Simultaneous production of alpha and beta amylase enzymes using separate gene bearing recombinant vectors in the same Escherichia coli cells

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dc.authorid0000-0002-2304-2794en_US
dc.contributor.authorÖzcan, Dilek
dc.contributor.authorSipahioǧlu, Hikmet Murat
dc.date.accessioned2021-06-15T07:53:05Z
dc.date.available2021-06-15T07:53:05Z
dc.date.issued2020en_US
dc.departmentMTÖ Üniversitesi, Ziraat Fakültesi, Bitki Koruma Bölümüen_US
dc.description.abstractThe present study describes the simultaneous expression of thermostable industrial alpha (alpha) and beta (beta) amylase enzymes that have been used widely in starch industry. Genomic DNA of Bacillus stearothermophilus DSM 22 strain for alpha amylase and, Thermoanaerobacterium (Clostridium) thermosulfurogenes DSM 2229 strain for beta amylase were used as gene sources. Both genes were ligated into pETDuet-1 expression vector separately and resulting recombinant vectors were transformed into Escherichia coli BL21 competent cells by electroporation. The cells were first transformed by pETDuet-1/ alpha Amy recombinant plasmid, then the competent cells carrying this plasmid were prepared for the transformation of pETDuet-1/beta Amy plasmid. Enzymatic activities of bacterial colonies were detected on LB agar staining with iodide. Both enzymes were more produced by IPTG induction in BL21 cells and were purified using Ni-NTA agarose column. SDS-PAGE and western blot analyses showed that the molecular weight of purified alpha and beta amylase to be approximately 60 kDa and 55kDa, respectively. The concentration of the purified alpha and beta amylase were calculated as 4.59 mu g/mL and 3.17 mu g/mL with IPTG as an inducer in LB medium. The present study proposes a novel and efficient method for the production of thermostable alpha and beta amylases at the same E coli cells containing separate engineered plasmid vectors.en_US
dc.identifier.citationÖzcan, D., & Sipahioğlu, H. M. ( 2020). Simultaneous production of alpha and beta amylase enzymes using separate gene bearing recombinant vectors in the same Escherichia coli cells. Turkish Journal of Biology, 44 (4), 201-207.en_US
dc.identifier.doi10.3906/biy-2001-71
dc.identifier.endpage207en_US
dc.identifier.issn1300-0152en_US
dc.identifier.issn1303-6092en_US
dc.identifier.issue4en_US
dc.identifier.scopus2-s2.0-85089657865en_US
dc.identifier.scopusqualityQ2en_US
dc.identifier.startpage201en_US
dc.identifier.urihttps://doi.org/doi:10.3906/biy-2001-71
dc.identifier.urihttps://hdl.handle.net/20.500.12899/256
dc.identifier.volume44en_US
dc.identifier.wosWOS:000582626100008en_US
dc.identifier.wosqualityQ3en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.institutionauthorSipahioǧlu, Hikmet Murat
dc.language.isoenen_US
dc.publisherTÜBİTAKen_US
dc.relation.ispartofTurkish Journal of Biologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectAlpha amylaseen_US
dc.subjectBeta amylaseen_US
dc.subjectDual gene expressionen_US
dc.subjectBeta amylaseen_US
dc.subjectPurification of recombinant enzymeen_US
dc.titleSimultaneous production of alpha and beta amylase enzymes using separate gene bearing recombinant vectors in the same Escherichia coli cellsen_US
dc.typeArticleen_US

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