Bacillus circulans Jordan 32352 was isolated from decaying organic matter in the New Jersey soil in the early 1930s. This soil-dwelling bacterium produced an enzyme capable of degrading the type 3 capsular polysaccharide (Pn3P) of Streptococcus pneumoniae (Spn). Early reports of this enzyme, Pn3Pase, demonstrated its inducibility by, and specificity for Pn3P. We set out to identify and clone this enzyme for its recombinant expression and characterization. We first sequenced the genome of this bacterial species, and reclassified the Pn3Pase producing bacterium as Paenibacillus species 32352. We identified the putative protein of Pn3Pase through mass spectrometry-based proteomics and cloned the gene for recombinant expression. We then characterized the oligosaccharide products generated upon the enzymatic depolymerization of Pn3P. Sequence analysis suggests that this glycoside hydrolase (GH) belongs to a new Carbohydrate-Active enzyme (CAZy) GH family. To our knowledge, this is the only enzyme to demonstrate Pn3P depolymerization activity.

中文翻译：

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类芽孢杆菌 32352 肺炎链球菌 3 型胶囊特异性糖苷水解酶的鉴定和表征

循环芽孢杆菌Jordan 32352 在 1930 年代初期从新泽西土壤中的腐烂有机物中分离出来。这种居住在土壤中的细菌产生了一种能够降解肺炎链球菌(Spn) 的 3 型荚膜多糖 (Pn3P) 的酶。这种酶 Pn3Pase 的早期报道证明了它对 Pn3P 的诱导性和特异性。我们着手鉴定和克隆这种酶，用于其重组表达和表征。我们首先对该细菌物种的基因组进行了测序，并将产生 Pn3Pase 的细菌重新分类为Paenibacillus species 32352. 我们通过基于质谱的蛋白质组学鉴定了推定的 Pn3Pase 蛋白，并克隆了该基因用于重组表达。然后，我们对 Pn3P 酶解聚产生的寡糖产物进行了表征。序列分析表明，这种糖苷水解酶（GH）属于一个新的碳水化合物活性酶（CAZy）GH家族。据我们所知，这是唯一表现出 Pn3P 解聚活性的酶。