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

Abstract:

[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.