Identification and characterization of the Streptococcus pneumoniae type 3 capsule-specific glycoside hydrolase of Paenibacillus species 32352
Identification and characterization of the Streptococcus pneumoniae type 3 capsule-specific glycoside hydrolase of Paenibacillus species 32352
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DOI：
10.1093/glycob/cwx097
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发表时间：
2018-02-01
期刊：
影响因子：
4.3
通讯作者：
Avci, Fikri Y.
中科院分区：
文献类型：
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作者：
Middleton, Dustin R.;Zhang, Xing;Avci, Fikri Y.
环状芽孢杆菌乔丹32352是20世纪30年代初从新泽西州腐烂的有机质中分离出来的。这种土壤细菌产生一种能够降解肺炎链球菌(Spn)3型衣壳多糖(Pn3P)的酶。关于这种酶Pn3Pase的早期报道表明，它可被Pn3P诱导，并对其具有特异性。我们着手对该酶进行鉴定和克隆，为其重组表达和鉴定奠定基础。我们首先对这种细菌的基因组进行了测序，并将Pn3Pase产生菌重新归类为拟青霉属32352种。我们通过蛋白质组学鉴定了Pn3Pase的可能蛋白，并克隆了该基因进行了重组表达。然后我们对Pn3P酶解聚产生的寡糖产物进行了表征。序列分析表明该糖苷水解酶属于一个新的碳水化合物活性酶GH家族。据我们所知，这是唯一一种显示Pn3P解聚活性的酶。
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 belongs to a new carbohydrate-active enzyme GH family. To our knowledge, this is the only enzyme to demonstrate Pn3P depolymerization activity.