Reconstruction of the carnitine biosynthesis pathway from Neurospora crassa in the yeast Saccharomyces cerevisiae

Verfasser / Beitragende:
[Jaco Franken, Anita Burger, Jan Swiegers, Florian Bauer]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/15(2015-08-01), 6377-6389
Format:
Artikel (online)
ID: 605506817
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024 7 0 |a 10.1007/s00253-015-6561-x  |2 doi 
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245 0 0 |a Reconstruction of the carnitine biosynthesis pathway from Neurospora crassa in the yeast Saccharomyces cerevisiae  |h [Elektronische Daten]  |c [Jaco Franken, Anita Burger, Jan Swiegers, Florian Bauer] 
520 3 |a Industrial synthesis of l-carnitine is currently performed by whole-cell biotransformation of industrial waste products, mostly d-carnitine and cronobetaine, through specific bacterial species. No comparable system has been established using eukaryotic microorganisms, even though there is a significant and growing international demand for either the pure compound or carnitine-enriched consumables. In eukaryotes, including the fungus Neurospora crassa, l-carnitine is biosynthesized through a four-step metabolic conversion of trimethyllysine to l-carnitine. In contrast, the industrial yeast, Saccharomyces cerevisiae lacks the enzymes of the eukaryotic biosynthesis pathway and is unable to synthesize carnitine. This study describes the cloning of all four of the N. crassa carnitine biosynthesis genes and the reconstruction of the entire pathway in S. cerevisiae. The engineered yeast strains were able to catalyze the synthesis of l-carnitine, which was quantified using hydrophilic interaction liquid chromatography electrospray ionization mass spectrometry (HILIC-ESI-MS) analyses, from trimethyllysine. Furthermore, the yeast threonine aldolase Gly1p was shown to effectively catalyze the second step of the pathway, fulfilling the role of a serine hydroxymethyltransferase. The analyses also identified yeast enzymes that interact with the introduced pathway, including Can1p, which was identified as the yeast transporter for trimethyllysine, and the two yeast serine hydroxymethyltransferases, Shm1p and Shm2p. Together, this study opens the possibility of using an engineered, carnitine-producing yeast in various industrial applications while providing insight into possible future strategies aimed at tailoring the production capacity of such strains. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Carnitine  |2 nationallicence 
690 7 |a Biosynthesis  |2 nationallicence 
690 7 |a Neurospora crassa  |2 nationallicence 
690 7 |a Saccharomyces cerevisiae  |2 nationallicence 
700 1 |a Franken  |D Jaco  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
700 1 |a Burger  |D Anita  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
700 1 |a Swiegers  |D Jan  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
700 1 |a Bauer  |D Florian  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/15(2015-08-01), 6377-6389  |x 0175-7598  |q 99:15<6377  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6561-x  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6561-x  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Franken  |D Jaco  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Burger  |D Anita  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Swiegers  |D Jan  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Bauer  |D Florian  |u Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, 7602, Matieland, South Africa  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/15(2015-08-01), 6377-6389  |x 0175-7598  |q 99:15<6377  |1 2015  |2 99  |o 253