Enhanced electrochemical measurement of β-galactosidase activity in whole cells by coexpression of lactose permease, LacY

Enhanced electrochemical measurement of β-galactosidase activity in whole cells by coexpression of lactose permease, LacY

Whole-cell biosensing links the sensing and computing capabilities of microbes to the generation of a detectable reporter. Whole cells enable dynamic biological computation (filtered noise, amplified signals, logic gating etc.). Enzymatic reporters enable in situ signal amplification. Electrochemica...

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Veröffentlicht in:BioTechniques. - 1988. - 73(2022), 5 vom: 08. Nov., Seite 233-237
1. Verfasser: VanArsdale, Eric (VerfasserIn)
Weitere Verfasser: Pitzer, Juliana, Wang, Sally, Stephens, Kristina, Chen, Chen-Yu, Payne, Gregory F, Bentley, William E
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2022
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Miller assay electrochemical sensing synthetic biology whole-cell β-galactosidase assay lactose permease 9068-45-5 Membrane Transport Proteins beta-Galactosidase mehr... EC 3.2.1.23 Galactose X2RN3Q8DNE Escherichia coli Proteins LacY protein, E coli Monosaccharide Transport Proteins Symporters
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500 |a Date Revised 04.12.2022 
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500 |a Citation Status MEDLINE 
520 |a Whole-cell biosensing links the sensing and computing capabilities of microbes to the generation of a detectable reporter. Whole cells enable dynamic biological computation (filtered noise, amplified signals, logic gating etc.). Enzymatic reporters enable in situ signal amplification. Electrochemical measurements are easily quantified and work in turbid environments. In this work we show how the coexpression of the lactose permease, LacY, dramatically improves electrochemical sensing of β-galactosidase (LacZ) expressed as a reporter in whole cells. The permease facilitates transport of the LacZ substrate, 4-aminophenyl β-d-galactopyranoside, which is converted to redox active p-aminophenol, which, in turn, is detected via cyclic voltammetry or chronocoulometry. We show a greater than fourfold improvement enabled by lacY coexpression in cells engineered to respond to bacterial signal molecules, pyocyanin and quorum-sensing autoinducer-2 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a Miller assay 
650 4 |a electrochemical sensing 
650 4 |a synthetic biology 
650 4 |a whole-cell β-galactosidase assay 
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650 7 |a 9068-45-5  |2 NLM 
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650 7 |a Galactose  |2 NLM 
650 7 |a X2RN3Q8DNE  |2 NLM 
650 7 |a Escherichia coli Proteins  |2 NLM 
650 7 |a LacY protein, E coli  |2 NLM 
650 7 |a Monosaccharide Transport Proteins  |2 NLM 
650 7 |a Symporters  |2 NLM 
700 1 |a Pitzer, Juliana  |e verfasserin  |4 aut 
700 1 |a Wang, Sally  |e verfasserin  |4 aut 
700 1 |a Stephens, Kristina  |e verfasserin  |4 aut 
700 1 |a Chen, Chen-Yu  |e verfasserin  |4 aut 
700 1 |a Payne, Gregory F  |e verfasserin  |4 aut 
700 1 |a Bentley, William E  |e verfasserin  |4 aut 
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773 1 8 |g volume:73  |g year:2022  |g number:5  |g day:08  |g month:11  |g pages:233-237 
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