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2.
Biosens Bioelectron ; 18(5-6): 547-53, 2003 May.
Article in English | MEDLINE | ID: mdl-12706561

ABSTRACT

A novel capacitance biosensor based on synthetic phytochelatins for sensitive detection of heavy metals is described. Synthetic phytochelatin (Glu-Cys)(20)Gly (EC20) fused to the maltose binding domain protein was expressed in Escherichia coli and purified for construction of the biosensor. The new biosensor was able to detect Hg(2+), Cd(2+), Pb(2+), Cu(2+) and Zn(2+) ions in concentration range of 100 fM-10 mM, and the order of sensitivity was S(Zn)>S(Cu)>S(Hg)>>S(Cd) congruent with S(Pb). The biological sensing element of the sensor could be regenerated using EDTA and the storage stability of the biosensor was 15 days.


Subject(s)
Biosensing Techniques , Electrochemistry/instrumentation , Equipment Failure Analysis , Metalloproteins/biosynthesis , Metalloproteins/chemistry , Metals, Heavy/analysis , Metals, Heavy/chemistry , Electric Capacitance , Electrochemistry/methods , Equipment Design , Escherichia coli/genetics , Escherichia coli/metabolism , Glutathione , Metalloproteins/genetics , Microchemistry/instrumentation , Microchemistry/methods , Phytochelatins , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/chemistry , Reproducibility of Results , Sensitivity and Specificity
3.
Biomacromolecules ; 3(3): 462-5, 2002.
Article in English | MEDLINE | ID: mdl-12005515

ABSTRACT

Heavy metals are major contributors to pollution of the biosphere, and their efficient removal from contaminated water is required. Biosorption is an emerging technology that has been shown to be effective in removing very low levels of heavy metal from wastewater. Although peptides such as metallothioneins or phytotchelatins are known to immobilize heavy metals, peptide-based biosorbents have not been extensively investigated. In this paper, we describe the construction and expression of bifunctional fusion proteins consisting of synthetic phytochelatin (EC20) linked to a Clostridium-derived cellulose-binding domain (CBD(clos)), enabling purification and immobilization of the fusions onto different cellulose materials in essentially a single step. The immobilized sorbents were shown to be highly effective in removing cadmium at parts per million levels. Repeated removal of cadmium was demonstrated in an immobilized column. The ability to genetically engineer biosorbents with precisely defined properties could provide an attractive strategy for developing high-affinity bioadsorbents suitable for heavy metal removal.


Subject(s)
Bacterial Proteins/metabolism , Carrier Proteins/metabolism , Metalloproteins/metabolism , Metals, Heavy/isolation & purification , Adsorption , Bacterial Proteins/genetics , Cadmium/isolation & purification , Cadmium/pharmacokinetics , Carrier Proteins/genetics , Cellulose/metabolism , Chelating Agents , Clostridium/chemistry , Glutathione , Metalloproteins/genetics , Metals, Heavy/pharmacokinetics , Phytochelatins , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Water Pollutants/isolation & purification , Water Pollutants/pharmacokinetics
4.
J Colloid Interface Sci ; 240(2): 525-532, 2001 Aug 15.
Article in English | MEDLINE | ID: mdl-11482961

ABSTRACT

Photocatalytic degradation of water pollutants using nanometersized semiconductor colloids is an emerging area of environmental remediation. The synthesis of semiconductor nanocrystals (NCs), however, can be costly and result in low product yields. For large-scale photocatalytic application in environmental remediation, cost-effective production of the semiconductor NCs would be ideal. Demonstrated in this report is the efficient photocatalytic degradation of p-nitrophenol (pNP) and Acid Orange 7 (AO7) using ZnS nanocrystals ( approximately 3 to 5 nm diameter) produced in gram quantities with >50% product yield. The pNP half-life in ZnS nanocrystal photocatalyzed reactions was about 1.95 to 2.45 min, whereas in comparable TiO(2) reactions, the pNP half-lives were in the range of 12 to 15 min. Absorption spectra of the photocatalysis reactions suggested the decolorization of pNP without any noticeable formation of phenolic intermediates, implying a mechanism that involves a pNP ring opening via a radical mediated attack. Likewise, the degradation of AO7 was suggested to occur via an oxidative pathway involving hydroxyl radicals formed at the photocatalyst/liquid interface. Optimum conditions for AO7 degradation such as pH, photocatalyst-to-AO7 ratio, and photocatalyst surface passivation were similar to those for pNP. By demonstrating efficient mineralization of these model pollutants using mass-produced ZnS nanocrystals, we hope to lay the foundations necessary for development of large-scale, field-applicable systems. Copyright 2001 Academic Press.

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