A functional screen for recovery of 4'-phosphopantetheinyl transferase and associated natural product biosynthesis genes from metagenome libraries

Environmental Microbiology
Jeremy G OwenDavid F Ackerley

Abstract

The single-module non-ribosomal peptide synthetase BpsA from Streptomyces lavendulae has the unique ability to autonomously synthesize a coloured product (indigoidine) from a single substrate (l-glutamine), conditional upon activation by a 4'-phosphopantetheinyl transferase (PPTase) partner. We show that bpsA can be expressed in an entD PPTase gene deleted mutant of Escherichia coli to yield a sensitive reporter strain for recovery of PPTase genes from metagenome libraries. We also show that recombinant bpsA constructs, generated by substitution of the native peptidyl carrier protein domain followed by directed evolution to restore function, can be used to increase the diversity of PPTase genes recovered from a sample. As PPTases are essential for activation of non-ribosomal peptide synthetase and polyketide synthase enzymes, they are frequently associated with secondary metabolite gene clusters. Nearly half of the PPTases recovered in our screening of two small-insert soil metagenome libraries were genetically linked to recognizable secondary metabolite biosynthetic genes, demonstrating that PPTase-targeting functional screens can be used for efficient recovery of natural product gene clusters from metagenome libraries. The pl...Continue Reading

References

Nov 1, 1996·Chemistry & Biology·R H LambalotC T Walsh
Oct 6, 1999·Genome Research·X Huang, A Madan
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Aug 30, 2002·Applied and Environmental Microbiology·Doreen E GillespieJo Handelsman
Oct 19, 2002·Journal of Biotechnology·Stefano DonadioAnna Maria Puglia
Dec 25, 2002·Chemistry & Biology·Richard H Baltz
Jan 4, 2003·Journal of Bacteriology·Guntram ChristiansenElke Dittmann
Apr 18, 2003·Journal of Bacteriology·David F AckerleyIain L Lamont
Oct 7, 2003·Annual Review of Microbiology·Michael S Rappé, Stephen J Giovannoni
Mar 26, 2004·Nature Reviews. Microbiology·Martin Keller, Karsten Zengler
Oct 21, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jun YinChristopher T Walsh
Dec 13, 2006·Nature Biotechnology·Jon ClardyChristopher T Walsh
Feb 21, 2007·Journal of Natural Products·David J Newman, Gordon M Cragg
Sep 23, 2008·Fungal Genetics and Biology : FG & B·Hans von Döhren
Oct 28, 2008·BMC Bioinformatics·Mohd Zeeshan AnsariDebasisa Mohanty
Oct 30, 2008·Journal of Molecular Microbiology and Biotechnology·Manuel FerrerPeter N Golyshin
Jul 11, 2009·Science·Jesse W-H Li, John C Vederas
Oct 24, 2009·Current Opinion in Biotechnology·Taku Uchiyama, Kentaro Miyazaki
Apr 7, 2011·The Biochemical Journal·Jeremy G OwenDavid F Ackerley
May 7, 2011·Biotechnology for Biofuels·Nádia Skorupa Parachin, Marie F Gorwa-Grauslund

❮ Previous
Next ❯

Citations

Sep 13, 2014·Biotechnology Letters·Mark J Calcott, David F Ackerley
Apr 13, 2013·The FEBS Journal·Teunke van RossumJohn van der Oost
Mar 30, 2016·Current Opinion in Microbiology·Eduardo SanteroFernando Govantes
Sep 23, 2014·Chemistry & Biology·Aleksandr MilshteynSean F Brady
Aug 14, 2015·BMC Microbiology·Mark J Calcott, David F Ackerley
Jun 5, 2013·Current Opinion in Biotechnology·Michael M Schofield, David H Sherman
Jul 8, 2014·Current Opinion in Microbiology·Zachary Charlop-PowersSean F Brady
Feb 11, 2015·Applied Microbiology and Biotechnology·Pengwei LiYihua Chen
Aug 20, 2016·Applied Microbiology and Biotechnology·M V TejesviA M Pirttilä
Mar 10, 2017·Expert Opinion on Drug Discovery·Mingzi M ZhangHuimin Zhao
May 24, 2018·Antibiotics·Joachim J HugRolf Müller
Jul 13, 2014·Applied and Environmental Microbiology·Mark J CalcottDavid F Ackerley
Oct 12, 2012·Natural Product Reports·Christopher T Walsh, Timothy A Wencewicz
Oct 15, 2013·Genomics & Informatics·Myung Hwan Lee, Seon-Woo Lee
Mar 30, 2017·Applied Microbiology and Biotechnology·Zhoujie XieYihua Chen
Dec 12, 2018·Nature Communications·Lorillee TallorinNathan C Gianneschi
Dec 10, 2019·Chemical Society Reviews·Ulrich MarkelUlrich Schwaneberg
Dec 10, 2019·Bioorganic & Medicinal Chemistry Letters·Christopher R VickeryMichael D Burkart

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.