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Functional characterization and crystal structure of thermostable amylase from Thermotoga petrophila, reveals high thermostability and an unusual form of dimerization.
Hameed, Uzma; Price, Ian; Ke, Ailong; Wilson, David B; Mirza, Osman.
Afiliación
  • Hameed U; Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan.
  • Price I; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
  • Ikram-Ul-Haq; Institute of Industrial Biotechnology, Government College University, Lahore, 54000, Pakistan.
  • Ke A; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
  • Wilson DB; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
  • Mirza O; Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark. Electronic address: om@sund.ku.dk.
Biochim Biophys Acta Proteins Proteom ; 1865(10): 1237-1245, 2017 Oct.
Article en En | MEDLINE | ID: mdl-28648523
Thermostable α-amylases have many industrial applications and are therefore continuously explored from novel sources. We present the characterization of a novel putative α-amylase gene product (Tp-AmyS) cloned from Thermotoga petrophila. The purified recombinant enzyme is highly thermostable and able to hydrolyze starch into dextrin between 90 and 100°C, with optimum activity at 98°C and pH8.5. The activity increased in the presence of Rb1+, K1+ and Ca2+ ions, whereas other ions inhibited activity. The crystal structure of Tp-AmyS at 1.7Å resolution showed common features of the GH-13 family, however was apparently found to be a dimer. Several residues from one monomer interacted with a docked acarbose, an inhibitor of Tp-AmyS, in the other monomer, suggesting catalytic cooperativity within the dimer. The most striking feature of the dimer was that it resembled the dimerization of salivary amylase from a previous crystal structure, and thus could be a functional feature of some amylases.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bacterias / Proteínas Bacterianas / Alfa-Amilasas Idioma: En Revista: Biochim Biophys Acta Proteins Proteom Año: 2017 Tipo del documento: Article País de afiliación: Pakistán Pais de publicación: Países Bajos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Bacterias / Proteínas Bacterianas / Alfa-Amilasas Idioma: En Revista: Biochim Biophys Acta Proteins Proteom Año: 2017 Tipo del documento: Article País de afiliación: Pakistán Pais de publicación: Países Bajos