Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible Promoter
This study focuses on the development of a xylose-inducible system to produce recombinant α-amylase from the bacterium Thermoactinomyces vulgaris 94-2A (amyTV) in the Bacillus expression system. To achieve this, a gene cassette was constructed using pWH1520 shuttle vector, containing a PxylA promote...
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| Format: | Article |
| Language: | English |
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North Carolina State University
2025-07-01
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| Series: | BioResources |
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| Online Access: | https://ojs.bioresources.com/index.php/BRJ/article/view/24573 |
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| author | Abd-El Aziem Farouk Iqra Aroob Quratulain Amjad Hala Mohamed Abdelmigid Ralf Greiner |
| author_facet | Abd-El Aziem Farouk Iqra Aroob Quratulain Amjad Hala Mohamed Abdelmigid Ralf Greiner |
| author_sort | Abd-El Aziem Farouk |
| collection | DOAJ |
| description | This study focuses on the development of a xylose-inducible system to produce recombinant α-amylase from the bacterium Thermoactinomyces vulgaris 94-2A (amyTV) in the Bacillus expression system. To achieve this, a gene cassette was constructed using pWH1520 shuttle vector, containing a PxylA promoter comprising glucose box and amyTV gene. The cassette was initially cloned in Escherichia coli for replication and subsequently, introduced into the Bacillus subtilis (GSB26) for expression. To optimize the secretion of α-amylase, different concentrations of xylose were used as inducers. It was found that the maximum amylase activity was achieved when 2% xylose was used as the inducer. Furthermore, using glucose alone and in combination with xylose as inducers indicated that glucose acted as a catabolic repressor for amyTV expression. Moreover, protein was efficiently secreted and did not accumulate in the cellular fractions, even at high expression levels. |
| format | Article |
| id | doaj-art-6ae2e346cdcc486995a85b5c41e51028 |
| institution | Kabale University |
| issn | 1930-2126 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | North Carolina State University |
| record_format | Article |
| series | BioResources |
| spelling | doaj-art-6ae2e346cdcc486995a85b5c41e510282025-08-20T03:43:55ZengNorth Carolina State UniversityBioResources1930-21262025-07-01203772877372872Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible PromoterAbd-El Aziem Farouk0https://orcid.org/0009-0007-0095-2270Iqra Aroob1https://orcid.org/0000-0002-2430-8646Quratulain Amjad2https://orcid.org/0009-0000-1597-8560Hala Mohamed Abdelmigid3https://orcid.org/0000-0002-2361-4959Ralf Greiner4https://orcid.org/0000-0002-0675-9849Department of Biotechnology, College of Science, Taif University, P.O. Box 1109 Taif 21944, Saudi ArabiaSchool of Allied Health Sciences, University of Child Health Sciences, The Children’s Hospital LahoreSchool of Biological Sciences, Faculty of Allied Health Sciences, Superior University, Raiwind Road, Lahore, PakistanDepartment of Biotechnology, College of Science, Taif University, P.O. Box 1109 Taif 21944, Saudi ArabiaDepartment of Food Technology and Bioprocess Engineering, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, Karlsruhe, GermanyThis study focuses on the development of a xylose-inducible system to produce recombinant α-amylase from the bacterium Thermoactinomyces vulgaris 94-2A (amyTV) in the Bacillus expression system. To achieve this, a gene cassette was constructed using pWH1520 shuttle vector, containing a PxylA promoter comprising glucose box and amyTV gene. The cassette was initially cloned in Escherichia coli for replication and subsequently, introduced into the Bacillus subtilis (GSB26) for expression. To optimize the secretion of α-amylase, different concentrations of xylose were used as inducers. It was found that the maximum amylase activity was achieved when 2% xylose was used as the inducer. Furthermore, using glucose alone and in combination with xylose as inducers indicated that glucose acted as a catabolic repressor for amyTV expression. Moreover, protein was efficiently secreted and did not accumulate in the cellular fractions, even at high expression levels.https://ojs.bioresources.com/index.php/BRJ/article/view/24573recombinant amylasebacillus expression systemscatabolic repression |
| spellingShingle | Abd-El Aziem Farouk Iqra Aroob Quratulain Amjad Hala Mohamed Abdelmigid Ralf Greiner Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible Promoter BioResources recombinant amylase bacillus expression systems catabolic repression |
| title | Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible Promoter |
| title_full | Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible Promoter |
| title_fullStr | Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible Promoter |
| title_full_unstemmed | Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible Promoter |
| title_short | Efficient Extracellular Production of α-Amylase in Bacillus subtilis under the Influence of Xylose-Inducible Promoter |
| title_sort | efficient extracellular production of α amylase in bacillus subtilis under the influence of xylose inducible promoter |
| topic | recombinant amylase bacillus expression systems catabolic repression |
| url | https://ojs.bioresources.com/index.php/BRJ/article/view/24573 |
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