Purification of sulfated fucoglucuronomannan lyase from bacterial strain of Fucobacter marina and study of appropriate conditions for its enzyme digestion
Purification of sulfated fucoglucuronomannan lyase from bacterial strain of Fucobacter marina and study of appropriate conditions for its enzyme digestion
复制标题
DOI：
10.1007/s10126-002-0083-0
复制
发表时间：
2003-07-01
影响因子：
3
通讯作者：
Kato, I
中科院分区：
文献类型：
--
作者：
Sakai, T;Kimura, H;Kato, I
一种海洋细菌菌株，海洋岩藻，产生胞外硫酸化岩藻葡糖醛酸甘露聚糖（SFGM）裂解酶的存在下培养的粗SFGM从岩藻聚糖硫酸酯的crassifolia（褐藻）通过氯化十六烷基吡啶分级分离。对于SFGM裂解酶测定，从K.以通过阴离子交换柱层析得到的厚叶褐藻糖胶为底物。胞外SFGM裂解酶经电泳凝胶纯化至均一，纯度为4240倍，产率为13.8%。由于凝胶过滤和十二烷基硫酸钠聚丙烯酰胺凝胶电泳给出相同的相对分子质量67，000，因此证明该酶是单体。该酶特异性地消化SFGM，但不消化任何其他测试的含糖醛酸的多糖。酶促反应的最适条件为pH7.5，43 ℃，0.4M NaCl浓度。该酶被CuCl_2和ZnCl_2强烈抑制，也被一些巯基试剂抑制。
A marine bacterial strain, Fucobacter marina, produced extracellular sulfated fucoglucuronomannan (SFGM) lyase when cultivated in the presence of crude SFGM obtained from fucoidan of Kjellmaniella crassifolia (brown algae) by cetyl pyridinium chloride fractionation. For the SFGM lyase assay, SFGM fraction separated from K. crassifolia fucoidan by anion exchange column chromatography was used as the substrate. The extracellular SFGM lyase was purified to homogeneity on an electrophoresis gel with 4240-fold purity at 13.8% yield. The enzyme proved to be a monomer, since gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis gave the same relative molecular mass of 67,000. The enzyme specifically digested SFGM but did not digest any other uronic-acid-containing polysaccharides tested. The optimum conditions for the enzyme reaction were around pH 7.5, 43 degreesC, and 0.4 M NaCl concentration. The enzyme was strongly inhibited by CuCl2 and ZnCl2, and also by some sulfhydryl reagents.