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Expression, Characterisation and Structural Homology Modelling of Recombinant Mercuric Reductase of Streptomyces sp. AS2

Journal of Tropical Biodiversity and Biotechnology · Vol. 9 No. 4 (2024)

DOI
10.22146/jtbb.88773
ISSN
2540-9581, 2540-9573
Terbit
2024-10-21
Penerbit
Universitas Gadjah Mada
Bahasa
eng
Lisensi
creativecommons.org/licenses/by-sa/4.0

Abstrak

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.

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  • Universitas Gadjah Mada
  • Universitas Muhammadiyah Pontianak
  • Muhammadiyah University of Yogyakarta
  • Universitas Islam Negeri Sultan Maulana Hasanuddin Banten
  • Universitas Halu Oleo
  • Universitas Muhammadiyah Semarang
  • National Research and Innovation Agency
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Rekan penulisOrganisasiArtikel bersama
Masashi KawaichiNara Institute of Science and Technology6
Ardaning NurilianiUniversitas Gadjah Mada4
Didik IndradewaUniversitas Gadjah Mada3
Charlie Ester de FretesNational Research and Innovation Agency3
Ari SunandarUniversitas Muhammadiyah Pontianak2
Nur Arfa YantiUniversitas Halu Oleo2
Sebastian MarginoUniversitas Gadjah Mada2
Widya AsmaraUniversitas Gadjah Mada2
Wayan Tunas ArtamaUniversitas Gadjah Mada2
Sri DarmawatiUniversitas Muhammadiyah Semarang2
Woro Anindito Sri TunjungUniversitas Gadjah Mada2
Radhiyah Mardhiyah HamidUniversitas Gadjah Mada2
Rifqi Zahroh JanatunaimUniversitas Gadjah Mada2
Rarastoeti PratiwiUniversitas Gadjah Mada2
Tri Rini NuringtyasUniversitas Gadjah Mada2
Donny WidiantoUniversitas Gadjah Mada2
Alfino SebastianUniversitas Gadjah Mada2
Yuni SineUniversitas Gadjah Mada2
Taryono TaryonoUniversitas Gadjah Mada2
Ratna Dewi EskundariUniversitas Bangun Nusantara2
Yoshihiko YanoKobe University Hospital1
Yoni AstutiMuhammadiyah University of Yogyakarta1
Anisa Zuliana UtamiMuhammadiyah University of Yogyakarta1
Agus SuhartoMuhammadiyah University of Yogyakarta1
Resti AprianiUniversitas Muhammadiyah Pontianak1
Friti May SandiUniversitas Muhammadiyah Pontianak1
Mahwar QurbaniahUniversitas Muhammadiyah Pontianak1
Nurhayati NurhayatiUniversitas Muhammadiyah Pontianak1
Bima MahendraUniversitas Gadjah Mada1
Nabila Ramiza PuteriUniversitas Gadjah Mada1
Sekar RahayuUniversitas Gadjah Mada1
Yuka OgakiAfiliasi tidak diketahui1
Hiroyuki TsujiAfiliasi tidak diketahui1
Ko ShimamotoAfiliasi tidak diketahui1
Syaiful AnwarDiponegoro University1

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Sumber metadata: OAI-PMH (metadata jurnal) https://jurnal.ugm.ac.id/jtbb/oai, pertama di-harvest 9 Nov 2026, terakhir terlihat 9 Nov 2026.
Metadata asli (oai_dc) 
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	<dc:title xml:lang="en-US">Expression, Characterisation and Structural Homology Modelling of Recombinant Mercuric Reductase of Streptomyces sp. AS2</dc:title>
	<dc:creator>Khasanah, Anis Uswatun</dc:creator>
	<dc:creator>Putri, Wahyu Aristyaning</dc:creator>
	<dc:creator>Rahayu, Hanum Mukti</dc:creator>
	<dc:creator>Sembiring, Langkah</dc:creator>
	<dc:creator>Purwestri, Yekti Asih</dc:creator>
	<dc:subject xml:lang="en-US">Microbiology, Bioinformatics, Biochemistry</dc:subject>
	<dc:subject xml:lang="en-US">E. coli BL-21; gene cloning; mercuric reductase; protein expression</dc:subject>
	<dc:description xml:lang="en-US">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. </dc:description>
	<dc:publisher xml:lang="en-US">Universitas Gadjah Mada</dc:publisher>
	<dc:contributor xml:lang="en-US"></dc:contributor>
	<dc:date>2024-10-21</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
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	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://journal.ugm.ac.id/jtbb/article/view/88773</dc:identifier>
	<dc:identifier>10.22146/jtbb.88773</dc:identifier>
	<dc:source xml:lang="en-US">Journal of Tropical Biodiversity and Biotechnology; Vol 9, No 4 (2024): December; jtbb88773</dc:source>
	<dc:source>2540-9581</dc:source>
	<dc:source>2540-9573</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>https://journal.ugm.ac.id/jtbb/article/view/88773/40132</dc:relation>
	<dc:relation>https://journal.ugm.ac.id/jtbb/article/downloadSuppFile/88773/28828</dc:relation>
	<dc:relation>https://journal.ugm.ac.id/jtbb/article/downloadSuppFile/88773/28829</dc:relation>
	<dc:coverage xml:lang="en-US">Sout East Asia</dc:coverage>
	<dc:coverage xml:lang="en-US"></dc:coverage>
	<dc:coverage xml:lang="en-US"></dc:coverage>
	<dc:rights xml:lang="en-US">Copyright (c) 2024 Journal of Tropical Biodiversity and Biotechnology</dc:rights>
	<dc:rights xml:lang="en-US">http://creativecommons.org/licenses/by-sa/4.0</dc:rights>
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