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Improvement of D‐Lactic acid production in Saccharomyces cerevisiae under acidic conditions by evolutionary and rational metabolic engineering

17:35 EDT 21 Jul 2017 | Wiley Biotechnology Journal

Microbial lactic acid (LA) production under acidic fermentation conditions is favorable to reduce the production cost, but circumventing LA toxicity is a major challenge. We generated D‐LA‐producing Saccharomyces cerevisiae strain JHY5610 by expressing D‐lactate dehydrogenase gene (Lm. ldhA) from Leuconostoc mesenteroides, while deleting genes involved in ethanol production (ADH1, ADH2, ADH3, ADH4, and ADH5), glycerol production (GPD1 and GPD2), and degradation of D‐LA (DLD1). Adaptive laboratory evolution of JHY5610 led to a strain JHY5710 having higher LA tolerance and D‐LA‐production capability. Genome sequencing of JHY5710 revealed that SUR1I245S mutation increases LA tolerance and D‐LA‐production, whereas a loss‐of‐function mutation of ERF2 only contributes to increasing D‐LA production. Introduction of both SUR1I245S and erf2Δ mutations into JHY5610 largely mimicked the D‐LA‐production capability of JHY5710, suggesting that these two mutations, which could modulate sphingolipid production and protein palmitoylation, are mainly responsible for the improved D‐LA production in JHY5710. JHY5710 was further improved by deleting PDC1 encoding pyruvate decarboxylase and additional integration of Lm. ldhA gene. The resulting strain JHY5730 produced up to 82.6 g/L of D‐LA with a yield of 0.83 g/g glucose and a productivity of 1.50 g/(L · h) in fed‐batch fermentation at pH 3.5.

Original Article: Improvement of D‐Lactic acid production in Saccharomyces cerevisiae under acidic conditions by evolutionary and rational metabolic engineering

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