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Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose

Sucrose is an important disaccharide used as a substrate in many industrial applications. It is a major component of molasses, a cheap by‐product of the sugar industry. Unfortunately, not all industrially relevant organisms, among them Pseudomonas putida, are capable of metabolizing sucrose. We chos...

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Autores principales: Löwe, Hannes, Schmauder, Lukas, Hobmeier, Karina, Kremling, Andreas, Pflüger‐Grau, Katharina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5552902/
https://www.ncbi.nlm.nih.gov/pubmed/28349670
http://dx.doi.org/10.1002/mbo3.473
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author Löwe, Hannes
Schmauder, Lukas
Hobmeier, Karina
Kremling, Andreas
Pflüger‐Grau, Katharina
author_facet Löwe, Hannes
Schmauder, Lukas
Hobmeier, Karina
Kremling, Andreas
Pflüger‐Grau, Katharina
author_sort Löwe, Hannes
collection PubMed
description Sucrose is an important disaccharide used as a substrate in many industrial applications. It is a major component of molasses, a cheap by‐product of the sugar industry. Unfortunately, not all industrially relevant organisms, among them Pseudomonas putida, are capable of metabolizing sucrose. We chose a metabolic engineering approach to circumvent this blockage and equip P. putida with the activities necessary to consume sucrose. Therefore, we constructed a pair of broad‐host range mini‐transposons (pSST – sucrose splitting transposon), carrying either cscA, encoding an invertase able to split sucrose into glucose and fructose, or additionally cscB, encoding a sucrose permease. Introduction of cscA was sufficient to convey sucrose consumption and the additional presence of cscB had no further effect, though the sucrose permease was built and localized to the membrane. Sucrose was split extracellularly by the activity of the invertase CscA leaking out of the cell. The transposons were also used to confer sucrose consumption to Cupriavidus necator. Interestingly, in this strain, CscB acted as a glucose transporter, such that C. necator also gained the ability to grow on glucose. Thus, the pSST transposons are functional tools to extend the substrate spectrum of Gram‐negative bacterial strains toward sucrose.
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spelling pubmed-55529022017-08-15 Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose Löwe, Hannes Schmauder, Lukas Hobmeier, Karina Kremling, Andreas Pflüger‐Grau, Katharina Microbiologyopen Original Research Sucrose is an important disaccharide used as a substrate in many industrial applications. It is a major component of molasses, a cheap by‐product of the sugar industry. Unfortunately, not all industrially relevant organisms, among them Pseudomonas putida, are capable of metabolizing sucrose. We chose a metabolic engineering approach to circumvent this blockage and equip P. putida with the activities necessary to consume sucrose. Therefore, we constructed a pair of broad‐host range mini‐transposons (pSST – sucrose splitting transposon), carrying either cscA, encoding an invertase able to split sucrose into glucose and fructose, or additionally cscB, encoding a sucrose permease. Introduction of cscA was sufficient to convey sucrose consumption and the additional presence of cscB had no further effect, though the sucrose permease was built and localized to the membrane. Sucrose was split extracellularly by the activity of the invertase CscA leaking out of the cell. The transposons were also used to confer sucrose consumption to Cupriavidus necator. Interestingly, in this strain, CscB acted as a glucose transporter, such that C. necator also gained the ability to grow on glucose. Thus, the pSST transposons are functional tools to extend the substrate spectrum of Gram‐negative bacterial strains toward sucrose. John Wiley and Sons Inc. 2017-03-27 /pmc/articles/PMC5552902/ /pubmed/28349670 http://dx.doi.org/10.1002/mbo3.473 Text en © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research
Löwe, Hannes
Schmauder, Lukas
Hobmeier, Karina
Kremling, Andreas
Pflüger‐Grau, Katharina
Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose
title Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose
title_full Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose
title_fullStr Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose
title_full_unstemmed Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose
title_short Metabolic engineering to expand the substrate spectrum of Pseudomonas putida toward sucrose
title_sort metabolic engineering to expand the substrate spectrum of pseudomonas putida toward sucrose
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5552902/
https://www.ncbi.nlm.nih.gov/pubmed/28349670
http://dx.doi.org/10.1002/mbo3.473
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