Simultaneous detection of analytes based on genetically engineered whole cell sensing systems

Suresh Shrestha, Ranjit S. Shetty, Sridhar Ramanathan, Sylvia Daunert

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Sensing systems for the simultaneous detection of analytes utilizing whole cells provide efficient ways to develop analytical assay systems with reduced cost. In this work, a whole cell-based sensing system was developed for the simultaneous detection of two model analytes, β-lactose and L-arabinose, using genetically-designed bacteria. Two variants of the green fluorescent protein (GFP), BFP2 and GFPuv, were used as the reporter protein for the detection of each analyte. The corresponding reporter genes were introduced into the bacteria in such a way that they can be co-expressed along with the other genes induced by the respective sugar. Each of these fluorescent proteins is expressed only in the presence of their respective analyte and their fluorescence emission can be monitored using the intact whole cell samples. By exciting the cells at 380 nm, emission for BFP2 and GFPuv can be collected at 440 and 509 nm, respectively. The fluorescence emission thus obtained can be correlated with the amount of sugars present in the sample. Calibration curves for β-lactose and L-arabinose were generated. It was observed that this system shows no significant response to other closely related sugars thus providing high selectivity for β-lactose and L-arabinose detection.

Original languageEnglish
Pages (from-to)251-260
Number of pages10
JournalAnalytica Chimica Acta
Volume444
Issue number2
DOIs
StatePublished - Oct 18 2001
Externally publishedYes

Fingerprint

Arabinose
Lactose
Sugars
sugar
Bacteria
Genes
Fluorescence
protein
fluorescence
Green Fluorescent Proteins
bacterium
gene
Assays
Proteins
Reporter Genes
Calibration
assay
calibration
Costs and Cost Analysis
detection

Keywords

  • Detection of analytes
  • Genetically-designed bacteria
  • Model analytes

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Spectroscopy
  • Environmental Chemistry

Cite this

Simultaneous detection of analytes based on genetically engineered whole cell sensing systems. / Shrestha, Suresh; Shetty, Ranjit S.; Ramanathan, Sridhar; Daunert, Sylvia.

In: Analytica Chimica Acta, Vol. 444, No. 2, 18.10.2001, p. 251-260.

Research output: Contribution to journalArticle

Shrestha, Suresh ; Shetty, Ranjit S. ; Ramanathan, Sridhar ; Daunert, Sylvia. / Simultaneous detection of analytes based on genetically engineered whole cell sensing systems. In: Analytica Chimica Acta. 2001 ; Vol. 444, No. 2. pp. 251-260.
@article{451c6160d37e4fd8a632f43a47979cef,
title = "Simultaneous detection of analytes based on genetically engineered whole cell sensing systems",
abstract = "Sensing systems for the simultaneous detection of analytes utilizing whole cells provide efficient ways to develop analytical assay systems with reduced cost. In this work, a whole cell-based sensing system was developed for the simultaneous detection of two model analytes, β-lactose and L-arabinose, using genetically-designed bacteria. Two variants of the green fluorescent protein (GFP), BFP2 and GFPuv, were used as the reporter protein for the detection of each analyte. The corresponding reporter genes were introduced into the bacteria in such a way that they can be co-expressed along with the other genes induced by the respective sugar. Each of these fluorescent proteins is expressed only in the presence of their respective analyte and their fluorescence emission can be monitored using the intact whole cell samples. By exciting the cells at 380 nm, emission for BFP2 and GFPuv can be collected at 440 and 509 nm, respectively. The fluorescence emission thus obtained can be correlated with the amount of sugars present in the sample. Calibration curves for β-lactose and L-arabinose were generated. It was observed that this system shows no significant response to other closely related sugars thus providing high selectivity for β-lactose and L-arabinose detection.",
keywords = "Detection of analytes, Genetically-designed bacteria, Model analytes",
author = "Suresh Shrestha and Shetty, {Ranjit S.} and Sridhar Ramanathan and Sylvia Daunert",
year = "2001",
month = "10",
day = "18",
doi = "10.1016/S0003-2670(01)01214-4",
language = "English",
volume = "444",
pages = "251--260",
journal = "Analytica Chimica Acta",
issn = "0003-2670",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Simultaneous detection of analytes based on genetically engineered whole cell sensing systems

AU - Shrestha, Suresh

AU - Shetty, Ranjit S.

AU - Ramanathan, Sridhar

AU - Daunert, Sylvia

PY - 2001/10/18

Y1 - 2001/10/18

N2 - Sensing systems for the simultaneous detection of analytes utilizing whole cells provide efficient ways to develop analytical assay systems with reduced cost. In this work, a whole cell-based sensing system was developed for the simultaneous detection of two model analytes, β-lactose and L-arabinose, using genetically-designed bacteria. Two variants of the green fluorescent protein (GFP), BFP2 and GFPuv, were used as the reporter protein for the detection of each analyte. The corresponding reporter genes were introduced into the bacteria in such a way that they can be co-expressed along with the other genes induced by the respective sugar. Each of these fluorescent proteins is expressed only in the presence of their respective analyte and their fluorescence emission can be monitored using the intact whole cell samples. By exciting the cells at 380 nm, emission for BFP2 and GFPuv can be collected at 440 and 509 nm, respectively. The fluorescence emission thus obtained can be correlated with the amount of sugars present in the sample. Calibration curves for β-lactose and L-arabinose were generated. It was observed that this system shows no significant response to other closely related sugars thus providing high selectivity for β-lactose and L-arabinose detection.

AB - Sensing systems for the simultaneous detection of analytes utilizing whole cells provide efficient ways to develop analytical assay systems with reduced cost. In this work, a whole cell-based sensing system was developed for the simultaneous detection of two model analytes, β-lactose and L-arabinose, using genetically-designed bacteria. Two variants of the green fluorescent protein (GFP), BFP2 and GFPuv, were used as the reporter protein for the detection of each analyte. The corresponding reporter genes were introduced into the bacteria in such a way that they can be co-expressed along with the other genes induced by the respective sugar. Each of these fluorescent proteins is expressed only in the presence of their respective analyte and their fluorescence emission can be monitored using the intact whole cell samples. By exciting the cells at 380 nm, emission for BFP2 and GFPuv can be collected at 440 and 509 nm, respectively. The fluorescence emission thus obtained can be correlated with the amount of sugars present in the sample. Calibration curves for β-lactose and L-arabinose were generated. It was observed that this system shows no significant response to other closely related sugars thus providing high selectivity for β-lactose and L-arabinose detection.

KW - Detection of analytes

KW - Genetically-designed bacteria

KW - Model analytes

UR - http://www.scopus.com/inward/record.url?scp=0035909434&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035909434&partnerID=8YFLogxK

U2 - 10.1016/S0003-2670(01)01214-4

DO - 10.1016/S0003-2670(01)01214-4

M3 - Article

AN - SCOPUS:0035909434

VL - 444

SP - 251

EP - 260

JO - Analytica Chimica Acta

JF - Analytica Chimica Acta

SN - 0003-2670

IS - 2

ER -