scholarly journals Identification of a Novel Biosynthetic Gene Cluster in Aspergillus niger Using Comparative Genomics

2021 ◽  
Vol 7 (5) ◽  
pp. 374
Author(s):  
Gregory Evdokias ◽  
Cameron Semper ◽  
Montserrat Mora-Ochomogo ◽  
Marcos Di Falco ◽  
Thi Truc Minh Nguyen ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Aspergillus Niger ◽  
Gene Cluster ◽  
Protein A ◽  
Biosynthetic Gene Cluster ◽  
Binding Domain ◽  
Spectrometric Analysis ◽  
Nonribosomal Peptides ◽  
Biosynthetic Gene ◽  
Uncharacterized Protein

Previously, DNA microarrays analysis showed that, in co-culture with Bacillus subtilis, a biosynthetic gene cluster anchored with a nonribosomal peptides synthetase of Aspergillus niger is downregulated. Based on phylogenetic and synteny analyses, we show here that this gene cluster, NRRL3_00036-NRRL3_00042, comprises genes predicted to encode a nonribosomal peptides synthetase, a FAD-binding domain-containing protein, an uncharacterized protein, a transporter, a cytochrome P450 protein, a NAD(P)-binding domain-containing protein and a transcription factor. We overexpressed the in-cluster transcription factor gene NRRL3_00042. The overexpression strain, NRRL3_00042OE, displays reduced growth rate and production of a yellow pigment, which by mass spectrometric analysis corresponds to two compounds with masses of 409.1384 and 425.1331. We deleted the gene encoding the NRRL3_00036 nonribosomal peptides synthetase in the NRRL3_00042OE strain. The resulting strain reverted to the wild-type phenotype. These results suggest that the biosynthetic gene cluster anchored by the NRRL3_00036 nonribosomal peptides synthetase gene is regulated by the in-cluster transcriptional regulator gene NRRL3_00042, and that it is involved in the production of two previously uncharacterized compounds.

scholarly journals Transcription Factor Repurposing Offers Insights into Evolution of Biosynthetic Gene Cluster Regulation

mBio ◽  
2021 ◽  
Author(s):  
Wenjie Wang ◽  
Milton Drott ◽  
Claudio Greco ◽  
Dianiris Luciano-Rosario ◽  
Pinmei Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Secondary Metabolites ◽  
Gene Cluster ◽  
Gene Clusters ◽  
Biosynthetic Gene Cluster ◽  
The Other ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters ◽  
Metabolite Production ◽  
Fungal Secondary Metabolites

Fungal secondary metabolites (SMs) are an important source of pharmaceuticals on one hand and toxins on the other. Efforts to identify the biosynthetic gene clusters (BGCs) that synthesize SMs have yielded significant insights into how variation in the genes that compose BGCs may impact subsequent metabolite production within and between species.

scholarly journals Identification of a Biosynthetic Gene Cluster and the Six Associated Lipopeptides Involved in Swarming Motility of Pseudomonas syringae pv. tomato DC3000

2007 ◽  
Vol 189 (17) ◽  
pp. 6312-6323 ◽  
Author(s):  
Andrew D. Berti ◽  
Nathan J. Greve ◽  
Quin H. Christensen ◽  
Michael G. Thomas
Keyword(s):  
Secondary Metabolites ◽  
Gene Cluster ◽  
Pseudomonas Syringae ◽  
Biosynthetic Gene Cluster ◽  
Nonribosomal Peptides ◽  
Biosynthetic Gene ◽  
Tomato Dc3000 ◽  
Metabolic Potential ◽  
Swarming Motility ◽  
Nonribosomal Peptide

ABSTRACT Pseudomonas species are known to be prolific producers of secondary metabolites that are synthesized wholly or in part by nonribosomal peptide synthetases. In an effort to identify additional nonribosomal peptides produced by these bacteria, a bioinformatics approach was used to “mine” the genome of Pseudomonas syringae pv. tomato DC3000 for the metabolic potential to biosynthesize previously unknown nonribosomal peptides. Herein we describe the identification of a nonribosomal peptide biosynthetic gene cluster that codes for proteins involved in the production of six structurally related linear lipopeptides. Structures for each of these lipopeptides were proposed based on amino acid analysis and mass spectrometry analyses. Mutations in this cluster resulted in the loss of swarming motility of P. syringae pv. tomato DC3000 on medium containing a low percentage of agar. This phenotype is consistent with the loss of the ability to produce a lipopeptide that functions as a biosurfactant. This work gives additional evidence that mining the genomes of microorganisms followed by metabolite and phenotypic analyses leads to the identification of previously unknown secondary metabolites.

Frequency of three mutations in the fumonisin biosynthetic gene cluster of Fusarium fujikuroi that are predicted to block fumonisin production

2021 ◽  
Vol 14 (1) ◽  
pp. 49-59
Author(s):  
S. Sultana ◽  
W.X. Bao ◽  
M. Shimizu ◽  
K. Kageyama ◽  
H. Suga
Keyword(s):  
Transcription Factor ◽  
Secondary Metabolites ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Pcr Analysis ◽  
Fusarium Fujikuroi ◽  
Biosynthetic Gene ◽  
Transcription Factor Gene ◽  
Factor Gene ◽  
Group Strain

Fusarium fujikuroi is the most prominent pathogen found in rice. In addition to gibberellin, F. fujikuroi produces various secondary metabolites, including the polyketide mycotoxins, fumonisins. Fumonisin production is conferred by the fumonisin biosynthetic gene (FUM) cluster consisting of 15-17 genes. F. fujikuroi is phylogenetically subclassified into one group with fumonisin production (F-group) and another group in which fumonisin production is undetectable (G-group). In a previous study, a G-to-T substitution (FUM21_G2551T) in the FUM cluster transcription factor gene, FUM21, was identified as a cause of fumonisin-non-production in a G-group strain. In the current study, further analysis of G-group strains identified two additional mutations that involved FUM-cluster genes essential for fumonisin production: (1) a 22.4-kbp deletion in the FUM10-FUM19 region; and (2) a 1.4-kbp insertion in FUM6. PCR analysis of 44 G-group strains, indicated that 84% had the FUM21_G2551T mutation, 50% had the 22.4-kbp FUM10-FUM19 deletion, and 32% had the 1.4-kbp insertion in FUM6, and some strains had two or all the mutations. None of the mutations were detected in the 51 F-group strains examined. Each of the three mutations alone could account for the lack of fumonisin production in G-group strains. However, one G-group strain did not have any of the mutations. Therefore, another mutation(s) is likely responsible for the lack of fumonisin production in some G-group strains of F. fujikuroi.

Anacyclamide D8P, a prenylated cyanobactin from a Sphaerospermopsis sp. cyanobacterium

2017 ◽  
Author(s):  
Joana Martins ◽  
Niina Leikoski ◽  
Matti Wahlsten ◽  
Joana Azevedo ◽  
Jorge Antunes ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Cyclic Peptides ◽  
Biosynthetic Gene Cluster ◽  
The Novel ◽  
Tyrosine Residue ◽  
Biosynthetic Gene ◽  
Post Translational Modification ◽  
Biological Assays ◽  
Mass Spectrometric Data ◽  
Spectrometric Data

Cyanobactins are a family of linear and cyclic peptides produced through the post-translational modification of short precursor peptides. Anacyclamides are macrocyclic cyanobactins with a highly diverse sequence that are common in the genus <i>Anabaena</i>. A mass spectrometry-based screening of potential cyanobactin producers led to the discovery of a new prenylated member of this family of compounds, anacyclamide D8P (<b>1</b>), from <i>Sphaerospermopsis</i> sp. LEGE 00249. The anacyclamide biosynthetic gene cluster (<i>acy</i>) encoding the novel macrocyclic prenylated cyanobactin, was sequenced. Heterologous expression of the acy gene cluster in <i>Escherichia</i> <i>coli</i> established the connection between genomic and mass spectrometric data. Unambiguous establishment of the type and site of prenylation required the full structural elucidation of <b>1</b> using Nuclear Magnetic Resonance (NMR), which demonstrated that a forward prenylation occurred on the tyrosine residue. Compound <b>1</b> was tested in pharmacologically or ecologically relevant biological assays and revealed moderate antimicrobial activity towards the fouling bacterium <i>Halomonas aquamarina</i> CECT 5000.<br>

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