tailieunhanh - Báo cáo khoa học: LmbE proteins from Bacillus cereus are de-N-acetylases with broad substrate specificity and are highly similar to proteins in Bacillus anthracis

The genomes ofBacillus cereusand its closest relative Bacillus anthracis each contain two LmbE protein family homologs: BC1534 (BA1557) and BC3461 (BA3524). Only a few members of this family have been biochemi-cally characterized including N-acetylglucosaminylphosphatidyl inositol (GlcNAc-PI), 1-d-myo-inosityl-2-acetamido-2-deoxy-a-d-glucopyranoside (GlcNAc-Ins), N,N¢-diacetylchitobiose (GlcNAc2) and lipoglycopeptide antibiotic de-N-acetylases. | LmbE proteins from Bacillus cereus are de-N-acetylases with broad substrate specificity and are highly similar to proteins in Bacillus anthracis Alexandra Deli1 Dimitrios Koutsioulis2 Vasiliki E. Fadouloglou2 Panagiota Spiliotopoulou1 Stavroula Balomenou1 Sofia Arnaouteli1 Maria Tzanodaskalaki2 Konstantinos Mavromatis3 Michalis Kokkinidis1 2 and Vassilis Bouriotis1 2 1 Department of Biology Enzyme Biotechnology Group University of Crete Greece 2 Institute of Molecular Biology and Biotechnology Heraklion Crete Greece 3 Department of Energy Joint Genome Institute Genome Biology Program Walnut Creek CA USA Keywords Bacillus anthracis de-N-acetylase glucosamine LmbE mutationalanalysis Correspondence V. Bouriotis Department of Biology Enzyme Biotechnology Group University of Crete PO Box 2208 Vasilika Vouton 714 09 Heraklion Crete Greece Fax 30 2810 394055 Tel 30 2810 394375 E-mail bouriotis@ Present address Department of Biochemistry University of Cambridge UK Received 22 October 2009 revised 15 March 2010 accepted 20 April 2010 doi The genomes of Bacillus cereus and its closest relative Bacillus anthracis each contain two LmbE protein family homologs BC1534 BA1557 and BC3461 BA3524 . Only a few members of this family have been biochemically characterized including N-acetylglucosaminylphosphatidyl inositol GlcNAc-PI 1-D-myo-inosityl-2-acetamido-2-deoxy-a-D-glucopyranoside GlcNAc-Ins -diacetylchitobiose GlcNAc2 and lipoglycopeptide antibiotic de-N-acetylases. All these enzymes share a common feature in that they de-N-acetylate the N-acetyl-D-glucosamine GlcNAc moiety of their substrates. The bc1534 gene has previously been cloned and expressed in Escherichia coli. The recombinant enzyme was purified and its 3D structure determined. In this study the bc3461 gene from B. cereus ATCC14579 was cloned and expressed in E. coli. The recombinant enzymes BC1534 EC and BC3461 were biochemically characterized. The enzymes .