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来自罗西亚乳杆菌DSM 15814(T)的β-D-木糖苷酶的克隆、表达及特性分析

Cloning, expression and characterization of a β-D-xylosidase from Lactobacillus rossiae DSM 15814(T).

作者信息

Pontonio Erica, Mahony Jennifer, Di Cagno Raffaella, O'Connell Motherway Mary, Lugli Gabriele Andrea, O'Callaghan Amy, De Angelis Maria, Ventura Marco, Gobbetti Marco, van Sinderen Douwe

机构信息

Department of Soil, Plant and Food Science, University of Bari Aldo Moro, Via G. Amendola 165/A, 70126, Bari, Italy.

School of Microbiology, University College Cork, Cork, Ireland.

出版信息

Microb Cell Fact. 2016 May 3;15:72. doi: 10.1186/s12934-016-0473-z.

Abstract

BACKGROUND

Among the oligosaccharides that may positively affect the gut microbiota, xylo-oligosaccharides (XOS) and arabinoxylan oligosaccharides (AXOS) possess promising functional properties. Ingestion of XOS has been reported to contribute to anti-oxidant, anti-bacterial, immune-modulatory and anti-diabetic activities. Because of the structural complexity and chemical heterogeneity, complete degradation of xylan-containing plant polymers requires the synergistic activity of several enzymes. Endo-xylanases and β-D-xylosidases, collectively termed xylanases, represent the two key enzymes responsible for the sequential hydrolysis of xylan. Xylanase cocktails are used on an industrial scale for biotechnological purposes. Lactobacillus rossiae DSM 15814(T) can utilize an extensive set of carbon sources, an ability that is likely to contribute to its adaptive ability. In this study, the capacity of this strain to utilize XOS, xylan, D-xylose and L-arabinose was investigated.

RESULTS

Genomic and transcriptomic analyses revealed the presence of two gene clusters, designated xyl and ara, encoding proteins predicted to be responsible for XOS uptake and hydrolysis and D-xylose utilization, and L-arabinose metabolism, respectively. The deduced amino acid sequence of one of the genes of the xyl gene cluster, LROS_1108 (designated here as xylA), shows high similarity to (predicted) β-D-xylosidases encoded by various lactic acid bacteria, and belongs to glycosyl hydrolase family 43. Heterologously expressed XylA was shown to completely hydrolyse XOS to xylose and showed optimal activity at pH 6.0 and 40 °C. Furthermore, β-D-xylosidase activity of L. rossiae DSM 15814(T) was also measured under sourdough conditions.

CONCLUSIONS

This study highlights the ability of L. rossiae DSM 15814(T) to utilize XOS, which is a very useful trait when selecting starters with specific metabolic performances for sourdough fermentation or as probiotics.

摘要

背景

在可能对肠道微生物群产生积极影响的低聚糖中,木寡糖(XOS)和阿拉伯木聚糖寡糖(AXOS)具有良好的功能特性。据报道,摄入XOS有助于抗氧化、抗菌、免疫调节和抗糖尿病活性。由于含木聚糖的植物聚合物结构复杂且化学性质不均一,其完全降解需要几种酶的协同作用。内切木聚糖酶和β-D-木糖苷酶统称为木聚糖酶,是负责木聚糖顺序水解的两种关键酶。木聚糖酶混合物在工业规模上用于生物技术目的。罗西亚乳杆菌DSM 15814(T)能够利用多种碳源,这种能力可能有助于其适应能力。在本研究中,对该菌株利用XOS、木聚糖、D-木糖和L-阿拉伯糖的能力进行了研究。

结果

基因组和转录组分析揭示了存在两个基因簇,分别命名为木糖(xyl)和阿拉伯糖(ara),它们编码的蛋白质预计分别负责XOS的摄取和水解、D-木糖的利用以及L-阿拉伯糖的代谢。木糖基因簇中一个基因LROS_1108(此处命名为xylA)推导的氨基酸序列与各种乳酸菌编码的(预测)β-D-木糖苷酶具有高度相似性,属于糖基水解酶家族43。异源表达的XylA被证明能将XOS完全水解为木糖,在pH 6.0和40℃时表现出最佳活性。此外,还在酸面团条件下测定了罗西亚乳杆菌DSM 15814(T)的β-D-木糖苷酶活性。

结论

本研究突出了罗西亚乳杆菌DSM 15814(T)利用XOS的能力,这在选择具有特定代谢性能的发酵剂用于酸面团发酵或作为益生菌时是一个非常有用的特性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c116/4855831/1ac1f619817c/12934_2016_473_Fig1_HTML.jpg

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