Bandyopadhyay, Sushobhan;
Pavlika, Vasos;
Bracewell, Daniel;
Nesbeth, Darren;
(2023)
A biological OR(XNOR) logic gate couples carbon source and transgene expression switching in a Komagatella phaffii (Pichia pastoris) strain co-producing process-enhancing lipase and a virus-like particle (VLP) vaccine.
ACS Synthetic Biology
, 12
(3)
pp. 657-663.
10.1021/acssynbio.2c00342.
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Abstract
We constructed a three-input biological logic gate: S OR (G XNOR M), where S is sorbitol, G is glycerol, and M is methanol, to optimize co-expression of two transgenes in Komagataella phaffii using batch-mode carbon source switching (CSS). K. phaffii was engineered to harbor transgenes encoding a Candida rugosa triacylglycerol lipase, which can enhance downstream processing by removing host cell lipids from homogenates, and the hepatitis B virus surface antigen (HBsAg), a protein that self-assembles into a virus-like particle (VLP) vaccine. Using the native alcohol oxidase 1 (PAOX1) and enolase 1 (PENO1) promoters to direct VLP vaccine and lipase expression, respectively, successfully provided an OR(XNOR) gate function with double-repression as the output. This logic gate functionality enabled use of CSS to ensure that approximately 80% of total VLP yield was accumulated before cells were burdened with lipase expression in 250 mL DasGip bioreactor cultivation.
| Type: | Article |
|---|---|
| Title: | A biological OR(XNOR) logic gate couples carbon source and transgene expression switching in a Komagatella phaffii (Pichia pastoris) strain co-producing process-enhancing lipase and a virus-like particle (VLP) vaccine |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1021/acssynbio.2c00342 |
| Publisher version: | https://doi.org/10.1021/acssynbio.2c00342 |
| Language: | English |
| Additional information: | Copyright © 2023 The Authors. Published under the Creative Commons CC BY public use license. |
| Keywords: | Triacylglycerol lipase, logic gate, virus-like particle, vaccine, bioreactor, carbon source |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10164604 |
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