期刊
METABOLIC ENGINEERING
卷 54, 期 -, 页码 293-300出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2019.05.004
关键词
Itaconate; Morphology; Metabolic engineering; Ustilago maydis; Ustilago cynodontis; pH-tolerance
资金
- German Federal Ministry of Food and Agriculture (BMEL), through the Specialist agency renewable raw materials e. V. (FNR) as part of the ERA-IB project TTRAFFIC [FKZ 22030515]
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy within the Cluster of Excellence [TMFB 236, Exzellenzcluster 2186]
Besides Aspergillus terreus and Ustilago maydis, Ustilago cynodontis is also known as a natural itaconate producer. U. cynodontis was reported as one of the best itaconate producing species in the family of the Ustilaginaceae, featuring a relatively high pH tolerance in comparison to other smut fungi. However, in contrast to U. maydis, it readily displays filamentous growth under sub-optimal growth conditions. In this study, U. cynodontis is established as efficient pH-tolerant itaconic acid producer through a combination of morphological and metabolic engineering. Deletions of the genes ras2, fuz7, and ubc3 abolished the filamentous growth of U. cynodontis, leading to a stable yeast-like growth under a range of stress-inducing conditions. The yeast-like morphology was also maintained in a pulsed fed batch production of 21 g L-1 itaconic acid and 9.3 g L-1 (S)-2-hydroxyparaconate at a pH of 3.8. The genetic and metabolic basis of itaconic acid production in U. cynodontis was characterized through comparative genomics and gene deletion studies. A hyper-producer strain was metabolically engineered using this knowledge resulting in a 6.5-fold improvement of titer.
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