Please use this identifier to cite or link to this item: https://hdl.handle.net/11147/10376
Title: Identification and characterization of novel thermostable alpha-amylase from Geobacillus sp. GS33
Authors: Burhanoğlu, Tülin
Sürmeli, Yusuf
Şanlı Mohamed, Gülşah
Keywords: alpha-Amylase
Geobacillus
Thermostability
Issue Date: 2020
Publisher: Elsevier Ltd.
Abstract: In this study, the heterologous expression and biochemical characterization of a thermostable alpha-amylase from Geobacillus sp. GS33 was investigated. The recombinant alpha-amylase was overexpressed in Escherichia coli BL21 (lambda DE) and purified via anion exchange and size-exclusion chromatography. The purified alpha-amylase had a molecular weight of about 60 kDa, and was active in a broad range of pH 3-10 and temperature (40-90 degrees C) withmaximumactivity at pH 7-8 and 60 degrees C. The enzyme retained 50% residual activity at 65 degrees C, but only 20% at 85 degrees C after 16 h. At pH 9 and pH 7, the residual activity at 65 degrees C was 50% and 30%, respectively. The enzymewas 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 alpha-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 alpha-amylase from Geobacillus sp. GS33. The temperature stability and neutral pH optimum suggest that the enzyme may be useful for industrial applications. (C) 2020 Elsevier B.V. All rights reserved.
Description: PubMed: 32693140
URI: https://doi.org/10.1016/j.ijbiomac.2020.07.171
https://hdl.handle.net/10376
ISSN: 0141-8130
1879-0003
Appears in Collections:PubMed İndeksli Yayınlar Koleksiyonu / PubMed Indexed Publications Collection
Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection

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