来自岛苹果螺（Pomacea insularus (de Ordigny)）的内切-1,4-β-D-甘露聚糖酶的纯化及某些性质

Purification and some properties of endo-1,4-beta-D-mannanase from a mud snail, Pomacea insularus (de Ordigny).

作者信息

Yamaura I, Matsumoto T

机构信息

Department of Applied Microbial Technology, Kumamoto Institute of Technology, Japan.

出版信息

Biosci Biotechnol Biochem. 1993 Aug;57(8):1316-9. doi: 10.1271/bbb.57.1316.

DOI:10.1271/bbb.57.1316

PMID:7764016

Abstract

Endo-1,4-beta-D-mannanase (1,4-beta-D-mannanohydrolase, EC 3.2.1.78) was purified from viscera of a mud snail, Pomacea insularus (de Ordigny). The purified enzyme gave a single protein band in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of the purified enzyme was estimated to be 44,000. The amino-terminal sequence was H.Gly-X-Leu-Arg-Arg-Gln- Gly-Thr-Asn-Ile-Val-Asp-Ser-His-Gly-His-Lys-Val-Phe-Leu-Ser-Gly-Ala-Asn- Thr-Ala-Trp-Val-Ala-Tyr-Gly-Tyr-Asp-. The enzyme was stable from pH about 5.0 to about 10.5 and had its maximum activity at pH about 5.5. The purified enzyme produced M2, M3, M4, and M5 from beta-1,4-mannan. Enzyme activity was greatly inhibited by Ag+, Hg2+, Cu2+, and dithiothreitol at 1 mM concentration. In addition, N-bromosuccinimide completely inhibited the enzyme activity.

摘要

从福寿螺（Pomacea insularus (de Ordigny)）的内脏中纯化出了内切 - 1,4 - β - D - 甘露聚糖酶（1,4 - β - D - 甘露聚糖水解酶，EC 3.2.1.78）。纯化后的酶在十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS - PAGE）中呈现出单一的蛋白条带。纯化酶的分子量估计为44,000。其氨基末端序列为H.Gly - X - Leu - Arg - Arg - Gln - Gly - Thr - Asn - Ile - Val - Asp - Ser - His - Gly - His - Lys - Val - Phe - Leu - Ser - Gly - Ala - Asn - Thr - Ala - Trp - Val - Ala - Tyr - Gly - Tyr - Asp -。该酶在pH约5.0至约10.5范围内稳定，在pH约5.5时具有最大活性。纯化后的酶能从β - 1,4 - 甘露聚糖产生M2、M3、M4和M5。在1 mM浓度下，Ag⁺、Hg²⁺、Cu²⁺和二硫苏糖醇对酶活性有极大抑制作用。此外，N - 溴代琥珀酰亚胺完全抑制了酶的活性。