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![[Figure 9]](jc5057fig9mag.jpg)
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Figure 9
Time-course TLC analysis of the reaction products. Time-course analysis of soluble oxidized oligosaccharide production from Avicel PH-101 cellulose (a), colloidal α-chitin from shrimp (b) and β-chitin from squid pen (c). The reaction was performed in 20 mM Tris–HCl pH 7.5 containing 25 mg ml−1 colloidal chitin, 1.0 mM AscA and 10 µM purified VhLPMO10A. The reaction mixture was incubated at 30°C for different times from 0.5 to 48 h and the reaction was stopped by boiling 100 µl aliquots of the reaction mixture. After centrifugation, 1 µl aliquots of each reaction supernatant were applied 20 times to a silica TLC plate (9 × 10 cm) and then chromatographed twice (1 h each) in the mobile phase. The TLC plates were dried in air and the degraded products were visualized by baking at 180°C for 3–5 min after spraying with a solution consisting of 0.4 g α-diphenylamine, 0.4 ml aniline, 20 ml acetone in 3 ml 85% orthophosphoric acid. Lanes: STD, non-oxidized standard chitooligosaccharides (GlcNAc)1–6; Chito mix, non-oxidized chitooligosaccharides; (Chito mix)Ox, chitooligosaccharide mixture oxidized by CuSO4 in Fehling's solution; cellulose (a), shrimp chitin (b) and squid pen chitin (c) incubated with VhLPMO10A for 48 h but in the absence of 1 mM AscA; the sugar products obtained at 0.5, 1, 3, 6, 12, 24 and 48 h of reaction. |
![[Figure 9]](jc5057fig9.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
