In Vivo Characterization of Phosphotransferase-Encoding Genes istP and forP as Interchangeable Launchers of the C3��,4��-Dideoxygenation Biosynthetic Pathway of 1,4-Diaminocyclitol Antibiotics

Hương, Nguyễn Lan; Lee, Na Joon; Hwang, Hyun Ha; Son, Hye Bin; Kim, Hye-Ji; Seo, Eun Gyo; Hoang, Nguyen Huu; Park, Je Won

doi:10.4014/jmb.1809.09021

“…AMEs exist in both resistant pathogens and AGA producers [ 33 ]. It has been speculated that these enzymes may perform other metabolic functions [ 22 ]. In the present study, we have demonstrated that gentamicin producers evolved a smart strategy to evade APH(3′) deactivation by pathogens.…”

Section: Discussionmentioning

confidence: 99%

“…CAF31545.1), while ForP can complement mutants in the dehydroxylation process of the fortimicin A biosynthesis pathway. Therefore, it is speculated that GenP can phosphorylate JI-20A, JI-20Ba, and JI-20B, which are the starting compounds of the C-3′,4′-dideoxygenation biosynthetic process [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

Pyridoxal-5′-phosphate-dependent enzyme GenB3 Catalyzes C-3′,4′-dideoxygenation in gentamicin biosynthesis

Zhou

¹

,

Chen

²

,

Ni

³

et al. 2021

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Background The C-3′,4′-dideoxygenation structure in gentamicin can prevent deactivation by aminoglycoside 3′-phosphotransferase (APH(3′)) in drug-resistant pathogens. However, the enzyme catalyzing the dideoxygenation step in the gentamicin biosynthesis pathway remains unknown. Results Here, we report that GenP catalyzes 3′ phosphorylation of the gentamicin biosynthesis intermediates JI-20A, JI-20Ba, and JI-20B. We further demonstrate that the pyridoxal-5′-phosphate (PLP)-dependent enzyme GenB3 uses these phosphorylated substrates to form 3′,4′-dideoxy-4′,5′-ene-6′-oxo products. The following C-6′-transamination and the GenB4-catalyzed reduction of 4′,5′-olefin lead to the formation of gentamicin C. To the best of our knowledge, GenB3 is the first PLP-dependent enzyme catalyzing dideoxygenation in aminoglycoside biosynthesis. Conclusions This discovery solves a long-standing puzzle in gentamicin biosynthesis and enriches our knowledge of the chemistry of PLP-dependent enzymes. Interestingly, these results demonstrate that to evade APH(3′) deactivation by pathogens, the gentamicin producers evolved a smart strategy, which utilized their own APH(3′) to activate hydroxyls as leaving groups for the 3′,4′-dideoxygenation in gentamicin biosynthesis.

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Two Cryptic Self‐Resistance Mechanisms in Streptomyces tenebrarius Reveal Insights into the Biosynthesis of Apramycin

Zhang

¹

,

Chi

²

,

Wu

³

et al. 2021

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Apramycin is aclinically promising aminoglycoside antibiotic (AGA). To date,m echanisms underlying the biosynthesis and self-resistance of apramycin remain largely unknown. Here we report that apramycin biosynthesis proceeds through unexpected phosphorylation, deacetylation, and dephosphorylation steps,i nw hich an ovel aminoglycoside phosphotransferase (AprU), ap utative creatinine amidohydrolase (AprP), and an alkaline phosphatase (AprZ) are involved. Biochemical characterization revealed that AprU specifically phosphorylates 5-OH of ap seudotrisaccharide intermediate,w hose N-7' acetyl group is subsequently hydrolyzed by AprP.A prZ is located extracellularly where it removes the phosphate group from ap seudotetrasaccharide intermediate,leading to the maturation of apramycin. Intriguingly,7'-N-acetylated and 5-O-phosphorylated apramycin that were accumulated in DaprU and DaprZ respectively exhibited significantly reduced antibacterial activities,i mplying Streptomyces tenebrarius employs C-5 phosphorylation and N-7' acetylation as two strategies to avoid auto-toxicity.S ignificantly,t his study provides insight into the design of new generation AGAs to circumvent the emergence of drugresistant pathogens.

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Two Cryptic Self‐Resistance Mechanisms in Streptomyces tenebrarius Reveal Insights into the Biosynthesis of Apramycin

Zhang

¹

,

Chi

²

,

Wu

³

et al. 2021

View full text Add to dashboard Cite

Apramycin is aclinically promising aminoglycoside antibiotic (AGA). To date,m echanisms underlying the biosynthesis and self-resistance of apramycin remain largely unknown. Here we report that apramycin biosynthesis proceeds through unexpected phosphorylation, deacetylation, and dephosphorylation steps,i nw hich an ovel aminoglycoside phosphotransferase (AprU), ap utative creatinine amidohydrolase (AprP), and an alkaline phosphatase (AprZ) are involved. Biochemical characterization revealed that AprU specifically phosphorylates 5-OH of ap seudotrisaccharide intermediate,w hose N-7' acetyl group is subsequently hydrolyzed by AprP.A prZ is located extracellularly where it removes the phosphate group from ap seudotetrasaccharide intermediate,leading to the maturation of apramycin. Intriguingly,7'-N-acetylated and 5-O-phosphorylated apramycin that were accumulated in DaprU and DaprZ respectively exhibited significantly reduced antibacterial activities,i mplying Streptomyces tenebrarius employs C-5 phosphorylation and N-7' acetylation as two strategies to avoid auto-toxicity.S ignificantly,t his study provides insight into the design of new generation AGAs to circumvent the emergence of drugresistant pathogens.

show abstract

In Vivo Characterization of Phosphotransferase-Encoding Genes istP and forP as Interchangeable Launchers of the C3��,4��-Dideoxygenation Biosynthetic Pathway of 1,4-Diaminocyclitol Antibiotics

Cited by 5 publications

References 14 publications

Pyridoxal-5′-phosphate-dependent enzyme GenB3 Catalyzes C-3′,4′-dideoxygenation in gentamicin biosynthesis

Pyridoxal-5′-phosphate-dependent enzyme GenB3 Catalyzes C-3′,4′-dideoxygenation in gentamicin biosynthesis

Two Cryptic Self‐Resistance Mechanisms in Streptomyces tenebrarius Reveal Insights into the Biosynthesis of Apramycin

Two Cryptic Self‐Resistance Mechanisms in Streptomyces tenebrarius Reveal Insights into the Biosynthesis of Apramycin

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