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Effects of Purification on the Crystallization of LysozymeWe have additionally purified a commercial lysozyme preparation by cation exchange chromatography, followed by recrystallization. This material is 99.96% pure with respect to macromolecular impurities. At basic pH, the purified lysozyme gave only tetragonal crystals at 20 C. Protein used directly from the bottle, prepared by dialysis against distilled water, or which did not bind to the cation exchange column had considerably altered crystallization behavior. Lysozyme which did not bind to the cation exchange column was subsequently purified by size exclusion chromatography. This material gave predominately bundles of rod-shaped crystals with some small tetragonal crystals at lower pHs. The origin of the bundled rod habit was postulated to be a thermally dependent tetragonal- orthorhombic change in the protein structure. This was subsequently ruled out on the basis of crystallization behavior and growth rate experiments. This suggests that heterogeneous forms of lysozyme may be responsible. These results demonstrate three classes of impurities: (1) small molecules, which may be removed by dialysis; (2) macromolecules, which are removable by chromatographic techniques; and (3) heterogeneous forms of the protein, which can be removed in this case by cation exchange chromatography. Of these, heterogeneous forms of the lysozyme apparently have the greatest affect on its crystallization behavior.
Document ID
19970023115
Acquisition Source
Marshall Space Flight Center
Document Type
Reprint (Version printed in journal)
External Source(s)
doi:10.1016/0022-0248(95)00933-7
Authors
Ewing, Felecia L. (Universities Space Research Association Huntsville, AL United States)
Forsythe, Elizabeth L. (Universities Space Research Association Huntsville, AL United States)
Van Der Woerd, Mark (Alabama Univ. Birmingham, AL United States)
Pusey, Marc L. (NASA Marshall Space Flight Center Huntsville, AL United States)
Date Acquired
September 6, 2013
Publication Date
January 1, 1996
Publication Information
Publication: Journal of Crystal Growth
Publisher: Elsevier Science B. V.
Volume: 160
ISSN: 0022-0248
Subject Category
Solid-State Physics
Report/Patent Number
NASA-TM-112499
NAS 1.15:112499
Accession Number
97N23512
Distribution Limits
Public
Copyright
Work of the US Gov. Public Use Permitted.

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