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Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach

BACKGROUND: 3-Phenylpropanol with a pleasant odor is widely used in foods, beverages and cosmetics as a fragrance ingredient. It also acts as the precursor and reactant in pharmaceutical and chemical industries. Currently, petroleum-based manufacturing processes of 3-phenypropanol is environmentally...

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Autores principales: Liu, Zhenning, Zhang, Xue, Lei, Dengwei, Qiao, Bin, Zhao, Guang-Rong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8237410/
https://www.ncbi.nlm.nih.gov/pubmed/34176467
http://dx.doi.org/10.1186/s12934-021-01615-1
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author Liu, Zhenning
Zhang, Xue
Lei, Dengwei
Qiao, Bin
Zhao, Guang-Rong
author_facet Liu, Zhenning
Zhang, Xue
Lei, Dengwei
Qiao, Bin
Zhao, Guang-Rong
author_sort Liu, Zhenning
collection PubMed
description BACKGROUND: 3-Phenylpropanol with a pleasant odor is widely used in foods, beverages and cosmetics as a fragrance ingredient. It also acts as the precursor and reactant in pharmaceutical and chemical industries. Currently, petroleum-based manufacturing processes of 3-phenypropanol is environmentally unfriendly and unsustainable. In this study, we aim to engineer Escherichia coli as microbial cell factory for de novo production of 3-phenypropanol via retrobiosynthesis approach. RESULTS: Aided by in silico retrobiosynthesis analysis, we designed a novel 3-phenylpropanol biosynthetic pathway extending from l-phenylalanine and comprising the phenylalanine ammonia lyase (PAL), enoate reductase (ER), aryl carboxylic acid reductase (CAR) and phosphopantetheinyl transferase (PPTase). We screened the enzymes from plants and microorganisms and reconstructed the artificial pathway for conversion of 3-phenylpropanol from l-phenylalanine. Then we conducted chromosome engineering to increase the supply of precursor l-phenylalanine and combined the upstream l-phenylalanine pathway and downstream 3-phenylpropanol pathway. Finally, we regulated the metabolic pathway strength and optimized fermentation conditions. As a consequence, metabolically engineered E. coli strain produced 847.97 mg/L of 3-phenypropanol at 24 h using glucose-glycerol mixture as co-carbon source. CONCLUSIONS: We successfully developed an artificial 3-phenylpropanol pathway based on retrobiosynthesis approach, and highest titer of 3-phenylpropanol was achieved in E. coli via systems metabolic engineering strategies including enzyme sources variety, chromosome engineering, metabolic strength balancing and fermentation optimization. This work provides an engineered strain with industrial potential for production of 3-phenylpropanol, and the strategies applied here could be practical for bioengineers to design and reconstruct the microbial cell factory for high valuable chemicals. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01615-1.
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spelling pubmed-82374102021-06-29 Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach Liu, Zhenning Zhang, Xue Lei, Dengwei Qiao, Bin Zhao, Guang-Rong Microb Cell Fact Research BACKGROUND: 3-Phenylpropanol with a pleasant odor is widely used in foods, beverages and cosmetics as a fragrance ingredient. It also acts as the precursor and reactant in pharmaceutical and chemical industries. Currently, petroleum-based manufacturing processes of 3-phenypropanol is environmentally unfriendly and unsustainable. In this study, we aim to engineer Escherichia coli as microbial cell factory for de novo production of 3-phenypropanol via retrobiosynthesis approach. RESULTS: Aided by in silico retrobiosynthesis analysis, we designed a novel 3-phenylpropanol biosynthetic pathway extending from l-phenylalanine and comprising the phenylalanine ammonia lyase (PAL), enoate reductase (ER), aryl carboxylic acid reductase (CAR) and phosphopantetheinyl transferase (PPTase). We screened the enzymes from plants and microorganisms and reconstructed the artificial pathway for conversion of 3-phenylpropanol from l-phenylalanine. Then we conducted chromosome engineering to increase the supply of precursor l-phenylalanine and combined the upstream l-phenylalanine pathway and downstream 3-phenylpropanol pathway. Finally, we regulated the metabolic pathway strength and optimized fermentation conditions. As a consequence, metabolically engineered E. coli strain produced 847.97 mg/L of 3-phenypropanol at 24 h using glucose-glycerol mixture as co-carbon source. CONCLUSIONS: We successfully developed an artificial 3-phenylpropanol pathway based on retrobiosynthesis approach, and highest titer of 3-phenylpropanol was achieved in E. coli via systems metabolic engineering strategies including enzyme sources variety, chromosome engineering, metabolic strength balancing and fermentation optimization. This work provides an engineered strain with industrial potential for production of 3-phenylpropanol, and the strategies applied here could be practical for bioengineers to design and reconstruct the microbial cell factory for high valuable chemicals. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01615-1. BioMed Central 2021-06-27 /pmc/articles/PMC8237410/ /pubmed/34176467 http://dx.doi.org/10.1186/s12934-021-01615-1 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Liu, Zhenning
Zhang, Xue
Lei, Dengwei
Qiao, Bin
Zhao, Guang-Rong
Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach
title Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach
title_full Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach
title_fullStr Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach
title_full_unstemmed Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach
title_short Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach
title_sort metabolic engineering of escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8237410/
https://www.ncbi.nlm.nih.gov/pubmed/34176467
http://dx.doi.org/10.1186/s12934-021-01615-1
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