scholarly journals Pentamycin biosynthesis in PhilippineStreptomycessp. S816: Cytochrome P450-catalysed installation of the C-14 hydroxyl group

2019 ◽  
Author(s):  
Shanshan Zhou ◽  
Lijiang Song ◽  
Joleen Masschelein ◽  
Felaine A.M. Sumang ◽  
Irene A. Papa ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Vaginal Candidiasis ◽  
Hydroxyl Group ◽  
Biosynthetic Gene ◽  
Mangrove Soil ◽  
Genes Encoding ◽  
Biosynthetic Engineering ◽  
High Degree

ABSTRACTPentamycin is a polyene antibiotic, registered in Switzerland for the treatment of vaginal candidiasis, trichomo-niasis and mixed infections. Chemical instability has hindered its wide-spread application and development as a drug. Here we report the identification ofStreptomycessp. S816, isolated from Philippine mangrove soil, as a pentamycin producer. Genome sequence analysis identified the putative pentamycin biosynthetic gene cluster, which shows a high degree of similarity to the gene cluster responsible for filipin III biosynthesis. TheptnJgene, which is absent from the filipin III biosynthetic gene cluster, was shown to encode a cytochrome P450 capable of converting filipin III to pentamycin. This confirms that the cluster directs pentamycin biosynthesis, paving the way for biosynthetic engineering approaches to the production of pentamycin analogues. Several otherStreptomycesgenomes were found to containptnJorthologues clustered with genes encoding polyketide synthases that appear to have similar architectures to those responsible for the assembly of filipin III and pentamycin, suggesting pentamycin production may be common inStreptomycesspecies.

scholarly journals Biosynthetic Investigations of Lactonamycin and Lactonamycin Z: Cloning of the Biosynthetic Gene Clusters and Discovery of an Unusual Starter Unit

2007 ◽  
Vol 52 (2) ◽  
pp. 574-585 ◽  
Author(s):  
Xiujun Zhang ◽  
Lawrence B. Alemany ◽  
Hans-Peter Fiedler ◽  
Michael Goodfellow ◽  
Ronald J. Parry
Keyword(s):  
Gene Cluster ◽  
Genetic Locus ◽  
Gene Clusters ◽  
Biosynthetic Gene Cluster ◽  
Open Reading Frames ◽  
Antibacterial Drug ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters ◽  
Starter Unit ◽  
High Degree

ABSTRACT The antibiotics lactonamycin and lactonamycin Z provide attractive leads for antibacterial drug development. Both antibiotics contain a novel aglycone core called lactonamycinone. To gain insight into lactonamycinone biosynthesis, cloning and precursor incorporation experiments were undertaken. The lactonamycin gene cluster was initially cloned from Streptomyces rishiriensis. Sequencing of ca. 61 kb of S. rishiriensis DNA revealed the presence of 57 open reading frames. These included genes coding for the biosynthesis of l-rhodinose, the sugar found in lactonamycin, and genes similar to those in the tetracenomycin biosynthetic gene cluster. Since lactonamycin production by S. rishiriensis could not be sustained, additional proof for the identity of the S. rishiriensis cluster was obtained by cloning the lactonamycin Z gene cluster from Streptomyces sanglieri. Partial sequencing of the S. sanglieri cluster revealed 15 genes that exhibited a very high degree of similarity to genes within the lactonamycin cluster, as well as an identical organization. Double-crossover disruption of one gene in the S. sanglieri cluster abolished lactonamycin Z production, and production was restored by complementation. These results confirm the identity of the genetic locus cloned from S. sanglieri and indicate that the highly similar locus in S. rishiriensis encodes lactonamycin biosynthetic genes. Precursor incorporation experiments with S. sanglieri revealed that lactonamycinone is biosynthesized in an unusual manner whereby glycine or a glycine derivative serves as a starter unit that is extended by nine acetate units. Analysis of the gene clusters and of the precursor incorporation data suggested a hypothetical scheme for lactonamycinone biosynthesis.

scholarly journals Utilization of the Methoxymalonyl-Acyl Carrier Protein Biosynthesis Locus for Cloning the Oxazolomycin Biosynthetic Gene Cluster from Streptomyces albus JA3453

2006 ◽  
Vol 188 (11) ◽  
pp. 4142-4147 ◽  
Author(s):  
Chunhua Zhao ◽  
Jianhua Ju ◽  
Steven D. Christenson ◽  
Wyatt C. Smith ◽  
Danfeng Song ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Acyl Carrier Protein ◽  
Protein Biosynthesis ◽  
Biosynthetic Gene Cluster ◽  
Carrier Protein ◽  
Biosynthetic Gene ◽  
Degenerate Primers ◽  
Streptomyces Albus ◽  
Genes Encoding ◽  
Antiviral Activities

ABSTRACT Oxazolomycin (OZM), a hybrid peptide-polyketide antibiotic, exhibits potent antitumor and antiviral activities. Using degenerate primers to clone genes encoding methoxymalonyl-acyl carrier protein (ACP) biosynthesis as probes, a 135-kb DNA region from Streptomyces albus JA3453 was cloned and found to cover the entire OZM biosynthetic gene cluster. The involvement of the cloned genes in OZM biosynthesis was confirmed by deletion of a 12-kb DNA fragment containing six genes for methoxymalonyl-ACP biosynthesis from the specific region of the chromosome, as well as deletion of the ozmC gene within this region, to generate OZM-nonproducing mutants.

scholarly journals Biosynthetic engineering of the antifungal, anti-MRSA auroramycin

2019 ◽  
Author(s):  
Wan Lin Yeo ◽  
Elena Heng ◽  
Lee Ling Tan ◽  
Yi Wee Lim ◽  
Kuan Chieh Ching ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
Genome Editing ◽  
Gene Cluster ◽  
Biosynthetic Pathway ◽  
Biosynthetic Gene Cluster ◽  
Combinatorial Biosynthesis ◽  
Biosynthetic Gene ◽  
Antifungal Activities ◽  
Biosynthetic Engineering

AbstractUsing an established CRISPR-Cas mediated genome editing technique for streptomycetes, we explored the combinatorial biosynthesis potential of the auroramycin biosynthetic gene cluster in Streptomyces roseoporous. Auroramycin is a potent anti-MRSA polyene macrolactam. In addition, it also displays antifungal activities, which is unique among structurally similar polyene macrolactams, such as incednine and silvalactam. In this work, we employed different engineering strategies to target glycosylation and acylation biosynthetic machineries within its recently elucidated biosynthetic pathway. Six auroramycin analogs with variations in C-, N-methylation, hydroxylation and extender units incorporation were produced and characterized. By comparing the bioactivity profiles of these analogs, we determined that unique disaccharide motif of auroramycin is essential for its antimicrobial bioactivity. We further demonstrated that C-methylation of the 3, 5-epi-lemonose unit, which is unique among structurally similar polyene macrolactams, is key to its antifungal activity.

scholarly journals Heterologous Biosynthesis and Genomics-Driven Derivatization of Fungal Bioactive Sesterterpenoid Variecolin

2021 ◽  
Author(s):  
Dexiu Yan ◽  
Jemma Arakelyan ◽  
Teng Wan ◽  
Tsz Ki Chan ◽  
Dohyun Ahn ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Anticancer Activity ◽  
Gene Cluster ◽  
Aspergillus Oryzae ◽  
Biosynthetic Gene Cluster ◽  
Heterologous Production ◽  
Biosynthetic Gene ◽  
Aspergillus Aculeatus ◽  
Heterologous Biosynthesis

The biosynthetic gene cluster of fungal bioactive sesterterpenoids, variecolin (1) and variecolactone (2), was identified in Aspergillus aculeatus ATCC 16872. Heterologous production of 1 and 2 was achieved in Aspergillus oryzae by expressing the sesterterpene synthase VrcA and the cytochrome P450 VrcB. Intriguingly, the replacement of VrcB with homologous P450s from other fungal terpenoid pathways yielded three new variecolin analogues, one of which exhibited potent anticancer activity comparable to that of 1.

scholarly journals Analysis of the Tunicamycin Biosynthetic Gene Cluster ofStreptomyces chartreusisReveals New Insights into Tunicamycin Production and Immunity

2018 ◽  
Vol 62 (8) ◽  
Author(s):  
David Widdick ◽  
Sylvain F. Royer ◽  
Hua Wang ◽  
Natalia M. Vior ◽  
Juan Pablo Gomez-Escribano ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Streptomyces Coelicolor ◽  
Biosynthetic Gene Cluster ◽  
Transcriptional Analysis ◽  
Efficient Production ◽  
Primary Metabolism ◽  
Biosynthetic Gene ◽  
Content Type ◽  
High Degree ◽  
Single Operon

ABSTRACTThe tunicamycin biosynthetic gene cluster ofStreptomyces chartreusisconsists of 14 genes (tunAtotunN) with a high degree of apparent translational coupling. Transcriptional analysis revealed that all of these genes are likely to be transcribed as a single operon from two promoters,tunp1 andtunp2. In-frame deletion analysis revealed that just six of these genes (tunABCDEH) are essential for tunicamycin production in the heterologous hostStreptomyces coelicolor, while five (tunFGKLN) with likely counterparts in primary metabolism are not necessary, but presumably ensure efficient production of the antibiotic at the onset of tunicamycin biosynthesis. Three genes are implicated in immunity, namely,tunIandtunJ, which encode a two-component ABC transporter presumably required for export of the antibiotic, andtunM, which encodes a putativeS-adenosylmethionine (SAM)-dependent methyltransferase. Expression oftunIJortunMinS. coelicolorconferred resistance to exogenous tunicamycin. The results presented here provide new insights into tunicamycin biosynthesis and immunity.

scholarly journals Characterization of two cytochrome P450 monooxygenase genes of the pyripyropene biosynthetic gene cluster from Penicillium coprobium

2011 ◽  
Vol 64 (3) ◽  
pp. 221-227 ◽  
Author(s):  
Jie Hu ◽  
Hiroto Okawa ◽  
Kentaro Yamamoto ◽  
Kazuhiko Oyama ◽  
Masaaki Mitomi ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Cytochrome P450 Monooxygenase ◽  
Biosynthetic Gene ◽  
P450 Monooxygenase

scholarly journals Expansion within the CYP71D subfamily drives the heterocyclization of tanshinones synthesis in Salvia miltiorrhiza

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ying Ma ◽  
Guanghong Cui ◽  
Tong Chen ◽  
Xiaohui Ma ◽  
Ruishan Wang ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gene Cluster ◽  
Genome Assembly ◽  
Salvia Miltiorrhiza ◽  
Gene Array ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene ◽  
Chinese Medicinal Herb ◽  
Gene Expansion ◽  
Homozygous Genotype

AbstractTanshinones are the bioactive nor-diterpenoid constituents of the Chinese medicinal herb Danshen (Salvia miltiorrhiza). These groups of chemicals have the characteristic furan D-ring, which differentiates them from the phenolic abietane-type diterpenoids frequently found in the Lamiaceae family. However, how the 14,16-epoxy is formed has not been elucidated. Here, we report an improved genome assembly of Danshen using a highly homozygous genotype. We identify a cytochrome P450 (CYP71D) tandem gene array through gene expansion analysis. We show that CYP71D373 and CYP71D375 catalyze hydroxylation at carbon-16 (C16) and 14,16-ether (hetero)cyclization to form the D-ring, whereas CYP71D411 catalyzes upstream hydroxylation at C20. In addition, we discover a large biosynthetic gene cluster associated with tanshinone production. Collinearity analysis indicates a more specific origin of tanshinones in Salvia genus. It illustrates the evolutionary origin of abietane-type diterpenoids and those with a furan D-ring in Lamiaceae.

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