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All Journal HAYATI Journal of Biosciences Reaktor International Journal of Renewable Energy Development JURNAL KIMIA SAINS DAN APLIKASI Microbiology Indonesia Jurnal Matematika & Sains Journal of Engineering and Technological Sciences Indonesian Journal of Biotechnology EduChemia: Jurnal Kimia dan Pendidikan INDONESIAN JOURNAL OF PHARMACY Indonesian Journal of Chemistry Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal Indonesian Journal of Biotechnology and Biodiversity International Journal of Renewable Energy Development
Enny Ratnaningsih
Biochemistry Research Division, Faculty Of Mathematics And Natural Sciences, Institut Teknologi Bandung
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Control & Systems Engineering Earth & Planetary Sciences Education Electrical & Electronics Engineering Energy Engineering Environmental Science Immunology & microbiology Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Other

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Optimization of Monochloroacetic Acid Biodegradation by Recombinant E. coli BL21 (DE3)/pET-bcfd1 Carrying Haloacid Dehalogenase Gene from Bacillus cereus IndB1 Enny Ratnaningsih; Rachmad Ade; Rindia Maharani Putri; Idris Idris
International Journal of Renewable Energy Development Vol 10, No 4 (2021): November 2021
Publisher : Center of Biomass & Renewable Energy, Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/ijred.2021.38887

Abstract

In recent years, attention to microbial dehalogenase has continually increased due to its potential application, both in bioremediation and in the biosynthesis of fine chemicals. Many microbial recombinant strains carrying dehalogenase gene have been developed, particularly to increase the dehalogenase production and its quality. In this study, we aimed to find the optimum condition for the production of active haloacid dehalogenase by E. coli BL21 (DE3) harboring recombinant plasmid pET-bcfd1 that carried haloacid dehalogenase gene from Bacillus cereus IndB1 local strain. This would be examined by assessing the ability of whole cell life culture to degrade monochloroacetic acid (MCA) and quantifying the chloride ion released into the medium. Several variables were evaluated to find this optimal condition. We found that the best condition for MCA biodegradation using this recombinant clone was at 0.2 mM MCA, 10 μM of isopropyl β-D-1-thiogalactopyranoside (IPTG), 6 hours of pre-induction incubation at 37ºC with shaking, 2 hours IPTG induction at 30ºC with shaking, at pH 7 in Luria Bertani (LB) liquid medium without NaCl, which produced about 0.056 mM chloride ions. Inducer concentration, pre-induction incubation time and temperature, as well as induction time and temperature were apparent to be associated with the expression of the protein, while the MCA concentration and the pH of the medium influenced the ability of the recombinant E. coli BL21 (DE3)/pET-bcfd1 to grow in toxic environment. Our findings laid the foundation for exploration of dehalogenases from local Bacillus strains through genetic engineering for MCA biodegradation
Co-Authors . WISJNUPRAPTO Ade, Rachmad Aidah Nur Fithriah Anggraeni, Vina Juliana Anita K. Wardani DEBBIE SOEFIE RETNONINGRUM Debbie Sofie Retnoningrum DESSY NATALIA EDWAN KARDENA Endang Purwantini Hartiwi Diastuti Hartiwi Diastuti I Gede Wenten Idris Idris Idris Idris Idris Idris Idris Idris Ira Oktavia Kadja, Grandprix Thomryes Marth Khoiruddin Khoiruddin Lina Herlina Lina Herlina Lousiana Dwinta Utami Muhaimin Muhaimin Nanik Wijayati Ni Putu Yuliastri Nicholaus Prasetya Nora, Adri Nur Umriani Permatasari Nurlaida Nurlaida Nurul F. Himma Oei Ban Liang OEI BAN LIANG Pujawati Suryatmana Putri, Rindia Maharani Putu T.P. Aryanti Rachmad Ade Ridani Rino Anggoro Rindia Maharani Putri Rindia Maharani Putri Rindia Maharani Putri RUKMAN HERTADI Sadiah Achmad Sadijah Achmad Sadijah Achmad Sulistiya Nirta Sunaryo Supartono - ZEILY NURACHMAN Zuhdina Sabiqoh