Identification and characterization of novel thermostable ?-amylase from Geobacillus sp. GS33

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dc.authorscopusid57218225937
dc.authorscopusid55758999000
dc.authorscopusid36680469600
dc.contributor.authorBurhanoğlu, T.
dc.contributor.authorSürmeli, Yusuf
dc.contributor.authorŞanlı Mohamed, Gülşah
dc.date.accessioned2022-05-11T14:07:11Z
dc.date.available2022-05-11T14:07:11Z
dc.date.issued2020
dc.departmentFakülteler, Ziraat Fakültesi, Tarımsal Biyoteknoloji Bölümü
dc.description.abstractIn 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.
dc.description.sponsorshipThe authors would like to thank Biotechnology & Bioengineering Research Center at ?zmir Institute of Technology for the facilities and technical support.
dc.identifier.doi10.1016/j.ijbiomac.2020.07.171
dc.identifier.endpage585
dc.identifier.issn0141-8130
dc.identifier.pmid32693140
dc.identifier.scopus2-s2.0-85088394880
dc.identifier.scopusqualityQ1
dc.identifier.startpage578
dc.identifier.urihttps://doi.org/10.1016/j.ijbiomac.2020.07.171
dc.identifier.urihttps://hdl.handle.net/20.500.11776/5003
dc.identifier.volume164
dc.identifier.wosWOS:000588093700054
dc.identifier.wosqualityQ1
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.institutionauthorSürmeli, Yusuf
dc.language.isoen
dc.publisherElsevier B.V.
dc.relation.ispartofInternational Journal of Biological Macromolecules
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectGeobacillus
dc.subjectThermostability
dc.subject?-Amylase
dc.subjectacetone
dc.subjectalcohol
dc.subjectamylase
dc.subjectbenzylsulfonyl fluoride
dc.subjectcalcium ion
dc.subjectcobalt
dc.subjectcopper ion
dc.subjectdithiothreitol
dc.subjectdodecyl sulfate sodium
dc.subjecthexane
dc.subjectmagnesium ion
dc.subjectmethanol
dc.subjectpolysorbate 20
dc.subjecttriton x 100
dc.subjectamylase
dc.subjectbacterial protein
dc.subjectamino acid sequence
dc.subjectanion exchange chromatography
dc.subjectArticle
dc.subjectcontrolled study
dc.subjectenzyme activation
dc.subjectenzyme active site
dc.subjectenzyme activity
dc.subjectenzyme inhibition
dc.subjectenzyme stability
dc.subjectenzyme structure
dc.subjectGeobacillus
dc.subjectGeobacillus thermoleovorans
dc.subjectmolecular phylogeny
dc.subjectmolecular weight
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectpH
dc.subjectsequence alignment
dc.subjectsize exclusion chromatography
dc.subjecttemperature
dc.subjectthermostability
dc.subjectchemistry
dc.subjectenzymology
dc.subjectgenetics
dc.subjectGeobacillus
dc.subjectheat
dc.subjection exchange chromatography
dc.subjectmetabolism
dc.subjectmolecular cloning
dc.subjectmolecular evolution
dc.subjectmolecular model
dc.subjectphylogeny
dc.subjectprocedures
dc.subjectprotein conformation
dc.subjectthermodynamics
dc.subjectalpha-Amylases
dc.subjectAmino Acid Sequence
dc.subjectBacterial Proteins
dc.subjectChromatography, Gel
dc.subjectChromatography, Ion Exchange
dc.subjectCloning, Molecular
dc.subjectEnzyme Stability
dc.subjectEvolution, Molecular
dc.subjectGeobacillus
dc.subjectHot Temperature
dc.subjectHydrogen-Ion Concentration
dc.subjectModels, Molecular
dc.subjectMolecular Weight
dc.subjectPhylogeny
dc.subjectProtein Conformation
dc.subjectThermodynamics
dc.titleIdentification and characterization of novel thermostable ?-amylase from Geobacillus sp. GS33
dc.typeArticle

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