Planta Piloto de Procesos Industriales Microbiológicos (PROIMI), CONICET, Tucumán, Argentina.
Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Tucumán, Argentina.
Prep Biochem Biotechnol. 2021;51(9):871-880. doi: 10.1080/10826068.2020.1870136. Epub 2021 Jan 13.
sp. AR03 have been described as an important producer of carbohydrate-active enzymes (CAZymes) when growing in a peptone-based medium supplemented with simple sugars and/or carboxymethyl cellulose (CMC) as carbon sources. This work aimed to identify the extracellular enzymatic cocktails through shotgun proteomics. The proteomic analysis showed that enzymes involved in cellulose and xylan degradation were among the most abundant proteins. These enzymes included an endo-glucanase GH5_2 and a glucuronoxylanase GH30_8, which were found in all conditions. In addition, several proteins were differentially expressed in the three evaluated culture media, indicating microbial metabolic changes due to the different supplied carbon sources, particularly, in the presence of CMC. Finally, the capability of the crude enzymatic cocktails from culture media to degrade birchwood xylan was assessed, which produced mostly xylooligosaccharides containing among 3-5 xylose units. Consequently, this work shows the potential of the extracellular enzymes from sp. AR03 for producing emergent prebiotics.
当在含有简单糖和/或羧甲基纤维素（CMC）作为碳源的蛋白胨培养基中生长时，AR03 被描述为碳水化合物活性酶（CAZymes）的重要生产者。本研究旨在通过鸟枪法蛋白质组学鉴定细胞外酶混合物。蛋白质组学分析表明，参与纤维素和木聚糖降解的酶是最丰富的蛋白质之一。这些酶包括内切葡聚糖酶 GH5_2 和木聚糖酶 GH30_8，它们在所有条件下都存在。此外，在三种评估的培养基中，有几种蛋白质的表达存在差异，这表明由于不同的碳源供应，微生物代谢发生了变化，特别是在存在 CMC 的情况下。最后，评估了培养基中粗酶混合物对桦木木聚糖的降解能力，主要产生含有 3-5 个木糖单元的木二糖寡糖。因此，这项工作展示了 sp. AR03 细胞外酶产生新兴益生元的潜力。
