Production of a Pseudomonas lipase in n-alkane substrate and its isolation using an improved ammonium sulfate precipitation technique

• Published in: Bioresource technology 2002-09, Vol.84 (3), p.207-211
• Main Authors: Kanwar, Lambit, Gogoi, Binod Kumar, Goswami, Pranab
• Format: Article
• Language: English
• Subjects: Alkanes - chemistry; ammonium sulfate; Ammonium Sulfate - pharmacology; Antifoaming Agents - pharmacology; Biological and medical sciences; Biotechnology; Biotechnology - methods; carbon; Carbon - chemistry; culture media; Dose-Response Relationship, Drug; Enzyme engineering; Fermentation; fractionation; Fundamental and applied biological sciences. Psychology; hexadecane; Hydrogen-Ion Concentration; iron; Lipase; Lipase - chemistry; Lipase - isolation & purification; Methods. Procedures. Technologies; n-Alkane; Production of selected enzymes; Pseudomonas; Pseudomonas - enzymology; silicone; Silicones - pharmacology; Temperature; Time Factors; tributyrin
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/S0960-8524(02)00061-5

Among the various lipidic and non-lipidic substances, normal alkanes within the chain lengths of C-12 to C-20 served as the best carbon substrates for the production of extracellular lipase by Pseudomonas species G6. Maximum lipase production of 25 U/ml of the culture broth was obtained by using n-hexadecane as the sole carbon substrate. The optimum pH of 8 and temperature of 34±1 °C were demonstrated for the production of lipase in n-hexadecane substrate. The optimum concentration of iron, which played a critical role on the lipase production, was found to be 0.25 mg/l. Lipase production could be enhanced to nearly 2.4-fold by using tributyrin at a concentration of 0.05% (v/v) in the culture medium. High recovery of the lipase protein (83%) from the culture broth was achieved by treating the culture supernatant with Silicone 21 Defoamer followed by ammonium sulfate (60% saturation) fractionation.