The DsbA signal peptide-mediated secretion of a highly efficient raw-starch-digesting, recombinant alpha-amylase from Bacillus licheniformis ATCC 9945a
Authorized Users Only
2013
Authors
Božić, NatašaPuertas, Juan-Miguel
Lončar, Nikola
Sans, Duran Cristina
Lopez-Santin, Josep
Vujčić, Zoran
Article (Published version)
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Show full item recordAbstract
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.
Keywords:
alpha-Amylase / Bacillus licheniformis / Escherichia coli / DsbA signal peptide / Raw starch hydrolysisSource:
Process Biochemistry, 2013, 48, 3, 438-442Publisher:
- Elsevier Sci Ltd, Oxford
Funding / projects:
- Production, purification and characterization of enzymes and small molecules and their application as soluble or immobilized in food biotechnology, biofuels production and environmental protection (RS-172048)
- Federation of European Biochemical Societies
- Joint Serbian-Spanish Action
- Spanish MICINN [CTQ2011-28398-CO2-01]
DOI: 10.1016/j.procbio.2013.01.016
ISSN: 1359-5113
WoS: 000318262900008
Scopus: 2-s2.0-84875906938
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IHTMTY - JOUR AU - Božić, Nataša AU - Puertas, Juan-Miguel AU - Lončar, Nikola AU - Sans, Duran Cristina AU - Lopez-Santin, Josep AU - Vujčić, Zoran PY - 2013 UR - https://cer.ihtm.bg.ac.rs/handle/123456789/1368 AB - 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. PB - Elsevier Sci Ltd, Oxford T2 - Process Biochemistry T1 - The DsbA signal peptide-mediated secretion of a highly efficient raw-starch-digesting, recombinant alpha-amylase from Bacillus licheniformis ATCC 9945a VL - 48 IS - 3 SP - 438 EP - 442 DO - 10.1016/j.procbio.2013.01.016 ER -
@article{ author = "Božić, Nataša and Puertas, Juan-Miguel and Lončar, Nikola and Sans, Duran Cristina and Lopez-Santin, Josep and Vujčić, Zoran", year = "2013", abstract = "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.", publisher = "Elsevier Sci Ltd, Oxford", journal = "Process Biochemistry", title = "The DsbA signal peptide-mediated secretion of a highly efficient raw-starch-digesting, recombinant alpha-amylase from Bacillus licheniformis ATCC 9945a", volume = "48", number = "3", pages = "438-442", doi = "10.1016/j.procbio.2013.01.016" }
Božić, N., Puertas, J., Lončar, N., Sans, D. C., Lopez-Santin, J.,& Vujčić, Z.. (2013). The DsbA signal peptide-mediated secretion of a highly efficient raw-starch-digesting, recombinant alpha-amylase from Bacillus licheniformis ATCC 9945a. in Process Biochemistry Elsevier Sci Ltd, Oxford., 48(3), 438-442. https://doi.org/10.1016/j.procbio.2013.01.016
Božić N, Puertas J, Lončar N, Sans DC, Lopez-Santin J, Vujčić Z. The DsbA signal peptide-mediated secretion of a highly efficient raw-starch-digesting, recombinant alpha-amylase from Bacillus licheniformis ATCC 9945a. in Process Biochemistry. 2013;48(3):438-442. doi:10.1016/j.procbio.2013.01.016 .
Božić, Nataša, Puertas, Juan-Miguel, Lončar, Nikola, Sans, Duran Cristina, Lopez-Santin, Josep, Vujčić, Zoran, "The DsbA signal peptide-mediated secretion of a highly efficient raw-starch-digesting, recombinant alpha-amylase from Bacillus licheniformis ATCC 9945a" in Process Biochemistry, 48, no. 3 (2013):438-442, https://doi.org/10.1016/j.procbio.2013.01.016 . .