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The ornithine-urea cycle involves fumaric acid biosynthesis in Aureobasidium pullulans var. aubasidani, a green and eco-friendly process for fumaric acid production.

ABSTRACT: The current petroleum chemical methods for fumaric acid production can cause heavy pollution and global warming. In this study, the engineered strains of A. pullulans var. aubasidani were found to be suitable for green fumaric acid producer. Removal and complementation of the relevant genes showed only the ornithine-urea cycle (OUC) was involved in high level fumarate biosynthesis which was controlled by the Ca²⁺ signaling pathway. Removal of both the GOX gene encoding glucose oxidase and the PKS1 gene encoding the polyketide synthase for 3,5-dihydroxydecanoic acid biosynthesis and overexpression of the PYC gene encoding pyruvate carboxylase made the strain e-PYC produce 88.1 ± 4.3 g/L of fumarate at flask level and 93.9 ± 0.8 g/L of fumarate during the fed-batch fermentation. As a yeast-like fungal strain, it was very easy to cultivate A. pullulans var. aubasidani DH177 and their mutants in the bioreactor and to edit its genomic DNAs to enhance fumarate production. It was found that 2 mol of CO₂ could be fixed during a maximal theoretical yield of 2 mol of fumarate per mole of glucose consumed in the OUC. Therefore, the OUC-mediated fumarate biosynthesis pathway in A. pullulans var. aubasidani was a green and eco-friendly process for the global sustainable development and carbon neutrality.

SUBMITTER: Wei X

PROVIDER: S-EPMC9647333 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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The ornithine-urea cycle involves fumaric acid biosynthesis in Aureobasidium pullulans var. aubasidani, a green and eco-friendly process for fumaric acid production.

Wei Xin X Zhang Miao M Wang Guang-Yuan GY Liu Guang-Lei GL Chi Zhen-Ming ZM Chi Zhe Z

Synthetic and systems biotechnology 20221019 1

The current petroleum chemical methods for fumaric acid production can cause heavy pollution and global warming. In this study, the engineered strains of A. pullulans var. aubasidani were found to be suitable for green fumaric acid producer. Removal and complementation of the relevant genes showed only the ornithine-urea cycle (OUC) was involved in high level fumarate biosynthesis which was controlled by the Ca2+ signaling pathway. Removal of both the GOX gene enc ...[more]

PMID: 36381963

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Project description:Extracellular vesicles (EVs) are increasingly recognized as an important mechanism for cell-cell interactions. Their role in fungi is still poorly understood and they have been isolated from only a handful of species. Here, we isolated and characterized EVs from Aureobasidium pullulans, a biotechnologically important black yeast-like fungus that is increasingly used for biocontrol of phytopathogenic fungi and bacteria. After optimization of the isolation protocol, characterization of EVs from A. pullulans by transmission electron microscopy (TEM) revealed a typical cup-shaped morphology and different subpopulations of EVs. These results were confirmed by nanoparticle tracking analysis (NTA), which revealed that A. pullulans produced 6.1 × 10^8 nanoparticles per milliliter of culture medium. Proteomic analysis of EVs detected 642 proteins. A small fraction of them had signal peptides for secretion and transmembrane domains. Proteins characteristic of different synthesis pathways were found, suggesting that EVs are synthesized by multiple pathways in A. pullulans. Enrichment analysis using Gene Ontology showed that most of the proteins found in the EVs were associated with primary metabolism. When sequencing the small RNA fraction of A. pullulans EVs, we found two hypothetical novel mil-RNAs. Finally, we tested the biocontrol potential of EVs from A. pullulans. The EVs did not inhibit the germination of spores of three important phytopathogenic fungi – Botrytis cinerea, Colletotrichum acutatum, and Penicillium expansum. However, exposure of grown cultures of C. acutatum and P. expansum to A. pullulans EVs resulted in visible changes in morphology of colonies. These preliminary results suggest that EVs may be part of the antagonistic activity of A. pullulans, which is so far only partially understood. Thus, the first isolation and characterization of EVs from A. pullulans provides a starting point for further studies of EVs in the biotechnologically important traits of the biocontrol black fungus A. pullulans in particular and in the biological role of fungal EVs in general.

Dataset Information

The ornithine-urea cycle involves fumaric acid biosynthesis in Aureobasidium pullulans var. aubasidani, a green and eco-friendly process for fumaric acid production.

Publications

The ornithine-urea cycle involves fumaric acid biosynthesis in <i>Aureobasidium pullulans</i> var. <i>aubasidani</i>, a green and eco-friendly process for fumaric acid production.

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