Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in Fermentation

<i>Pseudomonas putida</i> PCL1760 is a well-studied bacterium that can be used for a wide range of biotechnological applications. In our study we investigated the effect of deletion of the genes involved in alginate accumulation (<i>alg</i>A), flagellar export (<i>flh&l...

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Main Authors: Mikhail Frolov, Galim Alimzhanovich Kungurov, Emil Elmirovich Valiakhmetov, Artur Sergeyevich Gogov, Natalia Viktorovna Trachtmann, Shamil Zavdatovich Validov
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Fermentation
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Online Access:https://www.mdpi.com/2311-5637/10/12/606
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author Mikhail Frolov
Galim Alimzhanovich Kungurov
Emil Elmirovich Valiakhmetov
Artur Sergeyevich Gogov
Natalia Viktorovna Trachtmann
Shamil Zavdatovich Validov
author_facet Mikhail Frolov
Galim Alimzhanovich Kungurov
Emil Elmirovich Valiakhmetov
Artur Sergeyevich Gogov
Natalia Viktorovna Trachtmann
Shamil Zavdatovich Validov
author_sort Mikhail Frolov
collection DOAJ
description <i>Pseudomonas putida</i> PCL1760 is a well-studied bacterium that can be used for a wide range of biotechnological applications. In our study we investigated the effect of deletion of the genes involved in alginate accumulation (<i>alg</i>A), flagellar export (<i>flh</i>A), and pili formation <i>pil</i>Q on the behavior of the strain in bioreactors. We obtained the knockout mutant P. putida LN6160 with the deletion of these genes and showed that the absence of these genes reduces mobility and biofilm formation (40% lower after 72 h) in the mutant. At the same time, we noted the positive effect of these deletions on the growth of the mutant strain on rich medium (1.39 × 10<sup>10</sup> CFU/mL in the mutant and 6.4 × 10<sup>9</sup> CFU/mL in the wild type) and on mineral medium (6.11 × 10<sup>9</sup> CFU/mL in the mutant and 1.36 × 10<sup>9</sup> CFU/mL in the wild type) by growing them in small-volume bioreactors. A significant decrease in the biofilm and the foam formation was also observed for LN6160 in a small-volume bioreactor. Most probably, the rapid growth of the deletion strain occurs due to a decrease in the energy load on the bacterial apparatus.
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institution Kabale University
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publishDate 2024-11-01
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series Fermentation
spelling doaj-art-ee90e13f3a2b452cb75a5947d4d1be8d2024-12-27T14:25:24ZengMDPI AGFermentation2311-56372024-11-01101260610.3390/fermentation10120606Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in FermentationMikhail Frolov0Galim Alimzhanovich Kungurov1Emil Elmirovich Valiakhmetov2Artur Sergeyevich Gogov3Natalia Viktorovna Trachtmann4Shamil Zavdatovich Validov5Laboratory of Molecular Genetics and Microbiology Methods, Kazan Scientific Center of Russian Academy of Sciences, Orenburgskiy Tract 20A, 420059 Kazan, RussiaLaboratory of Molecular Genetics and Microbiology Methods, Kazan Scientific Center of Russian Academy of Sciences, Orenburgskiy Tract 20A, 420059 Kazan, RussiaLaboratory of Molecular Genetics and Microbiology Methods, Kazan Scientific Center of Russian Academy of Sciences, Orenburgskiy Tract 20A, 420059 Kazan, RussiaNational Research Center Kurchatov Institute, po. Akademika Kurchatova, v. 1, 123182 Moscow, RussiaLaboratory of Molecular Genetics and Microbiology Methods, Kazan Scientific Center of Russian Academy of Sciences, Orenburgskiy Tract 20A, 420059 Kazan, RussiaLaboratory of Molecular Genetics and Microbiology Methods, Kazan Scientific Center of Russian Academy of Sciences, Orenburgskiy Tract 20A, 420059 Kazan, Russia<i>Pseudomonas putida</i> PCL1760 is a well-studied bacterium that can be used for a wide range of biotechnological applications. In our study we investigated the effect of deletion of the genes involved in alginate accumulation (<i>alg</i>A), flagellar export (<i>flh</i>A), and pili formation <i>pil</i>Q on the behavior of the strain in bioreactors. We obtained the knockout mutant P. putida LN6160 with the deletion of these genes and showed that the absence of these genes reduces mobility and biofilm formation (40% lower after 72 h) in the mutant. At the same time, we noted the positive effect of these deletions on the growth of the mutant strain on rich medium (1.39 × 10<sup>10</sup> CFU/mL in the mutant and 6.4 × 10<sup>9</sup> CFU/mL in the wild type) and on mineral medium (6.11 × 10<sup>9</sup> CFU/mL in the mutant and 1.36 × 10<sup>9</sup> CFU/mL in the wild type) by growing them in small-volume bioreactors. A significant decrease in the biofilm and the foam formation was also observed for LN6160 in a small-volume bioreactor. Most probably, the rapid growth of the deletion strain occurs due to a decrease in the energy load on the bacterial apparatus.https://www.mdpi.com/2311-5637/10/12/606<i>Pseudomonas putida</i>bioreactorfermentationbiofilm formationmotilitypilQ
spellingShingle Mikhail Frolov
Galim Alimzhanovich Kungurov
Emil Elmirovich Valiakhmetov
Artur Sergeyevich Gogov
Natalia Viktorovna Trachtmann
Shamil Zavdatovich Validov
Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in Fermentation
Fermentation
<i>Pseudomonas putida</i>
bioreactor
fermentation
biofilm formation
motility
pilQ
title Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in Fermentation
title_full Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in Fermentation
title_fullStr Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in Fermentation
title_full_unstemmed Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in Fermentation
title_short Construction of the <i>Pseudomonas putida</i> Strain with Low Motility and Reduced Biofilm Formation for Application in Fermentation
title_sort construction of the i pseudomonas putida i strain with low motility and reduced biofilm formation for application in fermentation
topic <i>Pseudomonas putida</i>
bioreactor
fermentation
biofilm formation
motility
pilQ
url https://www.mdpi.com/2311-5637/10/12/606
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AT emilelmirovichvaliakhmetov constructionoftheipseudomonasputidaistrainwithlowmotilityandreducedbiofilmformationforapplicationinfermentation
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