Document Detail


A single-use luciferase-based mercury biosensor using Escherichia coli HB101 immobilized in a latex copolymer film.
MedLine Citation:
PMID:  10455499     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A single-use Hg(II) patch biosensor has been developed consisting of 1.25-cm diameter patches of two acrylic vinyl acetate copolymer layers coated on polyester. The top layer copolymer was 47 microm thick whereas the bottom layer of copolymer plus E. coli cells was 30 microm thick. The immobilized E. coli HB101 cells harbored a mer-lux plasmid construct and produced a detectable light signal when exposed to Hg(II). The immobilized-cell Hg(II) biosensor had a sensitivity similar to that of suspended cells but a significantly larger detection range. The levels of mercury detected by the patches ranged from 0.1 nM to 10 000 nM HgCl2 in pyruvate buffer, and luciferase induction as a function of Hg(II) concentration was sigmoidal. Luciferase activity was detected in immobilized cells for more than 78 h after exposure of the cells to HgCl2. Addition of 1 mM D-cysteine to the pyruvate buffer increased luciferase induction more than 100-fold in the immobilized cell patches and 3.5-fold in a comparable suspension culture. The copolymer patches with immobilized cells were stable at -20 degrees C for at least 3 months, and the Hg(II)-induced luciferase activity after storage was similar to that of samples assayed immediately after coating. Patches stored desiccated at room temperature for 2 weeks showed lower mercury-induced luciferase activity when compared to freshly prepared patches, but they still had a considerable detection range of 1 to 10 000 nM HgCl2.
Authors:
O K Lyngberg; D J Stemke; J L Schottel; M C Flickinger
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of industrial microbiology & biotechnology     Volume:  23     ISSN:  1367-5435     ISO Abbreviation:  J. Ind. Microbiol. Biotechnol.     Publication Date:  1999 Jul 
Date Detail:
Created Date:  2002-04-11     Completed Date:  2002-05-09     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9705544     Medline TA:  J Ind Microbiol Biotechnol     Country:  England    
Other Details:
Languages:  eng     Pagination:  668-76     Citation Subset:  IM    
Affiliation:
Biological Process Technology Institute, University of Minnesota, St Paul, MN 55108, USA.
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MeSH Terms
Descriptor/Qualifier:
Biosensing Techniques / methods*
Cysteine / pharmacology
Escherichia coli / drug effects*,  enzymology
Kinetics
Luciferases / analysis*,  metabolism
Mercuric Chloride / analysis*,  pharmacology
Microspheres
Plasmids
Polyvinyls
Grant Support
ID/Acronym/Agency:
T32GM08347/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Polyvinyls; 0/pioloform; 52-90-4/Cysteine; 7487-94-7/Mercuric Chloride; EC 1.13.12.-/Luciferases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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