A rapid inhibitive enzyme assay for monitoring heavy metals pollution in the Juru Industrial Estate

Authors

  • Mohd Izuan Effendi Halmi Department of Land Management, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
  • Baskaran Gunasekaran Faculty of Applied Sciences, UCSI University Kuala Lumpur (South Wing), No.1, Jalan Menara Gading, UCSI Heights 56000 Cheras, Kuala Lumpur, Malaysia
  • Ahmad Razi Othman Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia 43400 UPM Serdang, Selangor, Malaysia.
  • Kamarulzaman Kamaruddin Nanotechnology Directorate, Ministry of Science, Technology & Innovation, Malaysia.
  • Farrah Aini Dahalan School of Environmental Engineering, Kompleks Pusat Pengajian Jejawi 3, Universiti Malaysia Perlis, 02600 Arau, Perlis, Malaysia.
  • Naimah Ibrahim School of Environmental Engineering, Kompleks Pusat Pengajian Jejawi 3, Universiti Malaysia Perlis, 02600 Arau, Perlis, Malaysia.
  • Mohd Yunus Shukor Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia 43400 UPM Serdang, Selangor, Malaysia.

DOI:

https://doi.org/10.54987/bstr.v3i2.290

Keywords:

Inhibitive assay, biomonitoring, molybdenum-reducing enzyme, heavy metals, Juru industrial estate

Abstract

The volume of contaminated rivers in Malaysia continues to keep rising through the years. The cost of instrumental monitoring is uneconomical and prohibits schedule monitoring of contaminants particularly heavy metals. In this work, a rapid enzyme assay utilizing the molybdenum-reducing enzyme as an inhibitive assay, prepared in crude form from the molybdenum-reducing bacterium Serratia sp. strain DRY5 has been developed for monitoring the heavy metals mercury, silver, copper and chromium in contaminated waters in the Juru Industrial Estate. The crude enzyme extract transformed soluble molybdenum (phosphomolybdate) into a deep blue solution, which is inhibited by heavy metals such as mercury, silver, copper and chromium. The IC50 and Limits of Detection (LOD) values for mercury, copper, silver and cadmium were 0.245, 0.298, 0.367, 0.326, and 0.124, 0.086, 0.088 and 0.094 mg L-1, respectively. The assay is rapid, and can be carried out in less than 10 minutes. In addition, the assay can be carried out at ambient temperature. The IC50 values for these heavy metals are more sensitive than several established assays. Water samples from various locations in the month of November from the Juru Industrial Estate (Penang) were tested for the presence of heavy metals using the developed assay. Enzyme activity was nearly inhibited for water samples from several locations. The presence of heavy metals was confirmed instrumentally using Atomic Emission Spectrometry and a Flow Injection Mercury System. The assay is rapid and simple and can be used as a first screening method for large scale monitoring of heavy metals.

References

Poli A, Salerno A, Laezza G, Di Donato P, Dumontet S, Nicolaus B. Heavy metal resistance of some thermophiles: potential use of α-amylase from Anoxybacillus amylolyticus as a microbial enzymatic bioassay. Res Microbiol. 2009;160(2):99–106.

Bondici VF, Lawrence JR, Khan NH, Hill JE, Yergeau E, Wolfaardt GM, et al. Microbial communities in low permeability, high pH uranium mine tailings: Characterization and potential effects. J Appl Microbiol. 2013;114(6):1671–86.

Fathi HB., Othman MS., Mazlan AG., Arshad A., Amin SMN., Simon KD. Trace metals in muscle, liver and gill tissues of marine fishes from Mersing, eastern coast of peninsular Malaysia: Concentration and assessment of human health risk. Asian J Anim Vet Adv. 2013;8(2):227–36.

Phuong NTK, Khoa NC. Evaluation of heavy metals in tissue of shellfish from Can Gio coastline in Ho Chi Minh city, Vietnam. Asian J Chem. 2013;25(15):8552–6.

Sabullah MK, Sulaiman MR, Shukor MYA, Shamaan NA, Khalid A, Ahmad SA. In vitro and in vivo effects of Puntius javanicus cholinesterase by copper. Fresenius Environ Bull. 2015;24(12B):4615–21.

Mohammad Ali BN, Lin CY, Cleophas F, Abdullah MH, Musta B. Assessment of heavy metals contamination in Mamut river sediments using sediment quality guidelines and geochemical indices. Environ Monit Assess. 2015;187(1).

Shukor Y, Baharom NA, Rahman FA, Abdullah MP, Shamaan NA, Syed MA. Development of a heavy metals enzymatic-based assay using papain. Anal Chim Acta. 2006;566(2):283–9.

Shukor MY, Masdor N, Baharom NA, Jamal JA, Abdullah MPA, Shamaan NA, et al. An inhibitive determination method for heavy metals using bromelain, a cysteine protease. Appl Biochem Biotechnol. 2008;144(3):283–91.

DOE. Malaysia Environmental Quality Report 2014. Department of Environment, Ministry of Natural Resources and Environment, Malaysia; 2015.

Yap CK, Ismail A, Tan SG. Heavy metal (Cd, Cu, Pb and Zn) concentrations in the green-lipped mussel Perna viridis (Linnaeus) collected from some wild and aquacultural sites in the west coast of Peninsular Malaysia. Food Chem. 2004;84(4):569–75.

Shaari H, Mohamad Azmi SNH, Sultan K, Bidai J, Mohamad Y, Shaari H, et al. Spatial distribution of selected heavy metals in surface sediments of the EEZ of the east coast of Peninsular Malaysia. Int J Oceanogr Int J Oceanogr. 2015;2015, 2015:e618074.

Hsieh C-Y, Tsai M-H, Ryan DK, Pancorbo OC. Toxicity of the 13 priority pollutant metals to Vibrio fisheri in the Microtox® chronic toxicity test. Sci Total Environ. 2004;320(1):37–50.

Halmi MIE, Jirangon H, Johari WLW, Abdul Rachman AR, Shukor MY, Syed MA. Comparison of Microtox and Xenoassay light as a near real time river monitoring assay for heavy metals. Sci World J. 2014;2014.

Botsford JL. A simple assay for toxic chemicals using a bacterial indicator. World J Microbiol Biotechnol. 1998;14(3):369–76.

Isa HWM, Johari WLW, Syahir A, Shukor MYA, Nor A, Shaharuddin NA, et al. Development of a bacterial-based tetrazolium dye (MTT) assay for monitoring of heavy metals. Int J Agric Biol. 2014;16(6):1123–8.

Jung K, Bitton G, Koopman B. Assessment of urease inhibition assays for measuring toxicity of environmental samples. Water Res. 1995;29(8):1929–33.

Shukor MY, Baharom NA, Masdor NA, Abdullah MPA, Shamaan NA, Jamal JA, et al. The development of an inhibitive determination method for zinc using a serine protease. J Environ Biol. 2009;30(1):17–22.

Baskaran G, Masdor NA, Syed MA, Shukor MY. An inhibitive enzyme assay to detect mercury and zinc using protease from Coriandrum sativum. Sci World J [Internet]. 2013;2013. Available from: http://www.scopus.com/inward/record.url?eid=2-s2.0-84886463804&partnerID=40&md5=56fe4ef11ba50ff5275953a2612962c1

Sahlani MZ, Halmi MIE, Masdor NA, Gunasekaran B, Wasoh H, Syed MA, et al. A rapid inhibitive assay for the determination of heavy metals using α-chymotrypsin; a serine protease. Nanobio Bionano. 2014;1(2):41–6.

Shukor MY, Anuar N, Halmi MIE, Masdor NA. Near real-time inhibitive assay for heavy metals using achromopeptidase. Indian J Biotechnol. 2014;13(3):398–403.

Aidil MS, Sabullah MK, Halmi MIE, Sulaiman R, Shukor MS, Shukor MY, et al. Assay for heavy metals using an inhibitive assay based on the acetylcholinesterase from Pangasius hypophthalmus (Sauvage, 1878). Fresenius Environ Bull. 2013;22(12):3572–6.

Shukor MY, Tham LG, Halmi MIE, Khalid I, Begum G, Syed MA. Development of an inhibitive assay using commercial Electrophorus electricus acetylcholinesterase for heavy metal detection. J Environ Biol. 2013;34(5):967–70.

Sabullah MK, Sulaiman MR, Shukor MS, Yusof MT, Johari WLW, Shukor MY, et al. Heavy metals biomonitoring via inhibitive assay of acetylcholinesterase from Periophthalmodon schlosseri. Rendiconti Lincei. 2015;26(2):151–8.

Shukor MY, Bakar NA, Othman AR, Yunus I, Shamaan NA, Syed MA. Development of an inhibitive enzyme assay for copper. J Environ Biol. 2009;30(1):39–44.

Ahmad SA, Halmi MIE, Wasoh MH, Johari WLW, Shukor MY, Syed, M.A. The development of a specific inhibitive enzyme assay for the heavy metal, lead. J Environ Bioremediation Toxicol. 2013;1(1):9–13.

Othman AR, Ahmad SA, Baskaran G, Halmi MIE, Shamaan NA, Syed M, et al. River monitoring of mercury using a novel molybdenum-reducing enzyme assay. Bull Environ Sci Manag. 2014;2(1):30–35.

Gunasekaran B, Ahmad SA, Halmi MIE. A rapid inhibitive enzyme assay for monitoring heavy metals pollution in a river running through a UNESCO heritage site. J Biochem Microbiol Biotechnol. 2015;3(2):21–5.

Rahman MFA, Shukor MY, Suhaili Z, Mustafa S, Shamaan NA, Syed MA. Reduction of Mo(VI) by the bacterium Serratia sp. strain DRY5. J Environ Biol. 2009;30(1):65–72.

Shukor MY, Rahman MFA, Shamaan NA, Lee CH, Karim MIA, Syed MA. An improved enzyme assay for molybdenum-reducing activity in bacteria. Appl Biochem Biotechnol. 2008;144(3):293–300.

Shukor MY, Lee CH, Omar I, Karim MIA, Syed MA, Shamaan NA. Isolation and characterization of a molybdenum-reducing enzyme in Enterobacter cloacae strain 48. Pertanika J Sci Technol. 2003;11(2):261–72.

Shukor MY, Halmi MIE, Rahman MFA, Shamaan NA, Syed MA. Molybdenum reduction to molybdenum blue in Serratia sp. strain DRY5 is catalyzed by a novel molybdenum-reducing enzyme. BioMed Res Int. 2014;2014.

Shukor MY, Habib SHM, Rahman MFA, Jirangon H, Abdullah MPA, Shamaan NA, et al. Hexavalent molybdenum reduction to molybdenum blue by S. marcescens strain Dr. Y6. Appl Biochem Biotechnol. 2008;149(1):33–43.

Shukor MY, Rahman MF, Shamaan NA, Syed MS. Reduction of molybdate to molybdenum blue by Enterobacter sp. strain Dr.Y13. J Basic Microbiol. 2009;49(SUPPL. 1):S43–54.

Yunus SM, Hamim HM, Anas OM, Aripin SN, Arif SM. Mo (VI) reduction to molybdenum blue by Serratia marcescens strain Dr. Y9. Pol J Microbiol. 2009;58(2):141–7.

Shukor MY, Rahman MF, Suhaili Z, Shamaan NA, Syed MA. Bacterial reduction of hexavalent molybdenum to molybdenum blue. World J Microbiol Biotechnol. 2009;25(7):1225–34.

Shukor MY, Ahmad SA, Nadzir MMM, Abdullah MP, Shamaan NA, Syed MA. Molybdate reduction by Pseudomonas sp. strain DRY2. J Appl Microbiol. 2010;108(6):2050–8.

Shukor MY, Rahman MF, Suhaili Z, Shamaan NA, Syed MA. Hexavalent molybdenum reduction to Mo-blue by Acinetobacter calcoaceticus. Folia Microbiol (Praha). 2010;55(2):137–43.

Lim HK, Syed MA, Shukor MY. Reduction of molybdate to molybdenum blue by Klebsiella sp. strain hkeem. J Basic Microbiol. 2012;52(3):296–305.

Abo-Shakeer LKA, Ahmad SA, Shukor MY, Shamaan NA, Syed MA. Isolation and characterization of a molybdenum-reducing Bacillus pumilus strain lbna. J Environ Microbiol Toxicol. 2013;1(1):9–14.

Othman AR, Bakar NA, Halmi MIE, Johari WLW, Ahmad SA, Jirangon H, et al. Kinetics of molybdenum reduction to molybdenum blue by Bacillus sp. strain A.rzi. BioMed Res Int. 2013;2013:Article number 371058.

Khan A, Halmi MIE, Shukor MY. Isolation of Mo-reducing bacterium in soils from Pakistan. J Environ Microbiol Toxicol. 2014;2(1):38–41.

Mansur R, Gusmanizar N, Dahalan FA, Masdor NA, Ahmad SA, Shukor MS, et al. Isolation and characterization of a molybdenum-reducing and amide-degrading Burkholderia cepacia strain neni-11 in soils from west Sumatera, Indonesia. IIOAB. 2016;7(1):28–40.

Halmi MIE, Wasoh H, Sukor S, Ahmad SA, Yusof MT, Shukor MY. Bioremoval of molybdenum from aqueous solution. Int J Agric Biol. 2014;16(4):848–50.

Shukor MS, Shukor MY. Bioremoval of toxic molybdenum using dialysis tubing. Chem Eng Res Bull. 2015;18(1):6–11.

Koutras GA, Schneider AS, Hattori M, Valentine WN. Studies on chromated erythrocytes. Mechanisms of chromate inhibition of glutathione reductase. Br J Haematol. 1965;11:360–9.

Sabullah MK, Ahmad SA, Hussin J, Gansau AJ, Sulaiman MR. Acute effect of copper on Puntius javanicus survival and a current opinion for future biomarker development. J Environ Bioremediation Toxicol. 2014;2(1):28–32.

Schenker N, Gentleman JF. On judging the significance of differences by examining the overlap between confidence intervals. Am Stat. 2001;55(3):182–6.

Mat I, Maah MJ, Johari A. Trace metal geochemical associations in sediments from the culture- bed of Anadara granosa. Mar Pollut Bull. 1994;28(5):319–23.

Mohd Tawil N, Irfan Che Ani SA, Hamid SMY, Mihd Radzuan NA, Khalil N, Husin HN, et al. 2nd International Building Control ConferenceSustainable Environment: Issues and Solutions from the Perspective of Facility Managers. Procedia Eng. 2011;20:458–65.

Downloads

Published

2015-12-15

How to Cite

Halmi, M. I. E., Gunasekaran, B., Othman, A. R., Kamaruddin, K., Dahalan, F. A., Ibrahim, N., & Shukor, M. Y. (2015). A rapid inhibitive enzyme assay for monitoring heavy metals pollution in the Juru Industrial Estate. Bioremediation Science and Technology Research, 3(2), 7–12. https://doi.org/10.54987/bstr.v3i2.290

Issue

Vol. 3 No. 2 (2015)

Section

Articles

License

Authors who publish with this journal agree to the following terms:

  1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
  2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
  3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).