PKS–NRPS Enzymology and Structural Biology: Considerations in Protein Production

Skiba, Meredith A.; Maloney, Finn P.; Dan, Qingyun; Fraley, Amy E.; Aldrich, Courtney C.; Smith, Janet L.; Brown, William Clay

doi:10.1016/bs.mie.2018.01.035

Cited by 15 publications

(13 citation statements)

References 161 publications

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“…Meanwhile, NRPSs are composed of a modular set of repeating enzyme domains for the activation and incorporation of amino acids (Park and Yoon, 2015). The modular NRPSs typically consist of a condensation domain, adenylation domain, and a thiolation domain, while type I PKSs generally contain a ketosynthase domain, acyltransferase domain, and an acyl carrier protein (Komaki et al, 2015; Skiba et al, 2018). Natural product structures can be modified by mixing and matching the megasynthases at the subunit, module, and domain levels.…”

Section: Efforts In Product Improvements and Generation Of New Analogsmentioning

confidence: 99%

A Review of the Microbial Production of Bioactive Natural Products and Biologics

et al. 2019

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A variety of organisms, such as bacteria, fungi, and plants, produce secondary metabolites, also known as natural products. Natural products have been a prolific source and an inspiration for numerous medical agents with widely divergent chemical structures and biological activities, including antimicrobial, immunosuppressive, anticancer, and anti-inflammatory activities, many of which have been developed as treatments and have potential therapeutic applications for human diseases. Aside from natural products, the recent development of recombinant DNA technology has sparked the development of a wide array of biopharmaceutical products, such as recombinant proteins, offering significant advances in treating a broad spectrum of medical illnesses and conditions. Herein, we will introduce the structures and diverse biological activities of natural products and recombinant proteins that have been exploited as valuable molecules in medicine, agriculture and insect control. In addition, we will explore past and ongoing efforts along with achievements in the development of robust and promising microorganisms as cell factories to produce biologically active molecules. Furthermore, we will review multi-disciplinary and comprehensive engineering approaches directed at improving yields of microbial production of natural products and proteins and generating novel molecules. Throughout this article, we will suggest ways in which microbial-derived biologically active molecular entities and their analogs could continue to inspire the development of new therapeutic agents in academia and industry.

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Section: Efforts In Product Improvements and Generation Of New Analogsmentioning

confidence: 99%

A Review of the Microbial Production of Bioactive Natural Products and Biologics

et al. 2019

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“…For example, exchanging the dehydratase domain or linker domain for combinatorial biosynthesis of polyketides (PKs) often generates insoluble aggregates of enzymes (Liew et al, 2012;Cai and Zhang, 2018). In addition, uncoupling transcription and translation may lead to insolubility due to misfolding, which requires optimization of the transcription and translation rate for heterologous expression (Skiba et al, 2018). Codon usage has a significant impact on the translation rate of GC-rich BGC sequences that are likely to be translated inefficiently at low-use codons in E. coli (Ke and Yoshikuni, 2020).…”

Section: Streptomyces As a Heterologous Expression Host Of Secondary Metabolite Bgcsmentioning

confidence: 99%

Streptomyces as Microbial Chassis for Heterologous Protein Expression

Hwang¹,

Lee²,

Kim³

et al. 2021

Front. Bioeng. Biotechnol.

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Heterologous production of recombinant proteins is gaining increasing interest in biotechnology with respect to productivity, scalability, and wide applicability. The members of genus Streptomyces have been proposed as remarkable hosts for heterologous production due to their versatile nature of expressing various secondary metabolite biosynthetic gene clusters and secretory enzymes. However, there are several issues that limit their use, including low yield, difficulty in genetic manipulation, and their complex cellular features. In this review, we summarize rational engineering approaches to optimizing the heterologous production of secondary metabolites and recombinant proteins in Streptomyces species in terms of genetic tool development and chassis construction. Further perspectives on the development of optimal Streptomyces chassis by the design-build-test-learn cycle in systems are suggested, which may increase the availability of secondary metabolites and recombinant proteins.

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“…AprA apo-ACP (residues 1058-1138, GenBank: WP_075900460) was produced and purified as described previously (Skiba et al, 2017). A trace metals mix was included in the growth medium to inhibit addition of the phosphopantetheinyl post translational modification by endogenous E. coli enzymes (Skiba et al, 2018a). CurA ACP (residues 444-519, GenBank: AEE88289.1) was produced and purified identically to AprA ACP.…”

Section: Declaration Of Interestsmentioning

confidence: 99%