Publication: Improved X-ray diffraction from Bacillus megaterium penicillin G acylase crystals through long cryosoaking dehydration
Issued Date
2011-12-01
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ISSN
17443091
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2-s2.0-83055165596
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Mahidol University
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SCOPUS
Bibliographic Citation
Acta Crystallographica Section F: Structural Biology and Crystallization Communications. Vol.67, No.12 (2011), 1570-1574
Suggested Citation
Catleya Rojviriya, Thunyaluck Pratumrat, Mark A. Saper, Jirundon Yuvaniyama Improved X-ray diffraction from Bacillus megaterium penicillin G acylase crystals through long cryosoaking dehydration. Acta Crystallographica Section F: Structural Biology and Crystallization Communications. Vol.67, No.12 (2011), 1570-1574. doi:10.1107/S1744309111040462 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/11413
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Title
Improved X-ray diffraction from Bacillus megaterium penicillin G acylase crystals through long cryosoaking dehydration
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Abstract
Penicillin G acylase from Bacillus megaterium (BmPGA) is currently used in the pharmaceutical industry as an alternative to PGA from Escherichia coli (EcPGA) for the hydrolysis of penicillin G to produce 6-aminopenicillanic acid (6-APA), a penam nucleus for semisynthetic penicillins. Despite the significant differences in amino-acid sequence between PGAs from Gram-positive and Gram-negative bacteria, a representative PGA structure of Gram-positive origin has never been reported. In this study, crystallization and diffraction studies of BmPGA are described. Poor diffraction patterns with blurred spots at higher resolution were typical for BmPGA crystals cryocooled after a brief immersion in cryoprotectant solution. Overnight soaking in the same cryo-solution substantially improved both the mosaicity and resolution limit through the establishment of a new crystal-packing equilibrium. A crystal of BmPGA diffracted X-rays to 2.20 Å resolution and belonged to the monoclinic space group P21 with one molecule of BmPGA in the asymmetric unit. © 2011 International Union of Crystallography. All rights reserved.