Biosynthesis of Aminoglycoside Antibiotics

Kudo, Fumitaka

doi:10.1016/b978-0-12-409547-2.14619-0

Cited by 6 publications

(10 citation statements)

References 171 publications

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“…By contrast, KasH is literally the aminoglycoside-modifying enzyme transferring the acetyl group from AcCoA specific to KSM to form 2-N'-acetyl KSM. Finally, the isotope-feeding study confirmed that 13 C, 15 N-glucosamine/UDP-GlcNH 2 rather than glucose/UDP-Glc is the direct precursor in the formation of KSM. To this end, we have addressed several long-standing unresolved issues, which should be conducive toward mapping out the correct KSM biosynthetic pathway.…”

Section: Discussionmentioning

confidence: 69%

“…1 H and 13 C NMR spectra were recorded on a Bruker 600 spectrometer with tetramethylsilane as the internal reference. 31 P NMR was performed in Bruker Avance 500 AV NMR.…”

Section: General Informationmentioning

confidence: 99%

“…1 H NMR (600 MHz, CDCl 3 ) δ 6.60 (s, 1H), 5.56 (d, J = 4.8 Hz, 1H), 5.25 (m, 1H), 4.51 (m, 1H), 4.47 (m, 1H), 4.19 (dd, J = 12.0, 3.0 Hz, 1H), 2.09 (s, 6H), 2.01 (s, 3H). 13 C NMR (150 MHz, CDCl 3 ) δ 170.7, 170.0, 169.7, 148.0, 113.5, 74.9, 67.4, 67.1, 61.1, 20.9, 20.8. To a glucal III (0.600 g, 2.09 mmol), dry methanol (14 mL) was added and dissolved, then sodium methoxide was added (pH ~8.5) and the mixture was stirred for up to 2 h. The mixture was neutralized with Amberlite IR-120 (H+), filtered, and concentrated.…”

Section: General Informationmentioning

confidence: 99%

“…The following supporting information can be downloaded at: https: //www.mdpi.com/article/10.3390/biomedicines10020212/s1, Figure S1 13 C NMR spectra, Figure S35: 1 H and 13 C NMR spectra, Figure S36: 1 H and 13 C NMR spectra, Table S1: Thermodynamic parameters of KasH for its binding affinity with testing ligands, Table S2: Kinetics of KasQ_WT in reaction with UDP-GlcNAc and MgCl 2 , Table S3: Thermodynamic parameters of KasQ mutants in the binding affinity assay.…”

Section: Supplementary Materialsmentioning

confidence: 99%

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KasQ an Epimerase Primes the Biosynthesis of Aminoglycoside Antibiotic Kasugamycin and KasF/H Acetyltransferases Inactivate Its Activity

et al. 2022

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Kasugamycin (KSM), an aminoglycoside antibiotic, is composed of three chemical moieties: D-chiro-inositol, kasugamine and glycine imine. Despite being discovered more than 50 years ago, the biosynthetic pathway of KSM remains an unresolved puzzle. Here we report a structural and functional analysis for an epimerase, KasQ, that primes KSM biosynthesis rather than the previously proposed KasF/H, which instead acts as an acetyltransferase, inactivating KSM. Our biochemical and biophysical analysis determined that KasQ converts UDP-GlcNAc to UDP-ManNAc as the initial step in the biosynthetic pathway. The isotope-feeding study further confirmed that 13C,15N-glucosamine/UDP-GlcNH2 rather than glucose/UDP-Glc serves as the direct precursor for the formation of KSM. Both KasF and KasH were proposed, respectively, converting UDP-GlcNH2 and KSM to UDP-GlcNAc and 2-N’-acetyl KSM. Experimentally, KasF is unable to do so; both KasF and KasH are instead KSM-modifying enzymes, while the latter is more specific and reactive than the former in terms of the extent of resistance. The information gained here lays the foundation for mapping out the complete KSM biosynthetic pathway.

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Section: Discussionmentioning

confidence: 69%

“…1 H and 13 C NMR spectra were recorded on a Bruker 600 spectrometer with tetramethylsilane as the internal reference. 31 P NMR was performed in Bruker Avance 500 AV NMR.…”

Section: General Informationmentioning

confidence: 99%

Section: General Informationmentioning

confidence: 99%

Section: Supplementary Materialsmentioning

confidence: 99%

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KasQ an Epimerase Primes the Biosynthesis of Aminoglycoside Antibiotic Kasugamycin and KasF/H Acetyltransferases Inactivate Its Activity

et al. 2022

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“…Base on bioinformatical analysis, the genes for the common intermediates 2-deoxystreptamine and paromamine can be easily identi ed both in apramycin and tobramycin biosynthetic gene clusters. 8 Lividamine is the 3' deoxygenated product of paromamine under the function of AprD3 and AprD4. [9][10][11][12] Then, lividamine was oxidized by AprQ to form 6'-oxolividamine which may be the intermediate of bicyclic 3'-deoxyoctose moiety.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Methyltransferase from Apramycin Biosynthetic Gene Cluster and Improvement Production of Apramycin in Engineered Streptoalloteichus Tenebrarius

Sun

Gao

Yan

et al. 2021

Preprint

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BackgroundApramycin is a structurally unique aminoglycoside, used in veterinary medicine or the treatment of Salmonella, Escherichia coli and Pasteurella multocida infections in farm. Although discovered and used many years ago, many biosynthetic steps of apramycin are still obscure. ResultsIn this study, we identified a HemK family methyltransferase, aprI, involved in apramycin biosynthesis. The function of aprI was studied by using gene disruption and biochemical experiments, and a new aminoglycoside antibiotic demethyl-apramycin was purified from aprI disruption strain. Experiments proved that AprI converted demethyl-aprosamine to aprosamine in vitro. Based on this, the apramycin production strain was improved by overexpression the AprI to decrease the impurity production. ConclusionsWe have identified aprI is a 7’-N-methyltransferase gene in apramycin biosynthesis and confirmed the substrate of methyltransferase. Engineering of aprI resulted in a strain producing a new aminoglycoside demethyl-apramycin and apramycin mono-producing strain with less impurity production. Finally, the yield of demethyl-apramycin in apramycin mono-producing strain decreased from 196±36 mg/L to 51±9 mg/L, and the yield of apramycin increased from 2227±320 mg/L to 2331±210 mg/L.

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Stepwise Post‐glycosylation Modification of Sugar Moieties in Kanamycin Biosynthesis

et al. 2021

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Kanamycin A is the major 2‐deoxystreptamine (2DOS)‐containing aminoglycoside antibiotic produced by Streptomyces kanamyceticus. The 2DOS moiety is linked with 6‐amino‐6‐deoxy‐d‐glucose (6ADG) at O‐4 and 3‐amino‐3‐deoxy‐d‐glucose at O‐6. Because the 6ADG moiety is derived from d‐glucosamine (GlcN), deamination at C‐2 and introduction of C‐6‐NH2 are required in the biosynthesis. A dehydrogenase, KanQ, and an aminotransferase, KanB, are presumed to be responsible for the introduction of C‐6‐NH2, although the substrates have not been identified. Here, we examined the substrate specificity of KanQ to better understand the biosynthetic pathway. It was found that KanQ oxidized kanamycin C more efficiently than the 3′′‐deamino derivative. Furthermore, the substrate specificity of an oxygenase, KanJ, that is responsible for deamination at C‐2 of the GlcN moiety was examined, and the crystal structure of KanJ was determined. It was found that C‐6‐NH2 is important for substrate recognition by KanJ. Thus, the modification of the GlcN moiety occurs after pseudo‐trisaccharide formation, followed by the introduction of C‐6‐NH2 by KanQ/KanB and deamination at C‐2 by KanJ.

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Biosynthesis of Aminoglycoside Antibiotics

Cited by 6 publications

References 171 publications

KasQ an Epimerase Primes the Biosynthesis of Aminoglycoside Antibiotic Kasugamycin and KasF/H Acetyltransferases Inactivate Its Activity

KasQ an Epimerase Primes the Biosynthesis of Aminoglycoside Antibiotic Kasugamycin and KasF/H Acetyltransferases Inactivate Its Activity

Characterization of Methyltransferase from Apramycin Biosynthetic Gene Cluster and Improvement Production of Apramycin in Engineered Streptoalloteichus Tenebrarius

Stepwise Post‐glycosylation Modification of Sugar Moieties in Kanamycin Biosynthesis

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