Natural and engineered biosynthesis of nucleoside antibiotics in <i>Actinomycetes</i>

Chen, Wenqing; Jing, Qi; Wu, Pan; Wan, Dan; Jin, Liu; Feng, Xuan; Deng, Zixin

doi:10.1007/s10295-015-1636-3

Cited by 41 publications

(29 citation statements)

References 97 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nucleoside antibiotics constitute a large family of important microbial natural products bearing diverse bioactivities and unusual structural features (Chen et al, 2016;Isono, 1988). Their biosynthesis usually involves a succinct logic by sequential modification of nucleoside or nucleotides originating from primary metabolism.…”

Section: Introductionmentioning

confidence: 99%

Deciphering Carbamoylpolyoxamic Acid Biosynthesis Reveals Unusual Acetylation Cycle Associated with Tandem Reduction and Sequential Hydroxylation

Jing

Wan

et al. 2016

Cell Chemical Biology

Self Cite

View full text Add to dashboard Cite

Polyoxin, produced by Streptomcyes cacaoi var. asoensis and Streptomyces aureochromogenes, contains two non-proteinogenic amino acids, carbamoylpolyoxamic acid (CPOAA) and polyoximic acid. Although the CPOAA moiety is highly unusual, its biosynthetic logic has remained enigmatic for decades. Here, we address CPOAA biosynthesis by reconstitution of its pathway. We demonstrated that its biosynthesis is initiated by a versatile N-acetyltransferase, PolN, catalyzing L-glutamate (1) to N-acetyl glutamate (2). Remarkably, we verified that PolM, a previously annotated dehydrogenase, catalyzes an unprecedented tandem reduction of acyl-phosphate to aldehyde, and subsequently to alcohol. We also unveiled a distinctive acetylation cycle catalyzed by PolN to synthesize α-amino-δ-hydroxyvaleric acid (6). Finally, we report that PolL is capable of converting a rare sequential hydroxylation of α-amino-δ-carbamoylhydroxyvaleric acid (7) to CPOAA. PolL represents an intriguing family of Fe(II)-dependent α-ketoglutarate dioxygenase with a cupin fold. These data illustrate several novel enzymatic reactions, and also set a foundation for rational pathway engineering for polyoxin production.

show abstract

Section: Introductionmentioning

confidence: 99%

Deciphering Carbamoylpolyoxamic Acid Biosynthesis Reveals Unusual Acetylation Cycle Associated with Tandem Reduction and Sequential Hydroxylation

Jing

Wan

et al. 2016

Cell Chemical Biology

Self Cite

View full text Add to dashboard Cite

show abstract

“…LC-HRMS/MS was operated in an ESI-ion trap mass spectrometer under the positive-ion mode, with drying gas at 275°C, 10 liters/ml, and nebulizer pressure of 30 lb/in. 2 . The activity assays were performed with the general protocol of the EnzChek pyrophosphate assay kit (Thermo).…”

mentioning

confidence: 99%

Biosynthesis of 2′-Chloropentostatin and 2′-Amino-2′-Deoxyadenosine Highlights a Single Gene Cluster Responsible for Two Independent Pathways in Actinomadura sp. Strain ATCC 39365

Gao

et al. 2017

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

2=-Chloropentostatin (2=-Cl PTN, 2=-chloro-2=-deoxycoformycin) and 2=-amino-2=-deoxyadenosine (2=-amino dA) are two adenosine-derived nucleoside antibiotics coproduced by Actinomadura sp. strain ATCC 39365. 2=-Cl PTN is a potent adenosine deaminase (ADA) inhibitor featuring an intriguing 1,3-diazepine ring, as well as a chlorination at C-2= of ribose, and 2=-amino dA is an adenosine analog showing bioactivity against RNA-type virus infection. However, the biosynthetic logic of them has remained poorly understood. Here, we report the identification of a single gene cluster (ada) essential for the biosynthesis of 2=-Cl PTN and 2=-amino dA. Further systematic genetic investigations suggest that 2=-Cl PTN and 2=-amino dA are biosynthesized by independent pathways. Moreover, we provide evidence that a predicted cation/H ϩ antiporter, AdaE, is involved in the chlorination step during 2=-Cl PTN biosynthesis. Notably, we demonstrate that 2=-amino dA biosynthesis is initiated by a Nudix hydrolase, AdaJ, catalyzing the hydrolysis of ATP. Finally, we reveal that the host ADA (designated ADA1), capable of converting adenosine/2=-amino dA to inosine/2=-amino dI, is not very sensitive to the powerful ADA inhibitor pentostatin. These findings provide a basis for the further rational pathway engineering of 2=-Cl PTN and 2=-amino dA production.IMPORTANCE 2=-Cl PTN/PTN and 2=-amino dA have captivated the great interests of scientists, owing to their unusual chemical structures and remarkable bioactivities. However, the precise logic for their biosynthesis has been elusive for decades. Actually, the identification and elucidation of their biosynthetic pathways not only enrich the biochemical repertoire of novel enzymatic reactions but may also lay solid foundations for the pathway engineering and combinatorial biosynthesis of this family of purine nucleoside antibiotics to generate novel hybrid analogs with improved features.KEYWORDS 2=-chloropentostatin, 2=-amino-2=-deoxyadenosine, nucleoside antibiotics, biosynthesis, Nudix hydrolase, adenosine deaminase N ucleoside antibiotics are a large family of important microbial natural products harboring wide-range biological properties and distinctive structural features (1-3). Their biosynthesis generally follows a succinct logic by sequential enzymatic modification of nucleosides or nucleotides originating from primary metabolisms (1). Usually, nucleosides and nucleotides play pleiotropic roles in most fundamental cellular metabolisms; therefore, nucleoside antibiotics are able to target the biosynthesis of diverse biomacromolecules, including nucleic acids, proteins, and glycans (1).

show abstract

“…Carbocyclic natural products include many interesting chemotypes, including the antitumor antibiotic pactamycin, the chitinase inhibitor allosamidin, and the carbocyclic purine nucleoside antibiotics (13). Generally, purine nucleoside natural products are synthesized by a nonmodular assembly line (2), which complicates the identification of their biosynthetic gene clusters. In this respect, the discovery of new ARM and COF producer strains (see Fig.…”

Section: Discussionmentioning

confidence: 99%

“…HPLC was performed with an elution gradient of 5% to 20% methanol-15 mM TFA over 20 min at a flow rate of 0.5 ml/min. LC-MS/MS analysis was conducted in an electrospray ionization (ESI) trap mass spectrometer in the positive-ion mode with the following parameters: drying gas at 275°C, 10 liters/ml, and nebulizer pressure at 30 lb/in 2 .…”

Section: Methodsmentioning

confidence: 99%

Coordinated Biosynthesis of the Purine Nucleoside Antibiotics Aristeromycin and Coformycin in Actinomycetes

Kong

Gong

et al. 2018

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

Purine nucleoside antibiotic pairs, concomitantly produced by a single strain, are an important group of microbial natural products. Here, we report a target-directed genome mining approach to elucidate the biosynthesis of the purine nucleoside antibiotic pair aristeromycin (ARM) and coformycin (COF) in DSM 45626 (a new producer for ARM and COF) and NBRC 13005 (a new COF producer). We also provide biochemical data that MacI and MacT function as unusual phosphorylases, catalyzing an irreversible reaction for the tailoring assembly of neplanocin A (NEP-A) and ARM. Moreover, we demonstrate that MacQ is shown to be an adenosine-specific deaminase, likely relieving the potential "excess adenosine" for producing cells. Finally, we report that MacR, an annotated IMP dehydrogenase, is actually an NADPH-dependent GMP reductase, which potentially plays a salvage role for the efficient supply of the precursor pool. Hence, these findings illustrate a fine-tuned pathway for the biosynthesis of ARM and also open the way for the rational search for purine antibiotic pairs. ARM and COF are well known for their prominent biological activities and unusual chemical structures; however, the logic of their biosynthesis has long been poorly understood. Actually, the new insights into the ARM and COF pathway will not only enrich the biochemical repertoire for interesting enzymatic reactions but may also lay a solid foundation for the combinatorial biosynthesis of this group of antibiotics via a target-directed genome mining strategy.

show abstract

Natural and engineered biosynthesis of nucleoside antibiotics in Actinomycetes

Cited by 41 publications

References 97 publications

Deciphering Carbamoylpolyoxamic Acid Biosynthesis Reveals Unusual Acetylation Cycle Associated with Tandem Reduction and Sequential Hydroxylation

Deciphering Carbamoylpolyoxamic Acid Biosynthesis Reveals Unusual Acetylation Cycle Associated with Tandem Reduction and Sequential Hydroxylation

Biosynthesis of 2′-Chloropentostatin and 2′-Amino-2′-Deoxyadenosine Highlights a Single Gene Cluster Responsible for Two Independent Pathways in Actinomadura sp. Strain ATCC 39365

Coordinated Biosynthesis of the Purine Nucleoside Antibiotics Aristeromycin and Coformycin in Actinomycetes

Contact Info

Product

Resources

About