Identification of a novel aminoglycoside O-nucleotidyltransferase AadA33 in Providencia vermicola

Chunlin Feng; Mengdi Gao; Weiyan Jiang; Weina Shi; Anqi Li; Shuang Liu; Lei Zhang; Xueya Zhang; Qiaoling Li; Hailong Lin; Junwan Lu; Kewei Li; Hailin Zhang; Yunliang Hu; Qiyu Bao; Xi Lin

doi:10.3389/fmicb.2022.990739

Identification of a novel aminoglycoside O-nucleotidyltransferase AadA33 in Providencia vermicola

Front Microbiol. 2022 Sep 13:13:990739. doi: 10.3389/fmicb.2022.990739. eCollection 2022.

Authors

Chunlin Feng^{1

2}, Mengdi Gao², Weiyan Jiang¹, Weina Shi², Anqi Li², Shuang Liu², Lei Zhang², Xueya Zhang¹, Qiaoling Li¹, Hailong Lin¹, Junwan Lu^{2

3}, Kewei Li², Hailin Zhang¹, Yunliang Hu¹, Qiyu Bao^{1

2

3}, Xi Lin^{1

2}

Affiliations

¹ The Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University, Wenzhou, China.
² Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, China.
³ Medical Molecular Biology Laboratory, School of Medicine, Jinhua Polytechnic, Jinhua, China.

Abstract

A novel chromosome-encoded aminoglycoside O-nucleotidyltransferase AadA33 was identified in Providencia vermicola strain P13. The AadA33 shares the highest amino acid identity of 51.28% with the function characterized AadA31. Antibiotic susceptibility testing and enzyme kinetics analysis revealed that the function of AadA33 is to mediate spectinomycin and streptomycin resistance. The recombinant strain harboring aadA33 (pUCP20-aadA33/Escherichia coli DH5α) displayed >256- and 128-fold increases in the minimum inhibitory concentration levels to spectinomycin and streptomycin, respectively, compared with the control strains pUCP20/DH5α. Enzyme kinetic parameters manifested the substrate of AadA33 including spectinomycin and streptomycin, with k _cat/K _m of 3.28 × 10⁴ (M^-1 s^-1) and 3.37 × 10⁴ (M^-1 s^-1), respectively. Bioinformatics analysis revealed its structural mechanism of antimicrobial resistance, genetic context, and phylogenetic relationship with other aminoglycoside O-nucleotidyltransferases. This study of AadA33 contributed to understanding the function and resistance mechanism of aminoglycoside O-nucleotidyltransferase.

Keywords: AadA33; Providencia vermicola; aminoglycoside O-nucleotidyltransferase; enzyme kinetics; novel antimicrobial resistance gene.