Dissecting tunicamycin biosynthesis by genome mining: cloning and heterologous expression of a minimal gene cluster

2010 ◽  
Vol 1 (5) ◽  
pp. 581 ◽  
Author(s):  
Filip J. Wyszynski ◽  
Andrew R. Hesketh ◽  
Mervyn J. Bibb ◽  
Benjamin G. Davis
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Genome Mining

scholarly journals Genome Mining and Heterologous Expression of Lipoamides and Amicoumacins From Compost-Associated Bacillus subtilis fmb60

2021 ◽  
Author(s):  
jie yang ◽  
Qingzheng Zhu ◽  
Feng Xu ◽  
Ming Yang ◽  
Hechao Du ◽  
...  
Keyword(s):  
Natural Products ◽  
Bacillus Subtilis ◽  
Secondary Metabolites ◽  
Heterologous Expression ◽  
Gene Cluster ◽  
Broad Spectrum ◽  
Genome Mining ◽  
Antibacterial Activities ◽  
Antimicrobial Activities ◽  
First Time

Abstract: Bacillus subtilis fmb60, which has broad-spectrum antimicrobial activities was isolated from plant straw compost. A hybrid NRPS/PKS cluster was screened from the genome. Sixteen secondary metabolites produced by the gene cluster were isolated and identified using LC-HRMS and NMR. Three lipoamides D–F (1-3) and two amicoumacin derivatives, amicoumacins D, E (4, 5), were identified, and are reported here for the first time. Lipoamides D–F exhibited strong antibacterial activities against harmful foodborne bacteria, with the MIC ranging from 6.25 to 25 µg/mL. Amicoumacin E scavenged 38.8% of ABTS+ radicals at 1 mg/mL. Direct cloning and heterologous expression of the NRPS/PKS and ace gene cluster identified its importance for the biosynthesis of amicoumacins. This study demonstrated that there is a high potential for biocontrol utilization of B. subtilis fmb60, and genome mining for clusters of secondary metabolites of B. subtilis fmb60 has revealed a greater biosynthetic potential for the production of novel natural products than previously anticipated.

scholarly journals Comparative genome mining and heterologous expression of an orphan NRPS gene cluster direct the production of ashimides

2019 ◽  
Vol 10 (10) ◽  
pp. 3042-3048 ◽  
Author(s):  
Jing Shi ◽  
Ying Jie Zeng ◽  
Bo Zhang ◽  
Fen Li Shao ◽  
Yan Chi Chen ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Genome Mining ◽  
Nrps Gene ◽  
Comparative Genome

Genome mining and heterologous expression of an orphan cluster led to the discovery of ashimides featuring an unusual cyclization mechanism.

scholarly journals Bonsecamin: A New Cyclic Pentapeptide Discovered through Heterologous Expression of a Cryptic Gene Cluster

2021 ◽  
Vol 9 (8) ◽  
pp. 1640
Author(s):  
Constanze Lasch ◽  
Marc Stierhof ◽  
Marta Rodríguez Estévez ◽  
Maksym Myronovskyi ◽  
Josef Zapp ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Gene Deletion ◽  
Genome Mining ◽  
Structural Complexity ◽  
Bioinformatic Analysis ◽  
Nrps Gene ◽  
Minimal Set ◽  
Streptomyces Albus ◽  
Important Basis

The intriguing structural complexity of molecules produced by natural organisms is uncontested. Natural scaffolds serve as an important basis for the development of molecules with broad applications, e.g., therapeutics or agrochemicals. Research in recent decades has demonstrated that by means of classic metabolite extraction from microbes only a small portion of natural products can be accessed. The use of genome mining and heterologous expression approaches represents a promising way to discover new natural compounds. In this paper we report the discovery of a novel cyclic pentapeptide called bonsecamin through the heterologous expression of a cryptic NRPS gene cluster from Streptomyces albus ssp. chlorinus NRRL B-24108 in Streptomyces albus Del14. The new compound was successfully isolated and structurally characterized using NMR. The minimal set of genes required for bonsecamin production was determined through bioinformatic analysis and gene deletion experiments. A biosynthetic route leading to the production of bonsecamin is proposed in this paper.

scholarly journals Identification and Verification of the Prodigiosin Biosynthetic Gene Cluster (BGC) in Pseudoalteromonas rubra S4059

2021 ◽  
Author(s):  
Xiyan Wang ◽  
Thomas Isbrandt ◽  
Emil Ørsted Christensen ◽  
Jette Melchiorsen ◽  
Thomas Ostenfeld Larsen ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Heterologous Expression ◽  
Gene Cluster ◽  
Genome Mining ◽  
Gene Clusters ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters ◽  
Pseudoalteromonas Rubra

Pigmented Pseudoalteromonas strains are renowned for their production of secondary metabolites, and genome mining has revealed a high number of biosynthetic gene clusters (BGCs) for which the chemistry is unknown. Identification of those BGCs is a prerequisite for linking products to gene clusters and for further exploitation through heterologous expression.

scholarly journals Identification, Cloning and Heterologous Expression of the Gene Cluster Directing RES-701-3, -4 Lasso Peptides Biosynthesis from a Marine Streptomyces Strain

Marine Drugs ◽  
2020 ◽  
Vol 18 (5) ◽  
pp. 238 ◽  
Author(s):  
Daniel Oves-Costales ◽  
Marina Sánchez-Hidalgo ◽  
Jesús Martín ◽  
Olga Genilloud
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Fermentation Broth ◽  
Genome Mining ◽  
Biosynthetic Gene Cluster ◽  
Open Reading Frames ◽  
Tryptophan Residue ◽  
Post Translational Modification ◽  
Lasso Peptides ◽  
Genes Encoding

RES-701-3 and RES-701-4 are two class II lasso peptides originally identified in the fermentation broth of Streptomyces sp. RE-896, which have been described as selective endothelin type B receptor antagonists. These two lasso peptides only differ in the identity of the C-terminal residue (tryptophan in RES-701-3, 7-hydroxy-tryptophan in RES-701-4), thus raising an intriguing question about the mechanism behind the modification of the tryptophan residue. In this study, we describe the identification of their biosynthetic gene cluster through the genome mining of the marine actinomycete Streptomyces caniferus CA-271066, its cloning and heterologous expression, and show that the seven open reading frames (ORFs) encoded within the gene cluster are sufficient for the biosynthesis of both lasso peptides. We propose that ResE, a protein lacking known putatively conserved domains, is likely to play a key role in the post-translational modification of the C-terminal tryptophan of RES-701-3 that affords RES-701-4. A BLASTP search with the ResE amino acid sequence shows the presence of homologues of this protein in the genomes of eight other Streptomyces strains, which also harbour the genes encoding the RES-701-3, -4 precursor peptide, split-B proteins and ATP-dependent lactam synthetase required for the biosynthesis of these compounds.

scholarly journals Genome Mining, Heterologous Expression, Antibacterial and Antioxidant Activities of Lipoamides and Amicoumacins from Compost-Associated Bacillus subtilis fmb60

Molecules ◽  
2021 ◽  
Vol 26 (7) ◽  
pp. 1892
Author(s):  
Jie Yang ◽  
Qingzheng Zhu ◽  
Feng Xu ◽  
Ming Yang ◽  
Hechao Du ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Secondary Metabolites ◽  
Heterologous Expression ◽  
Gene Cluster ◽  
Broad Spectrum ◽  
Antioxidant Activities ◽  
Genome Mining ◽  
Antibacterial Activities ◽  
Antimicrobial Activities ◽  
First Time

Bacillus subtilis fmb60, which has broad-spectrum antimicrobial activities, was isolated from plant straw compost. A hybrid NRPS/PKS cluster was screened from the genome. Sixteen secondary metabolites produced by the gene cluster were isolated and identified using LC-HRMS and NMR. Three lipoamides D–F (1–3) and two amicoumacin derivatives, amicoumacins D, E (4, 5), were identified, and are reported here for the first time. Lipoamides D–F exhibited strong antibacterial activities against harmful foodborne bacteria, with the MIC ranging from 6.25 to 25 µg/mL. Amicoumacin E scavenged 38.8% of ABTS+ radicals at 1 mg/mL. Direct cloning and heterologous expression of the NRPS/PKS and ace gene cluster identified its importance for the biosynthesis of amicoumacins. This study demonstrated that there is a high potential for biocontrol utilization of B. subtilis fmb60, and genome mining for clusters of secondary metabolites of B. subtilis fmb60 has revealed a greater biosynthetic potential for the production of novel natural products than previously anticipated.

Direct Cloning, Genetic Engineering, and Heterologous Expression of the Syringolin Biosynthetic Gene Cluster inE. colithrough Red/ET Recombineering

ChemBioChem ◽  
2012 ◽  
Vol 13 (13) ◽  
pp. 1946-1952 ◽  
Author(s):  
Xiaoying Bian ◽  
Fan Huang ◽  
Francis A. Stewart ◽  
Liqiu Xia ◽  
Youming Zhang ◽  
...  
Keyword(s):  
Genetic Engineering ◽  
Heterologous Expression ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene ◽  
Direct Cloning

Discovery of Polycyclic Macrolide Shuangdaolides by Heterologous Expression of a Cryptic trans-AT PKS Gene Cluster

2021 ◽  
Author(s):  
Yang Liu ◽  
Haibo Zhou ◽  
Qiyao Shen ◽  
Guangzhi Dai ◽  
Fu Yan ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster

Identification of the kinanthraquinone biosynthetic gene cluster by expression of an atypical response regulator

2021 ◽  
Vol 85 (3) ◽  
pp. 714-721
Author(s):  
Risa Takao ◽  
Katsuyuki Sakai ◽  
Hiroyuki Koshino ◽  
Hiroyuki Osada ◽  
Shunji Takahashi
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Secondary Metabolite ◽  
Response Regulator ◽  
Bac Library ◽  
Gene Clusters ◽  
Biosynthetic Gene Cluster ◽  
Gene Organization ◽  
Biosynthetic Gene ◽  
Streptomyces Sp

ABSTRACT Recent advances in genome sequencing have revealed a variety of secondary metabolite biosynthetic gene clusters in actinomycetes. Understanding the biosynthetic mechanism controlling secondary metabolite production is important for utilizing these gene clusters. In this study, we focused on the kinanthraquinone biosynthetic gene cluster, which has not been identified yet in Streptomyces sp. SN-593. Based on chemical structure, 5 type II polyketide synthase gene clusters were listed from the genome sequence of Streptomyces sp. SN-593. Among them, a candidate gene cluster was selected by comparing the gene organization with grincamycin, which is synthesized through an intermediate similar to kinanthraquinone. We initially utilized a BAC library for subcloning the kiq gene cluster, performed heterologous expression in Streptomyces lividans TK23, and identified the production of kinanthraquinone and kinanthraquinone B. We also found that heterologous expression of kiqA, which belongs to the DNA-binding response regulator OmpR family, dramatically enhanced the production of kinanthraquinones.

scholarly journals Mining of a streptothricin gene cluster from Streptomyces sp. TP-A0356 genome via heterologous expression

2013 ◽  
Vol 56 (7) ◽  
pp. 619-627 ◽  
Author(s):  
JinE Li ◽  
ZhengYan Guo ◽  
Wei Huang ◽  
XiangXi Meng ◽  
GuoMin Ai ◽  
...  
Keyword(s):  
Heterologous Expression ◽  
Gene Cluster ◽  
Streptomyces Sp
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