Characterization of a new endo-type alginate lyase from Vibrio sp. W13.
Characterization of a new endo-type alginate lyase from Vibrio sp. W13.
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DOI：
10.1016/j.ijbiomac.2015.01.053
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发表时间：
2015-04
影响因子：
8.2
通讯作者：
Benwei Zhu;Haidong Tan;Yuqi Qin;Qingsong Xu;Yuguang Du;H. Yin
中科院分区：
文献类型：
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作者：
Benwei Zhu;Haidong Tan;Yuqi Qin;Qingsong Xu;Yuguang Du;H. Yin
从海洋细菌弧菌中鉴定并克隆了编码新藻酸盐裂解酶 Algb 的基因。 W13。对重组藻酸盐裂解酶进行表征，然后在 Ni-NTA Sepharose 上纯化。它在 pH 8.0 和 30 °C 时表现出最高活性（457 U/mg）。有趣的是，Algb 具有更广泛的底物特异性。它对polyM（聚β-d-甘露糖醛酸）和polyG（聚α-l-古洛糖醛酸）均表现出活性。此外，Algb 对藻酸盐 (0.67 mg/ml) 和 PolyMG (0.50 mg/ml) 的 Km 值低于对 PolyG (1.04 mg/ml) 和 PolyM (6.90 mg/ml) 的 Km 值。 TLC和ESI-MS分析表明Algb主要以内溶方式从四种底物中释放DP为2-5的寡糖。因此，它可能是生产低DP海藻酸寡糖的有效工具。
A gene, encoding a new alginate lyase Algb, was identified and cloned from marine bacteriumVibriosp. W13. The recombinant alginate lyase was characterized followed by being purified on Ni-NTA Sepharose. It exhibited the highest activity (457 U/mg) at pH 8.0 and 30 °C. Interestingly, Algb possessed broader substrate specificity. It showed activities toward both polyM (poly β-d-mannuronate) and polyG (poly α-l-guluronate). Furthermore,Kmvalues of Algb toward alginate (0.67 mg/ml) and polyMG (0.50 mg/ml) are lower than those toward polyG (1.04 mg/ml) and polyM (6.90 mg/ml). The TLC and ESI-MS analysis suggested that Algb mainly released oligosaccharides with DP of 2–5 from the four kinds of substrates in an endolytic manner. Therefore, it may be a potent tool to produce alginate oligosaccharides with low DP.