Abstract
A comparative metabolomic analysis was conducted on recombinant Saccharomyces cerevisiae strain producing β-carotene and the parent strain cultivated with glucose as carbon source using gas chromatography-mass spectrometry (GC-MS), high performance liquid chromatography-mass spectrometry (HPLC-MS) and ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) based approach. The results showed that most of the central intermediates associated with amino acids, carbohydrates, glycolysis and TCA cycle intermediates (acetic acid, glycerol, citric acid, pyruvic acid and succinic acid), fatty acids, ergosterol and energy metabolites were produced in a lower amount in recombinant strain, as compared to the parent strain. To increase β-carotene production in recombinant strain, a strategy that exogenous addition of acetate (10 g/l) in exponential phase was developed, which could enhance most intracellular metabolites levels and result in 39.3% and 14.2% improvement of β-carotene concentration and production, respectively, which was accompanied by the enhancement of acetyl-CoA, fatty acids, ergosterol and ATP contents in cells. These results indicated that the amounts of intracellular metabolites in engineered strain are largely consumed by carotenoid formation. Therefore, maintaining intracellular metabolites pool at normal levels is essential for carotenoid biosynthesis. To relieve this limitation, rational supplementation of acetate could be a potential way because it can partially restore the levels of intracellular metabolites and improve the production of carotenoid compounds in recombinant S. cerevisiae.
Highlights
Carotenoids are natural pigments synthesized by plants and microorganisms [1]
The cell growth (Fig 2A) and glucose consumption (Fig 2B) of recombinant strain were negatively influenced by β-carotene biosynthesis compared to the parent strain due to metabolic burden, which was consistent with previous literature data [22,25]
The obtained results showed that heterologous biosynthesis of carotenoids in S. cerevisiae significantly reduce intracellular metabolites pool compared to the parent strain, especially in the rapid synthesis phase of carotenoids (Figs 3 and 4)
Summary
Carotenoids are natural pigments synthesized by plants and microorganisms [1]. Beta-carotene is an orange-colored carotenoid that possesses powerful free radical quenching activity, which has lots of applications in pharmaceuticals, neutraceuticals, cosmetics and foods [2]. Improving β-carotene production with acetate supplementation by comparative metabolomics
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
