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J. Biochem, 1970, Vol. 68, No. 1 53-62
© 1970 Japanese Biochemical Society

research-article

Studies on the Substrate Specificity of Egg White Lysozyme

III. The Mode of the Enzymatic Action on Partially O-Hydroxyethylated Chitin (Glycol Chitin)

Saburo HARA, Yoshiko NAKAGAWA* and Yoshio MATSUSHIMA

The Department of Chemistry, Osaka University College of Science Toyonaka

*Present address: Aichi Prefectural University, Faculty of Literature, 3–28 Takada-cho, Mizuho-ku, Nagoya.

1. Gas-liquid chromatography was applied to the analyses of N-acetylamino sugars and N-acetylamino sugar alcohols derived from glycol chitin. N-Acetyl-3-O-(2'-hydroxyethyl)-D-glucosamine, N-acetyl-6-O-(2'-hydroxyethyl)-D-glucosamine and the corresponding sugar alcohols were prepared as reference compounds. N-Acetyl-D-glucosamine, N-acetyl-3-O-(2'-hydroxyethyl)-D-glucosamine and N-acetyl-6-O-(2'-hydroxyethyl)-D-glucosamine were identified as the constituents of glycol chitin.

2. The gas-liquid chromatographic analysis showed that by the digestion of glycol chitin with hen egg white lysozyme [EC 3. 2.17], only N-acetyl-D-glucosamine was found to be liberated as a monosaccharide fraction.

3. For the purpose of elucidating the attacking points of the lysozyme on glycol chitin, reducting end groups were analyzed by converting them into sugar alcohols by borohydride reduction. N-Acetyl-D-glucosamine and N-acetyl-3-O-(2'-hydroxyethyl)-D-glu-cosamine were identified as reducing ends.

4. Oligosaacharides in the enzyme digests were separated by gel filtration on Sephadex G-15, Bio-Gel P-2, and paper chromatography. The structure of the saccharides so far identified were: N-acetyl-D-glucosamine, N, N-diacetryl-chitobiose, N, N-diacetyl -3-o-(2'-hydroxyethyl)-chitobiose, N, N-diacetyl-6'-O-(2'-hydroxyethyl)-chitobiose, N, N-diacetyl-3, 6'-di-O-(2'-hydroxyethyl)-chitobiose.


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