Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage

Filamentous marine cyanobacteria are extraordinarily rich sources of structurally novel, biomedically relevant natural products. To understand their biosynthetic origins as well as produce increased supplies and analog molecules, access to the clustered biosynthetic genes that encode for the assembl...

Full description

Bibliographic Details
Main Authors:	Grindberg, Rashel V., Ishoey, Thomas, Brinza, Dumitru, Esquenazi, Eduardo, Coates, R. Cameron, Liu, Wei-ting, Gerwick, Lena, Dorrestein, Pieter C., Pevzner, Pavel, Lasken, Roger, Gerwick, William H.
Format:	Online
Language:	English
Published:	Public Library of Science 2011
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3075265/

id	pubmed-3075265
recordtype	oai_dc
spelling	pubmed-30752652011-04-29 Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage Grindberg, Rashel V. Ishoey, Thomas Brinza, Dumitru Esquenazi, Eduardo Coates, R. Cameron Liu, Wei-ting Gerwick, Lena Dorrestein, Pieter C. Pevzner, Pavel Lasken, Roger Gerwick, William H. Research Article Filamentous marine cyanobacteria are extraordinarily rich sources of structurally novel, biomedically relevant natural products. To understand their biosynthetic origins as well as produce increased supplies and analog molecules, access to the clustered biosynthetic genes that encode for the assembly enzymes is necessary. Complicating these efforts is the universal presence of heterotrophic bacteria in the cell wall and sheath material of cyanobacteria obtained from the environment and those grown in uni-cyanobacterial culture. Moreover, the high similarity in genetic elements across disparate secondary metabolite biosynthetic pathways renders imprecise current gene cluster targeting strategies and contributes sequence complexity resulting in partial genome coverage. Thus, it was necessary to use a dual-method approach of single-cell genomic sequencing based on multiple displacement amplification (MDA) and metagenomic library screening. Here, we report the identification of the putative apratoxin. A biosynthetic gene cluster, a potent cancer cell cytotoxin with promise for medicinal applications. The roughly 58 kb biosynthetic gene cluster is composed of 12 open reading frames and has a type I modular mixed polyketide synthase/nonribosomal peptide synthetase (PKS/NRPS) organization and features loading and off-loading domain architecture never previously described. Moreover, this work represents the first successful isolation of a complete biosynthetic gene cluster from Lyngbya bouillonii, a tropical marine cyanobacterium renowned for its production of diverse bioactive secondary metabolites. Public Library of Science 2011-04-12 /pmc/articles/PMC3075265/ /pubmed/21533272 http://dx.doi.org/10.1371/journal.pone.0018565 Text en Grindberg et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Grindberg, Rashel V. Ishoey, Thomas Brinza, Dumitru Esquenazi, Eduardo Coates, R. Cameron Liu, Wei-ting Gerwick, Lena Dorrestein, Pieter C. Pevzner, Pavel Lasken, Roger Gerwick, William H.
spellingShingle	Grindberg, Rashel V. Ishoey, Thomas Brinza, Dumitru Esquenazi, Eduardo Coates, R. Cameron Liu, Wei-ting Gerwick, Lena Dorrestein, Pieter C. Pevzner, Pavel Lasken, Roger Gerwick, William H. Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage
author_facet	Grindberg, Rashel V. Ishoey, Thomas Brinza, Dumitru Esquenazi, Eduardo Coates, R. Cameron Liu, Wei-ting Gerwick, Lena Dorrestein, Pieter C. Pevzner, Pavel Lasken, Roger Gerwick, William H.
author_sort	Grindberg, Rashel V.
title	Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage
title_short	Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage
title_full	Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage
title_fullStr	Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage
title_full_unstemmed	Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage
title_sort	single cell genome amplification accelerates identification of the apratoxin biosynthetic pathway from a complex microbial assemblage
description	Filamentous marine cyanobacteria are extraordinarily rich sources of structurally novel, biomedically relevant natural products. To understand their biosynthetic origins as well as produce increased supplies and analog molecules, access to the clustered biosynthetic genes that encode for the assembly enzymes is necessary. Complicating these efforts is the universal presence of heterotrophic bacteria in the cell wall and sheath material of cyanobacteria obtained from the environment and those grown in uni-cyanobacterial culture. Moreover, the high similarity in genetic elements across disparate secondary metabolite biosynthetic pathways renders imprecise current gene cluster targeting strategies and contributes sequence complexity resulting in partial genome coverage. Thus, it was necessary to use a dual-method approach of single-cell genomic sequencing based on multiple displacement amplification (MDA) and metagenomic library screening. Here, we report the identification of the putative apratoxin. A biosynthetic gene cluster, a potent cancer cell cytotoxin with promise for medicinal applications. The roughly 58 kb biosynthetic gene cluster is composed of 12 open reading frames and has a type I modular mixed polyketide synthase/nonribosomal peptide synthetase (PKS/NRPS) organization and features loading and off-loading domain architecture never previously described. Moreover, this work represents the first successful isolation of a complete biosynthetic gene cluster from Lyngbya bouillonii, a tropical marine cyanobacterium renowned for its production of diverse bioactive secondary metabolites.
publisher	Public Library of Science
publishDate	2011
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3075265/
_version_	1611449178392625152

Single Cell Genome Amplification Accelerates Identification of the Apratoxin Biosynthetic Pathway from a Complex Microbial Assemblage

Similar Items