Genomic insight into O-demethylation of 4-methoxybenzoate by a two-component system from Amycolatopsis magusensis KCCM40447

Bashu Dev Pardhe; Lakshan Paudel; So-Ra Han; Tae-Jin Oh

doi:10.1016/j.heliyon.2024.e25083

Genomic insight into O-demethylation of 4-methoxybenzoate by a two-component system from Amycolatopsis magusensis KCCM40447

Heliyon. 2024 Jan 21;10(3):e25083. doi: 10.1016/j.heliyon.2024.e25083. eCollection 2024 Feb 15.

Authors

Bashu Dev Pardhe¹, Lakshan Paudel¹, So-Ra Han², Tae-Jin Oh^{1

2

3}

Affiliations

¹ Department of Life Science and Biochemical Engineering, Sun Moon University, Asan, Republic of Korea.
² Genome-based BioIT Convergence Institute, Asan, Republic of Korea.
³ Department of Pharmaceutical Engineering and Biotechnology, Sun Moon University, Asan, Republic of Korea.

Abstract

Cytochrome P450 monooxygenases perform a multitude of roles, including the generation of hydroxylated aromatic compounds that might be utilized by microorganisms for their survival. WGS data of Amycolatopsis magusensis KCCM40447 revealed a complete circular genome of 9,099,986 base pairs and functionally assigned 8601 protein-encoding genes. Genomic analysis confirmed that the gene for 4-methoxybenzoate monoxygenase (CYP199A35) was conserved in close proximity to the gene for 4-hydroxybenzoate transporter (PcaK). The co-localized genes encoding CYP199A35, and ferredoxin-NAD(P) reductase (Mbr) represent a two-component system for electron transfer. CYP199A35 was specific for O-demethylation of para O-methyl substituted benzoic acid derivatives, 4-methoxybenzoate (4 MB), and 4-methoxycinnamic acid (4MCA) using the native redox partner (Mbr); two-component system and non-physiological redox partners (Pdr/Pdx); three-component system. The catalytic efficiency for O-demethylation of 4 MB using Mbr and Pdr/Pdx was 0.02 ± 0.006 min^-1 μM^-1 and 0.07 ± 0.02 min^-1 μM^-1 respectively. Further, sequence annotation and function prediction by RAST and KEEG analysis revealed a complete catabolic pathway for the utilization of 4 MB by strain KCCM40447, which was also proved experimentally.

Keywords: 4-Methoxybenzoate; Amycolatopsis magusensis KCCM40447; Cytochrome P450; Electron transfer redox system; Genomic environment; O-demethylation.