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Int. J. Mol. Sci. 2013, 14(9), 17608-17617; doi:10.3390/ijms140917608
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Capillary-Seeding Crystallization and Preliminary Crystallographic Analysis of a Solvent-Tolerant Elastase from Pseudomonas aeruginosa Strain K

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1 Enzyme and Microbial Technology Research Centre, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang UPM 43400, Selangor, Malaysia 2 Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang UPM 43400, Selangor, Malaysia 3 Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang UPM 43400, Selangor, Malaysia 4 Department of Cell & Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang UPM 43400, Selangor, Malaysia 5 Faculty of Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia
* Author to whom correspondence should be addressed.
Received: 9 July 2013 / Revised: 19 July 2013 / Accepted: 22 July 2013 / Published: 28 August 2013
(This article belongs to the Section Material Sciences and Nanotechnology)
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Abstract

Seeding is a versatile method for optimizing crystal growth. Coupling this technique with capillary counter diffusion crystallization enhances the size and diffraction quality of the crystals. In this article, crystals for organic solvent-tolerant recombinant elastase strain K were successfully produced through microseeding with capillary counter-diffusion crystallization. This technique improved the nucleation success rate with a low protein concentration (3.00 mg/mL). The crystal was grown in 1 M ammonium phosphate monobasic and 0.1 M sodium citrate tribasic dihydrate pH 5.6. The optimized crystal size was 1 × 0.1 × 0.05 mm3. Elastase strain K successfully diffracted up to 1.39 Å at SPring-8, Japan, using synchrotron radiation for preliminary data diffraction analysis. The space group was determined to be monoclinic space group P1211 with unit cell parameters of a = 38.99 Ǻ, b = 90.173 Å and c = 40.60 Å.
Keywords: elastase strain K; organic solvent tolerant; seeding technique; capillary counter diffusion elastase strain K; organic solvent tolerant; seeding technique; capillary counter diffusion
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Ali, M.S.M.; Said, Z.S.A.M.; Rahman, R.N.Z.R.A.; Chor, A.L.T.; Basri, M.; Salleh, A.B. Capillary-Seeding Crystallization and Preliminary Crystallographic Analysis of a Solvent-Tolerant Elastase from Pseudomonas aeruginosa Strain K. Int. J. Mol. Sci. 2013, 14, 17608-17617.

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