An inhibitive determination method for heavy metals using bromelain, a cysteine protease

M. Y. Shukor, N. Masdor, N. A. Baharom, Jamia Azdina Jamal, M. P A Abdullah, N. A. Shamaan, M. A. Syed

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

A heavy-metal assay has been developed using bromelain, a protease. The enzyme is assayed using casein as a substrate with Coomassie dye to track completion of hydrolysis of casein. In the absence of inhibitors, casein is hydrolysed to completion, and the solution is brown. In the presence of metal ions such as Hg2+ and Cu2+, the hydrolysis of casein is inhibited, and the solution remains blue. Exclusion of sulfhydryl protective agent and ethylenediaminetetraacetic in the original assay improved sensitivity to heavy metals several fold. The assay is sensitive to Hg2+ and Cu2+, exhibiting a dose-response curve with an IC50 of 0.15 mg l-1 for Hg2+ and a one-phase binding curve with an IC50 of 0.23 mg l-1 for Cu2+. The IC 50 value for Hg2+ is found to be lower to several other assays such as immobilized urease and papain assay, whilst the IC50 value for Cu2+ is lower than immobilized urease, 15-min Microtox™, and rainbow trout.

Original languageEnglish
Pages (from-to)283-291
Number of pages9
JournalApplied Biochemistry and Biotechnology
Volume144
Issue number3
DOIs
Publication statusPublished - Mar 2008

Fingerprint

Bromelain
Bromelains
Cysteine Proteases
Heavy Metals
Caseins
Casein
Heavy metals
Assays
Inhibitory Concentration 50
Urease
Hydrolysis
Protective Agents
Papain
Oncorhynchus mykiss
Peptide Hydrolases
Coloring Agents
Metals
Metal ions
Ions
Enzymes

Keywords

  • Bromelain
  • Cysteine protease
  • Inhibitive determination method

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Bioengineering

Cite this

Shukor, M. Y., Masdor, N., Baharom, N. A., Jamal, J. A., Abdullah, M. P. A., Shamaan, N. A., & Syed, M. A. (2008). An inhibitive determination method for heavy metals using bromelain, a cysteine protease. Applied Biochemistry and Biotechnology, 144(3), 283-291. https://doi.org/10.1007/s12010-007-8063-5

An inhibitive determination method for heavy metals using bromelain, a cysteine protease. / Shukor, M. Y.; Masdor, N.; Baharom, N. A.; Jamal, Jamia Azdina; Abdullah, M. P A; Shamaan, N. A.; Syed, M. A.

In: Applied Biochemistry and Biotechnology, Vol. 144, No. 3, 03.2008, p. 283-291.

Research output: Contribution to journalArticle

Shukor, M. Y. ; Masdor, N. ; Baharom, N. A. ; Jamal, Jamia Azdina ; Abdullah, M. P A ; Shamaan, N. A. ; Syed, M. A. / An inhibitive determination method for heavy metals using bromelain, a cysteine protease. In: Applied Biochemistry and Biotechnology. 2008 ; Vol. 144, No. 3. pp. 283-291.
@article{a45e0b6ecc154efda7ac8b1270bfe13e,
title = "An inhibitive determination method for heavy metals using bromelain, a cysteine protease",
abstract = "A heavy-metal assay has been developed using bromelain, a protease. The enzyme is assayed using casein as a substrate with Coomassie dye to track completion of hydrolysis of casein. In the absence of inhibitors, casein is hydrolysed to completion, and the solution is brown. In the presence of metal ions such as Hg2+ and Cu2+, the hydrolysis of casein is inhibited, and the solution remains blue. Exclusion of sulfhydryl protective agent and ethylenediaminetetraacetic in the original assay improved sensitivity to heavy metals several fold. The assay is sensitive to Hg2+ and Cu2+, exhibiting a dose-response curve with an IC50 of 0.15 mg l-1 for Hg2+ and a one-phase binding curve with an IC50 of 0.23 mg l-1 for Cu2+. The IC 50 value for Hg2+ is found to be lower to several other assays such as immobilized urease and papain assay, whilst the IC50 value for Cu2+ is lower than immobilized urease, 15-min Microtox™, and rainbow trout.",
keywords = "Bromelain, Cysteine protease, Inhibitive determination method",
author = "Shukor, {M. Y.} and N. Masdor and Baharom, {N. A.} and Jamal, {Jamia Azdina} and Abdullah, {M. P A} and Shamaan, {N. A.} and Syed, {M. A.}",
year = "2008",
month = "3",
doi = "10.1007/s12010-007-8063-5",
language = "English",
volume = "144",
pages = "283--291",
journal = "Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology",
issn = "0273-2289",
publisher = "Humana Press",
number = "3",

}

TY - JOUR

T1 - An inhibitive determination method for heavy metals using bromelain, a cysteine protease

AU - Shukor, M. Y.

AU - Masdor, N.

AU - Baharom, N. A.

AU - Jamal, Jamia Azdina

AU - Abdullah, M. P A

AU - Shamaan, N. A.

AU - Syed, M. A.

PY - 2008/3

Y1 - 2008/3

N2 - A heavy-metal assay has been developed using bromelain, a protease. The enzyme is assayed using casein as a substrate with Coomassie dye to track completion of hydrolysis of casein. In the absence of inhibitors, casein is hydrolysed to completion, and the solution is brown. In the presence of metal ions such as Hg2+ and Cu2+, the hydrolysis of casein is inhibited, and the solution remains blue. Exclusion of sulfhydryl protective agent and ethylenediaminetetraacetic in the original assay improved sensitivity to heavy metals several fold. The assay is sensitive to Hg2+ and Cu2+, exhibiting a dose-response curve with an IC50 of 0.15 mg l-1 for Hg2+ and a one-phase binding curve with an IC50 of 0.23 mg l-1 for Cu2+. The IC 50 value for Hg2+ is found to be lower to several other assays such as immobilized urease and papain assay, whilst the IC50 value for Cu2+ is lower than immobilized urease, 15-min Microtox™, and rainbow trout.

AB - A heavy-metal assay has been developed using bromelain, a protease. The enzyme is assayed using casein as a substrate with Coomassie dye to track completion of hydrolysis of casein. In the absence of inhibitors, casein is hydrolysed to completion, and the solution is brown. In the presence of metal ions such as Hg2+ and Cu2+, the hydrolysis of casein is inhibited, and the solution remains blue. Exclusion of sulfhydryl protective agent and ethylenediaminetetraacetic in the original assay improved sensitivity to heavy metals several fold. The assay is sensitive to Hg2+ and Cu2+, exhibiting a dose-response curve with an IC50 of 0.15 mg l-1 for Hg2+ and a one-phase binding curve with an IC50 of 0.23 mg l-1 for Cu2+. The IC 50 value for Hg2+ is found to be lower to several other assays such as immobilized urease and papain assay, whilst the IC50 value for Cu2+ is lower than immobilized urease, 15-min Microtox™, and rainbow trout.

KW - Bromelain

KW - Cysteine protease

KW - Inhibitive determination method

UR - http://www.scopus.com/inward/record.url?scp=46049117104&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=46049117104&partnerID=8YFLogxK

U2 - 10.1007/s12010-007-8063-5

DO - 10.1007/s12010-007-8063-5

M3 - Article

VL - 144

SP - 283

EP - 291

JO - Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology

JF - Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology

SN - 0273-2289

IS - 3

ER -