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Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin

Tobramycin is a broad-spectrum aminoglycoside antibiotic agent. The compound is obtained from the base-catalyzed hydrolysis of carbamoyltobramycin (CTB), which is naturally produced by the actinomycete Streptoalloteichus tenebrarius. However, the strain uses the same precursors to synthesize several...

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Autores principales: Mitousis, Lena, Maier, Hannes, Martinovic, Luka, Kulik, Andreas, Stockert, Sigrid, Wohlleben, Wolfgang, Stiefel, Alfred, Musiol-Kroll, Ewa M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8304502/
https://www.ncbi.nlm.nih.gov/pubmed/34299618
http://dx.doi.org/10.3390/molecules26144343
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author Mitousis, Lena
Maier, Hannes
Martinovic, Luka
Kulik, Andreas
Stockert, Sigrid
Wohlleben, Wolfgang
Stiefel, Alfred
Musiol-Kroll, Ewa M.
author_facet Mitousis, Lena
Maier, Hannes
Martinovic, Luka
Kulik, Andreas
Stockert, Sigrid
Wohlleben, Wolfgang
Stiefel, Alfred
Musiol-Kroll, Ewa M.
author_sort Mitousis, Lena
collection PubMed
description Tobramycin is a broad-spectrum aminoglycoside antibiotic agent. The compound is obtained from the base-catalyzed hydrolysis of carbamoyltobramycin (CTB), which is naturally produced by the actinomycete Streptoalloteichus tenebrarius. However, the strain uses the same precursors to synthesize several structurally related aminoglycosides. Consequently, the production yields of tobramycin are low, and the compound’s purification is very challenging, costly, and time-consuming. In this study, the production of the main undesired product, apramycin, in the industrial isolate Streptoalloteichus tenebrarius 2444 was decreased by applying the fermentation media M10 and M11, which contained high concentrations of starch and dextrin. Furthermore, the strain was genetically engineered by the inactivation of the aprK gene (∆aprK), resulting in the abolishment of apramycin biosynthesis. In the next step of strain development, an additional copy of the tobramycin biosynthetic gene cluster (BGC) was introduced into the ∆aprK mutant. Fermentation by the engineered strain (∆aprK_1-17L) in M11 medium resulted in a 3- to 4-fold higher production than fermentation by the precursor strain (∆aprK). The phenotypic stability of the mutant without selection pressure was validated. The use of the engineered S. tenebrarius 2444 facilitates a step-saving, efficient, and, thus, more sustainable production of the valuable compound tobramycin on an industrial scale.
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spelling pubmed-83045022021-07-25 Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin Mitousis, Lena Maier, Hannes Martinovic, Luka Kulik, Andreas Stockert, Sigrid Wohlleben, Wolfgang Stiefel, Alfred Musiol-Kroll, Ewa M. Molecules Article Tobramycin is a broad-spectrum aminoglycoside antibiotic agent. The compound is obtained from the base-catalyzed hydrolysis of carbamoyltobramycin (CTB), which is naturally produced by the actinomycete Streptoalloteichus tenebrarius. However, the strain uses the same precursors to synthesize several structurally related aminoglycosides. Consequently, the production yields of tobramycin are low, and the compound’s purification is very challenging, costly, and time-consuming. In this study, the production of the main undesired product, apramycin, in the industrial isolate Streptoalloteichus tenebrarius 2444 was decreased by applying the fermentation media M10 and M11, which contained high concentrations of starch and dextrin. Furthermore, the strain was genetically engineered by the inactivation of the aprK gene (∆aprK), resulting in the abolishment of apramycin biosynthesis. In the next step of strain development, an additional copy of the tobramycin biosynthetic gene cluster (BGC) was introduced into the ∆aprK mutant. Fermentation by the engineered strain (∆aprK_1-17L) in M11 medium resulted in a 3- to 4-fold higher production than fermentation by the precursor strain (∆aprK). The phenotypic stability of the mutant without selection pressure was validated. The use of the engineered S. tenebrarius 2444 facilitates a step-saving, efficient, and, thus, more sustainable production of the valuable compound tobramycin on an industrial scale. MDPI 2021-07-18 /pmc/articles/PMC8304502/ /pubmed/34299618 http://dx.doi.org/10.3390/molecules26144343 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Mitousis, Lena
Maier, Hannes
Martinovic, Luka
Kulik, Andreas
Stockert, Sigrid
Wohlleben, Wolfgang
Stiefel, Alfred
Musiol-Kroll, Ewa M.
Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin
title Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin
title_full Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin
title_fullStr Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin
title_full_unstemmed Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin
title_short Engineering of Streptoalloteichus tenebrarius 2444 for Sustainable Production of Tobramycin
title_sort engineering of streptoalloteichus tenebrarius 2444 for sustainable production of tobramycin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8304502/
https://www.ncbi.nlm.nih.gov/pubmed/34299618
http://dx.doi.org/10.3390/molecules26144343
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