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Antimicrobial resistance (AMR) represents one of the main challenges in Tuberculosis (TB) treatment. Investigating the genes involved in AMR and the underlying mechanisms holds promise for developing alternative treatment strategies. The results indicate that dehydroquinate synthase (DHQS) regulates the susceptibility of Mycobacterium bovis BCG to first-line anti-TB drug streptomycin. Perturbation of the expression of aroB encoding DHQS affects the susceptibility of M. bovis BCG to streptomycin. Purified DHQS impairs in vitro antibacterial activity of streptomycin, but did not hydrolyze or modify streptomycin. DHQS directly binds to streptomycin while retaining its own catalytic activity. Computationally modeled structure analysis of DHQS–streptomycin complex reveals that DHQS binds to streptomycin without disturbing native substrate binding. In addition, streptomycin treatment significantly induces the expression of DHQS, thus resulting in DHQS-mediated susceptibility. Our findings uncover the additional function of DHQS in AMR and provide an insight into a non-canonical resistance mechanism by which protein hijacks antibiotic to reduce the interaction between antibiotic and its target with normal protein function retained.

Additional file 2: Supplemental Table 1. GTDB genomes with detected equol gene clusters. Supplemental Table 2: Bacterial strains from the Coriobacteriales that have been reported to produce equol in previous studies. Supplemental Table 3: Metagenome Sequencing Projects with Detected Contigs Containing Putative Equol Gene Clusters.

Additional file1: Supplemental Fig. 1. Presence of Equol Gene Clusters in Eggerthellaceae Genomes. A phylogenetic tree showing the presence of the equol gene cluster in the Coriobacteriales order. The Coriobacteriaceae, Atopobiaceae, and UMGS124 families were collapsed into a single leaf shown as a gray triangle. Species where equol production gene clusters were detected are highlighted in yellow and the genera containing these species are represented by shaded areas and colored branches in the tree. Supplemental Fig. 2: Conserved regions upstream and downstream of the equol gene clusters. Gene synteny plots showing the conserved genes upstream and downstream of the equol gene clusters in A) A. equolifaciens , B) S. faecalis, and C) Slackia_A isoflavoniconvertens. Colored connections are shown between similar genes if the genes have greater than 75% amino acid identity. The equol gene clusters are highlighted with black boxes when present. Positions on the genome sequence are provided foreach of the visualized regions below the labels to the left of each plot. Supplemental Fig. 3: Adlercreutzia Contig Equol Gene Clusters. Equol production gene clusters from metagenomic contigs assigned to A. equolifaciens, A. mucosicola, or the Adlercreutzia genus. Gene clusters are shown at the same scale and detected gene clusters from published A. equolifaceins and A. mucosicola genomes are included for reference. Supplemental Fig. 4: Slackia_A Contig Equol Gene Clusters. Equol production gene clusters from metagenomic contigs assigned to Slackia_A isoflavoniconvertens or the Slackia_A genus. Gene clusters are shown at the same scale and detected gene clusters from published Slackia_A.isoflavoniconvertens and Slackia_A equolifaciens genomes are included for reference. Supplemental Fig. 5: Senegalimassilia and Enteroscipio Contig Equol Gene Clusters. Equol production gene clusters from metagenomic contigs assigned to Senegalimassilia faecalis and to Enteroscipio sp000270285 or Slackia_A equolifaciens. Gene clusters are shown at the same scale and detected gene clusters from published Senegalimassilia faecalis or Enteroscipio sp000270285 genomes are included for reference.

Xiaofang Jiang,1,* Lin Zhou,2,3,* Lihua Zuo,2,3 Xiaohui Wang,4 Yingying Shi,2,3 Xiangyu Du,5 Jun Zhang,2,3 Liwei Liu,2,3 Zhuolun Li,2,3 Lianping Xue,2,3 Xin Liu,1 Zhi Sun2,3 1Department of Traditional Chinese Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province 450052, People’s Republic of China; 2Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province 450052, People’s Republic of China; 3Henan Key Laboratory of Precision Clinical Pharmacy, Zhengzhou, Henan Province 450052, People’s Republic of China; 4Department of Ultrasonography, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province 450052, People’s Republic of China; 5School of Basic Medical Science, Henan University, Kaifeng, Henan Province 475001, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhi Sun; Lin ZhouDepartment of Pharmacy, The First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Erqi District, Zhengzhou 450052, Henan Province, People’s Republic of ChinaTel/Fax +86 371 66862570Email sunzhi2013@163.com; fcczhoul@zzu.edu.cnPurpose: Shenkang injection, a traditional Chinese herbal prescription, had been widely used in renal disease due to its perfect curative effect. In this research, a novel, sensitive, accurate and rapid liquid chromatography-tandem mass spectrometric method was developed to simultaneously detect the seven active ingredients in rat plasma of Shenkang injection and investigate its pharmacokinetic behaviors with metabolism profiling meanwhile.Methods: For accurate pharmacokinetic quantitation, a WATERS ACQUITY UPLC® BEH C18 column was used to perform a separation and acetonitrile-water (0.1% formic acid) was selected as mobile phase for gradient elution with a flow rate of 0.20 mL/min. A heated electrospray ionization with selective reaction monitoring mode was used to monitor the precursor-product ion transitions for all the analytes and IS.Results: They all showed good linearity over a wide concentration range (r> 0.996 3) and the lower limit of quantification (LLOQ) was 0.1– 1.0 ng/mL for analytes. The validation parameters were all within the acceptable limits. Furthermore, for metabolism profiling study, metabolites of the seven ingredients were identified from the rat plasma based on the accurate mass and fragment ions. The metabolic pathways mainly focus on reduction, dehydration and conjugation.Conclusion: This study provided an overview of disposition of Shenkang injection, which is highly instructive for better understanding the effectiveness and toxicity of this drug.Keywords: Shenkang injection, pharmacokinetics, metabolism profiling, UHPLC-MS/MS, UHPLC-Q-orbitrap HRMS