奥利万星中间体A82846B产生菌拟孢囊菌CCTCC M2020063全基因组序列分析与mbtH类基因的验证

目的] 分析荒漠拟孢囊菌CCTCC M2020063中A82846B合成的代谢途径和关键基因。方法] 使用Illumina二代测序和PacBio三代测序技术对荒漠拟孢囊菌CCTCC M2020063进行全基因组测序，利用Glimmer预测编码序列，使用HPLC和LCMS鉴定次级代谢产物，使用antiSMASH 5.0软件预测次级代谢产物合成基因簇。利用Geneious软件对A82846B合成相关基因进行分析，对其中的mbtH类基因着重分析。结果] 本实验菌株鉴定为荒漠拟孢囊菌（Kibdelosporangium aridum），基因组中有一条线性染色体，无质粒，序列全长12475688 bp，GC含量为66.27%，有11900个开放阅读框，共有47个基因簇。该菌株具有合成A82846B的能力，且生物合成相关基因位于Cluster32，包含33个基因，mbtH类基因gene07864的过表达促进A82846的合成，提升了26.42%，卤化酶基因为gene07859，与万古霉素、巴利霉素的卤化酶相似度较高。结论] 本研究对荒漠拟孢囊菌CCTCC M2020063进行了基因组序列分析，获得了A82846B生物合成相关的功能基因信息，为A82846B的代谢途径和工程改造提供了强有力的基础。

Oritavancin intermediate A82846B-producing strain Kibdelosporangium aridum CCTCC M2020063 whole genome sequence analysis and mbtH-like gene verification

Objective] We analyzed the metabolic pathways and key genes of A82846B synthesis of Kibdelosporangium aridum CCTCC M2020063.Methods] We used the second-generation sequencing technology Illumina and the third-generation sequencing technology PacBio to perform whole-genome sequencing of Kibdelosporangium aridum CCTCC M2020063. We also used Glimmer to predict the coding sequence. To identify the secondary metabolite, high performance liquid chromatography and liquid chromatograph mass spectrometer was applied. To predict the secondary metabolite synthesis gene cluster, AntiSMASH 5.0 software was applied as well. Besides, focusing on the analysis of its mbtH-like gene, geneious software was adopted in this paper to analyze A82846B synthesis-related genes. Results] Sequence analysis showed that the strain belonged to Kibdelosporangium aridum, containing a plasmid-free linear genome chromosome with the full-length gene sequence of 12475688 bp and the GC content of 66.27%. This linear genome chromosome had 11900 open reading frames and a total of 47 gene clusters. This strain was capable to synthesize A82846B, and the biosynthesis-related genes were located in Cluster32, which contained 33 genes. Overexpression of the mbtH-like gene gene07864 promoted the synthesis of A82846 by 26.42%. The halogenase gene was gene07859, being highly similar to the halogenase of vancomycin and balimycin. Conclusion] In this study, we carried out the genome sequence analysis of Kibdelosporangium aridum CCTCC M2020063, and we also obtained the information of A82846B biosynthesis-related functional genes, which provided a strong basis for the metabolic pathway and engineering modification of A82846B.