A highly efficient transcriptome-based biosynthesis of non-ethanol chemicals in Crabtree negative Saccharomyces cerevisiae
Journal article, 2023

Background: Owing to the Crabtree effect, Saccharomyces cerevisiae produces a large amount of ethanol in the presence of oxygen and excess glucose, leading to a loss of carbon for the biosynthesis of non-ethanol chemicals. In the present study, the potential of a newly constructed Crabtree negative S. cerevisiae, as a chassis cell, was explored for the biosynthesis of various non-ethanol compounds. Results: To understand the metabolic characteristics of Crabtree negative S. cerevisiae sZJD-28, its transcriptional profile was compared with that of Crabtree positive S. cerevisiae CEN.PK113-11C. The reporter GO term analysis showed that, in sZJD-28, genes associated with translational processes were down-regulated, while those related to carbon metabolism were significantly up-regulated. To verify a potential increase in carbon metabolism for the Crabtree negative strain, the production of non-ethanol chemicals, derived from different metabolic nodes, was then undertaken for both sZJD-28 and CEN.PK113-11C. At the pyruvate node, production of 2,3-butanediol and lactate in sZJD-28-based strains was remarkably higher than that of CEN.PK113-11C-based ones, representing 16.8- and 1.65-fold increase in titer, as well as 4.5-fold and 0.65-fold increase in specific titer (mg/L/OD), respectively. Similarly, for shikimate derived p-coumaric acid, the titer of sZJD-28-based strain was 0.68-fold higher than for CEN.PK113-11C-based one, with a 0.98-fold increase in specific titer. While farnesene and lycopene, two acetoacetyl-CoA derivatives, showed 0.21- and 1.88-fold increases in titer, respectively. From malonyl-CoA, the titer of 3-hydroxypropionate and fatty acids in sZJD-28-based strains were 0.19- and 0.76-fold higher than that of CEN.PK113-11C-based ones, respectively. In fact, yields of products also improved by the same fold due to the absence of residual glucose. Fed-batch fermentation further showed that the titer of free fatty acids in sZJD-28-based strain 28-FFA-E reached 6295.6 mg/L with a highest reported specific titer of 247.7 mg/L/OD in S. cerevisiae. Conclusions: Compared with CEN.PK113-11C, the Crabtree negative sZJD-28 strain displayed a significantly different transcriptional profile and obvious advantages in the biosynthesis of non-ethanol chemicals due to redirected carbon and energy sources towards metabolite biosynthesis. The findings, therefore, suggest that a Crabtree negative S. cerevisiae strain could be a promising chassis cell for the biosynthesis of various chemicals.

Crabtree negative

Fatty acids

Microbial production

Chassis strain

Crabtree effect

Saccharomyces cerevisiae

Author

Zhen Yao

Tianjin Institute of Industrial Biotechnology

National Technology Innovation Center of Synthetic Biology

Yufeng Guo

Tianjin Institute of Industrial Biotechnology

National Technology Innovation Center of Synthetic Biology

Huan Wang

Chongqing University

Yun Chen

Chalmers, Life Sciences, Systems and Synthetic Biology

Qinhong Wang

National Technology Innovation Center of Synthetic Biology

Tianjin Institute of Industrial Biotechnology

Jens B Nielsen

Chalmers, Life Sciences, Systems and Synthetic Biology

Beijing University of Chemical Technology

Zongijie Dai

National Technology Innovation Center of Synthetic Biology

Tianjin Institute of Industrial Biotechnology

Biotechnology for Biofuels and Bioproducts

27313654 (eISSN)

Vol. 16 1 37

Subject Categories

Biochemistry and Molecular Biology

Microbiology

Other Industrial Biotechnology

DOI

10.1186/s13068-023-02276-5

More information

Latest update

4/12/2023