<i>De Novo</i> Production of Hydroxytyrosol by Metabolic Engineering of <i>Saccharomyces cerevisiae</i>
Yingjie Liu, Han Liu, Haitao Hu, Kuan Rei Ng, Ruijin Yang, Xiaomei Lyu
Journal of Agricultural and Food Chemistry
Abstract
Hydroxytyrosol is an olive-derived phenolic compound of increasing commercial interest due to its health-promoting properties. In this study, a high-yield hydroxytyrosol-producing <i>Saccharomyces cerevisiae</i> cell factory was established via a comprehensive metabolic engineering scheme. First, <i>de novo</i> biosynthetic pathway of hydroxytyrosol was constructed in yeast by gene screening and overexpression of different phenol hydroxylases, among which paHD (from <i>Pseudomonas aeruginosa</i>) displayed the best catalytic performance. Next, hydroxytyrosol precursor supply was enhanced via a multimodular engineering approach: elimination of tyrosine feedback inhibition through genomic integration of <i>aro4</i><sup><i>K229L</i></sup> and <i>aro7</i><sup><i>G141S</i></sup>, construction of an aromatic aldehyde synthase (AAS)-based tyrosine metabolic pathway, and redistribution of metabolic flux between glycolytic pathway and pentose phosphate pathway (PPP) by introducing the exogenous gene <i>Bbxfpk</i><sup><i>opt</i></sup>. As a result, the titer of hydroxytyrosol was improved by 6.88-fold. Finally, a glucose-responsive dynamic regulation system based on <i>GAL80</i> deletion was implemented, resulting in the final hydroxytyrosol yields of 308.65 mg/L and 167.98 mg/g cell mass, the highest known from <i>de novo</i> production in <i>S. cerevisiae</i> to date.
Extracted Claims
2 claims extracted from this paper into the knowledge graph
hydroxytyrosol has improved yield 308.65 mg/L and 167.98 mg/g cell mass
“resulting in the final hydroxytyrosol yields of 308.65 mg/L and 167.98 mg/g cell mass, the highest known from de novo production in S. cerevisiae to date.”
hydroxytyrosol is produced by metabolic engineering of Saccharomyces cerevisiae
“a high-yield hydroxytyrosol-producing Saccharomyces cerevisiae cell factory was established via a comprehensive metabolic engineering scheme.”