Highly efficient neutralizer-free L-malic acid production using engineered Saccharomyces cerevisiae
Bioresource Technology(2023)
摘要
In industrial bioproduction of organic acids, numerous neutralizers are required which substantially increases production costs and burdens the environment. To address this challenge, a Saccharomyces cerevisiae mutant (named TAMC) with a low pH tolerance (pH 2.3) was isolated by adaptive laboratory evolution. Taking the synthesis of L-malic acid as an example, the malate dehydrogenase 3 without signal peptide (MDH Delta SKL) and pyruvate carboxylase 2 (PYC2) were overexpressed in cytoplasmic synthesis pathway, and the L-malic acid titer increased 5.6-fold. Subsequently, the malic acid transporter SpMae1 was designed, and the extracellular L-malic acid titer was increased from 7.3 to 73.6 g/L. Furthermore, by optimizing the synthesis of the precursor pyruvate, the titer reached 81.8 g/L. Finally, without any neutralizer, the titer in the 3-L bioreactor reached 232.9 g/L, the highest L-malic acid titer reported to date. Herein, the engineered L-malic acid overproducer paves the way for the large-scale green production of L-malic acid.
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关键词
Acid tolerance,Saccharomyces cerevisiae,Transporter engineering,L-malic acid,Neutralizer-free bioproduction
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