Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile Dye
The aim of the present research was the efficient degradation of industrial textile wastewater dyes using a very active cloned laccase enzyme. For this purpose, potent laccase-producing bacteria were isolated from soil samples collected from wastewater-replenished textile sites in Punjab, Pakistan....
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MDPI AG
2024-11-01
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| author | Zannara Mustafa Ikram ul Haq Ali Nawaz Abdulrahman H. Alessa Muhammad Nauman Aftab Ahmad A. Alsaigh Aziz ur Rehman |
| author_facet | Zannara Mustafa Ikram ul Haq Ali Nawaz Abdulrahman H. Alessa Muhammad Nauman Aftab Ahmad A. Alsaigh Aziz ur Rehman |
| author_sort | Zannara Mustafa |
| collection | DOAJ |
| description | The aim of the present research was the efficient degradation of industrial textile wastewater dyes using a very active cloned laccase enzyme. For this purpose, potent laccase-producing bacteria were isolated from soil samples collected from wastewater-replenished textile sites in Punjab, Pakistan. The laccase gene from locally isolated strain LI-81, identified as <i>Bacillus megaterium</i>, was cloned into vector pET21a, which was further transformed into <i>E. coli</i> BL21 codon plus. The optimized conditions for the increased production of laccase include fermentation in a 2% glucose, 5% yeast extract and 250 mg/L CuSO<sub>4</sub> medium with pH 7.5; inoculation with 5% inoculum; induction with 0.1 mM IPTG at 0.5 O.D.; and incubation for 36 h at 37 °C. The crude enzyme produced was employed for the removal of commercially used textile dyes. The dyes were quickly precipitated under optimized reaction conditions. Rose bengal, brilliant green, brilliant blue G, Coomassie brilliant blue R and methylene blue were precipitated at rates of 10.69, 54.47, 84.04, 78.99 and 7.40%, respectively. The FTIR and UV–Vis spectroscopic analyses of dyes before and after confirmed the chemical changes brought about by the cloned laccase that led to the dye removal. |
| format | Article |
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| institution | DOAJ |
| issn | 1420-3049 |
| language | English |
| publishDate | 2024-11-01 |
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| spelling | doaj-art-5fa7776e618d455da5f56a8d4f5181d82025-08-20T02:50:38ZengMDPI AGMolecules1420-30492024-11-012923551410.3390/molecules29235514Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile DyeZannara Mustafa0Ikram ul Haq1Ali Nawaz2Abdulrahman H. Alessa3Muhammad Nauman Aftab4Ahmad A. Alsaigh5Aziz ur Rehman6Dr. Ikram ul Haq Institute of Industrial Biotechnology, GC University, Lahore 54600, PakistanDr. Ikram ul Haq Institute of Industrial Biotechnology, GC University, Lahore 54600, PakistanDr. Ikram ul Haq Institute of Industrial Biotechnology, GC University, Lahore 54600, PakistanDepartment of Biology, Faculty of Science, University of Tabuk, Tabuk 47512, Saudi ArabiaDr. Ikram ul Haq Institute of Industrial Biotechnology, GC University, Lahore 54600, PakistanDepartment of Biology, Faculty of Science, Umm Al-Qura University, Makkah 24382, Saudi ArabiaDr. Ikram ul Haq Institute of Industrial Biotechnology, GC University, Lahore 54600, PakistanThe aim of the present research was the efficient degradation of industrial textile wastewater dyes using a very active cloned laccase enzyme. For this purpose, potent laccase-producing bacteria were isolated from soil samples collected from wastewater-replenished textile sites in Punjab, Pakistan. The laccase gene from locally isolated strain LI-81, identified as <i>Bacillus megaterium</i>, was cloned into vector pET21a, which was further transformed into <i>E. coli</i> BL21 codon plus. The optimized conditions for the increased production of laccase include fermentation in a 2% glucose, 5% yeast extract and 250 mg/L CuSO<sub>4</sub> medium with pH 7.5; inoculation with 5% inoculum; induction with 0.1 mM IPTG at 0.5 O.D.; and incubation for 36 h at 37 °C. The crude enzyme produced was employed for the removal of commercially used textile dyes. The dyes were quickly precipitated under optimized reaction conditions. Rose bengal, brilliant green, brilliant blue G, Coomassie brilliant blue R and methylene blue were precipitated at rates of 10.69, 54.47, 84.04, 78.99 and 7.40%, respectively. The FTIR and UV–Vis spectroscopic analyses of dyes before and after confirmed the chemical changes brought about by the cloned laccase that led to the dye removal.https://www.mdpi.com/1420-3049/29/23/5514recombinant laccasetextiledyeoptimizationeffluentprecipitation |
| spellingShingle | Zannara Mustafa Ikram ul Haq Ali Nawaz Abdulrahman H. Alessa Muhammad Nauman Aftab Ahmad A. Alsaigh Aziz ur Rehman Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile Dye Molecules recombinant laccase textile dye optimization effluent precipitation |
| title | Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile Dye |
| title_full | Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile Dye |
| title_fullStr | Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile Dye |
| title_full_unstemmed | Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile Dye |
| title_short | Laccase Production Optimization from Recombinant <i>E. coli</i> BL21 Codon Plus Containing Novel Laccase Gene from <i>Bacillus megaterium</i> for Removal of Wastewater Textile Dye |
| title_sort | laccase production optimization from recombinant i e coli i bl21 codon plus containing novel laccase gene from i bacillus megaterium i for removal of wastewater textile dye |
| topic | recombinant laccase textile dye optimization effluent precipitation |
| url | https://www.mdpi.com/1420-3049/29/23/5514 |
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