Extraction of lipase from Burkholderia cepacia by PEG/Phosphate ATPS and its biochemical characterization

Abstract

This work aimed to study the partitioning of a lipase produced by Burkholderia cepacia in PEG/Phosphate aqueous two phase system (ATPS) and its characterization. Lipase was produced by B. cepacia strains in a fermenter. Enzyme partitioning occurred at pH 6.0 and 8.0, using PEG 1500 and 6000 on two tie lines. Metal ions, pH and temperature effects on enzyme activity were evaluated. Five milliliter of 7.5% olive oil emulsion with 2.5% gumarabic in 0.1M sodium phosphate buffer at pH 8.0 and 37ºC were used for the activity determinations. Results showed that crude stratum from B. cepacia was partitioned by PEG1500/phosphate ATPS at pH 6.0 or 8.0 for, which the partitioning coefficients were 108-and 209-folds. Lipase presented optimal activity conditions at 37 o C and pH 8.0; it showed pH-stability for 4 h of incubation at different pH values at 37 o C. Metal ions such as Mn 2+ , Co 2+ , Iand Ca 2+ sustained enzymatic activities; however, it was inhibited by the presence of Fe 2+ , Hg 2+ and Al 3+. K m and V max values were 0.258 U/mg and 43.90 g/L, respectively. A molecular weight of 33 kDa and an isoelectric point at pH 5.0 were determined by SDS-PAGE and IFS electrophoresis, respectively.

Evaluating Self-buffering Ionic Liquids for Biotechnological Applications

“…As aforementioned, a commercial lipase from P. cepacia was applied as a model enzyme and the respective SDS-PAGE analysis is reported in SI ( Figure S3). It is clearly seen the presence of the target lipase from P. cepacia at circa 33 kDa, which is consistent with the literature, 60 and several less pronounced bands representing the contaminant proteins. Since the lipase used in this study is of commercial purity, the expectable purification in the GB-ILs-based ABS is limited.…”

Lipase and their different industrial applications: A review

“…In 1985, Candida rugosa is used by Japan for the production of fatty acids from castor bean (Macedo et al, 2003). Earlier it has been reported for use of microbial lipase in the flavoring of milk and creams production (Padilha et al, 2012). Organolephtic has proved that every lipase strains develop a characteristic flavor (Padilha et al, 2012).…”

“…Earlier it has been reported for use of microbial lipase in the flavoring of milk and creams production (Padilha et al, 2012). Organolephtic has proved that every lipase strains develop a characteristic flavor (Padilha et al, 2012). Saccharomyces, Torulospora, Kluyveromyces, Candida, Pseudozyma, Pischia, Lachancea and Zygosaccharomyces are the lipase producing yeast belong to different genera (Rajendran et al, 2008).…”

Response surface methodology assisted biodiesel production from waste cooking oil using encapsulated mixed enzyme

“…Lipase activity was determined according to Burkert et al (2004) and Padilha et al (2012). The immobilization was carried by the encapsulation method using 2% sodium alginate.…”