Keyword: Serine protease, DegP, halophilic bacteria.1. 1 Seminar Nasional Bioteknologi, âBioteknologi untuk mendukung kualitas hidup. Bangsa. 2014.
SERINE PROTEASE DegP CHROMOHALOBACTER SALEXIGENS-‐BKL-‐5: CLONING AND EXPRESSION M. Saifur Rohman1,2, Dewi Fitriani2, Irfan D. Prijambada1,2 1Laboratory of Agricultural Microbiology, Faculty of Agriculture, Universitas Gadjah Mada; 2Graduate Program in Biotechnology, Universitas Gadjah Mada, Yogyakarta-‐INDONESIA Abstract Serine protease DegP plays an important role in protein quality control. To cope its important role, DegP carries out dual unique functions that seem to be opposing to each other. DegP conveys chaperon and protease functions in a single entity. The expression of DegP is upregulated in response to various environmental stress conditions, such as high temperature, high salt concentration, low and high pH. Halophilic bacterium, a group of bacteria that able to grow in such high salt concentration, has their own mechanism to protect from osmotic stress. The common mechanism is by accumulation of compatible solute such as highly water soluble sugar, alcohols, amino acids, betaines, ectoines or their derivatives. In regard to the DegP, it is not known whether this protein is involved in the salt tolerance mechanism of halophilic bacterium. In order to understand of the role of DegP, it is necessary to clone and overexpress the DegP gene from halophilic bacterium. Therefore the objective of this work is to clone and overexpress the DegP gene from Chromohalobacter salexigens-‐BKL-‐ 5, a halophilic bacterium that was previously isolated from Bledug Kuwu in our laboratory. In this report, we have successfully cloned and over expressed the DegP gene from Chromohalobacter salexignes-‐BKL-‐5 in E. coli BL-‐ 21(DE3)CodonPlus. Analysis of the DegP encoding gene indicated that the gene consist of 1356 bp length and the overexpressed recombinant protein had molecular weight of 49 kDa. Solubility check analysis indicated that the protein was overexpressed in soluble form. Bioinformatics analysis showed that the proteins has similar characteristic as halophilic proteins. Keyword: Serine protease, DegP, halophilic bacteria.1
1 Seminar Nasional Bioteknologi, “Bioteknologi untuk mendukung kualitas hidup Bangsa. 2014. Sekolah Pascasarjana UGM