A High-Yield Streptomyces Transcription-Translation Toolkit for Synthetic Biology and Natural Product Applications

10.3791/63012 ◽  
2021 ◽  
Author(s):  
Ming Toh ◽  
Kameshwari Chengan ◽  
Tanith Hanson ◽  
Paul S. Freemont ◽  
Simon J. Moore
Keyword(s):  
Synthetic Biology ◽  
Natural Product ◽  
High Yield

Discovery of Arabidopsis UGT73C1 as a steviol-catalyzing UDP-glycosyltransferase with chemical probes

2018 ◽  
Vol 54 (52) ◽  
pp. 7179-7182 ◽  
Author(s):  
Yiqing Zhou ◽  
Weichao Li ◽  
Wenjing You ◽  
Zhengao Di ◽  
Mingli Wang ◽  
...  
Keyword(s):  
Synthetic Biology ◽  
Natural Product ◽  
Chemical Probes

A strategy for rapidly mining biological parts from plants for synthetic biology utilizing natural product-derived chemical probes has been reported.

scholarly journals mSphere of Influence: Synthetic Biology of Natural Product Biosynthesis

mSphere ◽  
2020 ◽  
Vol 5 (1) ◽  
Author(s):  
Mark C. Walker
Keyword(s):  
Natural Products ◽  
Synthetic Biology ◽  
Natural Product ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene ◽  
Natural Product Biosynthesis ◽  
Antibiotic Compounds ◽  
Approaches To Study

ABSTRACT Mark Walker studies the biosynthesis and engineering of bacterial natural products with the long-term goal of identifying new antibiotic compounds. In this mSphere of Influence, he reflects on how “Direct cloning and refactoring of a silent lipopeptide biosynthetic gene cluster yields the antibiotic taromycin A” by K. Yamanaka, K. A. Reynolds, R. D. Kersten, K. S. Ryan, et al. (Proc Natl Acad Sci USA 111:1957–1962, 2014, https://doi.org/10.1073/pnas.1319584111) impacted his thinking on using synthetic biology approaches to study natural product biosynthesis.

scholarly journals Natural product biosyntheses in cyanobacteria: A treasure trove of unique enzymes

2011 ◽  
Vol 7 ◽  
pp. 1622-1635 ◽  
Author(s):  
Jan-Christoph Kehr ◽  
Douglas Gatte Picchi ◽  
Elke Dittmann
Keyword(s):  
Natural Products ◽  
Synthetic Biology ◽  
Natural Product ◽  
Bioactive Compounds ◽  
Polyketide Synthase ◽  
Biosynthetic Pathways ◽  
Peptide Synthetase ◽  
Biochemical Studies ◽  
Terrestrial Habitats ◽  
The Individual

Cyanobacteria are prolific producers of natural products. Investigations into the biochemistry responsible for the formation of these compounds have revealed fascinating mechanisms that are not, or only rarely, found in other microorganisms. In this article, we survey the biosynthetic pathways of cyanobacteria isolated from freshwater, marine and terrestrial habitats. We especially emphasize modular nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) pathways and highlight the unique enzyme mechanisms that were elucidated or can be anticipated for the individual products. We further include ribosomal natural products and UV-absorbing pigments from cyanobacteria. Mechanistic insights obtained from the biochemical studies of cyanobacterial pathways can inspire the development of concepts for the design of bioactive compounds by synthetic-biology approaches in the future.

scholarly journals Reinvigorating natural product combinatorial biosynthesis with synthetic biology

2015 ◽  
Vol 11 (9) ◽  
pp. 649-659 ◽  
Author(s):  
Eunji Kim ◽  
Bradley S Moore ◽  
Yeo Joon Yoon
Keyword(s):  
Synthetic Biology ◽  
Natural Product ◽  
Combinatorial Biosynthesis

scholarly journals Making drugs out of sunlight and ‘thin air’: an emerging synergy of synthetic biology and natural product chemistry

2020 ◽  
Vol 42 (4) ◽  
pp. 34-39
Author(s):  
Michael J. Stephenson ◽  
Anne Osbourn
Keyword(s):  
Natural Products ◽  
Synthetic Biology ◽  
Natural Product ◽  
Large Scale ◽  
Production Systems ◽  
Biological Activities ◽  
Structural Complexity ◽  
Scale Production ◽  
Natural Product Chemistry ◽  
Product Chemistry

Nature has long served as a rich source of structurally diverse small organic molecules with medicinally relevant biological activities. Despite the historical success of these so-called natural products, the enthusiasm of big pharma to explore these compounds as leads in drug design has waxed and waned. A major contributor to this is their often inherent structural complexity. Such compounds are difficult (often impossible) to access synthetically, a hurdle that can stifle lead development and hinder sustainable large-scale production of promising leads for clinical evaluation. However, in recent years, an emerging synergy between synthetic biology and natural product chemistry offers the potential for a renaissance in our ability to access natural products for drug discovery and development. Advances in genome sequencing, bioinformatics and the maturing of heterologous expression platforms are increasing, enabling the study, and ultimately, the manipulation of plant biosynthetic pathways. The triterpenes are one of the most structurally diverse families of natural products and arguably one of the most underrepresented in the clinic. The plant kingdom is the richest source of triterpene diversity, with >20,000 triterpenes reported so far. Transient expression of genes for candidate enzymes and pathways in amenable plant species is emerging as a powerful and rapid means of investigating and harnessing the plant enzymes involved in generating this diversity. Such platforms also have the potential to serve as production systems in their own right, with the possibility of upscaling these discoveries into commercially useful products using the same overall basic procedure. Ultimately, the carbon source for generation of high-value compounds in plants is photosynthesis. Therefore, we could, with the help of plants, be producing new medicines out of sunlight and ‘thin air’ in green factories in the not too distant future.

scholarly journals A Streptomyces venezuelae Cell-Free Toolkit for Synthetic Biology

2020 ◽  
Author(s):  
Simon J Moore ◽  
Hung-En Lai ◽  
Soo-Mei Chee ◽  
Ming Toh ◽  
Seth Coode ◽  
...  
Keyword(s):  
Synthetic Biology ◽  
Gene Clusters ◽  
Test Tube ◽  
High Yield ◽  
Proof Of Concept ◽  
Biosynthetic Gene Clusters ◽  
Reaction Conditions ◽  
One Pot ◽  
Free System ◽  
High Level

AbstractProkaryotic cell-free coupled transcription-translation (TX-TL) systems are emerging as a powerful tool to examine natural product biosynthetic pathways in a test-tube. The key advantages of this approach are the reduced experimental timescales and controlled reaction conditions. In order to realise this potential, specialised cell-free systems in organisms enriched for biosynthetic gene clusters, with strong protein production and well-characterised synthetic biology tools, is essential. The Streptomyces genus is a major source of natural products. To study enzymes and pathways from Streptomyces, we originally developed a homologous Streptomyces cell-free system to provide a native protein folding environment, a high G+C (%) tRNA pool and an active background metabolism. However, our initial yields were low (36 μg/mL) and showed a high level of batch-to-batch variation. Here, we present an updated high-yield and robust Streptomyces TX-TL protocol, reaching up to yields of 266 μg/mL of expressed recombinant protein. To complement this, we rapidly characterise a range of DNA parts with different reporters, express high G+C (%) biosynthetic genes and demonstrate an initial proof of concept for combined transcription, translation and biosynthesis of Streptomyces metabolic pathways in a single ‘one-pot’ reaction.

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