scholarly journals Analysis of the Indole Diterpene Gene Cluster for Biosynthesis of the Epoxy-Janthitrems in Epichloë Endophytes

2019 ◽  
Vol 7 (11) ◽  
pp. 560 ◽  
Author(s):  
Emma J. Ludlow ◽  
Simone Vassiliadis ◽  
Piyumi N. Ekanayake ◽  
Inoka K. Hettiarachchige ◽  
Priyanka Reddy ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Gene Cluster ◽  
Perennial Ryegrass ◽  
Biosynthetic Pathway ◽  
Whole Genome Sequence ◽  
In Planta ◽  
Biosynthesis Gene Cluster ◽  
Biosynthesis Gene ◽  
Lolitrem B ◽  
Epichloë Endophytes

Epoxy-janthitrems are a class of indole diterpenes with structural similarity to lolitrem B. Two taxa of asexual Epichloë endophytes have been reported to produce epoxy-janthitrems, LpTG-3 (Lolium perenne Taxonomic Group 3; e.g., NEA12) and LpTG-4 (e.g., E1). Epichloë epoxy-janthitrems are not well understood, the biosynthetic pathway and associated gene complement have not been described and while the literature suggests they are associated with superior protection against pasture insect pests and are tremorgenic in grazing mammals, these properties have not been confirmed using isolated and purified compounds. Whole genome sequence analysis was used to identify candidate genes for epoxy-janthitrem biosynthesis that are unique to epoxy-janthitrem producing strains of Epichloë. A gene, jtmD, was identified with homology to aromatic prenyl transferases involved in synthesis of indole diterpenes. The location of the epoxy-janthitrem biosynthesis gene cluster (JTM locus) was determined in the assembled nuclear genomes of NEA12 and E1. The JTM locus contains cluster 1 and cluster 2 of the lolitrem B biosynthesis gene cluster (LTM locus), as well as four genes jtmD, jtmO, jtm01, and jtm02 that are unique to Epichloë spp. that produce epoxy-janthitrems. Expression of each of the genes identified was confirmed using transcriptome analysis of perennial ryegrass-NEA12 and perennial ryegrass-E1 symbiota. Sequence analysis confirmed the genes are functionally similar to those involved in biosynthesis of related indole diterpene compounds. RNAi silencing of jtmD and in planta assessment in host-endophyte associations confirms the role of jtmD in epoxy-janthitrem production. Using LCMS/MS technologies, a biosynthetic pathway for the production of epoxy-janthitrems I–IV in Epichloë endophytes is proposed.

The Blasticidin S Biosynthesis Gene Cluster from Streptomyces griseochromogenes: Sequence Analysis, Organization, and Initial Characterization

ChemBioChem ◽  
2003 ◽  
Vol 4 (9) ◽  
pp. 821-828 ◽  
Author(s):  
Martha C. Cone ◽  
Xihou Yin ◽  
Laura L. Grochowski ◽  
Morgan R. Parker ◽  
T. Mark Zabriskie
Keyword(s):  
Sequence Analysis ◽  
Gene Cluster ◽  
Biosynthesis Gene Cluster ◽  
Biosynthesis Gene ◽  
Initial Characterization ◽  
Blasticidin S

scholarly journals Genes for Production of the Enediyne Antitumor Antibiotic C-1027 in Streptomyces globisporus Are Clustered with the cagA Gene That Encodes the C-1027 Apoprotein

2000 ◽  
Vol 44 (2) ◽  
pp. 382-392 ◽  
Author(s):  
Wen Liu ◽  
Ben Shen
Keyword(s):  
Sequence Analysis ◽  
Gene Cluster ◽  
Antibiotic Biosynthesis ◽  
Antitumor Antibiotic ◽  
Caga Gene ◽  
Biosynthesis Gene Cluster ◽  
Starting Point ◽  
Biosynthesis Gene ◽  
Cloned Gene

ABSTRACT C-1027, the most potent member of the enediyne antitumor antibiotic family, is produced by Streptomyces globisporus C-1027 and consists of an apoprotein (encoded by the cagA gene) and a nonpeptidic chromophore. The C-1027 chromophore could be viewed as being derived biosynthetically from a benzoxazolinate, a deoxyamino hexose, a β-amino acid, and an enediyne core. By adopting a strategy for cloning of the C-1027 biosynthesis gene cluster by mapping a putative dNDP-glucose 4,6-dehydratase (NGDH) gene to cagA, we have localized 75 kb of contiguous DNA from S. globisporus. DNA sequence analysis of two regions of the cloned gene cluster revealed two genes, sgcA and sgcB, that encode an NGDH enzyme and a transmembrane efflux protein, respectively, and confirmed that the cagA gene resides approximately 14 kb upstream of the sgcAB locus. The involvement of the cloned gene cluster in C-1027 biosynthesis was demonstrated by disrupting the sgcA gene to generate C-1027-nonproducing mutants and by complementing the sgcAmutants in vivo to restore C-1027 production. These results represent the first cloning of a gene cluster for enediyne antitumor antibiotic biosynthesis and provide a starting point for future genetic and biochemical investigations of C-1027 biosynthesis.

The Serratia gene cluster encoding biosynthesis of the red antibiotic, prodigiosin, shows species- and strain-dependent genome context variation

Microbiology ◽  
2004 ◽  
Vol 150 (11) ◽  
pp. 3547-3560 ◽  
Author(s):  
Abigail K. P. Harris ◽  
Neil R. Williamson ◽  
Holly Slater ◽  
Anthony Cox ◽  
Sophia Abbasi ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Serratia Marcescens ◽  
Gene Cluster ◽  
Streptomyces Coelicolor ◽  
Polyketide Synthases ◽  
Gene Products ◽  
Biosynthesis Gene Cluster ◽  
Biosynthesis Gene ◽  
Genome Context ◽  
Strain Dependent

The prodigiosin biosynthesis gene cluster (pig cluster) from two strains of Serratia (S. marcescens ATCC 274 and Serratia sp. ATCC 39006) has been cloned, sequenced and expressed in heterologous hosts. Sequence analysis of the respective pig clusters revealed 14 ORFs in S. marcescens ATCC 274 and 15 ORFs in Serratia sp. ATCC 39006. In each Serratia species, predicted gene products showed similarity to polyketide synthases (PKSs), non-ribosomal peptide synthases (NRPSs) and the Red proteins of Streptomyces coelicolor A3(2). Comparisons between the two Serratia pig clusters and the red cluster from Str. coelicolor A3(2) revealed some important differences. A modified scheme for the biosynthesis of prodigiosin, based on the pathway recently suggested for the synthesis of undecylprodigiosin, is proposed. The distribution of the pig cluster within several Serratia sp. isolates is demonstrated and the presence of cryptic clusters in some strains shown. The pig cluster of Serratia marcescens ATCC 274 is flanked by cueR and copA homologues and this configuration is demonstrated in several S. marcescens strains, whilst these genes are contiguous in strains lacking the pig cluster.

scholarly journals Characterization of the Biosynthesis Gene Cluster for the Pyrrole Polyether Antibiotic Calcimycin (A23187) inStreptomyces chartreusisNRRL 3882

2010 ◽  
Vol 55 (3) ◽  
pp. 974-982 ◽  
Author(s):  
Qiulin Wu ◽  
Jingdan Liang ◽  
Shuangjun Lin ◽  
Xiufen Zhou ◽  
Linquan Bai ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Mutational Analysis ◽  
Divalent Cations ◽  
Biological Effects ◽  
Biosynthetic Gene Cluster ◽  
Type I ◽  
Biosynthesis Gene Cluster ◽  
Biosynthesis Gene ◽  
Hydroxyanthranilic Acid

ABSTRACTThe pyrrole polyether antibiotic calcimycin (A23187) is a rare ionophore that is specific for divalent cations. It is widely used as a biochemical and pharmacological tool because of its multiple, unique biological effects. Here we report on the cloning, sequencing, and mutational analysis of the 64-kb biosynthetic gene cluster fromStreptomyces chartreusisNRRL 3882. Gene replacements confirmed the identity of the gene cluster, andin silicoanalysis of the DNA sequence revealed 27 potential genes, including 3 genes for the biosynthesis of the α-ketopyrrole moiety, 5 genes that encode modular type I polyketide synthases for the biosynthesis of the spiroketal ring, 4 genes for the biosynthesis of 3-hydroxyanthranilic acid, anN-methyltransferase tailoring gene, a resistance gene, a type II thioesterase gene, 3 regulatory genes, 4 genes with other functions, and 5 genes of unknown function. We propose a pathway for the biosynthesis of calcimycin and assign the genes to the biosynthesis steps. Our findings set the stage for producing much desired calcimycin derivatives using genetic modification instead of chemical synthesis.

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