Effects of acetoacetyl-CoA synthase expression on production of farnesene in Saccharomyces cerevisiae
Journal article, 2017

Efficient production of sesquiterpenes in Saccharomyces cerevisiae requires a high flux through the mevalonate pathway. To achieve this, the supply of acetyl-CoA plays a crucial role, partially because nine moles of acetyl-CoA are necessary to produce one mole of farnesyl diphosphate, but also to overcome the thermodynamic constraint imposed on the first reaction, in which acetoacetyl-CoA is produced from two moles of acetyl-CoA by acetoacetyl-CoA thiolase. Recently, a novel acetoacetyl-CoA synthase (nphT7) has been identified from Streptomyces sp. strain CL190, which catalyzes the irreversible condensation of malonyl-CoA and acetyl-CoA to acetoacetyl-CoA and, therefore, represents a potential target to increase the flux through the mevalonate pathway. This study investigates the effect of acetoacetyl-CoA synthase on growth as well as the production of farnesene and compares different homologs regarding their efficiency. While plasmid-based expression of nphT7 did not improve final farnesene titers, the construction of an alternative pathway, which exclusively relies on the malonyl-CoA bypass, was detrimental for growth and farnesene production. The presented results indicate that the overall functionality of the bypass was limited by the efficiency of acetoacetyl-CoA synthase (nphT7). Besides modulation of the expression level, which could be used as a means to partially restore the phenotype, nphT7 from Streptomyces glaucescens showed clearly higher efficiency compared to Streptomyces sp. strain CL190. © 2017, The Author(s).

Mevalonate pathway

Biofuels

Metabolic engineering

Isoprenoids

Yeast

Author

Stefan Tippmann

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Raphael Ferreira

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Verena Siewers

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Jens B Nielsen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Yun Chen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Journal of Industrial Microbiology and Biotechnology

1367-5435 (ISSN) 1476-5535 (eISSN)

Vol. 44 6 911-922

Subject Categories

Biochemistry and Molecular Biology

Microbiology

Infrastructure

Chalmers Infrastructure for Mass spectrometry

Areas of Advance

Life Science Engineering (2010-2018)

DOI

10.1007/s10295-017-1911-6

PubMed

28185100

More information

Created

10/7/2017