Production of a Pseudomonas lipase in n-alkane substrate and its isolation using an improved ammonium sulfate precipitation technique.
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 degrees 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.
Biochemistry Division, Regional Research Laboratory, Jorhat, Assam, India.
Lambit
L
Kanwar
Binod Kumar
BK
Gogoi
Pranab
P
Goswami
D000473:Alkanes / Q000737:chemistry*; D000645:Ammonium Sulfate / Q000494:pharmacology*; D000934:Antifoaming Agents / Q000494:pharmacology; D001709:Biotechnology / Q000379:methods; D002244:Carbon / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry* / Q000302:isolation & purification*; D011549:Pseudomonas / Q000201:enzymology*; D012828:Silicones / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
D000473:Alkanes; D000934:Antifoaming Agents; D012828:Silicones; D002244:Carbon; D008049:Lipase; D000645:Ammonium Sulfate