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  • Optimization of process parameters influencing the submerged fermentation of extracellular lipases from Pseudomonas aeruginosa, candida albicans and Aspergillus flavus.

Optimization of process parameters influencing the submerged fermentation of extracellular lipases from Pseudomonas aeruginosa, candida albicans and Aspergillus flavus.

Pakistan journal of biological sciences : PJBS (2012-04-21)
Jigita Padhiar, Arijit Das, Sourav Bhattacharya
ABSTRACT

The present study was aimed at optimization, production and partial purification of lipases from Pseudomonas aeruginosa, Candida albicans and Aspergillus flavus. Various nutritional and physical parameters affecting lipase production such as carbon and nitrogen supplements, pH, temperature, agitation speed and incubation time were studied. Refined sunflower oil (1% v/v) and tryptone at a pH of 6.2 favored maximum lipase production in Pseudomonas at 30 degrees C and 150 rpm, when incubated for 5 days. In C. albicans refined sunflower oil (3% v/v) and peptone resulted in maximum lipase production at pH 5.2, 30 degrees C and 150 rpm, when incubated for 5 days. In A. flavus coconut oil (3% v/v) and peptone yielded maximum lipase at pH 6.2, 37 degrees C, 200 rpm after an incubation period of 5 days. The lipases were partially purified by ammonium sulphate precipitation and dialysis. In P. aeruginosa enzyme activity of the dialyzed fraction was found to be 400 U mL-' and for C. albicans 410 U mL(-1). The dialysed lipase fraction from A. flavus demonstrated an activity of 460 U mL(-1). The apparent molecular weights of the dialyzed lipases were determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The dialyzed lipase fraction obtained from P. aeruginosa revealed molecular weights of 47, 49 and 51 kDa, whereas, lipases from C. albicans and A. flavus demonstrated 3 bands (16.5, 27 and 51 kDa) and one band (47 kDa), respectively. These extracellular lipases may find wide industrial applications.

MATERIALS
Product Number
Brand
Product Description

Millipore
Proteose Peptone, Enzymatic hydrolysate
Millipore
Peptone from casein and other animal proteins, suitable for microbiology
Millipore
Peptone from milk solids, Refined hydrolysate
Sigma-Aldrich
Peptone from animal tissue, from meat, BioReagent, suitable for cell culture, suitable for plant cell culture
Millipore
Peptone Primatone® RL, suitable for microbiology
Millipore
Peptone from animal tissue, from meat