Cell surface display of synthetic phytochelatins using ice nucleation protein for enhanced heavy metal bioaccumulation.

Synthetic phytochelatins (ECs) composed of (Glu-Cys)nGly are protein analogs of phytochelatin that exhibit improved metal-binding capacity over metallothioneins (MTs). Expression of EC20 on the surface of E. coli using the Lpp-OmpA anchor resulted in improved bioaccumulation of cadmium and mercury, providing a new method for treating heavy metal contamination. To further improve the whole-cell accumulation of heavy metals, EC20 was expressed on the surface of Moraxella sp., a bacterium known to survive in contaminated environments, using the truncated ice nucleation protein (INPNC) anchor. Production of EC20 was approximately three-fold higher in Moraxella sp. than E. coli. As a consequence, the mercury-binding capacity of the recombinant Moraxella sp. was increased by more than 10-fold. Owing to the very high level of surface expression, the use of Moraxella sp. and INPNC anchor may prove to be useful for the remediation of other environmental contaminants.

[1]  P. Wolber Bacterial ice nucleation. , 1993, Advances in microbial physiology.

[2]  G. Gadd,et al.  Microbial treatment of metal pollution--a working biotechnology? , 1993, Trends in biotechnology.

[3]  D. Wilson,et al.  Construction and characterization of Escherichia coli genetically engineered for bioremediation of Hg(2+)-contaminated environments , 1997, Applied and environmental microbiology.

[4]  A Mulchandani,et al.  Cell Surface Display of Organophosphorus Hydrolase Using Ice Nucleation Protein , 2001, Biotechnology progress.

[5]  W. Bae,et al.  Metal-binding characteristics of a phytochelatin analog (Glu-Cys)2Gly , 1997 .

[6]  D. Gibson,et al.  Enzymatic oxidation of p-nitrophenol. , 1979, Biochemical and biophysical research communications.

[7]  Ashok Mulchandani,et al.  Biodegradation of organophosphorus pesticides by surface-expressed organophosphorus hydrolase , 1997, Nature Biotechnology.

[8]  M. Zenk Heavy metal detoxification in higher plants--a review. , 1996, Gene.

[9]  A. Mulchandani,et al.  Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins. , 2000, Biotechnology and bioengineering.

[10]  V. de Lorenzo,et al.  Analysis of Pseudomonas gene products using lacIq/Ptrp-lac plasmids and transposons that confer conditional phenotypes. , 1993, Gene.

[11]  V. de Lorenzo,et al.  Metalloadsorption by Escherichia coliCells Displaying Yeast and Mammalian Metallothioneins Anchored to the Outer Membrane Protein LamB , 1998, Journal of bacteriology.

[12]  Y. Kwak,et al.  Cell Surface Display of Human Immunodeficiency Virus Type 1 gp120 on Escherichia coli by Using Ice Nucleation Protein , 1999, Clinical Diagnostic Laboratory Immunology.

[13]  J. Nriagu,et al.  Quantitative assessment of worldwide contamination of air, water and soils by trace metals , 1988, Nature.

[14]  B. Gaber,et al.  Expression of the Neurospora crassa metallothionein gene in Escherichia coli and its effect on heavy-metal uptake , 1995, Applied Microbiology and Biotechnology.

[15]  Andrew G. Glen,et al.  APPL , 2001 .

[16]  R. Mehra,et al.  Metal ion resistance in fungi: Molecular mechanisms and their regulated expression , 1991, Journal of cellular biochemistry.

[17]  Chul-Joong Kim,et al.  Surface-displayed viral antigens on Salmonella carrier vaccine , 2000, Nature Biotechnology.