Acta Biologica Hungarica
Volume/Issue:
Volume 57: Issue 3
Effect of temperature on subsite map of Bacillus licheniformis a -amylase
Authors:
Lili Kandra Department of Biochemistry, Faculty of Sciences, University of Debrecen P.O. Box 55, H-4010 Debrecen, Hungary

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Judit Remenyik Research Group for Carbohydrates of the Hungarian Academy of Sciences P.O. Box 55, H-4010 Debrecen, Hungary

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Gyöngyi Gyémánt Department of Biochemistry, Faculty of Sciences, University of Debrecen P.O. Box 55, H-4010 Debrecen, Hungary

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A. Lipták Department of Biochemistry, Faculty of Sciences, University of Debrecen Research Group for Carbohydrates of the Hungarian Academy of Sciences P.O. Box 55, H-4010 Debrecen, Hungary P.O. Box 55, H-4010 Debrecen, Hungary

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A. Lipták Department of Biochemistry, Faculty of Sciences, University of Debrecen P.O. Box 55, H-4010 Debrecen, Hungary

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A. Lipták Department of Biochemistry, Faculty of Sciences, University of Debrecen P.O. Box 55, H-4010 Debrecen, Hungary

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Pages:
367–375
Online Publication Date:
14 Oct 2006
Publication Date:
01 Sep 2006
Article Category:
Research Article
DOI:
https://doi.org/10.1556/abiol.57.2006.3.10
Restricted access

To elucidate how temperature effects subsite mapping of a thermostable a-amylase from Bacillus licheniformis (BLA), a comparative study was performed by using 2-chloro-4-nitrophenyl (CNP) b-maltooligosides with degree of polymerisation (DP) 4-10 as model substrates. Action patterns, cleavage frequencies and subsite binding energies were determined at 50 °C, 80 °C and 100 °C. Subsite map at 80 °C indicates more favourable bindings compared to the hydrolysis at 50 °C. Hydrolysis at 100 °C resulted in a clear shift in the product pattern and suggests significant differences in the active site architecture. Two preferred cleavage modes were seen for all substrates in which subsite (+2) and (+3) were dominant, but CNP-G1 was never formed. In the preferred binding mode of shorter oligomers, CNP-G2 serves as the leaving group (79%, 50%, 59% and 62% from CNP-G4, CNP-G5, CNP-G6 and CNP-G7, respectively), while CNP-G3 is the dominant hydrolysis product from CNP-G8, CNP-G9, and CNP-G10 (62%, 68% and 64%, respectively). The high binding energy value (-17.5 kJ/mol) found at subsite (+2) is consistent with the significant formation of CNP-G2. Subsite mapping at 80 °C and 100 °C confirms that there are no further binding sites despite the presence of longer products.

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Cover Acta Biologica Hungarica
Acta Biologica Hungarica
Print ISSN:
0236-5383
Online ISSN:
1588-256X

Senior editors

Editor(s)-in-Chief: Elekes, Károly

Editorial Board

    1. Csányi, Vilmos (Göd)
    1. Dudits, Dénes (Szeged)
    1. Falus, András (Budapest)
    1. Fischer, Ernő (Pécs)
    1. Gábriel, Róbert (Pécs)
    1. Gulya, Károly (Szeged)
    1. Gulyás, Balázs (Stockholm)
    1. Hajós, Ferenc (Budapest)
    1. Hámori, József (Budapest)
    1. Heszky, László (Gödöllő)
    1. Hideg, Éva (Szeged)
    1. E. Ito (Sanuki)
    1. Janda, Tibor (Martonvásár)
    1. Kavanaugh, Michael P. (Missoula)
    1. Kása, Péter (Szeged)
    1. Klein, Éva (Stockholm)
    1. Kovács, János (Budapest)
    1. Brigitte Mauch-Mani (Neuchâtel)
    1. Nässel, Dick R. (Stockholm)
    1. Nemcsók, János (Szeged)
    1. Péczely, Péter (Gödöllő)
    1. Roberts, D. F. (Newcastle-upon-Tyne)
    1. Sakharov, Dimitri A. (Moscow)
    1. Singh, Meharvan (Fort Worth)
    1. Sipiczky, Mátyás (Debrecen)
    1. Szeberényi, József (Pécs)
    1. Székely, György (Debrecen)
    1. Tari, Irma (Szeged)
    1. Vágvölgyi, Csaba (Szeged),
    1. L. Zaborszky (Newark)

 

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Acta Biologica Hungarica
Language English
Size  
Year of
Foundation		 1950
Publication
Programme		 changed title
Volumes
per Year		  
Issues
per Year		  
Founder Magyar Tudományos Akadémia
Founder's
Address		 H-1051 Budapest, Hungary, Széchenyi István tér 9.
Publisher Akadémiai Kiadó
Publisher's
Address		 H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
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ISSN 0236-5383 (Print)
ISSN 1588-256X (Online)

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