Purification and Characterization of an Alkaline Lipase from Pseudomonas fluorescens AK102

• Published in: Bioscience, biotechnology, and biochemistry Vol. 58; no. 9; pp. 1564 - 1568
• Main Authors: Kojima, Yuzo, Yokoe, Masaaki, Mase, Tamio
• Format: Journal Article
• Language: English
• Published: Tokyo Taylor & Francis 01.09.1994; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: Amino Acid Sequence; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Bacteriology; Biological and medical sciences; Biotechnology; Electrophoresis, Polyacrylamide Gel; Endopeptidases - pharmacology; Enzyme Stability; Fundamental and applied biological sciences. Psychology; Hydrogen-Ion Concentration; Isoelectric Point; Lipase - chemistry; Lipase - isolation & purification; Lipase - metabolism; Metabolism. Enzymes; Microbiology; Molecular Sequence Data; Molecular Weight; Pseudomonas fluorescens - classification; Pseudomonas fluorescens - enzymology; Sequence Homology, Amino Acid; Sodium Dodecyl Sulfate; Substrate Specificity; Surface-Active Agents - pharmacology; Temperature; Ultrafiltration; Pseudomonadales; Purification; Enzymatic activity; Enzyme; Triacylglycerol lipase; Bacteria; Hydrolases; Pseudomonadaceae; Esterases; Pseudomonas fluorescens; Physicochemical properties; Carboxylic ester hydrolases
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.58.1564

An extracellular, novel alkaline lipase produced by Pseudomonas fluorescens AK102 was purified by ultrafiltration, ammonium sulfate precipitation, and DEAE-Toyopearl 650M and Phenyl-Toyopearl 650M column chromatographies. The purified enzyme was homogeneous on SDS-PAGE. The molecular weight was estimated to be about 33,000 by SDS-PAGE. The isoelectric point was pH 4.0 by isoelectric focusing. The pH stability was 4 to 10 and the optimum pH was 8 to 10. The optimum temperature was 55°C and the enzyme was stable below 50°C. The enzyme unspecifkally liberated short chain to long chain fatty acids from p-nitrophenyl esters, methyl esters, and triglycerides. In the presence of an anionic surfactant, the enzyme was characteristically stable. These results suggested that the enzyme can be used as a home laundry product ingredient.