title: Capillary-seeding crystallization and preliminary crystallographic analysis of a solvent-tolerant elastase from Pseudomonas aeruginosa strain K.
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 P12(1)1 with unit cell parameters of a = 38.99 Ǻ, b = 90.173 Å and c = 40.60 Å.
authors.affiliation: Enzyme and Microbial Technology Research Centre, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang UPM 43400, Selangor, Malaysia. shukuri@biotech.upm.edu.my
authors.forename: Mohd Shukuri Mohamad
authors.initials: MS
authors.lastname: Ali
authors.forename: Zatty Syamimi  Adura Mat
authors.initials: ZS
authors.lastname: Said
authors.forename: Raja Noor Zaliha Raja Abd
authors.initials: RN
authors.lastname: Rahman
authors.forename: Adam Leow Thean
authors.initials: AL
authors.lastname: Chor
authors.forename: Mahiran
authors.initials: M
authors.lastname: Basri
authors.forename: Abu Bakar
authors.initials: AB
authors.lastname: Salleh
mesh_terms: D003460:Crystallization / Q000379:methods*; D010196:Pancreatic Elastase / Q000737:chemistry*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D012997:Solvents / Q000737:chemistry*
chemical_list: D012997:Solvents; D010196:Pancreatic Elastase