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Precipitants and additives for membrane crystallization of lysozyme.

Published on Nov 1, 2006in Biotechnology Journal3.543
· DOI :10.1002/biot.200600088
Xinmiao Zhang2
Estimated H-index: 2
(Huada: Beijing University of Chemical Technology),
Mohamed S. El‐Bourawi1
Estimated H-index: 1
(Huada: Beijing University of Chemical Technology)
+ 2 AuthorsRunyu Ma8
Estimated H-index: 8
(Huada: Beijing University of Chemical Technology)
Abstract
Membrane crystallization is a newly developed crystallization technique that has proven to be superior in producing good crystal forms under operating conditions that are not appropriate to perform the crystallization process by other traditional techniques. In this work, static membrane crystallization was carried out on lysozyme, with hollow-fiber microporous hydrophobic membranes. Numerous precipitant and additive types and concentrations were employed in the crystallization processes in order to select the most appropriate precipitant and additive types and to find their corresponding concentration levels that can yield the best crystal forms. The crystallization processes were analyzed in two ways: firstly, by evaluation of the transmembrane fluxes obtained by using different precipitants and additives; secondly, by utilization of the images and results obtained from the micrography and IR spectra in comparisons and evaluations of the crystals formed under all kinds of conditions. Moreover, the size distributions of the crystals yielded under several typical crystallization conditions were analyzed, and turbidity and induction time periods obtained during typical crystallization experiments were also measured. Amongst the numerous precipitants and additives tested, the most appropriate precipitant type and additive were chosen and their concentrations were optimized. Good lysozyme crystals were obtained using a certain precipitant and additive. The obtained results from this work further support the advantages of utilizing the membrane crystallization technique for macromolecule crystallizations.
  • References (15)
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References15
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#1Efrem Curcio (University of Calabria)H-Index: 29
#1Efrem Curcio (University of Calabria)H-Index: 34
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Abstract The production of protein crystals with adequate size and high lattice perfection for X-ray analysis appears as the bottleneck for obtaining diffraction data at a resolution sufficient to establish 3D structure-function correlation. In previous investigations, microporous hydrophobic membranes have emerged as effective tools for macromolecular crystallization: polymeric supports have been used both to generate a controlled supersaturated environment, and to promote heterogeneous nucleat...
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#1Gianluca Di Profio (University of Calabria)H-Index: 27
#2Efrem Curcio (University of Calabria)H-Index: 29
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Abstract The relevant kinetic aspects related to an innovative method of biological macromolecules crystallization based on microporous hydrophobic membranes, used both as active surfaces to promote heterogeneous nucleation and as a mass-transfer apparatus to concentrate macromolecular solutions by solvent removal in vapour phase, have been evaluated. Polypropylene membranes, supplied in the form of hollow fibres, have been aligned in a versatile system, designed for an on-line spectrophotometri...
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Abstract The great importance of protein science both in industrial and scientific fields, in conjunction with the intrinsic difficulty to grow macromolecular crystals, stimulates the development of new observations and ideas that can be useful in initiating more systematic studies using novel approaches. In this regard, an innovative technique, based on the employment of microporous hydrophobic membranes in order to promote the formation of lysozyme crystals from supersaturated solutions, is in...
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