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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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Journal : reaktor

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Recombinant Production and One-Pot Purification for Enhancing Activity of Haloacid Dehalogenase from Bacillus cereus IndB1 Enny Ratnaningsih; Sulistiya Nirta Sunaryo; Idris Idris; Rindia Maharani Putri
Reaktor Volume 21 No. 2 June 2021
Publisher : Dept. of Chemical Engineering, Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (239.33 KB) | DOI: 10.14710/reaktor.21.2.59-64

Abstract

In recent years we have witnessed the emergence of organohalogen utilization in various chemical-based industries, particularly polymer-based, agricultural, and pharmaceutical sectors. Despite this, organohalogen compounds are actually very dangerous to the environment, as they are difficult to be naturally degraded and generally toxic to organisms. A green and biocompatible method to overcome this issue is by employing enzymes that could convert organohalogens into non-toxic compounds, such as the class of enzymes known as haloacid dehalogenases. To enhance the activity of haloacid dehalogenase isolated from local strains of Bacillus cereus IndB1, we have developed a recombinant expression system using pET-bcfd1 plasmid in E. coli BL21 (DE3) host cells. Following enzyme production, we also demonstrated a one-pot purification system for the expressed dehalogenase, harnessing the presence of His-tag in the recombinant clones. Purification was carried out using Ni-NTA affinity column chromatography, using imidazole eluent with a concentration gradient of 10 mM to 500 mM. The enzyme activity was tested against the monochloroacetic acid (MCA) substrate according to the Bergmann and Sanik method, and the protein content in the solution was measured using the Bradford method. The purity of the enzyme after one-pot purification was confirmed by SDS-PAGE analyses, showing a single band of 40 kDa in size. Remarkably, the purified haloacid dehalogenase specific activity was increased by 12-fold compared to its crude enzyme extract. Therefore, the expression and purification system developed in this study allow further exploration of dehalogenases from local strains as an efficient catalyst for MCA biodegradation.Keywords: recombinant expression, haloacid dehalogenase, monochloroacetic acid, enzyme purification
PRODUKSI ANTIBIOTIKA OLEH Bacillus subtilis M10 DALAM MEDIA UREA-SORBITOL Supartono Supartono; Nanik Wijayati; Lina Herlina; Enny Ratnaningsih
Reaktor Volume 13, Nomor 3, Juni 2011
Publisher : Dept. of Chemical Engineering, Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (348.974 KB) | DOI: 10.14710/reaktor.13.3.185-193

Abstract

PRODUCTION OF ANTIBIOTICS BY Bacillus subtilis M10 IN UREA-SORBITOL MEDIUM. Infection diseases still become the main health problems that suffered by people in Indonesia. Besides, there were many pathogen bacteria found to be resistant to the some antibiotics. Therefore, the efforts to get a new antibiotic require to be done continuously. A new local strain of Bacillus subtilis BAC4 has been known producing an antibiotic that inhibit Serratia marcescens ATCC 27117 growth. To make efficient the local strain, mutation on Bacillus subtilis BAC4 was done by using acridine orange and a mutant cell of Bacillus subtilis M10 that overproduction for producing antibiotic was obtained. Nevertheless, the production kinetics of antibiotic by this mutant has not been reported. The objective of this research was to study the production kinetics of antibiotic by Bacillus subtilis M10 mutant. The production of antibiotic was conducted using batch fermentation and antibiotic assay was performed with agar absorption method using Serratia marcescens ATCC 27117 as bacteria assay. Research result provided that Bacillus subtilis M10 mutant with overproduction of antibiotic produced an antibiotic since 8th hour’s fermentation and optimum of it production was at 14th hours after inoculation.  Penyakit infeksi masih menjadi masalah yang utama diderita oleh masyarakat Indonesia. Di samping itu, banyak bakteri patogen yang ditemukan resisten terhadap beberapa antibiotika. Oleh karena itu, upaya-upaya untuk mendapatkan antibiotika baru perlu dilakukan secara terus-menerus. Suatu galur lokal baru Bacillus subtilis BAC4 teridentifikasi memproduksi senyawa antibiotika yang menghambat pertumbuhan Serratia marcescens ATCC27117. Untuk memberdayakan galur tersebut, terhadap Bacillus subtilis BAC4 dilakukan mutasi dengan larutan akridin oranye dan diperoleh mutan Bacillus subtilis M10 yang memproduksi antibiotika berlebihan. Namun, kinetika produksi antibiotika oleh Bacillus subtilis M10 belum pernah dilaporkan. Penelitian ini bertujuan untuk mempelajari kinetika produksi antibiotika oleh mutan Bacillus subtilis M10. Bacillus subtilis M10 difermentasikan ke dalam media urea-sorbitol dan diamati kemampuan produksi antibiotikanya menggunakan Serratia marcescens ATCC 2711 sebagai bakteri uji. Hasil penelitian menunjukkan bahwa mutan Bacillus subtilis M10 memproduksi antibiotika sejak jam ke 8, dan produksi optimumnya terjadi pada jam ke 14 setelah inokulasi.
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 Grandprix T.M. Kadja Hartiwi Diastuti Hartiwi Diastuti I Gede Wenten Idris Idris Idris Idris Idris Idris Idris Idris Ira Oktavia 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