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All Journal Natural Science: Journal of Science and Technology EUGENIA Jurnal Natur Indonesia Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Indonesian Journal of Biotechnology Journal of Tropical Biodiversity and Biotechnology Prosiding SNPBS (Seminar Nasional Pendidikan Biologi dan Saintek)
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Laboratorium Mikrobiologi Fakultas Biologi Universitas Gadjah Mada
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Environmental Science Immunology & microbiology Materials Science & Nanotechnology

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Expression, Characterisation and Structural Homology Modelling of Recombinant Mercuric Reductase of Streptomyces sp. AS2 Khasanah, Anis Uswatun; Putri, Wahyu Aristyaning; Rahayu, Hanum Mukti; Sembiring, Langkah; Purwestri, Yekti Asih
Journal of Tropical Biodiversity and Biotechnology Vol 9, No 4 (2024): December
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.88773

Abstract

Mercury pollution poses a significant environmental challenge worldwide, prompting extensive efforts over the past two decades to combat its detrimental effects. Cloning merA from Streptomyces sp. AS2 (Accession numbers LC026157) into the expression vector pET-28c (+) marks a critical advancement in this field, necessitating further investigation into the expression and structural analysis of the resulting recombinant mercuric reductase protein. This study aimed to optimise the expression and characterise the structural MerA protein. The study involved the expression of merA from AS2 isolate in the host Escherichia coli BL21 and the measurement of mercuric reductase using SDS-PAGE. Induction of E. coli BL21 was optimized by adding IPTG concentration and incubation time. Purification of mercuric reductase was attempted using ammonium sulphate precipitation, dialysis, and column chromatography. Protein structural characterisation was conducted using computational modelling tools Swiss-Model and Phyre2. Expression of merA from AS2 isolate was successfully performed in E. coli BL21, with SDS-PAGE showing a dominant band in the 55-70 kDa range using IPTG concentration 1 and 1,2 mM and 18-hour incubation time. The specific activity of mercuric reductase was obtained at an enzyme concentration of 294.07 Unit/mg. Protein structural characterisation revealed homology with Lysinibacillus sphaericus (Swiss-Model) and similar folding to c5c1Yc, a known mercuric reductase from the same species using Phyre2. The successful expression of recombinant pET-28c (+)-MerA in E. coli BL21 offers new opportunities for bioremediation efforts targeting mercury contamination. 
Isolasi dan Uji Aktivitas Antibakteri Actinomycetes dari Rhizosfer Bakau di Hutan Bakau Torosiaje Gorontalo Retnowati, Yuliana; Sembiring, Langkah; Moeljopawiro, Sukarti; Djohan, Tjut S.; Soetarto, Endang S.
Prosiding SNPBS (Seminar Nasional Pendidikan Biologi dan Saintek) 2017: Prosiding SNPBS (Seminar Nasional Pendidikan Biologi dan Saintek)
Publisher : Universitas Muhammadiyah Surakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (268.963 KB)

Abstract

Actinomycetes penghasil antibiotik telah dieksplorasi dari berbagai sumber di lingkungan, terutama lingkungan ekstrim.Hutan bakau Torosiaje di Provinsi Gorontalo memiliki kondisi geomorfologi yang unik berupa ekosistem hutan bakau karsdengan dua tipe area yaitu tipe fringe dan overwash mangrove yang tersusun oleh jenis bakau yang bervariasi. Penelitian inidi desain untuk mendapatkan isolat Actinomycetes dari rhizosfer berbagai jenis bakau di hutan bakau Torosiaje Gorontalodan menganalisis aktifitas antibakteri melawan bakteri patogen. Sampel tanah dikoleksi dari rizosfer tujuh jenis pohon bakauyaitu Rhizophora mucronata dan Bruguiera gymnorhiza pada tipe hutan overwash, Rhizophora apiculata, Bruguieragymnorhiza pada zona middle tipe hutan Fringe, Avicenia marina, Xylocarpus sp, Ceriops tagal dan Soneratia alba padazona upper tipe hutan fringe. Pre-treatmen sampel tanah berdasarkan metode panas basah pada suhu 60oC selama 15 menit.Isolasi selektif Actinomycetes menggunakan medium Starch Casein Agar yang disuplementasi dengan cyclohexamide dannystatin. Seleksi isolat penghasil antibiotik berdasarkan metode agar blok menggunakan bakteri uji Eschericia coli,Staphylococcus aureus dan Bacillus subtillis. Aktifitas antibakteri ditandai dengan pembentukan zona hambat disekitarpertumbuhan actinomyctes. Diameter zona hambat dan diameter koloni Actinomycetes diukur untuk menentukan indekszona hambat. Hasil penelitian diperoleh sebanyak 167 isolat Actinomycetes yang terdistribusi pada rizosfer 7 jenis bakau. 77isolat Actinomycetes menunjukkan aktifitas antibakteri melawan bakteri patogen, terdiri dari 52 isolat melawan bakteriGram-positif (narraw spectrum) dan 25 isolat melawan bakteri Gram-positif dan Gram-negatif (broad spectrum). IsolatActinomycetes penghasil antibiotik memiliki karakter morfologi yang bervariasi yang didominasi oleh koloni berwana putihdan pigmen terdifusi berwarna kekuningan sampai coklat dan dikelompokkan kedalam 15 grup.
Co-Authors Ambarwati Artama, I Wayan Tunas Bambang Setiaji C. J. Soegihardjo Charlie Ester de Fretes Christina Salaki Dwi Suhartanti Elisa Nurnawati ENDANG S. SOETARTO Endang Sutriswati Rahayu Helen Joan Lawalata Jamhari Jamhari Jesmandt Situmorang Jusup Subagja, Jusup Khasanah, Novita Uswatun Laksono Trisnantoro Lela Susilawati Lisa Lisdiana Masashi Kawaichi, Masashi Niken Handayani Nurhayani H. Muhiddin, Nurhayani H. Orryani Lambui Putri, Wahyu Aristyaning Rahayu, Hanum Mukti S. Rahayu, S. Sebastian Margino Selvi Marcellia Sri Darmawati Subagja, Yusup Subagus Wahyuono Sukarti Moeljopawiro Syaiful Anwar Tanti Azizah S Tjut S. Djohan Tri Ardyati Wayan T. Artama Wayan T. Artama Widayati, Wiwik E. Widya Asmara Yekti Asih Purwestri Yuliana Retnowati Yurnaliza, Yurnaliza