scholarly journals The biosynthetic gene cluster of the C-nucleoside antibiotic pyrazomycin with a rare pyrazole moiety

2019 ◽  
Author(s):  
Guiyun Zhao ◽  
Shunyu Yao ◽  
Kristina W Rothchild ◽  
Tengfei Liu ◽  
Yu Liu ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Biochemical Characterization ◽  
Pyrazole Ring ◽  
Biosynthetic Gene Cluster ◽  
Bond Forming ◽  
Feeding Experiments ◽  
Naturally Occurring ◽  
Nitrogen Bond

AbstractPyrazomycin is a rare C-nucleoside antibiotic with a naturally occurring pyrazole ring, whose biosynthetic origin has remained obscure for decades. In this study, we report the identification of the gene cluster responsible for pyrazomycin biosynthesis in Streptomyces candidus NRRL 3601, revealing that StrR-family regulator PyrR is the cluster-situated transcriptional activator governing pyrazomycin biosynthesis. Furthermore, our results from in vivo reconstitution and stable-isotope feeding experiments support that PyrN is a new nitrogen-nitrogen bond forming enzyme linking the ε-NH2 nitrogen of l-N6-OH-lysine and α-NH2 nitrogen of l-glutamate. This study lays the foundation for further genetic and biochemical characterization of pyrazomycin pathway enzymes constructing the characteristic pyrazole ring.

scholarly journals The L-alanosine gene cluster encodes a pathway for diazeniumdiolate biosynthesis

2019 ◽  
Author(s):  
Tai L. Ng ◽  
Monica E. McCallum ◽  
Christine R. Zheng ◽  
Jennifer X. Wang ◽  
Kelvin J. Y. Wu ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Biochemical Characterization ◽  
Biosynthetic Gene Cluster ◽  
Structural Motif ◽  
Bioactive Metabolites ◽  
Anticancer Properties ◽  
Feeding Experiments ◽  
Wide Range ◽  
Diaminopropionic Acid

AbstractN-nitroso-containing natural products are bioactive metabolites with antibacterial and anticancer properties. In particular, compounds containing the diazeniumdiolate (N-nitrosohydroxylamine) group display a wide range of bioactivities ranging from cytotoxicity to metal chelation. Despite the importance of this structural motif, knowledge of its biosynthesis is limited. Herein, we describe the discovery of a biosynthetic gene cluster in Streptomyces alanosinicus ATCC 15710 responsible for producing the diazeniumdiolate natural product L-alanosine. Gene disruption and stable isotope feeding experiments identified essential biosynthetic genes and revealed the nitrogen source of the N-nitroso group. Additional biochemical characterization of the biosynthetic enzymes revealed that the non-proteinogenic amino acid L-2,3-diaminopropionic acid (L-Dap) is synthesized and loaded onto a peptidyl carrier protein (PCP) domain in L-alanosine biosynthesis, which we propose may be a mechanism of handling unstable intermediates generated en route to the diazeniumdiolate. This research framework will facilitate efforts to determine the biochemistry of diazeniumdiolate formation.

Genetic localization and in vivo characterization of a Monascus azaphilone pigment biosynthetic gene cluster

2013 ◽  
Vol 97 (14) ◽  
pp. 6337-6345 ◽  
Author(s):  
Bijinu Balakrishnan ◽  
Suman Karki ◽  
Shih-Hau Chiu ◽  
Hyun-Ju Kim ◽  
Jae-Won Suh ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene ◽  
Genetic Localization ◽  
In Vivo Characterization ◽  
Azaphilone Pigment

scholarly journals Parsing a multifunctional biosynthetic gene cluster from rice: Biochemical characterization of CYP71Z6 & 7

FEBS Letters ◽  
2011 ◽  
Vol 585 (21) ◽  
pp. 3446-3451 ◽  
Author(s):  
Yisheng Wu ◽  
Matthew L. Hillwig ◽  
Qiang Wang ◽  
Reuben J. Peters
Keyword(s):  
Gene Cluster ◽  
Biochemical Characterization ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene

scholarly journals Cloning and Characterization of the Tetrocarcin A Gene Cluster from Micromonospora chalcea NRRL 11289 Reveals a Highly Conserved Strategy for Tetronate Biosynthesis in Spirotetronate Antibiotics

2008 ◽  
Vol 190 (17) ◽  
pp. 6014-6025 ◽  
Author(s):  
Jie Fang ◽  
Yiping Zhang ◽  
Lijuan Huang ◽  
Xinying Jia ◽  
Qi Zhang ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Structural Diversity ◽  
Biosynthetic Gene Cluster ◽  
Combinatorial Biosynthesis ◽  
In Vitro Characterization ◽  
Fused Gene ◽  
Micromonospora Chalcea

ABSTRACT Tetrocarcin A (TCA), produced by Micromonospora chalcea NRRL 11289, is a spirotetronate antibiotic with potent antitumor activity and versatile modes of action. In this study, the biosynthetic gene cluster of TCA was cloned and localized to a 108-kb contiguous DNA region. In silico sequence analysis revealed 36 putative genes that constitute this cluster (including 11 for unusual sugar biosynthesis, 13 for aglycone formation, and 4 for glycosylations) and allowed us to propose the biosynthetic pathway of TCA. The formation of d-tetronitrose, l-amicetose, and l-digitoxose may begin with d-glucose-1-phosphate, share early enzymatic steps, and branch into different pathways by competitive actions of specific enzymes. Tetronolide biosynthesis involves the incorporation of a 3-C unit with a polyketide intermediate to form the characteristic spirotetronate moiety and trans-decalin system. Further substitution of tetronolide with five deoxysugars (one being a deoxynitrosugar) was likely due to the activities of four glycosyltransferases. In vitro characterization of the first enzymatic step by utilization of 1,3-biphosphoglycerate as the substrate and in vivo cross-complementation of the bifunctional fused gene tcaD3 (with the functions of chlD3 and chlD4) to ΔchlD3 and ΔchlD4 in chlorothricin biosynthesis supported the highly conserved tetronate biosynthetic strategy in the spirotetronate family. Deletion of a large DNA fragment encoding polyketide synthases resulted in a non-TCA-producing strain, providing a clear background for the identification of novel analogs. These findings provide insights into spirotetronate biosynthesis and demonstrate that combinatorial-biosynthesis methods can be applied to the TCA biosynthetic machinery to generate structural diversity.

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