Heterologous expression of an orphan NRPS gene cluster from Paenibacillus larvae in Escherichia coli revealed production of sevadicin

ABSTRACT In vivo reconstitution of the TDP-l-megosamine pathway from the megalomicin gene cluster of Micromonospora megalomicea was accomplished by the heterologous expression of its biosynthetic genes in Escherichia coli. Mass spectrometric analysis of the TDP-sugar intermediates produced from operons containing different sets of genes showed that the production of TDP-l-megosamine from TDP-4-keto-6-deoxy-d-glucose requires only five biosynthetic steps, catalyzed by MegBVI, MegDII, MegDIII, MegDIV, and MegDV. Bioconversion studies demonstrated that the sugar transferase MegDI, along with the helper protein MegDVI, catalyzes the transfer of l-megosamine to either erythromycin C or erythromycin D, suggesting two possible routes for the production of megalomicin A. Analysis in vivo of the hydroxylation step by MegK indicated that erythromycin C is the intermediate of megalomicin A biosynthesis.

Download Full-text

Heterologous Production of Cyanobacterial Mycosporine-Like Amino Acids Mycosporine-Ornithine and Mycosporine-Lysine in Escherichia coli

Applied and Environmental Microbiology ◽

10.1128/aem.01632-16 ◽

2016 ◽

Vol 82 (20) ◽

pp. 6167-6173 ◽

Cited By ~ 24

Author(s):

Meenu Katoch ◽

Rabia Mazmouz ◽

Rocky Chau ◽

Leanne A. Pearson ◽

Russell Pickford ◽

...

Keyword(s):

Escherichia Coli ◽

Amino Acids ◽

Heterologous Expression ◽

Gene Cluster ◽

Uv Radiation ◽

Active Ingredient ◽

Stress Factors ◽

Heterologous Production ◽

Content Type ◽

E Coli

ABSTRACTMycosporine-like amino acids (MAAs) are an important class of secondary metabolites known for their protection against UV radiation and other stress factors. Cyanobacteria produce a variety of MAAs, including shinorine, the active ingredient in many sunscreen creams. Bioinformatic analysis of the genome of the soil-dwelling cyanobacteriumCylindrospermum stagnalePCC 7417 revealed a new gene cluster with homology to MAA synthase fromNostoc punctiforme. This newly identified gene cluster is unusual because it has five biosynthesis genes (mylAtomylE), compared to the four found in other MAA gene clusters. Heterologous expression ofmylAtomylEinEscherichia coliresulted in the production of mycosporine-lysine and the novel compound mycosporine-ornithine. To our knowledge, this is the first time these compounds have been heterologously produced inE. coliand structurally characterized via direct spectral guidance. This study offers insight into the diversity, biosynthesis, and structure of cyanobacterial MAAs and highlights their amenability to heterologous production methods.IMPORTANCEMycosporine-like amino acids (MAAs) are significant from an environmental microbiological perspective as they offer microbes protection against a variety of stress factors, including UV radiation. The heterologous expression of MAAs inE. coliis also significant from a biotechnological perspective as MAAs are the active ingredient in next-generation sunscreens.

Download Full-text

Biochemical characterization and substrate specificity of the gene cluster for biosyntheses of K-252a and its analogs by in vitro heterologous expression system of Escherichia coli

Molecular BioSystems ◽

10.1039/b912395b ◽

2009 ◽

Vol 5 (10) ◽

pp. 1192 ◽

Cited By ~ 10

Author(s):

Hsien-Tai Chiu ◽

Yu-Chin Lin ◽

Meng-Na Lee ◽

Yi-Lin Chen ◽

Mei-Sin Wang ◽

...

Keyword(s):

Escherichia Coli ◽

Substrate Specificity ◽

Heterologous Expression ◽

Gene Cluster ◽

Biochemical Characterization ◽

Expression System ◽

Heterologous Expression System

Download Full-text

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

Chemical Science ◽

10.1039/c8sc05670f ◽

2019 ◽

Vol 10 (10) ◽

pp. 3042-3048 ◽

Cited By ~ 10

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.

Download Full-text

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

Microorganisms ◽

10.3390/microorganisms9081640 ◽

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.

Download Full-text