The effect of protein impurities on lysozyme crystal growth.
While bulk crystallization from impure solutions is used industrially as a purification step for a wide variety of materials, it is a technique that has rarely been used for proteins. Proteins have a reputation for being difficult to crystallize and high purity of the initial crystallization solution is considered paramount for success in the crystallization. Although little is written on the purifying capability of protein crystallization or of the effect of impurities on the various aspects of the crystallization process, recent published reports show that crystallization shows promise and feasibility as a purification technique for proteins. To further examine the issue of purity in macromolecule crystallization, this study investigates the effect of the protein impurities, avidin, ovalbumin, and conalbumin at concentrations up to 50%, on the solubility, crystal face growth rates, and crystal purity of the protein lysozyme. Solubility was measured in batch experiments while a computer controlled video microscope system was used to measure the ¿110¿ and ¿101¿ lysozyme crystal face growth rates. While little effect was observed on solubility and high crystal purity was obtained (>99.99%), the effect of the impurities on the face growth rates varied from no effect to a significant face specific effect leading to growth cessation, a phenomenon that is frequently observed in protein crystal growth. The results shed interesting light on the effect of protein impurities on protein crystal growth and strengthen the feasibility of using crystallization as a unit operation for protein purification.
Judge
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Alliance for Microgravity Material Science and Applications, NASA/Marshall Space Flight Center, Huntsville, Alabama 35812, USA.
Forsythe
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Pusey
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ML
D000818:Animals; D001360:Avidin / Q000378:metabolism; D002642:Chick Embryo; D003207:Conalbumin / Q000378:metabolism; D003461:Crystallography / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D009113:Muramidase / Q000737:chemistry* / Q000378:metabolism*; D010047:Ovalbumin / Q000378:metabolism; D016622:Silver Staining
D003207:Conalbumin; D001360:Avidin; D010047:Ovalbumin; D009113:Muramidase