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Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement

BACKGROUND: FK506 is an important immunosuppressant, which can be produced by Streptomyces tsukubaensis. However, the production capacity of the strain is very low. Hereby, a computational guided engineering approach was proposed in order to improve the intracellular precursor and cofactor availabil...

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Autores principales: Huang, Di, Li, Shanshan, Xia, Menglei, Wen, Jianping, Jia, Xiaoqiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3680238/
https://www.ncbi.nlm.nih.gov/pubmed/23705993
http://dx.doi.org/10.1186/1475-2859-12-52
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author Huang, Di
Li, Shanshan
Xia, Menglei
Wen, Jianping
Jia, Xiaoqiang
author_facet Huang, Di
Li, Shanshan
Xia, Menglei
Wen, Jianping
Jia, Xiaoqiang
author_sort Huang, Di
collection PubMed
description BACKGROUND: FK506 is an important immunosuppressant, which can be produced by Streptomyces tsukubaensis. However, the production capacity of the strain is very low. Hereby, a computational guided engineering approach was proposed in order to improve the intracellular precursor and cofactor availability of FK506 in S. tsukubaensis. RESULTS: First, a genome-scale metabolic model of S. tsukubaensis was constructed based on its annotated genome and biochemical information. Subsequently, several potential genetic targets (knockout or overexpression) that guaranteed an improved yield of FK506 were identified by the recently developed methodology. To validate the model predictions, each target gene was manipulated in the parent strain D852, respectively. All the engineered strains showed a higher FK506 production, compared with D852. Furthermore, the combined effect of the genetic modifications was evaluated. Results showed that the strain HT-ΔGDH-DAZ with gdhA-deletion and dahp-, accA2-, zwf2-overexpression enhanced FK506 concentration up to 398.9 mg/L, compared with 143.5 mg/L of the parent strain D852. Finally, fed-batch fermentations of HT-ΔGDH-DAZ were carried out, which led to the FK506 production of 435.9 mg/L, 1.47-fold higher than the parent strain D852 (158.7 mg/L). CONCLUSIONS: Results confirmed that the promising targets led to an increase in FK506 titer. The present work is the first attempt to engineer the primary precursor pathways to improve FK506 production in S. tsukubaensis with genome-scale metabolic network guided metabolic engineering. The relationship between model prediction and experimental results demonstrates the rationality and validity of this approach for target identification. This strategy can also be applied to the improvement of other important secondary metabolites.
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spelling pubmed-36802382013-06-25 Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement Huang, Di Li, Shanshan Xia, Menglei Wen, Jianping Jia, Xiaoqiang Microb Cell Fact Research BACKGROUND: FK506 is an important immunosuppressant, which can be produced by Streptomyces tsukubaensis. However, the production capacity of the strain is very low. Hereby, a computational guided engineering approach was proposed in order to improve the intracellular precursor and cofactor availability of FK506 in S. tsukubaensis. RESULTS: First, a genome-scale metabolic model of S. tsukubaensis was constructed based on its annotated genome and biochemical information. Subsequently, several potential genetic targets (knockout or overexpression) that guaranteed an improved yield of FK506 were identified by the recently developed methodology. To validate the model predictions, each target gene was manipulated in the parent strain D852, respectively. All the engineered strains showed a higher FK506 production, compared with D852. Furthermore, the combined effect of the genetic modifications was evaluated. Results showed that the strain HT-ΔGDH-DAZ with gdhA-deletion and dahp-, accA2-, zwf2-overexpression enhanced FK506 concentration up to 398.9 mg/L, compared with 143.5 mg/L of the parent strain D852. Finally, fed-batch fermentations of HT-ΔGDH-DAZ were carried out, which led to the FK506 production of 435.9 mg/L, 1.47-fold higher than the parent strain D852 (158.7 mg/L). CONCLUSIONS: Results confirmed that the promising targets led to an increase in FK506 titer. The present work is the first attempt to engineer the primary precursor pathways to improve FK506 production in S. tsukubaensis with genome-scale metabolic network guided metabolic engineering. The relationship between model prediction and experimental results demonstrates the rationality and validity of this approach for target identification. This strategy can also be applied to the improvement of other important secondary metabolites. BioMed Central 2013-05-24 /pmc/articles/PMC3680238/ /pubmed/23705993 http://dx.doi.org/10.1186/1475-2859-12-52 Text en Copyright © 2013 Huang et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Huang, Di
Li, Shanshan
Xia, Menglei
Wen, Jianping
Jia, Xiaoqiang
Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement
title Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement
title_full Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement
title_fullStr Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement
title_full_unstemmed Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement
title_short Genome-scale metabolic network guided engineering of Streptomyces tsukubaensis for FK506 production improvement
title_sort genome-scale metabolic network guided engineering of streptomyces tsukubaensis for fk506 production improvement
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3680238/
https://www.ncbi.nlm.nih.gov/pubmed/23705993
http://dx.doi.org/10.1186/1475-2859-12-52
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