Characterization of a novel alkaline arylsulfatase from Marinomonas sp FW-1 and its application in the desulfation of red seaweed agar
Characterization of a novel alkaline arylsulfatase from Marinomonas sp FW-1 and its application in the desulfation of red seaweed agar
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DOI：
10.1007/s10295-015-1625-6
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发表时间：
2015-10-01
影响因子：
3.4
通讯作者：
Fu, Xiaoting
中科院分区：
文献类型：
--
作者：
Wang, Xueyan;Duan, Delin;Fu, Xiaoting
从青岛沿海地区分离到一株能够降解对硝基苯硫酸酯(PNPs)和琼脂的硫酸酯键的细菌中国。根据其16S rRNA基因序列鉴定为Marinomonas，命名为Marinomonas sp.FW-1。用离子交换层析和凝胶过滤层析纯化了一种回收率为13%、倍数为12的芳基硫酸酯酶。经SDS-PAGE分析，该酶由一条相对分子质量为33 kDa的多肽链组成。芳基硫酸酯酶的最适pH为9.0，最适温度为45。芳基硫酸盐酶在pH值为8-11，温度低于55A时稳定，测定了该酶对PNPs的K(M)和V(Max)分别为13.73mU/m in和270.27 mU/m in。对琼脂的脱硫率分别为86.11%和89.61%。经芳基硫酸酯酶处理的琼脂糖凝胶的DNA电泳谱与商品琼脂糖凝胶上的DNA电泳谱没有差异。因此，这种新型的碱性芳基硫酸酯酶在琼脂的酶促转化为琼脂糖类中具有很大的应用潜力。
A bacterial strain capable of hydrolyzing sulfate ester bonds of p-nitrophenyl sulfate (pNPS) and agar was isolated from the coast area of Qingdao, China. It was identified as Marinomonas based on its 16S rRNA gene sequence and named as Marinomonas sp. FW-1. An arylsulfatase with a recovery of 13 % and a fold of 12 was purified to a homogeneity using ion exchange and gel filtration chromatographies. The enzyme was composed of a single polypeptide chain with the molecular mass of 33 kDa estimated using SDS-PAGE. The optimal pH and temperature of arylsulfatase were pH 9.0 and 45, respectively. Arylsulfatase was stable over pH 8-11 and at temperature below 55 A degrees C. The K (m) and V (max) of this enzyme for the hydrolysis of pNPS were determined to be 13.73 and 270.27 mu M/min, respectively. The desulfation ratio against agar from red seaweed Gelidium amansii and Gracilaria lemaneiformis were 86.11 and 89.61 %, respectively. There was no difference between the DNA electrophoresis spectrum on the gel of the arylsulfatase-treated G. amansii agar and that of the commercial agarose. Therefore, this novel alkaline arylsulfatase might have a great potential for application in enzymatic conversion of agar to agarose.