Saccharomyces cerevisiae OS303 expression of an alkaline protease from a newly isolated Bacillus subtilis D9

The aim of this study was to produce high yield a local bacterial alkaline protease in the yeast system because scientific involvement microorganisms enzyme production is still not given enough attention Saudi Arabia. Soil samples were collected from rhizosphere some desert plants Ninety-three producing isolates recovered on skimmed-milk agar at pH 9.4 and 45°C for 48 hr. Isolate D9 obtained Heliotropium digynum Dhahran City most potent isolate respect productivity (184.6 U/ml). full gene amplified showed expected size (1300 bp). Restriction enzymes analysis also verified integrity PCR product. sequence revealed an open reading frame 1329 nt correspond length encoding 443 aa protein. After ligation by TA cloning method, digestion with appropriate restriction confirmed cloned gene. insert prepared two PCRs that conducted pair primers specifically designed purpose. digested purified vector pRS426/GAL1p-207-Glu-MS ligated then transformed into various strains Saccharomyces cerevisiae via electroporation method. Maximum expression done recombinant OS303 galactose containing media (145.5 U/ml) approximately 2-fold increase when compared wild strain., may be due ability activate gal operon.