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PubMed2000

Can we produce a human corneal equivalent by tissue engineering?

Tissue engineering is progressing rapidly. Bioengineered substitutes are already available for experimental applications and some clinical purposes such as skin replacement. This review focuses on the development of reconstructed human cornea in vitro by tissue engineering. Key elements to consider in the corneal reconstruction, such as the source for epithelial cells and keratocytes, are discussed and the various steps of production are presented. Since one application of this human model is to obtain a better understanding of corneal wound healing, the mechanisms of this phenomenon as well as the function played both by membrane-bound integrins and components from the extracellular matrix have also been addressed. The analysis of integrins by immunohistofluorescence labelling of our reconstructed human cornea revealed that beta(1), alpha(3), alpha(5), and alpha(6) integrin subunits were expressed but alpha(4) was not. Laminin, type VII collagen and fibronectin were also detected. Finally, the future challenges of corneal reconstruction by tissue engineering are discussed and the tremendous applications of such tissue produced in vitro for experimental as well as clinical purposes are considered.

Biomedical Engineering↗
PubMedDate not supplied

Cell-based tissue engineering for the intervertebral disc: in vitro studies of human disc cell gene expression and matrix production within selected cell carriers.

BACKGROUND CONTEXT: Little is known about how disc cells attach, proliferate and form extracellular matrix (ECM) within carrier materials. Such information is needed to help formulate criteria for successful cell-carrier interactions in tissue engineering. PURPOSE: To compare proliferation, ECM production and gene expression in annulus cells cultured in a variety of cell carrier materials with potential application in tissue engineering of the disc. STUDY DESIGN: Human intervertebral disc cells from the annulus were used in a prospective study of proliferation, ECM production and gene expression within selected cell carriers. METHODS: Annulus cells from discs of 29 individuals were tested in collagen sponge, collagen gel, agarose, alginate or fibrin gel formulations. In situ hybridization assessed ECM gene expression of Types I and II collagen, aggrecan and chondroitin-6 sulfotransferase. Cell proliferation, cell shape, attachment and ECM production were evaluated. RESULTS: Collagen sponges provided the best microenvironment for disc cell ECM production and gene expression. Although collagen gels often could support good cell growth, such constructs did not result in either abundant ECM production or ECM gene expression, as shown by in situ hybridization. Growth and ECM production and gene expression in alginate, agarose and fibrin microenvironments were inferior. CONCLUSIONS: Tissue engineering techniques open new therapeutic possibilities for use of autologous disc cells, but fundamental questions on how these cells interact with cell carriers are unexplored. Results provide novel data on disc cell gene expression within diverse microenvironments. The collagen sponge proved to be a superior microenvironment.

Absorbable Implants↗
PubMed2006

Tissue engineering: current and future approaches to ocular surface reconstruction.

Although cells have been cultured outside the body for many years, research has only recently begun to develop complex three-dimensional tissue constructs that will, ideally, mature into fully functional tissues and organs. Tissue engineering is an emerging field in the area of biotechnology that combines the principles and methods of life sciences with those of engineering for the purpose of regenerating, repairing, or replacing diseased tissues. In this review, we describe the recent advances and current development of tissue engineering approaches as related to the ocular surface system, which comprises the three main integrated tissue units: conjunctiva, cornea and lacrimal glands.

Animals↗
PubMed2001

Enhanced fluorescence resonance energy transfer between spectral variants of green fluorescent protein through zinc-site engineering.

Although spectral variants of GFP should in theory be suited for fluorescence resonance energy transfer (FRET) and therefore suited for studies of protein-protein interactions, the unfavorable location of the fluorophore 15 A deep inside the GFP molecule has especially impaired this application. Here, metal-ion site engineering around the dimerization interface known from the X-ray structure of GFP is applied to the cyan and the yellow spectral variant of GFP to stabilize the heterodimeric form of these molecules and thereby increase FRET signaling. The FRET signal, determined as the ratio between the maximal emission for the yellow variant, 530 nm, and the cyan variant, 475 nm, during excitation of the cyan variant at 433 nm was increased up to 8-10-fold in the presence of 10(-4) M ZnCl2 by engineering of two symmetric metal-ion sites being either bidentate or tridentate. A similar increase in FRET signaling was however obtained in a pair of molecules in which a single bidentate metal-ion site was generated by introducing a zinc-binding residue in each of the two spectral variants of GFP and therefore creating an obligate heterodimeric pair. It is concluded that FRET signaling between spectral variants of GFP can be increased by stabilizing dimer formation and especially by favoring heterodimer formation in this case performed by metal-ion site engineering.

Amino Acid Sequence↗
PubMed2005

Structure-based engineering of internal cavities in coiled-coil peptides.

Cavities and clefts are frequently important sites of interaction between natural enzymes or receptors and their corresponding substrate or ligand molecules and exemplify the types of molecular surfaces that would facilitate engineering of artificial catalysts and receptors. Even so, structural characterizations of designed cavities are rare. To address this issue, we performed a systematic study of the structural effects of single-amino acid substitutions within the hydrophobic cores of tetrameric coiled-coil peptides. Peptides containing single glycine, serine, alanine, or threonine amino acid substitutions at the buried L9, L16, L23, and I26 hydrophobic core positions of a GCN4-based sequence were synthesized and studied by solution-phase and crystallographic techniques. All peptides adopt the expected tetrameric state and contain tunnels or internal cavities ranging in size from 80 to 370 A(3). Two closely related sequences containing an L16G substitution, one of which adopts an antiparallel configuration and one of which adopts a parallel configuration, illustrate that cavities of different volumes and shapes can be engineered from identical core substitutions. Finally, we demonstrate that two of the peptides (L9G and L9A) bind the small molecule iodobenzene when present during crystallization, leaving the general peptide quaternary structure intact but altering the local peptide conformation and certain superhelical parameters. These high-resolution descriptions of varied molecular surfaces within solvent-occluded internal cavities illustrate the breadth of design space available in even closely related peptides and offer valuable models for the engineering of de novo helical proteins.

Amino Acid Sequence↗
PubMed1996

Activation of myosin light chain kinase and nitric oxide synthase activities by engineered calmodulins with duplicated or exchanged EF hand pairs.

We have constructed three engineered calmodulins (CaMs) in which the two EF hand pairs have been substituted for one another or exchanged: CaMNN, the C-terminal EF hand pair (residues 82-148) has been replaced by a duplication of the N-terminal pair (residues 9-75); CaMCC, the N-terminal pair has been replaced by a duplication of the C-terminal pair; CaMCN, the two EF had pairs have been exchanged. Skeletal muscle myosin light chain kinase (skMLCK) activity is activated to 75% of the maximum level by CaMCC and to 45% of the maximum level by CaMCN and is not significantly activated by CaMNN; Kact or Ki values for the engineered CaMs are 2-3.5 nM. Smooth muscle myosin light chain kinase activity (gMLCK) is fully activated by CaMCN and is not significantly activated by either CaMNN or CaMCC; the Kact value for CaMCN is 2 nM and the Ki values for CaMNN and CaMCC are 10 and 40 nM, respectively. Cerebellar nitric oxide synthase activity (nNOS) is fully activated by CaMNN and CaMCN and is not significantly activated by CaMCC; the engineered CaMs have Kact or Ki values for this enzyme activity of 2-8 nM. These results indicate that the EF hand pairs contain distinct but overlapping sets of determinants for binding and activation of enzymes, with the greater degree of overlap in determinants for binding. Furthermore, while the structural changes associated with swapping the EF hand pairs do not affect activation of nNOS or gMLCK activities, they significantly reduce activation of skMLCK activity, indicating that this process requires specific determinants in CaM outside the EF hand pairs.

Animals↗
PubMed1997

Protein engineering of the HMG-CoA reductase of Pseudomonas mevalonii. Construction of mutant enzymes whose activity is regulated by phosphorylation and dephosphorylation.

The activity of Pseudomonas mevalonii HMG-CoA reductase (EC 1.1.1.88) is not regulated by phosphorylation, presumably due to the absence of a suitable target serine and protein kinase recognition motif. We have engineered P. mevalonii HMG-CoA reductase to a form whose activity, like that of mammalian HMG-CoA reductases, is regulated by phosphorylation/dephosphorylation. We substituted serine for arginine 387, the residue that corresponds to the regulatory serine of the HMG-CoA reductases of higher eukaryotes. A recognition motif for cAMP-dependent protein kinase was added by replacing leucine 384 by histidine (enzyme L384H/R387S) and also valine 391 by leucine (enzyme L384H/R387S/V391L). The activity of P. mevalonii HMG-CoA reductase mutant enzymes L384H/R387S and L384H/R387S/V391L was attenuated by phosphorylation. Restoration of activity accompanied subsequent dephosphorylation catalyzed by lambda protein phosphatase. Incorporation and subsequent release of phosphate paralleled the attenuation and restoration of catalytic activity. Incorporation of 0.5 mol of phosphate per subunit was accompanied by an approximately 50% decrease in initial activity. As in the analogous Syrian hamster mutant enzyme S871D, P. mevalonii mutant enzyme R387D exhibited 10% wild-type activity, suggesting that the attenuation of activity that accompanies phosphorylation results at least in part from the introduction of negative charge. Engineering of P. mevalonii HMG-CoA reductase to forms whose activity is reversibly regulated by phosphorylation/dephosphorylation provides an attractive model for future structure-based mechanistic studies. Solution of the X-ray structure of phosphorylated and dephosphorylated forms of engineered P. mevalonii HMG-CoA reductase should then reveal interactions of the active site phosphoseryl residue that result in attenuation of catalytic activity.

Animals↗
PubMed1999

The effects of an engineered cation site on the structure, activity, and EPR properties of cytochrome c peroxidase.

Earlier work [Bonagura et al. (1996) Biochemistry 35, 6107] showed that the K+ site found in the proximal pocket of ascorbate peroxidase (APX) could be engineered into cytochrome c peroxidase (CCP). Binding of K+ at the engineered site results in a loss in activity and destabilization of the CCP compound I Trp191 cationic radical owing to long-range electrostatic effects. The engineered CCP mutant crystal structure has been refined to 1.5 A using data obtained at cryogenic temperatures which provides a more detailed basis for comparison with the naturally occurring K+ site in APX. The characteristic EPR signal associated with the Trp191 radical becomes progressively weaker as K+ is added, which correlates well with the loss in enzyme activity as [K+] is increased. These results coupled with stopped-flow studies support our earlier conclusions that the loss in activity and EPR signal is due to destabilization of the Trp191 cationic radical.

Binding Sites↗
PubMedDate not supplied

Concentric cylinder bioreactor for production of tissue engineered cartilage: effect of seeding density and hydrodynamic loading on construct development.

A concentric cylinder bioreactor has been developed to culture tissue engineered cartilage constructs under hydrodynamic loading. This bioreactor operates in a low shear stress environment, has a large growth area for construct production, allows for dynamic seeding of constructs, and provides for a uniform loading environment. Porous poly-lactic acid constructs, seeded dynamically in the bioreactor using isolated bovine chondrocytes, were cultured for 4 weeks at three seeding densities (60, 80, 100 x 10(6) cells per bioreactor) and three different shear stresses (imposed at 19, 38, and 76 rpm) to characterize the effect of chondrocyte density and hydrodynamic loading on construct growth. Construct seeding efficiency with chondrocytes is greater than 95% within 24 h. Extensive chondrocyte proliferation and matrix deposition are achieved so that after 28 days in culture, constructs from bioreactors seeded at the highest cell densities contain up to 15 x 10(6) cells, 2 mg GAG, and 3.5 mg collagen per construct and exhibit morphology similar to that of native cartilage. Bioreactors seeded with 60 million chondrocytes do not exhibit robust proliferation or matrix deposition and do not achieve morphology similar to that of native cartilage. In cultures under different steady hydrodynamic loading, the data demonstrate that higher shear stress suppresses matrix GAG deposition and encourages collagen incorporation. In contrast, under dynamic hydrodynamic loading conditions, cartilage constructs exhibit robust matrix collagen and GAG deposition. The data demonstrate that the concentric cylinder bioreactor provides a favorable hydrodynamic environment for cartilage construct growth and differentiation. Notably, construct matrix accumulation can be manipulated by hydrodynamic loading. This bioreactor is useful for fundamental studies of construct growth and to assess the significance of cell density, nutrients, and hydrodynamic loading on cartilage development. In addition, studies of cartilage tissue engineering in the well-characterized, uniform environment of the concentric cylinder bioreactor will develop important knowledge of bioprocessing parameters critical for large-scale production of engineered tissues.

Adaptation, Physiological↗
PubMedDate not supplied

Problem-based learning biotechnology courses in chemical engineering.

We have developed a series of upper undergraduate/graduate lecture and laboratory courses on biotechnological topics to supplement existing biochemical engineering, bioseparations, and biomedical engineering lecture courses. The laboratory courses are based on problem-based learning techniques, featuring two- and three-person teams, journaling, and performance rubrics for guidance and assessment. Participants initially have found them to be difficult, since they had little experience with problem-based learning. To increase enrollment, we are combining the laboratory courses into 2-credit groupings and allowing students to substitute one of them for the second of our 2-credit chemical engineering unit operations laboratory courses.

Biotechnology↗
PubMedDate not supplied

Development of technologies aiding large-tissue engineering.

There are many clinical situations in which a large tissue mass is required to replace tissue lost to surgical resection (e.g., mastectomy). It is possible that autologous cell transplantation on biodegradable polymer matrices may provide a new therapy to engineer large tissue which can be used to treat these patients. A number of challenges must be met to engineer a large soft tissue mass. These include the design of (1) a structural framework to maintain a space for tissue development, (2) a space-filling matrix which provides for localization of transplanted cells, and (3) a strategy to enhance vascularization of the forming tissue. In this paper we provide an overview of several technologies which are under development to address these issues. Specifically, support matrices to maintain a space for tissue development have been fabricated from polymers of lactide and glycolide. The ability of these structures to resist compressive forces was regulated by the ratio of lactide to glycolide in the polymer. Smooth muscle cell seeding onto polyglycolide fiber-based matrices has been optimized to allow formation of new tissues in vitro and in vivo. Finally, polymer microsphere drug delivery technology is being developed to release vascular endothelial growth factor (VEGF), a potent angiogenic molecule, at the site of tissue formation. This strategy, which combines several different technologies, may ultimately allow for the engineering of large soft tissues.

Animals↗
PubMedDate not supplied

Hg2+ removal by genetically engineered Escherichia coli in a hollow fiber bioreactor.

Escherichia coli cells engineered to express an Hg2+ transport system and metallothionein accumulated Hg2+ effectively over a concentration range of 0.2-4 mg/L in batch systems. Bioaccumulation was selective against other metal ions and resistant to changes in ambient conditions such as pH, ionic strength, and the presence of common metal chelators or complexing agents (Chen, S.-L.; Wilson, D. B. Appl. Environ. Microbiol. 1997, 63, 2442-2445; Biodegradation 1997, 8, 97-103). Here we report the characterization of the bioaccumulation system based on its kinetics and an isotherm. Bioaccumulation was rapid and followed Michaelis-Menten kinetics. A hollow fiber bioreactor was constructed to retain the genetically engineered cells. The bioreactor was capable of removing and recovering Hg2+ effectively at low concentrations, reducing a 2 mg/L solution to about 5 microgram/L. A mathematical equation that quantitatively described Hg2+ removal by the bioreactor provides a basis for the optimization and extrapolation of the bioreactor. The genetically engineered E. colicells and the bioreactor system have excellent properties for bioremediation of Hg2+-contaminated environments.

Bioreactors↗
PubMedDate not supplied

Computational Science and Engineering Online (CSE-Online): a cyber-infrastructure for scientific computing.

With the expansion of the Internet and World Wide Web (or the Web), research environments have changed dramatically. As a result, the need to be able to efficiently and securely access information and resources from remote computer systems is becoming even more critical. This paper describes the development of an extendable integrated Web-accessible simulation environment for computational science and engineering called Computational Science and Engineering Online (CSE-Online; http://cse-online.net). CSE-Online is based on a unique client-server software architecture that can distribute the workload between the client and server computers in such a way as to minimize the communication between the client and server, thus making the environment less-sensitive to network instability. Furthermore, the new software architecture allows the user to access data and resources on one or more remote servers as well as on the computing grid while having the full capability of the Web-services collaborative environment. It can be accessed anytime and anywhere from a Web browser connected to the network by either a wired or wireless connection. It has different modes of operations to support different working environments and styles. CSE-Online is evolving into middleware that can provide a framework for accessing and managing remote data and resources including the computing grid for any domain, not necessarily just within computational science and engineering.

Engineering↗
PubMed2006

Status of worker exposure to asphalt paving fumes with the use of engineering controls.

Since 1996, industry, labor, and government have partnered to minimize workers' exposure to asphalt fumes using engineering controls. The objective of this study was to determine the use after some years of experience and to benchmark the effectiveness of the engineering controls as compared to the current exposure limits. To accomplish this objective, the current highway class pavers equipped with controls to reduce asphalt fumes, occupational exposure levels, and ventilation flow rates were monitored, and a user acceptance survey was conducted. Personal breathing-zone sampling was administered to determine concentrations of total particulate matter (TPM) and benzene soluble matter (BSM). Personal monitoring of workers yielded a BSM arithmetic mean of 0.13 mg/m3 (95% confidence limits (0.07, 0.43) mg/m3). All site average worker BSM values are below the American Conference of Governmental Industrial Hygienists (ACGIH) adopted threshold limit value (TLV) time weighted average (TWA) of 0.5 mg/m3 as benzene soluble inhalable particulate, although five sites contained 95% confidence limits slightly above the ACGIH TLV. The TPM arithmetic mean was 0.35 mg/m3 (95% confidence limits (0.27, 0.69) mg/m3). All sites showed average worker and area TPM values below NIOSH's recommended exposure limit for asphalt fumes (5 mg/m3, 15 min). One screed area sample and one operator area sample were also taken each day. Area samples followed a similar pattern to the worker breathing zone samples, but were generally slightly higher in TPM and BSM concentration. The effect of work practices and application temperatures appears to have an impact on the ability of the engineering controls to keep exposure below the TLV for BSM. To gain a better understanding of the aerodynamic properties of asphalt fumes, particle size and airborne concentrations were also monitored using a TSI model 3320 aerodynamic particle sizer spectrometer. The geometric mean particle size was between 0.64 and 0.98 micrometers for the worker breathing zone samples, with a geometric mean of 0.73 micrometers for all sites. Total airborne concentrations were typically higher for the asphalt fume exposed groups than for the background samples. During high fume events, four 15-minute samples were taken each day. Only one 15-minute sample was above the limit of quantification. Stack flow rates were measured, and results are discussed and compared to the manufacturers' nominal values. Survey results were generally positive, with recommendations discussed for continuous improvement.

Air Pollutants, Occupational↗
PubMed2025

Engineering Polyketide Stereocenters with Ketoreductase Domain Exchanges.

Polyketide synthases (PKSs) are versatile biosynthetic megasynthases capable of producing a diverse range of natural products with many applications, including in pharmaceuticals. The stereochemical precision of PKSs makes them a powerful tool for engineering tailored, unnatural polyketides; however, modifying the stereocenters of a PKS product while maintaining production levels remains a significant challenge. In this study, we systematically tested and evaluated strategies for ketoreductase (KR) domain exchanges, the domain responsible for setting stereocenters of polyketide products. After first optimizing the method for KR exchanges, we then performed 44 KR domain exchanges on three different PKSs to obtain high production of all four stereoisomers in vivo. By testing both one- and two-module PKS systems, we investigated how downstream modules process intermediates with altered stereochemistry and found that the configuration of the α-substituents was critical for gatekeeping by the ketosynthase (KS). To overcome this constraint, we investigated two different strategies for altering the KS domain, including introducing targeted mutations in the downstream KS, and exploring boundaries in exchanging the entire functional unit from the donor PKS. Both strategies successfully modified the KS stereocontrol with distinct trade-offs; the functional unit exchange resulted in higher titer improvements, though it was more likely to break the entire PKS. This study demonstrates a comprehensive approach to successfully engineering all four stereochemical configurations in multiple PKS systems, advancing our understanding of and ability to rationally modify polyketide stereochemistry through multiple engineering strategies.

Polyketides↗
PubMed2006

Proteome changes after metabolic engineering to enhance aerobic mineralization of cis-1,2-dichloroethylene.

Metabolically engineered Escherichia coli has previously been used to degrade cis-1,2-dichloroethylene (cis-DCE). The strains express the six genes of an evolved toluene ortho-monooxygenase from Burkholderia cepacia G4 (TOM-Green, which formed a reactive epoxide) with either (1) gamma-glutamylcysteine synthetase (GSHI, which forms glutathione) and the glutathione S-transferase IsoILR1 from Rhodococcus AD45 (which adds glutathione to the reactive cis-DCE epoxide) or (2) with an evolved epoxide hydrolase from Agrobacterium radiobacter AD1 (EchA F108L/I219L/C248I which converts the reactive cis-DCE epoxide to a diol). Here, the impact of this metabolic engineering for bioremediation was assessed by investigating the changes in the proteome through a quantitative shotgun proteomics technique (iTRAQ) by tracking the changes due to the sequential addition of TOM-Green, IsoILR1, and GSHI and due to adding the evolved EchA versus the wild-type enzyme to TOM-Green. For the TOM-Green/EchA system, 8 proteins out of 268 identified proteins were differentially expressed in the strain expressing EchA F108L/I219L/C248I relative to wild-type EchA (e.g., EchA, protein chain elongation factor EF-Ts, 50S ribosomal subunits L7/L12/L32/L29, cysteine synthase A, glycerophosphodiester phosphodiesterase, iron superoxide dismutase). For the TOM-Green/IsoILR1/GSHI system, the expression level of 49 proteins was changed out of 364 identified proteins. The induced proteins due to the addition of TOM-Green, IsoILR1, and GSHI were involved in the oxidative defense mechanism, pyruvate metabolism, and glutathione synthesis (e.g., 30S ribosomal subunit proteins S3 and S16, 50S ribosomal subunit protein L20, alkyl hydroperoxide reductase, lactate dehydrogenase, acetate kinase, cysteine synthase A). Enzymes involved in indole synthesis, fatty acid synthesis, gluconeogenesis, and the tricarboxylic acid cycle were repressed (e.g., tryptophanase, acetyl-CoA carboxylase, phosphoenolpyruvate carboxykinase, malate dehydrogenase). Hence, the metabolic engineering that leads to enhanced aerobic degradation of 1 mM cis-DCE (2.4-4-fold more chloride ions released) and reduced toxicity from cis-DCE epoxide results in enhanced synthesis of glutathione coupled with an induced stress response as well as repression of fatty acid synthesis, gluconeogenesis, and the tricarboxylic acid cycle.

Aerobiosis↗
PubMed2000

Growth factor delivery for tissue engineering.

A tissue-engineered implant is a biologic-biomaterial combination in which some component of tissue has been combined with a biomaterial to create a device for the restoration or modification of tissue or organ function. Specific growth factors, released from a delivery device or from co-transplanted cells, would aid in the induction of host parenchymal cell infiltration and improve engraftment of co-delivered cells for more efficient tissue regeneration or ameliorate disease states. The characteristic properties of growth factors are described to provide a biological basis for their use in tissue engineered devices. The principles of polymeric device development for therapeutic growth factor delivery in the context of tissue engineering are outlined. A review of experimental evidence illustrates examples of growth factor delivery from devices such as microparticles, scaffolds, and encapsulated cells, for their use in the application areas of musculoskeletal tissue, neural tissue, and hepatic tissue.

Animals↗
PubMed1997

Genetic engineering of bacteria and their potential for Hg2+ bioremediation.

Ion exchange or biosorptive processes for metal removal generally lack specificity in metal binding and are sensitive to ambient conditions, e.g. pH, ionic strength and the presence of metal chelators. In this study, cells of a genetically engineered Escherichia coli strain, JM109, which expresses metallothionein and a Hg2+ transport system after induction were evaluated for their selectivity for Hg2+ accumulation in the presence of sodium, magnesium, or cadmium ions and their sensitivity to pH or the presence of metal chelators during Hg2+ bioaccumulation. The genetically engineered E. coli cells in suspension accumulated Hg2+ effectively at low concentrations (0-20 microM) over a broad range of pH (3 to 11). The presence of 400 mM sodium chloride, 200 mM magnesium chloride, or 100 microM cadmium ions did not have a significant effect on the bioaccumulation of 5 microM Hg2+, indicating that this process is not sensitive to high ionic strength and is highly selective against sodium, magnesium, or cadmium ions. Metal chelators usually interfere with ion exchange or biosorptive processes. However, two common metal chelators, EDTA and citrate, had no significant effect on Hg2+ bioaccumulation by the genetically engineered strain. These results suggest that this E. coli strain could be used for selective removal of Hg2+ from waste water or from contaminated solutions which are resistant to common treatments. A second potential application would be to remove Hg2+ from Hg(2+)-contaminated soil, sediment, or particulates by washing them with a Hg2+ chelator and regenerating the chelator by passing the solution through a reactor containing the strain.

Cadmium↗