Journal
JOURNAL OF BIOTECHNOLOGY
Volume 358, Issue -, Pages 111-117Publisher
ELSEVIER
DOI: 10.1016/j.jbiotec.2022.09.006
Keywords
Itaconic acid; Schizosacchromyces pombe; Metabolic engineering; Mitochondria
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Funding
- Japan Society for the Promotion of Science (JSPS), Japan [19H02526]
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This study developed a metabolic engineering strategy to produce Itaconic acid (IA) from glucose in the fission yeast Schizosaccharomyces pombe. Through heterologous expression of the CAD gene and mitochondrial localization, as well as the construction of mutant strains, the production of IA was significantly increased.
The economical production of value-added chemicals from renewable biomass is a promising aspect of producing a sustainable economy. Itaconic acid (IA) is a high value-added compound that is expected to be an alternative to petroleum-based chemicals. In this study, we developed a metabolic engineering strategy for the large-scale production of IA from glucose using the fission yeast Schizosaccharomyces pombe. Heterologous expression of the cis-aconitic acid decarboxylase (CAD) gene from Aspergillus terreus, which encodes cis-aconitate decarboxylase in the cytosol, led to the production of 0.132 g/L of IA. We demonstrated that mitochondrial localization of CAD enhanced the production of IA. To prevent the leakage of carbon flux from the TCA cycle, we generated a strain in which the endogenous malate exporter, citrate lyase, and citrate transporter genes were disrupted. A titer of 1.110 g/L of IA was obtained from a culture of this strain started with 50 g/L of glucose. By culturing the multiple mutant strain at increased cell density, we succeeded in enhancing the IA production to 1.555 g/L. The metabolic engineering strategies presented in this study have the potential to improve the titer of the biosynthesis of derivatives of intermediates of the TCA cycle.
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