Crucial role of 4-deoxy-L-erythro-5-hexoseulose uronate reductase for alginate utilization revealed by adaptive evolution in engineered Saccharomyces cerevisiae
| Autor: | Makoto Hirayama, Fumiya Matsuoka, Hideki Tanaka, Takayuki Kashihara, Shigeyuki Kawai, Wataru Hashimoto, Kousaku Murata |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Alginates Science 030106 microbiology Saccharomyces cerevisiae Biology Reductase medicine.disease_cause Article 03 medical and health sciences medicine Mannitol Gene Hexoses Mutation Multidisciplinary Strain (chemistry) Ethanol Metabolism biology.organism_classification Carbon Kinetics 030104 developmental biology Biochemistry Fermentation Medicine NAD+ kinase Directed Molecular Evolution Genetic Engineering Oxidoreductases |
| Zdroj: | Scientific Reports Scientific Reports, Vol 7, Iss 1, Pp 1-11 (2017) |
| ISSN: | 2045-2322 |
| Popis: | In brown macroalgae, alginate and D-mannitol are promising carbohydrates for biorefinery. Saccharomyces cerevisiae is widely used as a microbial cell factory, but this budding yeast is unable to utilize either alginate or D-mannitol. Alginate can be depolymerized by both endo-type and exo-type alginate lyases, yielding a monouronate, 4-deoxy-L-erythro-5-hexoseulose uronate (DEH), a key intermediate in the metabolism of alginate. Here, we constructed engineered two S. cerevisiae strains that are able to utilize both DEH and D-mannitol on two different strain backgrounds, and we also improved their aerobic growth in a DEH liquid medium through adaptive evolution. In both evolved strains, one of the causal mutations was surprisingly identical, a c.50A > G mutation in the codon-optimized NAD(P)H-dependent DEH reductase gene, one of the 4 genes introduced to confer the capacity to utilize DEH. This mutation resulted in an E17G substitution at a loop structure near the coenzyme-binding site of this reductase, and enhanced the reductase activity and aerobic growth in both evolved strains. Thus, the crucial role for this reductase reaction in the metabolism of DEH in the engineered S. cerevisiae is demonstrated, and this finding provides significant information for synthetic construction of a S. cerevisiae strain as a platform for alginate utilization. 代謝改変酵母のアルギン酸モノマー代謝能向上メカニズムの解明--国産海洋バイオマス資源から代替ガソリンや合成ゴム原料を生産するための重要な第一歩--. 京都大学プレスリリース. 2017-06-26. |
| Databáze: | OpenAIRE |
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