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

Brazilian Archives of Biology and Technology

Giovana Da Silva Padilha²  José Carlos Curvelo Santana¹  Ranulfo Monte Alegre¹  Elias Basile Tambourgi² 

[1] ,Universidade Estadual de Campinas Faculdade de Engenharia Química Departamento de Engenharia de Sistemas QuímicosCampinas SP ,Brasil

关键词: lipase;    Burkholderia cepacia;    aqueous two phase system;    partitioning;    characterization;   

DOI  :  10.1590/S1516-89132012000100002

来源: SciELO

【 摘 要 】

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 37ºC and pH 8.0; it showed pH-stability for 4 h of incubation at different pH values at 37ºC. Metal ions such as Mn2+ , Co2+, I-and Ca2+ sustained enzymatic activities; however, it was inhibited by the presence of Fe2+, Hg2+ and Al3+ . Km and Vmax 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.

【 授权许可】

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