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TLC of restriction products by ChiA from various N-acetyl-chitooligosaccharides. The reaction mixture (50 μl) containing 0.7 mg of substrates in 70 mM sodium acetate buffer (pH 5.0) was incubated with enzyme (GlcNAc1–3, 0.45 μg; GlcNAc4–6, 0.9 μg) at 80°C. The reaction products at 0, 5, 15, 30, 60, and 120 min were analyzed. Lanes C, negative control incubated without enzyme at 80°C for 120 min; lanes S, standard N-acetyl-chitooligosaccharides ranging from GlcNAc (G1) to GlcNAc6 (G6).
