Isolation and Selection of Thermophilic Bacteria as Hexavalent Chromium Reducer from Batik Processing Waste Water

  • Wijiastuti Wijiastuti Institut Pertanian Bogor
  • I Made Artika Institut Pertanian Bogor
  • Novik Nurhidayat Institut Pertanian Bogor


Hexavalent chromium (Cr VI) in the oxidized state is carcinogenic in humans. Chromium is widely used in various industries, and therefore Cr (VI) can be found in wastes from the industry. Heavy metal Cr (VI) waste, is one type of hazardous wastes, due to the high toxicity of Cr (VI) which is much higher than that of Cr (III). This study was conducted to isolate naturally occurring bacteria from batik wastewater with ability to reduce Cr (VI). Identification of chromate reductase gene was carried out using qPCR method. Results showed that three isolates Bacillus sp.1a, Pseudomonas sp.1b, dan Geobacillus sp.1c have chromate reductase-coding genes. Based on the results of qPCR analysis, the isolate Bacillus sp.1a was predicted to have the highest reduction activity. Hence, this isolate was then subjected to Cr(VI) reduction activity test.

Author Biographies

Wijiastuti Wijiastuti, Institut Pertanian Bogor
Departemen Biokimia
I Made Artika, Institut Pertanian Bogor
Departemen Biokimia
Novik Nurhidayat, Institut Pertanian Bogor
Departemen Biokimia


American Public Health Association (APHA). 1999 . Standard Methods for the Examination of Water and Wastewater.

Bustin S.A., Benes V., Garson J.A., Hellemans J., Huggett J, Kubista M., Mueller R, Nolan T., Pfaffl MW., Shipley G.L. Vandesompele Jo,5 and Wittwer C.T. 2009. The MIQE Guidelines: Minimum Information for Publication of Quantitative Real-Time PCR Experiments. Clinical Chemistry 2009 55:4.

Cheng Y, Holman H, dan Lin Z. 2012. Remediation of Chromium and Uranium Contamination by Microbial Activity. Elements, VOL. 8, PP. 107–112 DOI: 10.2113/gselements.8.2.107.

Chaturvedi M.K. 2011. Studies on Chromate Removal by Chromium-Resistant Bacillus sp. Isolated from Tannery Effluent Journal of Environmental Protection, 2011(2):76-82. Doi:10.4236/jep.2011.21008.

Conceição D.P., dos Passos C.T., Jacques R.J.S., Bento F.M., Simonetti A.B., and Camargo F.A.O. 2009. A Novel Chromate Reductase from Bacillus sp. ES29: Characterization and Partial Purification. Revista Ciências Exatas e Naturais, 11(2): 237-256.

Factfish. 2014. Indonesia: Chromium, articles thereof, waste or scrap, powders, export value [internet]. [diacu 2014 April 22]. Tersedia dari :

Factfish, 2014. Indonesia: Chromium trioxide, import value [internet]. [diacu 2014 April 22]. Tersedia dari :

Fatmawati, Sajidan, Suranto. 2010. Potensi mikroorganisme sebagai agen bioremidiasi dalam menurunkan kadar Cr (VI) dalam Limbah cair tekstil hasil pewarnaan. Seminar Nasional Pendidikan Biologi, FKIP, UNS.

Jacobs JA., Testa SM. 2005. Overview of chromium (vi) in the environment: background and history. Chromium (VI) Handbook. ISBN 1-56670-608-4, CRC Press, New York.

Karamah EF, Setijo B, dan Simbolon HM. 2008. Pengaruh ozon dan konsentrasi zeolit terhadap kinerja proses pengolahan limbah cair yang mengandung logam dengan proses flotasi. Makara. Teknologi. 12(1):43-47. ISSN 1693–4393.

Holt J.G. and Krieg N.R. 199 4. Enrichment and Isolation. Methods for General and Molecular Bacteriology. American Society for Microbiology.

Life Technologies Corporation [LTC]. 2011. Real-Time PCR : Understanding Ct. Life technologies corporation [internet]. [diacu 2014 November 24]. Tersedia dari : Carlsbad

Occupational Safety & Health Administration [OSHA]. 2013. Chromium. [internet]. [diacu 2014 Februari 21]. Tersedia dari :

Opperman, Piater, dan van Heerden. 2008. A Novel Chromate Reductase from Thermus scotoductus SA-01 Related to Old Yellow Enzyme. Journal of bacteriology. 2008, 190(8) : 3076–3082. American Society for Microbiology.

Park C. H., Keyhan M., Wielinga B., Fendorf S., Matin A. Purification to Homogeneity and Characterization of a Novel Pseudomonas putida Chromate Reduktase. Appl. Environ. Microbiol. 2000, 66(5):1788. DOI: 10.1128/AEM.66.5.1788-1795.2000.

Pattanapipitpaisal P, Reakyai T. 2013. Cr(VI) reduction by cell-free extract of thermophillic Bacillus fusiformis NTR9. Songklanakarin J. Sci. Technol. 35 (4), 407-414.

Ramírez-Díaz MI, Díaz-Pérez C, Vargas E, Riveros-Rosas H, Campos-Garcia J, Cervantes C. 2008. Mechanisms of bacterial resistance to chromium compounds. BioMetals 21: 321-332

Sachse K., 2002. Specificity and Performance of Diagnostic PCR Assays. Methods in Molecular Biology, vol. 216: PCR Detection of Microbial Pathogens. Humana Press Inc., Totowa.

Slamet, Arbianti, Daryanto. 2005. Pengolahan limbah organik (fenol) dan logam berat (Cr6+ atau Pt4+) secara simultan dengan fotokatalis TiO2, ZnO-TiO2, dan CdS-TiO2. Makara. Teknologi. 9(2):66-71.

Sunardi. 2011. Penurunan kadar krom(vi) dengan Sargassum sp, Saccharomyces cerevisiae dan kombinasinya pada limbah cair industri batik. Ekosains. 3(1):27-32.

Walker N.J., Tech. Sight. A technique whose time has come, Science 296 (2002) 557-559