Isolation of heavy metal-resistantStaphylococcus epidermidisstrainTWSL_22and evaluation of heavy metal bioremediation potential of recombinantE. colicloned with isolatedcadD

Author:

Dissanayake D M D C1ORCID,Kumari W M N H2,Chandrasekharan N V3,Wijayarathna C D1ORCID

Affiliation:

1. Biotechnology Laboratory, Department of Chemistry, Faculty of Science, University of Colombo , PO Box 1490, Cumarathunga Munidasa Mawatha, Colombo 00300, Sri Lanka

2. Department of Molecular Biology, Durdans Hospitals , No 3 Alfred Road , Colombo 03, Sri Lanka

3. Sri Lanka Institute of Biotechnology , Thalagala road, Pitipana , Homagama, Sri Lanka

Abstract

AbstractA heavy metal-resistant bacterial strain, TWSL_22 was isolated from an industrial effluent sample and tested for heavy metal tolerance and resistance. The strain was molecularly characterized as Staphylococcus epidermidis based on 16S rDNA gene analysis and the sequence was deposited in the NCBI repository (accession number KT184893.1). Metal removal activity (P < .001) of TWSL_22 was 99.99 ± 0.001%, 74.43 ± 2.51%, and 51.16 ± 4.17% for Cd, Pb, and Cu, respectively. Highest MIC was observed for Cd. Antibiotic susceptibility assays revealed the strain TWSL_22 to be resistant to several antibiotics. The strain was screened for possible heavy metal-resistant genes and presence of cadA, copA, and cadD was confirmed by PCR. A DNA fragment containing complete sequence of cadD (618 bp) was isolated and cloned into pET 21a(+), transformed into E. coli BL21 and designated as E. coli/cadDET. E. coli/cadDET showed high metal tolerance capacity and could remove over 82% of heavy metals (Zn2+, Cd2+, Cu2+, and Cr3+) in the industrial effluent.

Funder

National Science Foundation of Sri Lanka

University of Colombo

Publisher

Oxford University Press (OUP)

Subject

Genetics,Molecular Biology,Microbiology

Reference55 articles.

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