Dear Soumya,

The following publications describe methods to determine the accumlation of heavy metal within a bacterial cell:

1-Appl Environ Microbiol. 1999 Mar; 65(3): 1092–1098.

PMCID: PMC91149

Enhanced Bioaccumulation of Heavy Metal Ions by Bacterial Cells Due to Surface Display of Short Metal Binding Peptides

Pavel Kotrba,1 Lucie Dolečková,2 Víctor de Lorenzo,3 and Tomas Ruml1,*

Author information ► Article notes ► Copyright and License information ►

This article has been cited by other articles in PMC.

Metal binding peptides of sequences Gly-His-His-Pro-His-Gly (named HP) and Gly-Cys-Gly-Cys-Pro-Cys-Gly-Cys-Gly (named CP) were genetically engineered into LamB protein and expressed in Escherichia coli. The Cd2+-to-HP and Cd2+-to-CP stoichiometries of peptides were 1:1 and 3:1, respectively. Hybrid LamB proteins were found to be properly folded in the outer membrane of E. coli. Isolated cell envelopes of E. colibearing newly added metal binding peptides showed an up to 1.8-fold increase in Cd2+ binding capacity. The bioaccumulation of Cd2+, Cu2+, and Zn2+ by E. coli was evaluated. Surface display of CP multiplied the ability of E. coli to bind Cd2+ from growth medium fourfold. Display of HP peptide did not contribute to an increase in the accumulation of Cu2+ and Zn2+. However, Cu2+ ceased contribution of HP for Cd2+accumulation, probably due to the strong binding of Cu2+ to HP. Thus, considering the cooperation of cell structures with inserted peptides, the relative affinities of metal binding peptide and, for example, the cell wall to metal ion should be taken into account in the rational design of peptide sequences possessing specificity for a particular metal.

To view the full publication, please use the following link:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC91149/

2-Advances in Bioscience and Biotechnology, 2015, 6, 131-138

Published Online February 2015 in SciRes. http://www.scirp.org/journal/abb

http://dx.doi.org/10.4236/abb.2015.62013

How to cite this paper: Banerjee, S., Gothalwal, R., Sahu, P.K. and Sao, S. (2015) Microbial Observation in Bioaccumulation

of Heavy Metals from the Ash Dyke of Thermal Power Plants of Chhattisgarh, India. Advances in Bioscience and Biotechnology,

6, 131-138. http://dx.doi.org/10.4236/abb.2015.62013

Microbial Observation in Bioaccumulation

of Heavy Metals from the Ash Dyke of

Thermal Power Plants of Chhattisgarh, India

Sonali Banerjee1, Ragini Gothalwal2, Pankaj K. Sahu3, Shweta Sao1

http://creativecommons.org/licenses/by/4.0/

Abstract

Several mechanisms are developed by the microorganisms to tolerate few high concentrations of heavy metals. One of these mechanisms dependent upon anabolic and catabolic energy of microorganisms is the bioaccumulation of heavy metals. In present work, approximately four varieties of bacteria have been isolated from the ash dyke sample of four thermal power plants of Chhattisgarh, i.e., Bharat Aluminium Company (BALCO), Chhattisgarh State Electricity Board (CSEB), Korba,

Thermal Power Cooperation (NTPC), Bilaspur and KSK Akaltara, Chattisgarh. Out of one hundred fifty isolates, three were capable to grow in varying concentration of heavy metals. The strains were tested for their tolerance against six different types of heavy metals dominant in the ash samples viz. Pb, Hg, Ni, Co, Cu, Mn. Their maximum resistance existed up to 0.6mM/ml of the above mentioned different metals under lab standard conditions. Three isolates are found suitable for

the multiple metal resistance ability viz SM2, SM3, and SM12. These are categorized as Bacillus cereus (SM2, SM3), and Bacillus subtilis (SM12) after performing 16S rDNA sequencing.

To view the full publication, please see attached file.

3- ACCUMULATION OF HEAVY METALS BY A BACTERIUM ISOLATED FROM ELECTROPLATING EFFLUENT

F. Malekzadeh1, A. Farazmand1, H. Ghafourian1, M. Shahamat2, M. Levin2, C. Grim2, and R.R. Colwell2

1Department of Biology, University of Tehran, Iran; 2Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, MD

SUMMARY

Pseudomonas MGF-48, a gram-negative, motile, oxidase negative, catalase positive, yellow pigmented bacterium, isolated from electroplating effluent, was found to accumulate heavy metals, especially uranium. Uptake of uranium was high, rapid and the amount increased in direct proportion to concentration, e.g., from 50 to 200 mg/l uranium. The largest amount of uranium uptake was 174 mg per gram dry weight bacterial biomass, observed to occur in stationary phase, when incubation was at 25C. Uptake was determined by flow injection analysis. Maximum uranium accumulation was obtained at pH 6.5, with 86% of the uranium being removed within 5 min of incubation. Release of uranium bound to the cells was accomplished by addition of sodium carbonate and EDTA solution (0.1 M). The solution was reusable, serving as a biosorbent. Cells immobilized in polyacrylamide gel yielded 90% uranium removal. Pseudomonas MGF-48 showed excellent efficiency in biosorbing uranium both when immobilized and as free cells. The results of this study indicates that the bacterium was capable of accumulating several metals. Accumulation of uranium was higher than other metals. We concluded that PseudomonasMGF-48 shows excellent potential for bioremediation of uranium-polluted aqueous effluents.

Many industries, including mining and electroplating, discharge aqueous effluents containing relatively high levels of heavy metals, e.g. uranium, cadmium, mercury, and copper. Untreated effluent from these manufacturing processes have an adverse impact on the environment (6,8,11,13,16,17). A specific problem associated with heavy metals in the environment is accumulation in the food chain and persistence in the environment.

Physical and chemical methods have been designed to remove metal ions from effluents, but, in general, these methods are commercially impractical, either because of high operating cost or difficulty in treating the solid wastes generated (6,18).

Bioremediation of industrial wastes containing heavy metals has been demonstrated by several biotechnology companies employing bioaccumulation (5,6). Biosorption, bioprecipitation, and uptake by purified biopolymers derived from microbial cells provide alternative and/or additive processes for conventional physical and chemical methods (18). Intact microbial cells, live or dead, and their products can be highly efficient bioaccumulators of both soluble and particulate forms of metals (3,12,18). The cell surfaces of all microorganisms are negatively charged owing to the presence of various anionic structures. This gives bacteria the ability to bind metal cations. Various microbial species, mainly Pseudomonas, have been shown to be relatively efficient in bioaccumulation of uranium, copper, lead, and other metal ions from polluted effluents, both as immobilized cells and in the mobilized state (5). We report here the bioaccumulation of several metals by a bacterium, identified as a Pseudomonas sp., strain MGF-48, isolated from the effluent of a metal melting factory in the south of Tehran.

http://www.isb.vt.edu/brarg/brasym96/malekzadeh96.htm

4- Brazilian Journal of Microbiology

On-line version ISSN 1678-4405

Braz. J. Microbiol. vol.32 no.1 São Paulo Jan./Mar. 2001

http://dx.doi.org/10.1590/S1517-83822001000100001 

BIOACCUMULATION OF COPPER, ZINC, CADMIUM AND LEAD BYBACILLUS SP., BACILLUS CEREUS, BACILLUS SPHAERICUS ANDBACILLUS SUBTILIS

  Antonio Carlos Augusto da Costa*; Flavia Pereira Duta

Universidade do Estado do Rio de Janeiro, Instituto de Química, Departamento de Tecnologia de Processos Bioquímicos, Maracanã, RJ, Brasil

Submitted: July 17, 1999; Returned to authors for corrections: January 13, 2000; Approved: March 03, 2001

 This work presents some results on the use of microbes from the genus Bacillus for uptake of cadmium, zinc, copper and lead ions. Maximum copper bioaccumulations were 5.6 mol/g biomass for B.sphaericus, 5.9 mol/g biomass for B. cereus and B. subtilis, and 6.4 mol/g biomass for Bacillus sp. Maximum zinc bioaccumulations were 4.3 mol/g biomass for B. sphaericus, 4.6 mol/g biomass for B. cereus, 4.8 mol/g biomass for Bacillus sp. and 5.0 mol/g biomass forB. subtilis. Maximum cadmium bioaccumulations were 8.0 mol/g biomass for B. cereus, 9.5 mol/g biomass for B. subtilis, 10.8 mol/g biomass for Bacillus sp. and 11.8 mol/g biomass for B. sphaericus. Maximum lead biomaccumulations were 0.7 mol/g biomass for B. sphaericus, 1.1 mol/g biomass for B. cereus, 1.4 mol/g biomass for Bacillus sp. and 1.8 mol/g biomass for B. subtilis. The different Bacillus strains tested presented distinct uptake capacities, and the best results were obtained for B. subtilis and B. cereus.

http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822001000100001

5-Full Length Research Paper Bioaccumulation of cadmium and lead by Shewanella oneidensis isolated from soil in Basra

governorate, Iraq

Received 9 January, 2016; Accepted 26 February, 2016

In the present study heavy metals resistant bacteria were isolated from soil collected from Al-Zubair district in Basra governorate south of Iraq. On the basis of morphological, biochemical, 16S rRNA gene sequencing and phylogeny analysis, the isolates were authentically identified as Shewanella oneidensis in addition to Bacillus thuringiensis and Deinococcus radiodurans. The minimal inhibitory

concentration (MIC) of isolates against cadmium (Cd) and lead (Pb) was determined on solid medium. S. oneidensis showed significant resistance to high concentrations of Cd (1000 mgl-1 ) and Pb (700 mgl-1 ).

The bioaccumilation capabilities of S. oneidensis for Cd and Pb were monitored at different ion concentrations and contact times. The transmission electron microscope (TEM) study confirmed the accumulation of Cd and Pb by S. oneindensis causing morphological changes.

To view the publication, please see attached file.

Hoping this will be helpful,

Rafik

Dear Soumya,

Following steps may be done...

-Cfg the cells

-Dry the cell pellets

-do the SEM EDS

You may confirm whether cells contain heavy metals or not in EDS profile

you can determine bioaccumulation by TEM, SEM - EDX and XRF.

SEM - EDX will be good

Thank you every one for your suggestion.

you can determined bio-accumulated heavy metal by TEM or EDX 

Thanks everyone for your contributions. However, how do I determine bioaccumulation factor in bacteria and what information can you infer from the value.

U can get the details from this paper

DOI:10.1007/s10867-020-09560-7

How can we differentiate and quantify intracellular metal accumulation as well as cell surface adsorption?