NOVEL LIPASE GENE FROM PSEUDOMONAS SP. S5

摘要

A BIOLOGICALLY PURE STRAIN OF PSEUDOMONAS SP. S5 CAPABLE OF PRODUCING ORGANIC SOLVENT TOLERANT LIPASE SECRETED BY BTEX DEGRADER. THE ORGANIC SOLVENT TOLERANT S5 LIPASE WAS PURIFIED BY AFFINITY CHROMATOGRAPHY AND ANION EXCHANGE CHROMATOGRAPHY. THE MOLECULAR MASS OF THE LIPASE WAS ESTIMATED TO BE 60 KDA WITH 387 PURIFICATION FOLD. THE OPTIMUM TEMPERATURE AND PH WAS 45°C AND PH 9.0, RESPECTIVELY. THE PURIFIED LIPASE WAS STABLE AT PH 6-9 AND 45°C. IT EXHIBITED HIGHEST STABILITY IN THE PRESENCE OF VARIOUS ORGANIC SOLVENTS SUCH AS N-DODECANE, 1-PENTANOL AND TOLUENE. CA²⁺ AND MG²⁺ STIMULATED LIPASE ACTIVITY, WHILE EDTA HAD NO EFFECT ON ITS ACTIVITY. THE S5 LIPASE EXHIBITED HIGHEST ACTIVITY IN THE PRESENCE OF PALM OIL AS A NATURAL OIL AND TRIOLEIN AS SYNTHETIC TRIGLYCERIDE. IT SHOWED RANDOM POSITIONAL SPECIFICITY ON THE THIN LAYER CHROMATOGRAPHY. AN ORGANIC SOLVENT TOLERANT LIPASE GENE S5 WAS CLONED VIA SHOTGUN CLONING AND EXPRESSED IN E. COLI. FIGURE 1