Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase
We have engineered strain BG-10 of the methylotrophic yeast Komagataella phaffii for use as an effective whole cell biocatalyst. We introduced into the yeast a transgene encoding a Chromobacterium violaceum ω-transaminase for transcription in response to methanol induction. The strain was then asses...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-08-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844019359985 |
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| author | Stephanie Braun-Galleani Maria-José Henríquez Darren N. Nesbeth |
| author_facet | Stephanie Braun-Galleani Maria-José Henríquez Darren N. Nesbeth |
| author_sort | Stephanie Braun-Galleani |
| collection | DOAJ |
| description | We have engineered strain BG-10 of the methylotrophic yeast Komagataella phaffii for use as an effective whole cell biocatalyst. We introduced into the yeast a transgene encoding a Chromobacterium violaceum ω-transaminase for transcription in response to methanol induction. The strain was then assessed with respect to its growth performance and biotransformation of a fed ketoalcohol substrate to an amino-alcohol. In the resultant strain, BG-TAM, methanol induction did not compromise cell growth. Successful bioconversion of fed substrates to the by-product, acetophenone, indicated transaminase activity in shake flask-cultivated BG-TAM cells. We then used bioreactor cultivation to exploit the high levels of biomass achievable by Komagataella phaffii. In a 900 μL reaction the BG-TAM strain at OD600 = 1024 achieved up to 0.41 mol mol−1 (molproduct molsubstrate−1) yield on substrate (Yp/s) for production of 1-methyl-3-phenylpropylamine and a space time yield (STY) of 0.29 g L−1 h−1 for production of 2-amino-1,3,4-butanetriol. We have shown that transamination, an important step for bespoke synthesis of small molecule medicines, is biologically realisable using enzymes with a broad substrate range, such as ω-transaminases, within living yeast cells that are fed low-cost substrates for bioconversion. |
| first_indexed | 2024-12-10T15:18:39Z |
| format | Article |
| id | doaj.art-0129627e97b64688bb83cca671248edd |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-12-10T15:18:39Z |
| publishDate | 2019-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-0129627e97b64688bb83cca671248edd2022-12-22T01:43:45ZengElsevierHeliyon2405-84402019-08-0158e02338Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminaseStephanie Braun-Galleani0Maria-José Henríquez1Darren N. Nesbeth2Department of Biochemical Engineering, University College London, Bernard Katz Building, London, WC1E 6BT, United KingdomDepartment of Biochemical Engineering, University College London, Bernard Katz Building, London, WC1E 6BT, United KingdomCorresponding author.; Department of Biochemical Engineering, University College London, Bernard Katz Building, London, WC1E 6BT, United KingdomWe have engineered strain BG-10 of the methylotrophic yeast Komagataella phaffii for use as an effective whole cell biocatalyst. We introduced into the yeast a transgene encoding a Chromobacterium violaceum ω-transaminase for transcription in response to methanol induction. The strain was then assessed with respect to its growth performance and biotransformation of a fed ketoalcohol substrate to an amino-alcohol. In the resultant strain, BG-TAM, methanol induction did not compromise cell growth. Successful bioconversion of fed substrates to the by-product, acetophenone, indicated transaminase activity in shake flask-cultivated BG-TAM cells. We then used bioreactor cultivation to exploit the high levels of biomass achievable by Komagataella phaffii. In a 900 μL reaction the BG-TAM strain at OD600 = 1024 achieved up to 0.41 mol mol−1 (molproduct molsubstrate−1) yield on substrate (Yp/s) for production of 1-methyl-3-phenylpropylamine and a space time yield (STY) of 0.29 g L−1 h−1 for production of 2-amino-1,3,4-butanetriol. We have shown that transamination, an important step for bespoke synthesis of small molecule medicines, is biologically realisable using enzymes with a broad substrate range, such as ω-transaminases, within living yeast cells that are fed low-cost substrates for bioconversion.http://www.sciencedirect.com/science/article/pii/S2405844019359985BioengineeringBiotechnologyChemical engineeringKomagataella phaffiiPichia pastorisTransaminase |
| spellingShingle | Stephanie Braun-Galleani Maria-José Henríquez Darren N. Nesbeth Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase Heliyon Bioengineering Biotechnology Chemical engineering Komagataella phaffii Pichia pastoris Transaminase |
| title | Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase |
| title_full | Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase |
| title_fullStr | Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase |
| title_full_unstemmed | Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase |
| title_short | Whole cell biosynthesis of 1-methyl-3-phenylpropylamine and 2-amino-1,3,4-butanetriol using Komagataella phaffii (Pichia pastoris) strain BG-10 engineered with a transgene encoding Chromobacterium violaceum ω-transaminase |
| title_sort | whole cell biosynthesis of 1 methyl 3 phenylpropylamine and 2 amino 1 3 4 butanetriol using komagataella phaffii pichia pastoris strain bg 10 engineered with a transgene encoding chromobacterium violaceum ω transaminase |
| topic | Bioengineering Biotechnology Chemical engineering Komagataella phaffii Pichia pastoris Transaminase |
| url | http://www.sciencedirect.com/science/article/pii/S2405844019359985 |
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