Modular pathway rewiring of Saccharomyces cerevisiae enables high-level production of L-ornithine
Journal article, 2015

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.

Author

J. Qin

Yongjin Zhou

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Anastasia Krivoruchko

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Mingtao Huang

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Lifang Liu

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Sakda Khoomrung

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Verena Siewers

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

B. Jiang

Jens B Nielsen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Nature Communications

2041-1723 (ISSN)

Vol. 6 Sept. Art. no. 8224-

Industrial Systems Biology of Yeast and A. oryzae (INSYSBIO)

European Commission (FP7), 2010-01-01 -- 2014-12-31.

Subject Categories

Industrial Biotechnology

Bioinformatics and Systems Biology

Areas of Advance

Life Science Engineering (2010-2018)

DOI

10.1038/ncomms9224

PubMed

26345617

More information

Created

10/7/2017