Structure of Bacillus halmapalus alpha-amylase crystallized with and without the substrate analogue acarbose and maltose

Publication: Research - peer-reviewJournal article – Annual report year: 2006

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@article{bb6b75d546d24759a77dc2c5a9a37bfd,
title = "Structure of Bacillus halmapalus alpha-amylase crystallized with and without the substrate analogue acarbose and maltose",
publisher = "Wiley-Blackwell Munksgaard",
author = "Louise Lyhne-Iversen and Hobley, {Timothy John} and Kaasgaard, {Svend G.} and Pernille Harris",
year = "2006",
doi = "10.1107/S174430910603096X",
volume = "62",
pages = "849--854",
journal = "Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online",
issn = "1744-3091",

}

RIS

TY - JOUR

T1 - Structure of Bacillus halmapalus alpha-amylase crystallized with and without the substrate analogue acarbose and maltose

A1 - Lyhne-Iversen,Louise

A1 - Hobley,Timothy John

A1 - Kaasgaard,Svend G.

A1 - Harris,Pernille

AU - Lyhne-Iversen,Louise

AU - Hobley,Timothy John

AU - Kaasgaard,Svend G.

AU - Harris,Pernille

PB - Wiley-Blackwell Munksgaard

PY - 2006

Y1 - 2006

N2 - Recombinant Bacillus halmapalus alpha-amylase (BHA) was studied in two different crystal forms. The first crystal form was obtained by crystallisation of BHA at room temperature in the presence of acarbose and maltose - data was collected at cryogenic temperatures to a resolution of 1.9 Å. It was found that the crystal belonged to the space group P212121 and had the unit cell dimensions: a = 47.0 Å, b = 73.5 Å and c = 151.1 Å. A maltose molecule was observed and found to bind to BHA and previous reports of the binding of a nonasaccharide were confirmed. The second crystal form was obtained by pH-induced crystallisation of BHA in a MES-HEPES-Boric acid-buffer (MHB-buffer) at 30oC - the solubility of BHA in MHB has a retrograde temperature dependency, and crystallisation of BHA was only possible by raising the temperature to at least 25oC. Data was collected at cryogenic temperatures to a resolution of 2.0 Å. The crystal belonged to the space group P212121 and had the unit cell dimensions: a = 38.6 Å, b = 59.0 Å and c = 209.8 Å. The structure was solved using molecular replacement. The maltose binding site is described and the two structures are compared. No significant changes were seen in the structure upon binding of the substrates.

AB - Recombinant Bacillus halmapalus alpha-amylase (BHA) was studied in two different crystal forms. The first crystal form was obtained by crystallisation of BHA at room temperature in the presence of acarbose and maltose - data was collected at cryogenic temperatures to a resolution of 1.9 Å. It was found that the crystal belonged to the space group P212121 and had the unit cell dimensions: a = 47.0 Å, b = 73.5 Å and c = 151.1 Å. A maltose molecule was observed and found to bind to BHA and previous reports of the binding of a nonasaccharide were confirmed. The second crystal form was obtained by pH-induced crystallisation of BHA in a MES-HEPES-Boric acid-buffer (MHB-buffer) at 30oC - the solubility of BHA in MHB has a retrograde temperature dependency, and crystallisation of BHA was only possible by raising the temperature to at least 25oC. Data was collected at cryogenic temperatures to a resolution of 2.0 Å. The crystal belonged to the space group P212121 and had the unit cell dimensions: a = 38.6 Å, b = 59.0 Å and c = 209.8 Å. The structure was solved using molecular replacement. The maltose binding site is described and the two structures are compared. No significant changes were seen in the structure upon binding of the substrates.

U2 - 10.1107/S174430910603096X

DO - 10.1107/S174430910603096X

JO - Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online

JF - Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online

SN - 1744-3091

VL - 62

SP - 849

EP - 854

ER -