The DsbA signal peptide-mediated secretion of a highly efficient raw-starch-digesting, recombinant α-amylase from Bacillus licheniformis ATCC 9945a
| Autor: | Nataša Božić, Josep López-Santín, Zoran Vujčić, Cristina Sans Duran, Juan Miguel Puertas, Nikola Lončar |
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| Rok vydání: | 2013 |
| Předmět: |
0106 biological sciences
Signal peptide Bioengineering medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology Biochemistry law.invention Gene product alpha-Amylase 03 medical and health sciences DsbA signal peptide law 010608 biotechnology Escherichia coli medicine Bacillus licheniformis Amylase 030304 developmental biology 0303 health sciences Expression vector biology Raw starch hydrolysis biology.organism_classification DsbA biology.protein Recombinant DNA |
| Zdroj: | Process Biochemistry |
| ISSN: | 1359-5113 |
| DOI: | 10.1016/j.procbio.2013.01.016 |
| Popis: | In this study, a new approach for extracellular production of recombinant alpha-amylase in Escherichia coli was investigated. A gene encoding a highly efficient raw-starch-digesting alpha-amylase from Bacillus licheniformis ATCC 9945a was cloned and expressed in E. coli. The gene encoding mature alpha-amylase was cloned into the pDAss expression vector, and secretion of the gene product was regulated by fusion to the signal peptide of DsbA, a well-characterized E. coli periplasmic protein. E. coli BL21 (DE3) carrying pDAss vector containing amylase gene had approximately 2.5-fold higher volumetric enzyme productivity than the natural system. The recombinant enzyme showed higher efficiency for digesting diverse raw starches when compared with the native enzyme and was similar to commercial alpha-amylase in its ability to hydrolyze raw starches. The properties of the recombinant enzyme demonstrate the potential of the DsbA signal peptide approach for the secretory production of the fully active, industrially important recombinant enzyme. (C) 2013 Elsevier Ltd. All rights reserved. |
| Databáze: | OpenAIRE |
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