Actinoplanes aureus sp. nov., a novel protease-producing actinobacterium isolated from soil
Actinoplanes aureus sp. nov., a novel protease-producing actinobacterium isolated from soil
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DOI：
10.1007/s10482-021-01617-4
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发表时间：
2021-04
期刊：
影响因子：
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通讯作者：
Jia Song;Xiujun Sun;Xianxian Luo;Chuan He;Zhenzhen Huang;Junwei Zhao;Beiru He;Xiaowen Du
中科院分区：
文献类型：
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作者：
Jia Song;Xiujun Sun;Xianxian Luo;Chuan He;Zhenzhen Huang;Junwei Zhao;Beiru He;Xiaowen Du
从内蒙古赤峰市敖汉旗的土壤中分离到一株新的产酶放线菌，命名为NEAU-A11T，并对其进行了多相鉴定。水解酶，如蛋白酶，通过降解蛋白质连接破坏细胞壁的完整性，在真菌的破坏中发挥关键作用。这表明该菌株可能是一个很好的生防菌候选菌株，用于控制真菌病害。16S rRNA基因序列分析表明，菌株neau-A11T属于放线菌属，与JCM 3194T的放线菌亲缘关系最近(98.9%)。细胞壁中含有中位二氨基甲酸作为诊断二氨基酸，全细胞糖为阿拉伯糖、木糖和葡萄糖。磷脂组成包括二磷脂酰甘油、磷脂酰乙醇胺、磷脂酰甘油、磷脂酰肌醇和两种磷脂酰肌醇甘露糖苷。以MK-9(H4)、MK-9(H6)和MK-9(H8)为主。主要脂肪酸为C18：0、C16：0、C18：1、ω9c、C17：0和C15：0。基因组测序显示基因组大小为10,742,096个碱基，G- + C含量为70.5%，9,514个蛋白质编码基因(CDS)，其中包括102个编码蛋白酶的基因。基因组分析表明，菌株NEAU-A11T含有255个糖苷水解酶(GHS)、152个糖基转移酶(GTS)、40个糖酯酶(CES)、26个多糖裂解酶(PL)和12个辅助活性(AAs)基因。使用antiSMASH 5.0进行的基因组挖掘分析导致了20个可能负责产生不同次生代谢物的基因簇的鉴定。16S rRNA基因序列的系统发育分析表明，该菌株与A形成了一个稳定的分支。JCM 3194型直线型放线飞机。全基因组系统发育分析表明，NEAU-A11T与放线放线菌DSM 45883T(97.6%)形成了一个稳定的亲缘关系。然而，NEAU-A11T株与其参考株A的全基因组平均核苷酸同源性较高。直线型JCM 3194TAND A。DSM 45883T分别为81.1%和81.6%。数字化DNA-DNA杂交率分别为24.6%(22.2-27.0%)和24.8%(22.5-27.3%)。这些值远低于dDDH和ANI的物种划分标准，分别为70%和95%-96%，这表明该分离物在遗传上与其关系密切的模式株不同。基因组中G- + -C含量为70.5%，在67-76%之间。此外，表型、化学分类和基因分型研究表明，菌株neau-A11Trea是放线菌属的一个新种，命名为金黄色放线菌。11月以NEAU-A11T(= CCTCC AA2019063T= JCM33971T)为模式菌株。
A novel protease-producing actinobacterium, designated strain NEAU-A11T, was isolated from soil collected from Aohan banner, Chifeng, Inner Mongolia Autonomous Region, China, and characterised using a polyphasic approach. The hydrolytic enzymes, such as proteases, played critical roles in destruction of fungi by degrading the protein linkages to disrupt integrity in the cell wall. This suggested that the isolate could be a good biocontrol candidate against pathogens to control fungal diseases. On the basis of 16S rRNA gene sequence analysis, strain NEAU-A11Twas indicated to belong to the genusActinoplanesand was most closely related toActinoplanes rectilineatusJCM 3194T(98.9%). Cell walls containedmeso-diaminopimelic acid as the diagnostic diamino acid and the whole-cell sugars were arabinose, xylose and glucose. The phospholipid profile contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and two phosphatidylinositol mannosides. The predominant menaquinones were MK-9(H4), MK-9(H6) and MK-9(H8). The major fatty acids were C18:0, C16:0, C18:1ω9c, C17:0and C15:0. Genome sequencing revealed a genome size of 10,742,096 bp, a G + C content of 70.5% and 9,514 protein-coding genes (CDS), including 102 genes coding for protease. Moreover, Genome analysis showed that strain NEAU-A11Tcontained 255 glycoside hydrolases (GHs), 152 glycosyl transferases (GTs), 40 carbohydrate esterases (CEs), 26 polysaccharide lyases (PLs), and 12 auxiliary activities (AAs) genes. Genome mining analysis using antiSMASH 5.0 led to the identification of 20 putative gene clusters responsible for the production of diverse secondary metabolites. Phylogenetic analysis using the 16S rRNA gene sequences showed that the strain formed a stable clade withA. rectilineatusJCM 3194Tin the genusActinoplanes. Whole-genome phylogeny showed strain NEAU-A11Tformed a stable phyletic line withActinoplanes lutulentusDSM 45883T(97.6%). However, whole-genome average nucleotide identity value between strain NEAU-A11Tand its reference strainsA. rectilineatusJCM 3194TandA. lutulentusDSM 45883Twere found to be 81.1% and 81.6%, respectively. The levels of digital DNA-DNA hybridization between them were 24.6% (22.2–27.0%) and 24.8% (22.5–27.3%), respectively. The values were well below the criteria for species delineation of 70% for dDDH and 95–96% for ANI, suggesting that the isolate differed genetically from its closely related type strain. The content of G + C in genomic DNA was 70.5%, within the range of 67–76%. In addition, evidences from phenotypic, chemotaxonomic and genotypic studies indicated that strain NEAU-A11Trepresents a novel species of the genusActinoplanes, for which the nameActinoplanes aureussp. nov. is proposed, with NEAU-A11T(= CCTCC AA 2019063T= JCM 33971T) as the type strain.