Generation of a highly attenuated strain of Pseudomonas aeruginosa for commercial production of alginate

Valentine, Meagan E.; Kirby, Brandon D.; Withers, Thomas R.; Johnson, Shannon L.; Long, Timothy E.; Hao, Youai; Lam, Joseph S.; Niles, Richard M.; Yu, Hongwei D.

doi:10.1111/1751-7915.13411

Title: Generation of a highly attenuated strain of Pseudomonas aeruginosa for commercial production of alginate

Journal Article · 31 December 2019 · Microbial Biotechnology (Online)

DOI: https://doi.org/10.1111/1751-7915.13411 · OSTI ID:1629309

Valentine, Meagan E. ^[1]; Kirby, Brandon D. ^[2]; Withers, Thomas R. ^[2];

^[3]; Long, Timothy E. ^[4]; Hao, Youai ^[5];

^[5]; Niles, Richard M. ^[2];

^[6]

Robert C. Byrd Biotechnology Science Center Progenesis Technologies, LLC, Huntington, WV (United States); DOE/OSTI
Robert C. Byrd Biotechnology Science Center Progenesis Technologies, LLC, Huntington, WV (United States)
Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Biosecurity and Public Health
Marshall Univ., Huntington, WV (United States). Research School of Pharmacy. Dept. of Pharmaceutical Science
Univ. of Guelph, ON (Canada). Dept. of Molecular and Cellular Biology
Robert C. Byrd Biotechnology Science Center Progenesis Technologies, LLC, Huntington, WV (United States); Marshall Univ., Huntington, WV (United States). Joan C. Edwards School of Medicine. Dept. of Pediatrics. Dept. of Biomedical Sciences

Alginate is an important polysaccharide that is commonly used as a gelling agent in foods, cosmetics and healthcare products. Currently, all alginate used commercially is extracted from brown seaweed. However, with environmental changes such as increasing ocean temperature and the increasing number of biotechnological uses of alginates with specific properties, there is an emerging need for more reliable and customizable sources of alginate. An alternative to seaweed for alginate production is Pseudomonas aeruginosa, a common Gram-negative bacterium that can form alginate containing biofilms. However, P. aeruginosa is an opportunistic pathogen that can cause life-threatening infections in immunocompromised patients. Therefore, we sought to engineer a non-pathogenic P. aeruginosa strain that is safe for commercial production of alginate. Using a homologous recombination strategy, we sequentially deleted five key pathogenicity genes from the P. aeruginosa chromosome, resulting in the marker-free strain PGN5. Intraperitoneal injection of mice with PGN5 resulted in 0% mortality, while injection with wild-type P. aeruginosa resulted in 95% mortality, providing evidence that the systemic virulence of PGN5 is highly attenuated. Importantly, PGN5 produces large amounts of alginate in response to overexpression of MucE, an activator of alginate biosynthesis. The alginate produced by PGN5 is structurally identical to alginate produced by wildtype P. aeruginosa, indicating that the alginate biosynthetic pathway remains functional in this modified strain. The genetic versatility of P. aeruginosa will allow us to further engineer PGN5 to produce alginates with specific chemical compositions and physical properties to meet different industrial and biomedical needs.

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Research Organization:: Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

Grant/Contract Number:: AC52-06NA25396

OSTI ID:: 1629309

Journal Information:: Microbial Biotechnology (Online), Journal Name: Microbial Biotechnology (Online) Journal Issue: 1 Vol. 13; ISSN 1751-7915

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Bacterial biopolymers: from pathogenesis to advanced materials Moradali, M. Fata; Rehm, Bernd H. A. Nature Reviews Microbiology, Vol. 18, Issue 4 https://doi.org/10.1038/s41579-019-0313-3	journal	January 2020
Training bacteria to produce environmentally friendly polymers of industrial and medical relevance Huertas, María José; Matilla, Miguel A. Microbial Biotechnology, Vol. 13, Issue 1 https://doi.org/10.1111/1751-7915.13470	journal	January 2020

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