dc.contributor.author | Burhanoğlu, T. | |
dc.contributor.author | Sürmeli, Yusuf | |
dc.contributor.author | Şanlı Mohamed, Gülşah | |
dc.date.accessioned | 2022-05-11T14:07:11Z | |
dc.date.available | 2022-05-11T14:07:11Z | |
dc.date.issued | 2020 | |
dc.identifier.issn | 0141-8130 | |
dc.identifier.uri | https://doi.org/10.1016/j.ijbiomac.2020.07.171 | |
dc.identifier.uri | https://hdl.handle.net/20.500.11776/5003 | |
dc.description.abstract | In this study, the heterologous expression and biochemical characterization of a thermostable ?-amylase from Geobacillus sp. GS33 was investigated. The recombinant ?-amylase was overexpressed in Escherichia coli BL21 (?DE) and purified via anion exchange and size-exclusion chromatography. The purified ?-amylase had a molecular weight of about 60 kDa, and was active in a broad range of pH 3–10 and temperature (40–90 °C) with maximum activity at pH 7–8 and 60 °C. The enzyme retained 50% residual activity at 65 °C, but only 20% at 85 °C after 16 h. At pH 9 and pH 7, the residual activity at 65 °C was 50% and 30%, respectively. The enzyme was remarkably activated by Co2+, Ca2+, Mg2+, PMSF, DTT, and Triton X-100, but partially inhibited by Cu2+, methanol, hexane, ethanol, acetone, SDS, and Tween 20. A molecular phylogeny analysis showed that the enzyme's amino acid sequence had the closest connection with an ?-amylase from Geobacillus thermoleovorans subsp. stromboliensis nov. 3D-structure-based amino acid sequence alignments revealed that the three catalytic residues (D217, E246, D314) and the four Ca2+ ion coordination residues (N143, E177, D186, H221) were conserved in ?-amylase from Geobacillus sp. GS33. The temperature stability and neutral pH optimum suggest that the enzyme may be useful for industrial applications. © 2020 Elsevier B.V. | en_US |
dc.description.sponsorship | The authors would like to thank Biotechnology & Bioengineering Research Center at ?zmir Institute of Technology for the facilities and technical support. | en_US |
dc.language.iso | eng | en_US |
dc.publisher | Elsevier B.V. | en_US |
dc.identifier.doi | 10.1016/j.ijbiomac.2020.07.171 | |
dc.rights | info:eu-repo/semantics/openAccess | en_US |
dc.subject | Geobacillus | en_US |
dc.subject | Thermostability | en_US |
dc.subject | ?-Amylase | en_US |
dc.subject | acetone | en_US |
dc.subject | alcohol | en_US |
dc.subject | amylase | en_US |
dc.subject | benzylsulfonyl fluoride | en_US |
dc.subject | calcium ion | en_US |
dc.subject | cobalt | en_US |
dc.subject | copper ion | en_US |
dc.subject | dithiothreitol | en_US |
dc.subject | dodecyl sulfate sodium | en_US |
dc.subject | hexane | en_US |
dc.subject | magnesium ion | en_US |
dc.subject | methanol | en_US |
dc.subject | polysorbate 20 | en_US |
dc.subject | triton x 100 | en_US |
dc.subject | amylase | en_US |
dc.subject | bacterial protein | en_US |
dc.subject | amino acid sequence | en_US |
dc.subject | anion exchange chromatography | en_US |
dc.subject | Article | en_US |
dc.subject | controlled study | en_US |
dc.subject | enzyme activation | en_US |
dc.subject | enzyme active site | en_US |
dc.subject | enzyme activity | en_US |
dc.subject | enzyme inhibition | en_US |
dc.subject | enzyme stability | en_US |
dc.subject | enzyme structure | en_US |
dc.subject | Geobacillus | en_US |
dc.subject | Geobacillus thermoleovorans | en_US |
dc.subject | molecular phylogeny | en_US |
dc.subject | molecular weight | en_US |
dc.subject | nonhuman | en_US |
dc.subject | nucleotide sequence | en_US |
dc.subject | pH | en_US |
dc.subject | sequence alignment | en_US |
dc.subject | size exclusion chromatography | en_US |
dc.subject | temperature | en_US |
dc.subject | thermostability | en_US |
dc.subject | chemistry | en_US |
dc.subject | enzymology | en_US |
dc.subject | genetics | en_US |
dc.subject | Geobacillus | en_US |
dc.subject | heat | en_US |
dc.subject | ion exchange chromatography | en_US |
dc.subject | metabolism | en_US |
dc.subject | molecular cloning | en_US |
dc.subject | molecular evolution | en_US |
dc.subject | molecular model | en_US |
dc.subject | phylogeny | en_US |
dc.subject | procedures | en_US |
dc.subject | protein conformation | en_US |
dc.subject | thermodynamics | en_US |
dc.subject | alpha-Amylases | en_US |
dc.subject | Amino Acid Sequence | en_US |
dc.subject | Bacterial Proteins | en_US |
dc.subject | Chromatography, Gel | en_US |
dc.subject | Chromatography, Ion Exchange | en_US |
dc.subject | Cloning, Molecular | en_US |
dc.subject | Enzyme Stability | en_US |
dc.subject | Evolution, Molecular | en_US |
dc.subject | Geobacillus | en_US |
dc.subject | Hot Temperature | en_US |
dc.subject | Hydrogen-Ion Concentration | en_US |
dc.subject | Models, Molecular | en_US |
dc.subject | Molecular Weight | en_US |
dc.subject | Phylogeny | en_US |
dc.subject | Protein Conformation | en_US |
dc.subject | Thermodynamics | en_US |
dc.title | Identification and characterization of novel thermostable ?-amylase from Geobacillus sp. GS33 | en_US |
dc.type | article | en_US |
dc.relation.ispartof | International Journal of Biological Macromolecules | en_US |
dc.department | Fakülteler, Ziraat Fakültesi, Tarımsal Biyoteknoloji Bölümü | en_US |
dc.identifier.volume | 164 | en_US |
dc.identifier.startpage | 578 | en_US |
dc.identifier.endpage | 585 | en_US |
dc.institutionauthor | Sürmeli, Yusuf | |
dc.relation.publicationcategory | Makale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı | en_US |
dc.authorscopusid | 57218225937 | |
dc.authorscopusid | 55758999000 | |
dc.authorscopusid | 36680469600 | |
dc.identifier.wos | WOS:000588093700054 | en_US |
dc.identifier.scopus | 2-s2.0-85088394880 | en_US |
dc.identifier.pmid | 32693140 | en_US |