期刊
BIORESOURCE TECHNOLOGY
卷 370, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2023.128580
关键词
Acid tolerance; Saccharomyces cerevisiae; Transporter engineering; L-malic acid; Neutralizer-free bioproduction
A low pH tolerance yeast mutant TAMC was obtained through adaptive laboratory evolution. By overexpressing enzymes and designing a transporter, the production of L-malic acid was significantly increased.
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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