 300 79 ± 5 10.3 ± 0.4 15 ± 3 1,450 ± 80 24 ± 1
Cellotriose kcat (min-1) 436 ± 14 533 ± 5 695 ± 6 465 ± 17 1,057 ± 15 124 ± 2 20.0 ± 0.3 1,032 ± 12 245 ± 3
  Km (mM) 0.08 ± 0.01 0.62 ± 0.02 1.17 ± 0.03 0.11 ± 0.01 6.6 ± 0.2 6.2 ± 0.2 0.57 ± 0.03 0.20 ± 0.01 4.6 ± 0.1
  kcat/Km (mM-1 min-1) 5,600 ± 600 860 ± 30 596 ± 15 4,300 ± 500 160 ± 5 20 ± 1 35 ± 2 5,000 ± 200 54 ± 2
Xylose kcat (min-1) 330 ± 8 - - 189 ± 5 - - - - -
  Km (mM) 118 ± 7 - - 167 ± 11 - - - - -
  kcat/Km (mM-1 min-1) 2.8 ± 0.2 - - 1.1 ± 0.1 - - - - -
Xylobiose kcat (min-1) 449 ± 5 (446 ± 9) 764 ± 17 536 ± 6 (670 ± 50) - - (332 ± 11) (115 ± 7)
  Km (mM) 0.099 ± 0.005 (10.3 ± 0.3) 7.0 ± 0.3 0.43 ± 0.02 (48 ± 4) - - (11 ± 1) (23 ± 2)
  kcat/Km (mM-1 min-1) 4,500 ± 200 (43 ± 2) 109 ± 5 1,250 ± 60 (14 ± 2) - - (30 ± 2) (5.0 ± 0.5)
Xylotriose kcat (min-1) 524 ± 10 (590 ± 30) 920 ± 40 620 ± 20 (1,500 ± 300) - - (260 ± 30) (270 ± 30)
  Km (mM) 0.10 ± 0.01 (11 ± 1) 6.1 ± 0.5 0.34 ± 0.04 (100 ± 20) - - (10 ± 2) (41 ± 6)
  kcat/Km (mM-1 min-1) 5,100 ± 400 (54 ± 6) 150 ± 14 1,800 ± 200 (15 ± 5) - - (27 ± 5) (7 ± 1)
  1. *Estimated data are in parenthesis since Km values exceeded the tested substrate concentrations.
  2. -Activity was not detected.
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