Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system.
This paper presents the evaluation of an aqueous two-phase system (ATPS) for extracting elastase produced by Bacillus sp. EL31410. The elastase and cell partition behavior in polyethylene glycol (PEG)/salt systems was investigated. The suitable system for elastase extraction was PEG/KH(2)PO(4)-K(2)HPO(4), in which elastase is mainly partitioned into the PEG-rich phase, while the cells remained in the other phase. The influence of defined system parameters (e.g. PEG molecular mass, pH, NaCl addition) on the partitioning behavior of elastase is described. The concentration of phase forming components, PEG and KH(2)PO(4)-K(2)HPO(4), was optimized for elastase recovery by means of response surface methodology, and it was found that they greatly influenced extraction recovery. The optimal ATPS was 23.1% (w/w) PEG 2 000 and 11.7% (w/w) KH(2)PO(4)-K(2)HPO(4). The predicted recovery was about 89.5%, so this process is suggested to be a rapid and convenient method for elastase extraction.
Xu
Ying
Y
Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China.
He
Guo-qing
GQ
Li
Jing-jun
JJ
D001407:Bacillus / Q000201:enzymology*; D019149:Bioreactors / Q000382:microbiology; D018929:Cell Culture Techniques / Q000379:methods*; D005591:Chemical Fractionation / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D005285:Fermentation / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D010196:Pancreatic Elastase / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification*; D044367:Phase Transition; D011092:Polyethylene Glycols / Q000737:chemistry*; D014867:Water / Q000737:chemistry*
D003470:Culture Media; D014867:Water; D011092:Polyethylene Glycols; D010196:Pancreatic Elastase