Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine

Baker’s yeast Saccharomyces cerevisiae is an attractive cell factory for production of chemicals and biofuels. Many different products have been produced in this cell factory by reconstruction of heterologous biosynthetic pathways; however, endogenous metabolism by itself involves many metabolites o...

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Main Authors: Qin, J., Zhou, Y., Krivoruchko, A., Huang, M., Liu, Lifang, Khoomrung, S., Siewers, V., Jiang, B., Nielsen, J.
Format: Journal Article
Published: Nature Publishing Group 2015
Online Access:http://hdl.handle.net/20.500.11937/19038
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author Qin, J.
Zhou, Y.
Krivoruchko, A.
Huang, M.
Liu, Lifang
Khoomrung, S.
Siewers, V.
Jiang, B.
Nielsen, J.
author_facet Qin, J.
Zhou, Y.
Krivoruchko, A.
Huang, M.
Liu, Lifang
Khoomrung, S.
Siewers, V.
Jiang, B.
Nielsen, J.
author_sort Qin, J.
building Curtin Institutional Repository
collection Online Access
description Baker’s yeast Saccharomyces cerevisiae is an attractive cell factory for production of chemicals and biofuels. Many different products have been produced in this cell factory by reconstruction of heterologous biosynthetic pathways; however, endogenous metabolism by itself involves many metabolites of industrial interest, and de-regulation of endogenous pathways to ensure efficient carbon channelling to such metabolites is therefore of high interest. Furthermore, many of these may serve as precursors for the biosynthesis of complex natural products, and hence strains overproducing certain pathway intermediates can serve as platform cell factories for production of such products. Here we implement a modular pathway rewiring (MPR) strategy and demonstrate its use for pathway optimization resulting in high-level production of L-ornithine, an intermediate of L-arginine biosynthesis and a precursor metabolite for a range of different natural products. The MPR strategy involves rewiring of the urea cycle, subcellular trafficking engineering and pathway re-localization, and improving precursor supply either through attenuation of the Crabtree effect or through the use of controlled fed-batch fermentations, leading to an L-ornithine titre of 1,041±47 mg l−1 with a yield of 67 mg (g glucose)−1 in shake-flask cultures and a titre of 5.1 g l−1 in fed-batch cultivations. Our study represents the first comprehensive study on overproducing an amino-acid intermediate in yeast, and our results demonstrate the potential to use yeast more extensively for low-cost production of many high-value amino-acid-derived chemicals.
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spelling curtin-20.500.11937-190382017-09-13T13:45:15Z Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine Qin, J. Zhou, Y. Krivoruchko, A. Huang, M. Liu, Lifang Khoomrung, S. Siewers, V. Jiang, B. Nielsen, J. Baker’s yeast Saccharomyces cerevisiae is an attractive cell factory for production of chemicals and biofuels. Many different products have been produced in this cell factory by reconstruction of heterologous biosynthetic pathways; however, endogenous metabolism by itself involves many metabolites of industrial interest, and de-regulation of endogenous pathways to ensure efficient carbon channelling to such metabolites is therefore of high interest. Furthermore, many of these may serve as precursors for the biosynthesis of complex natural products, and hence strains overproducing certain pathway intermediates can serve as platform cell factories for production of such products. Here we implement a modular pathway rewiring (MPR) strategy and demonstrate its use for pathway optimization resulting in high-level production of L-ornithine, an intermediate of L-arginine biosynthesis and a precursor metabolite for a range of different natural products. The MPR strategy involves rewiring of the urea cycle, subcellular trafficking engineering and pathway re-localization, and improving precursor supply either through attenuation of the Crabtree effect or through the use of controlled fed-batch fermentations, leading to an L-ornithine titre of 1,041±47 mg l−1 with a yield of 67 mg (g glucose)−1 in shake-flask cultures and a titre of 5.1 g l−1 in fed-batch cultivations. Our study represents the first comprehensive study on overproducing an amino-acid intermediate in yeast, and our results demonstrate the potential to use yeast more extensively for low-cost production of many high-value amino-acid-derived chemicals. 2015 Journal Article http://hdl.handle.net/20.500.11937/19038 10.1038/ncomms9224 Nature Publishing Group unknown
spellingShingle Qin, J.
Zhou, Y.
Krivoruchko, A.
Huang, M.
Liu, Lifang
Khoomrung, S.
Siewers, V.
Jiang, B.
Nielsen, J.
Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine
title Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine
title_full Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine
title_fullStr Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine
title_full_unstemmed Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine
title_short Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine
title_sort modular pathway rewiring of saccharomyces cerevisiae enables high-level production of l-ornithine
url http://hdl.handle.net/20.500.11937/19038