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Design, Development and Applications of Highly Sensitive Protein-Based Capacitive Biosensors

Bontidean, Ibolya LU (2002)
Abstract
Highly sensitive biosensors were developed for environmental and medical applications. The biosensors were based on bioengineered proteins as biorecognition elements coupled to a highly sensitive capacitive transducer.



Heavy metal binding proteins like the metallothionein SmtA, regulatory protein MerR, periplasmic protein MerP and the synthetic phytochelatin EC20 were used for designing, constructing, and characterising biosensors for the determination of various heavy metal ions, i.e. mercury, copper, cadmium, zinc and lead, in a wide concentration range (fM-mM). The developed heavy metal biosensors had a useful storage stability (about two weeks) and could be regenerated using EDTA. The developed and optimised... (More)
Highly sensitive biosensors were developed for environmental and medical applications. The biosensors were based on bioengineered proteins as biorecognition elements coupled to a highly sensitive capacitive transducer.



Heavy metal binding proteins like the metallothionein SmtA, regulatory protein MerR, periplasmic protein MerP and the synthetic phytochelatin EC20 were used for designing, constructing, and characterising biosensors for the determination of various heavy metal ions, i.e. mercury, copper, cadmium, zinc and lead, in a wide concentration range (fM-mM). The developed heavy metal biosensors had a useful storage stability (about two weeks) and could be regenerated using EDTA. The developed and optimised biosensors were assessed in two practical applications, namely monitoring mercury ions in contaminated soil samples, and measuring inducer molecules.



The SmtA based biosensor was successfully used to monitor heavy metals in soil samples originating from a contaminated agricultural site in Denmark. The obtained results were compared to those given by a bacterial biosensor, a plant sensor, and the total amount of mercury contained in the soil. Both the protein-based and the bacterial biosensor, proved to accurately monitor the ion content, giving results well in agreement with those shown by the total amount.



By using the same sensing principle, biosensors based on lac repressor protein were developed for monitoring of different inducer molecules, e.g. IPTG, ONPG, and lactose, or DNA. The biosensor could differentiate between lac operator DNA contained in linearized plasmid DNA and circular plasmid DNA, and genomic DNA. (Less)
Please use this url to cite or link to this publication:
author
supervisor
opponent
  • Prof., Belkin, Shimshon, The Fredy and Nadine Hermann Graduate School of Applied Science, The Hebrew University of Jerusalem, 91904-Jerusalem, Israel
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Bioteknik, Biotechnology, Contaminated soil, Heavy metals, Lac repressor, Phytochelatin, MerP, MerR, SmtA, Self-assembled monolayer, Biosensor, Capacitance
pages
174 pages
publisher
Ibolya Bontidean, Department of Biotechnology, Lund University
defense location
Center for Chemistry and Chemical Engineering, Lecture Hall C, Getingevägen 60, 221 00 Lund
defense date
2002-09-13 10:15:00
external identifiers
  • other:ISRN: LUTKDH/TKBT—02/1062--SE
ISBN
91-89627-10-5
language
English
LU publication?
yes
additional info
Article: Bontidean, I., Berggren, C., Johansson, G., Csöregi, E., Mattiasson, B., Lloyd, J.R., Jakeman K.J., and Brown, N.L. (1998) Detection of heavy metal ions at femtomolar levels using protein-based biosensors, Anal. Chem., 70, 4162-4169. Article: Bontidean, I., Loyd, J.R., Hobman, J.L., Wilson, J.R., Csöregi, E., Mattiasson B., and Brown, N.L. (2000) Bacterial metal resistance proteins and their use in biosensors for the detection of bioavailable heavy metals, J. Inorg. Biochem., 79, 225-229. Article: Bontidean, I., Ahlqvist, J., Mulchandani, A., Chen, W., Bae, W., Mehra, R.K., Mortari, A., and Csöregi, E. (2002) Novel synthetic phytochelatin-based biosensor for monitoring of heavy metal ions, Biosens. Bioelectron., accepted. Article: Brown, N.L., Lloyd, J.R., Jakeman, K.J., Hobman, J.L., Bontidean, I., Mattiasson, B., and Csöregi, E. (1998) Heavy metal resistance genes and proteins in bacteria and their application, Biochem. Soc. Transactions, 26, 662-665. Article: Bontidean, I., Mortari, A., Brown, N.L., Karlson, U., Vangronsveld, J., Corbisier, P., and Csöregi, E. (2002) Biosensors for detection of mercury in contaminated soils, Environ. Pollut., submitted. Article: Bontidean, I., Kumar, A., Csöregi, E., Galaev, I.Y., and Mattiasson, B. (2001) Highly sensitive novel biosensor based on an immobilized lac repressor, Angewandte Chem. Int. Ed., 40, 2676-2678. Article: List of annexes Article: Bontidean, I., Mattiasson, B., Brown, N.L., Lloyd, J.R., Hobman, J.L., and Csöregi, E. (2000) Study of bacterial metal resistance protein-based sensitive biosensors for heavy metal monitoring, in: Chemical and Biological Sensors for Environmental Monitoring, (A. Mulchandani and O. Sadik, Eds.), ACS Symp. Series, 762, pp. 102-112. Article: Bontidean, I., Csöregi, E., Corbisier, P., Lloyd, J.R., and Brown, N.L., (2002) Bacterial metal-responsive elements and their use in biosensors for monitoring of heavy metals, in: Heavy Metals in the Environment, (B. Sankar, Ed.), Marcel Dekker Inc., New York, pp 647-680.
id
975e4d22-d0a4-4ddf-908a-7bfd7ee9982c (old id 464837)
date added to LUP
2016-04-04 11:33:07
date last changed
2018-11-21 21:05:36
@phdthesis{975e4d22-d0a4-4ddf-908a-7bfd7ee9982c,
  abstract     = {Highly sensitive biosensors were developed for environmental and medical applications. The biosensors were based on bioengineered proteins as biorecognition elements coupled to a highly sensitive capacitive transducer.<br/><br>
<br/><br>
Heavy metal binding proteins like the metallothionein SmtA, regulatory protein MerR, periplasmic protein MerP and the synthetic phytochelatin EC20 were used for designing, constructing, and characterising biosensors for the determination of various heavy metal ions, i.e. mercury, copper, cadmium, zinc and lead, in a wide concentration range (fM-mM). The developed heavy metal biosensors had a useful storage stability (about two weeks) and could be regenerated using EDTA. The developed and optimised biosensors were assessed in two practical applications, namely monitoring mercury ions in contaminated soil samples, and measuring inducer molecules.<br/><br>
<br/><br>
The SmtA based biosensor was successfully used to monitor heavy metals in soil samples originating from a contaminated agricultural site in Denmark. The obtained results were compared to those given by a bacterial biosensor, a plant sensor, and the total amount of mercury contained in the soil. Both the protein-based and the bacterial biosensor, proved to accurately monitor the ion content, giving results well in agreement with those shown by the total amount.<br/><br>
<br/><br>
By using the same sensing principle, biosensors based on lac repressor protein were developed for monitoring of different inducer molecules, e.g. IPTG, ONPG, and lactose, or DNA. The biosensor could differentiate between lac operator DNA contained in linearized plasmid DNA and circular plasmid DNA, and genomic DNA.},
  author       = {Bontidean, Ibolya},
  isbn         = {91-89627-10-5},
  language     = {eng},
  publisher    = {Ibolya Bontidean, Department of Biotechnology, Lund University},
  school       = {Lund University},
  title        = {Design, Development and Applications of Highly Sensitive Protein-Based Capacitive Biosensors},
  year         = {2002},
}