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Title: Overexpression, purification, crystallization and preliminary diffraction studies of the Protaminobacter rubrum sucrose isomerase SmuA

Abstract

The P. rubrum sucrose isomerase SmuA, a key enzyme in the industrial production of isomaltulose, was crystallized and diffraction data were collected to 1.95 Å resolution. Palatinose (isomaltulose, α-d-glucosylpyranosyl-1,6-d-fructofuranose), a nutritional and acariogenic reducing sugar, is industrially obtained from sucrose by using immobilized cells of Protaminobacter rubrum that produce the sucrose isomerase SmuA. The isomerization of sucrose catalyzed by this enzyme also results in the formation of trehalulose (α-d-glucosylpyranosyl-1,1-d-fructofuranose) in smaller amounts and glucose, fructose and eventually isomaltose as by-products, which lower the yield of the reaction and complicate the recovery of palatinose. The determination of the three-dimensional structure of SmuA will provide a basis for rational protein-engineering studies in order to optimize the industrial production of palatinose. A recombinant form of the 67.3 kDa SmuA enzyme has been crystallized in the native state by the vapour-diffusion method. Crystals belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 61.6, b = 81.4, c = 135.6 Å, and diffract to 1.95 Å resolution on a synchrotron-radiation source.

Authors:
 [1];  [2];  [1];  [2];  [1]
  1. Laboratoire de BioCristallographie, Institut de Biologie et Chimie des Protéines, CNRS and Université Claude Bernard Lyon 1, UMR 5086, IFR 128 BioSciences Lyon-Gerland, F-69367 Lyon CEDEX 07 (France)
  2. Universität Stuttgart, Institut für Industrielle Genetik, Allmandring 31, D-70569 Stuttgart (Germany)
Publication Date:
OSTI Identifier:
22356235
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 1; Other Information: PMCID: PMC2150920; PMID: 16511267; PUBLISHER-ID: bw5123; OAI: oai:pubmedcentral.nih.gov:2150920; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; BY-PRODUCTS; CRYSTALLIZATION; CRYSTALS; DIFFRACTION; DIFFUSION; GLUCOSE; ISOMERIZATION; RESOLUTION; SACCHAROSE; SPACE GROUPS; YIELDS

Citation Formats

Ravaud, Stéphanie, Watzlawick, Hildegard, Haser, Richard, Mattes, Ralf, and Aghajari, Nushin, E-mail: n.aghajari@ibcp.fr. Overexpression, purification, crystallization and preliminary diffraction studies of the Protaminobacter rubrum sucrose isomerase SmuA. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309105041758.
Ravaud, Stéphanie, Watzlawick, Hildegard, Haser, Richard, Mattes, Ralf, & Aghajari, Nushin, E-mail: n.aghajari@ibcp.fr. Overexpression, purification, crystallization and preliminary diffraction studies of the Protaminobacter rubrum sucrose isomerase SmuA. United Kingdom. doi:10.1107/S1744309105041758.
Ravaud, Stéphanie, Watzlawick, Hildegard, Haser, Richard, Mattes, Ralf, and Aghajari, Nushin, E-mail: n.aghajari@ibcp.fr. Sun . "Overexpression, purification, crystallization and preliminary diffraction studies of the Protaminobacter rubrum sucrose isomerase SmuA". United Kingdom. doi:10.1107/S1744309105041758.
@article{osti_22356235,
title = {Overexpression, purification, crystallization and preliminary diffraction studies of the Protaminobacter rubrum sucrose isomerase SmuA},
author = {Ravaud, Stéphanie and Watzlawick, Hildegard and Haser, Richard and Mattes, Ralf and Aghajari, Nushin, E-mail: n.aghajari@ibcp.fr},
abstractNote = {The P. rubrum sucrose isomerase SmuA, a key enzyme in the industrial production of isomaltulose, was crystallized and diffraction data were collected to 1.95 Å resolution. Palatinose (isomaltulose, α-d-glucosylpyranosyl-1,6-d-fructofuranose), a nutritional and acariogenic reducing sugar, is industrially obtained from sucrose by using immobilized cells of Protaminobacter rubrum that produce the sucrose isomerase SmuA. The isomerization of sucrose catalyzed by this enzyme also results in the formation of trehalulose (α-d-glucosylpyranosyl-1,1-d-fructofuranose) in smaller amounts and glucose, fructose and eventually isomaltose as by-products, which lower the yield of the reaction and complicate the recovery of palatinose. The determination of the three-dimensional structure of SmuA will provide a basis for rational protein-engineering studies in order to optimize the industrial production of palatinose. A recombinant form of the 67.3 kDa SmuA enzyme has been crystallized in the native state by the vapour-diffusion method. Crystals belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 61.6, b = 81.4, c = 135.6 Å, and diffract to 1.95 Å resolution on a synchrotron-radiation source.},
doi = {10.1107/S1744309105041758},
journal = {Acta Crystallographica. Section F},
number = Pt 1,
volume = 62,
place = {United Kingdom},
year = {Sun Jan 01 00:00:00 EST 2006},
month = {Sun Jan 01 00:00:00 EST 2006}
}