Direct purification of Burkholderia Pseudomallei lipase from fermentation broth using aqueous two-phase systems

Abstract

An aqueous two-phase purification process was employed for the recovery of _ìêâÜçäÇÉêá~=éëÉìÇçã~ääÉá=lipase from fermentation broth. The partition behavior of _K=éëÉìÇçã~ääÉá=lipase was investigated with various parameters such as phase composition, tie-line length (TLL), volume ratio (V R ), sample loading, system pH, and addition of neutral salts. Optimum conditions for the purification of lipase were obtained in polyethylene glycol (PEG) 6000-potassium phosphate system using TLL of 42.2% (w/w), with V R of 2.70, and 1% (w/w) NaCl addition at pH 7 for 20% (w/w) crude load. Based on this system, the purification factor of lipase was enhanced to 12.42 fold, with a high yield of 93%. Hence, the simplicity and effectiveness of aqueous two-phase systems (ATPS) in the purification of lipase were proven in this study. © KSBB hÉóïçêÇëW=~èìÉçìë=íïçJéÜ~ëÉ=ëóëíÉãëI=äáé~ëÉI=éìêáÑáÅ~íáçåI=éçäóÉíÜóäÉåÉ=ÖäóÅçäI=êÉÅçîÉêó= = = = =

Direct recovery of cyclodextringlycosyltransferase from Bacillus cereus using aqueous two-phase flotation

“…Therefore, it is believed that the higher salt concentration causes a reduction in the solubility of the CGTase protein due to the higher salting-out ability of potassium phosphates. A similar observation has been reported in the previous microbial lipase and recombinant human interferon alpha-2b partitioning studies (13,14). The result also shows a correlation between the PEG molecular weight and the P FT of CGTase.…”

“…The effect of the PEG molecular weights and tie-line length (TLL) on lipase and recombinant human interferon alpha2b partition performance have been explored previously (13,14).…”

“…The TLL values of various PEG (6000, 8000 and 10,000) used in this part of study (Table 1) were referred to the previous publications (13,14). Here, the main aim of the study was to examine the effect of the PEG molecular weights and TLL on B. cereus CGTase partition.…”

Extractive bioconversion of poly-ϵ-caprolactone by Burkholderia cepacia lipase in an aqueous two-phase system

“…The process variables examined included the molecular weights of PEG, tie-line length (TLL), pH, bioconversion temperature, volume ratio (V R ), and the presence of a neutral salt additive (NaCl) in the system ( Table 1). The levels of the central (0), low (−1) and high (+1) values of the six factors were selected based on the preliminary and published data of a related study in our laboratory [10]. Analysis of variance (ANOVA) was used to determine the significance of each process variable in affecting the partitioning of the biomolecules.…”

“…Selected parameters that affect the partitioning process significantly were optimized for maximal recovery of the hydrolyzed PCL and lipase in a systematic order. Six process variables were identified to be significant based on previous literatures, namely molecular weight of PEG, pH, bioconversion temperature, tie-line length (TLL), volume ratio (V R ) and NaCl additive [10][11][12]. The molecular weight of PEG influences not only the polymeric conformation (and therefore, intramolecular hydrophobic interactions) of the biomolecule, but also interfacial tension, which has been reported to play an important role in influencing the partitioning behavior of biomolecules in ATPS [13].…”