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An investigation on acarbose inhibition and the number of active sites in an amylopullulanase (L14-APU) from an Iranian Bacillus sp.

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Abstract

An amylopullulanase (L14-APU) from an Iranian thermophilic bacterium was purified and the effect of acarbose, as a general inhibitor of α-amylases, on pullulan and starch hydrolysis catalyzed by L14-APU was investigated. The inhibition is a competitive type whereas inhibition constants for pullulan and starch are 99 µM and 72 µM, respectively. Investigation of the reaction rate in a system contains competitive substrates and the inhibition type of acarbose in presence of different substrates suggests that L14-APU possesses only one active site for two activities. The analysis of metal ions and other reagents effects has shown that Ca2+, Mg2+, Mn2+ and Co2+ enhanced both activities of the enzyme while N-bromosuccinimide treatment leads to the complete inactivation of the enzyme. The enzyme activity increased in the presence of low concentration of SDS as a surfactant.

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Abbreviations

CD:

cyclodextrin

EPS:

ethylidene-blocked 4-nitrophenyl-maltoheptaoside

GH:

glycoside hydrolase

HPA:

human pancreatic α-amylase

L14-APU:

L14 amylopullulanase

NBS:

N-bromosuccinimide

PPA:

pig pancreatic α-amylase

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Tarbiat Modares University, P.O Box 14115-175, Tehran, Iran

    Marzieh Ghollasi, Khosro Khajeh, Nasrin Mollania, Shekufeh Zareian & Hossein Naderi-Manesh

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  1. Marzieh Ghollasi
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  2. Khosro Khajeh
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  3. Nasrin Mollania
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  4. Shekufeh Zareian
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  5. Hossein Naderi-Manesh
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Correspondence to Khosro Khajeh.

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Ghollasi, M., Khajeh, K., Mollania, N. et al. An investigation on acarbose inhibition and the number of active sites in an amylopullulanase (L14-APU) from an Iranian Bacillus sp.. Biologia 63, 1051–1056 (2008). https://doi.org/10.2478/s11756-008-0174-0

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