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MALDI-TOF MS analysis of the extracellular polysaccharides released by the diatom Thalassiosira pseudonana

机译:硅藻Thalassiosira pseudonana释放的细胞外多糖的MALDI-TOF MS分析

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摘要

The extracellular polysaccharides (ECPS) released by diatoms have significant roles in marine ecosystems and have potential applications including drug-discovery and biopharmaceutical precursors. In this study, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology was used in the structural analysis of the ECPS released by Thalassiosira pseudonana (Bacillariophyta). Three different deproteinization methods, the Sevag method, the trichloroacetic acid (TCA) method, and the enzymolysis method, were compared in the purification of ECPS. Our results suggested that TCA was the best deproteinization method among the three methods for subsequent MALDI-TOF MS investigation because of its high ECPS yield, protein removal ability and reliable MALDI-TOF MS fingerprint. The degree of polymerization (d.p.) profiles, the molecular weight of the ECPS and the distribution pattern of the polymers with different molecular mass were described from the MALDI-TOF MS spectra. This work represents the whole-level composition of the ECPS released by the diatom and has improved our knowledge of the structural characterization of ECPS.
机译:硅藻释放的细胞外多糖(ECPS)在海洋生态系统中具有重要作用,并具有潜在的应用,包括药物发现和生物制药前体。在这项研究中,基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)技术用于拟真拟南芥(Bacillariophyta)释放的ECPS的结构分析。在ECPS的纯化中,比较了三种不同的脱蛋白方法:Sevag方法,三氯乙酸(TCA)方法和酶解方法。我们的结果表明,TCA是随后进行MALDI-TOF MS研究的三种方法中最好的脱蛋白方法,因为它具有很高的ECPS产量,蛋白质去除能力和可靠的MALDI-TOF MS指纹图谱。从MALDI-TOF MS光谱描述了聚合度(d.p.)分布,ECPS的分子量和具有不同分子量的聚合物的分布模式。这项工作代表了硅藻释放的ECPS的整个层次的组成,并提高了我们对ECPS结构表征的认识。

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