Enhanced production of taxadiene in Saccharomyces cerevisiae

Autor:	Lungang Liang, Behnaz Nowrouzi, Laura E. Walls, Albert Isaac Lerma-Escalera, Jay D. Keasling, Jose Ruben Morones-Ramirez, Nestor Jonguitud-Borrego, Rachel Li, Koray Malcı, Leo d'Espaux, Leonardo Rios-Solis
Jazyk:	angličtina
Rok vydání:	2020
Předmět:	0106 biological sciences lcsh:QR1-502 Applied Microbiology and Biotechnology 01 natural sciences lcsh:Microbiology chemistry.chemical_compound Bioreactors Solubility Isomerases Shake flask 0303 health sciences Strain (chemistry) biology Chemistry Taxadiene Temperature Yeast metabolic engineering Biochemistry Metabolic Engineering Taxadiene synthase Production (computer science) Taxol™ Diterpenes Biotechnology Paclitaxel Stereochemistry Minibioreactor Saccharomyces cerevisiae Paclitaxel Taxol™ Bioengineering Antineoplastic Agents Alkenes Microbiology Industrial Biotechnology 03 medical and health sciences SDG 3 - Good Health and Well-being 010608 biotechnology Bioreactor Escherichia coli Bioprocess 030304 developmental biology 010405 organic chemistry Research biology.organism_classification 0104 chemical sciences biology.protein Taxol™
Zdroj:	Microbial Cell Factories, Vol 19, Iss 1, Pp 1-12 (2020) Microbial Cell Factories Microbial cell factories, vol 19, iss 1 Nowrouzi, B, Li, R A, Walls, L E, d 'Espaux, L, Malci, K, Liang, L, Jonguitud Borrego, N, Lerma-Escalera, A I, Morones-Ramirez, J R, Keasling, J D & Rios Solis, L 2020, ' Enhanced production of taxadiene in Saccharomyces cerevisiae ', Microbial Cell Factories, vol. 19, 200 . https://doi.org/10.1101/2020.06.08.139600, https://doi.org/10.1186/s12934-020-01458-2 Nowrouzi, B, Li, R A, Walls, L E, d’Espaux, L, Malcı, K, Liang, L, Jonguitud-Borrego, N, Lerma-Escalera, A I, Morones-Ramirez, J R, Keasling, J D & Rios-Solis, L 2020, ' Enhanced production of taxadiene in Saccharomyces cerevisiae ', Microbial Cell Factories, vol. 19, no. 1, 200 . https://doi.org/10.1186/s12934-020-01458-2
ISSN:	1475-2859
DOI:	10.1186/s12934-020-01458-2
Popis:	Background Cost-effective production of the highly effective anti-cancer drug, paclitaxel (Taxol®), remains limited despite growing global demands. Low yields of the critical taxadiene precursor remains a key bottleneck in microbial production. In this study, the key challenge of poor taxadiene synthase (TASY) solubility in S. cerevisiae was revealed, and the strains were strategically engineered to relieve this bottleneck. Results Multi-copy chromosomal integration of TASY harbouring a selection of fusion solubility tags improved taxadiene titres 22-fold, up to 57 ± 3 mg/L at 30 °C at microscale, compared to expressing a single episomal copy of TASY. The scalability of the process was highlighted through achieving similar titres during scale up to 25 mL and 250 mL in shake flask and bioreactor cultivations, respectively at 20 and 30 °C. Maximum taxadiene titres of 129 ± 15 mg/L and 127 mg/L were achieved through shake flask and bioreactor cultivations, respectively, of the optimal strain at a reduced temperature of 20 °C. Conclusions The results of this study highlight the benefit of employing a combination of molecular biology and bioprocess tools during synthetic pathway development, with which TASY activity was successfully improved by 6.5-fold compared to the highest literature titre in S. cerevisiae cell factories.
Databáze:	OpenAIRE
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