Toward Understanding the Alginate Catabolism in Microbulbifer sp. ALW1 by Proteomics Profiling.
Toward Understanding the Alginate Catabolism in Microbulbifer sp. ALW1 by Proteomics Profiling.
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DOI：
10.3389/fbioe.2022.829428
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发表时间：
2022
影响因子：
5.7
通讯作者：
Zhu Y
中科院分区：
文献类型：
--
作者：
Li Z;Huang X;Guo Y;Zhang C;Yang L;Du X;Ni H;Wang X;Zhu Y
Microbulbifer sp. 的细菌菌株。 ALW1 已表现出明显的降解海带细胞壁的能力，并且已对某些单独的酶进行了生化表征以阐明其遗传基础。然而，菌株 ALW1 如何成功分解主要细胞壁成分藻酸盐多糖并在其海洋宿主上定殖，仍然是个谜。在本研究中，采用基于质谱的细胞外和细胞内蛋白质组定量分析来阐明 ALW1 菌株中的藻酸盐降解途径。质谱和生化分析表明，菌株ALW1可以在12小时内有效地将海藻酸多糖降解为二糖和三糖。蛋白质组分析分别鉴定了 156 种和 1,047 种蛋白质专门定位于细胞外和细胞内区室，其中 1,086 种蛋白质具有双重定位特征。所鉴定蛋白质的功能注释表明多种催化酶和非催化分子参与藻酸盐多糖的裂解和代谢。构建了一条简化的途径来演示海藻酸盐多糖及其片段寡糖的细胞外消化、主动转运和细胞内转化，描绘了ALW1菌株控制海藻酸盐分解代谢的基因座图。本研究旨在为细菌菌株ALW1的利用和遗传操作提供指导，以通过发酵高效生产海藻酸寡糖。
The bacterial strain of Microbulbifer sp. ALW1 has demonstrated visible ability of degrading the cell wall of Laminaria japonica, and biochemical characterization has been performed on some individual enzymes to elucidate its genetic basis. However, it still remains elusive how strain ALW1 successfully breaks down the major cell wall component alginate polysaccharide and colonizes on its marine host. In this study, a mass spectrometry-based quantitative analysis of the extracellular and intracellular proteomes was introduced to elucidate the alginate degradation pathway in ALW1 strain. Mass spectrometry and biochemical assays indicated that strain ALW1 could effectively degrade alginate polysaccharide into disaccharides and trisaccharides within 12 h. Proteome analysis identified 156 and 1,047 proteins exclusively localized in extracellular and intracellular compartments, respectively, with 1,086 protein identities of dual localization. Functional annotation of the identified proteins suggested the involvement of diverse catalytic enzymes and non-catalytic molecules for the cleavage and metabolism of alginate polysaccharide. A simplified pathway was constructed to demonstrate the extracellular digestion, active transport, and intracellular conversion of alginate polysaccharide and its fragmented oligosaccharides, casting a picture of genetic loci controlling alginate catabolism by ALW1 strain. This study aims to provide a guide for utilization and genetic manipulation of the bacterial strain ALW1 for efficient alginate oligosaccharides production by fermentation.
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DOI：
10.1007/978-1-4939-6899-2_20
发表时间：
2017-01-01
期刊：
PROTEIN-CARBOHYDRATE INTERACTIONS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Kunath, Benoit J.;Bremges, Andreas;Pope, Phillip B.
通讯作者：
Pope, Phillip B.
影响因子：
11.2
作者：
Jiang, Zedong;Zhang, Xiwen;Zhu, Yanbing
通讯作者：
Zhu, Yanbing
影响因子：
4.4
作者：
Dunne, Jonathan C.;Li, Dong;Jordan, T. William
通讯作者：
Jordan, T. William
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影响因子：
5.4
作者：
Li M;Shang Q;Li G;Wang X;Yu G
通讯作者：
Yu G
影响因子：
3
作者：
Choi, DuBok;Piao, Yu Lan;Cho, Hoon
通讯作者：
Cho, Hoon