Rational engineering of Kluyveromyces marxianus to create a chassis for the production of aromatic products

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Date
2020-11
Authors
Rajkumar, Arun S.
Morrissey, John P.
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Publisher
BioMed Central
Published Version
10.1186/s12934-020-01461-7
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Abstract
Background: The yeast Kluyveromyces marxianus offers unique potential for industrial biotechnology because of useful features like rapid growth, thermotolerance and a wide substrate range. As an emerging alternative platform, K. marxianus requires the development and validation of metabolic engineering strategies to best utilise its metabolism as a basis for bio-based production. Results: To illustrate the synthetic biology strategies to be followed and showcase its potential, we describe a comprehensive approach to rationally engineer a metabolic pathway in K. marxianus. We use the phenylalanine biosynthetic pathway both as a prototype and because phenylalanine is a precursor for commercially valuable secondary metabolites. First, we modify and overexpress the pathway to be resistant to feedback inhibition so as to overproduce phenylalanine de novo from synthetic minimal medium. Second, we assess native and heterologous means to increase precursor supply to the biosynthetic pathway. Finally, we eliminate branch points and competing reactions in the pathway and rebalance precursors to redirect metabolic flux to a specific product, 2-phenylethanol (2-PE). As a result, we are able to construct robust strains capable of producing over 800 mg L-1 2-PE from minimal medium. Conclusions: The strains we constructed are a promising platform for the production of aromatic amino acid-based biochemicals, and our results illustrate challenges with attempting to combine individually beneficial modifications in an integrated platform.
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Keywords
Amino-acid biosynthesis , P-coumaric acid , Saccharomyces-cerevisiae , Yeast , Pyruvate , Pathway , Glucose , Protein , Gene , 2-phenylethanol , Aromatic amino acid , Shikimate pathway , Pentose phosphate pathway , Phosphoketolase , PEP synthase
Citation
Rajkumar, A. S. and Morrissey, J. P. (2020) 'Rational engineering of Kluyveromyces marxianus to create a chassis for the production of aromatic products', Microbial Cell Factories, 19, 207 (19pp). doi: 10.1186/s12934-020-01461-7
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https://hdl.handle.net/10468/12913
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Microbiology - Journal Articles
APC Microbiome Ireland - Journal Articles
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© The Author(s), 2020. This 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://creativeco mmons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://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.