Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1
An endoglucanase gene from glycoside hydrolase family 5, had been isolated from Bacillus sp. RP1 and cloned into Escherichia coli. The cloned gene comprised the promoter, coding sequence and terminator of the gene. This gene encoded a protein with 499 amino acid residues (Mr=55.2 kDa) with a typica...
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| Format: | Article |
| Language: | English |
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Indonesian Society for Microbiology
2015-03-01
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| Series: | Microbiology Indonesia |
| Subjects: | |
| Online Access: | https://jurnal.permi.or.id/index.php/mionline/article/view/307 |
| _version_ | 1818901557649866752 |
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| author | MAELITA RAMDANI MOEIS DESSY NATALIA RAHMA WIDYA NINGRUM ARI DWIJAYANTI |
| author_facet | MAELITA RAMDANI MOEIS DESSY NATALIA RAHMA WIDYA NINGRUM ARI DWIJAYANTI |
| author_sort | MAELITA RAMDANI MOEIS |
| collection | DOAJ |
| description | An endoglucanase gene from glycoside hydrolase family 5, had been isolated from Bacillus sp. RP1 and cloned into Escherichia coli. The cloned gene comprised the promoter, coding sequence and terminator of the gene. This gene encoded a protein with 499 amino acid residues (Mr=55.2 kDa) with a typical Bacillus signal peptide. The recombinant endoglucanase (EG) had optimum activity at pH 5.0 and 50 °C. The recombinant EG was expressed in the extracellular, intracellular, and periplasmic fractions with the highest total activity (60.15%) in the intracellular fraction, measured at three hours after isopropyl-β-Dthiogalactopyranoside (IPTG) induction. Three hours after the addition of 1% carboxymethyl cellulose (CMC), there was a two-fold increase in intracellular EG specific activity compared to the uninduced cells. Three hours after the addition of 1 mM IPTG, 1% glucose, 1% galactose or 1% cellobiose the intracellular EG specific activity decreased compared to the uninduced cells. |
| first_indexed | 2024-12-19T20:21:39Z |
| format | Article |
| id | doaj.art-fe7fdb65af0d4835a778a85df0c12e86 |
| institution | Directory Open Access Journal |
| issn | 1978-3477 2087-8575 |
| language | English |
| last_indexed | 2024-12-19T20:21:39Z |
| publishDate | 2015-03-01 |
| publisher | Indonesian Society for Microbiology |
| record_format | Article |
| series | Microbiology Indonesia |
| spelling | doaj.art-fe7fdb65af0d4835a778a85df0c12e862022-12-21T20:06:59ZengIndonesian Society for MicrobiologyMicrobiology Indonesia1978-34772087-85752015-03-018410.5454/mi.8.4.4Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1MAELITA RAMDANI MOEISDESSY NATALIARAHMA WIDYA NINGRUMARI DWIJAYANTIAn endoglucanase gene from glycoside hydrolase family 5, had been isolated from Bacillus sp. RP1 and cloned into Escherichia coli. The cloned gene comprised the promoter, coding sequence and terminator of the gene. This gene encoded a protein with 499 amino acid residues (Mr=55.2 kDa) with a typical Bacillus signal peptide. The recombinant endoglucanase (EG) had optimum activity at pH 5.0 and 50 °C. The recombinant EG was expressed in the extracellular, intracellular, and periplasmic fractions with the highest total activity (60.15%) in the intracellular fraction, measured at three hours after isopropyl-β-Dthiogalactopyranoside (IPTG) induction. Three hours after the addition of 1% carboxymethyl cellulose (CMC), there was a two-fold increase in intracellular EG specific activity compared to the uninduced cells. Three hours after the addition of 1 mM IPTG, 1% glucose, 1% galactose or 1% cellobiose the intracellular EG specific activity decreased compared to the uninduced cells.https://jurnal.permi.or.id/index.php/mionline/article/view/307Bacillus sp. RP1recombinant endoglucanasenative promoternative signal peptide |
| spellingShingle | MAELITA RAMDANI MOEIS DESSY NATALIA RAHMA WIDYA NINGRUM ARI DWIJAYANTI Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1 Microbiology Indonesia Bacillus sp. RP1 recombinant endoglucanase native promoter native signal peptide |
| title | Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1 |
| title_full | Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1 |
| title_fullStr | Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1 |
| title_full_unstemmed | Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1 |
| title_short | Cloning and Expression of Endoglucanase Gene from Thermophilic Bacteria Bacillus sp. RP1 |
| title_sort | cloning and expression of endoglucanase gene from thermophilic bacteria bacillus sp rp1 |
| topic | Bacillus sp. RP1 recombinant endoglucanase native promoter native signal peptide |
| url | https://jurnal.permi.or.id/index.php/mionline/article/view/307 |
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