[WS4] APPA/PSSJワークショップ 蛋白質研究の世界連携に向けて

[WS4-06] STRUCTURE DETERMINATION AND IMMOBILIZATION OF AN ORGANIC SOLVENT-TOLERANT ELASTASE FROM PSEUDOMONAS SP.

Muhammad Syafiq Mohd Razib, Zatty Syamimi @ Adura Mat Said, Raja Noor Zaliha Raja Abd Rahman, Adam Leow Thean Chor, Abu Bakar Salleh, ^○Mohd Shukuri Mohamad Ali (Enzyme and Microbial Technology Research Centre, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia)

An organic solvent-tolerant elastase from Pseudomonas aeruginosa strain K was previously isolated and screened in our laboratory. Further investigations to characterize the purified recombinant elastase strain K showed its capability for activity in hydrophilic organic solvent media. Given the previous investigations, it is important to discern its structural features as an organic solvent-stable enzyme. The crystals for organic solvent-tolerant recombinant elastase strain K were successfully produced through microseeding with capillary counter-diffusion crystallization methods. This technique improved the nucleation success rate and reproducibility of the crystal. High quality protein crystals were obtained. Elastase crystal was successfully diffracted using synchrotron radiation facility. The organic solvent tolerant elastase was immobilized using CLEA method. The CLEA-elastase exhibit enhanced thermostability and was able to operate at higher temperature and were shown to retain its organic solvent tolerant feature. The understanding of protein-organic solvent interaction and the future development on enzyme stabilization could be useful for potential industrial and biotechnological applications.
Keywords: Elastase, Pseudomonas aeruginosa, Crystallization, CLEA, Organic solvent tolerant enzyme