Isolation and purification of alpha-glycerophosphate oxidase in a polyethylene glycol/(NH4 )2SO4 aqueous two-phase system.
- Author:
Yao MENG
;
Jiagui JIN
;
Shuangfeng LIU
;
Min YANG
;
Qinglian ZHANG
;
Li WAN
;
Kun TANG
- Publication Type:Journal Article
- MeSH:
Ammonium Sulfate;
chemistry;
Glycerolphosphate Dehydrogenase;
chemistry;
isolation & purification;
Molecular Weight;
Polyethylene Glycols;
chemistry;
Water
- From:
Journal of Biomedical Engineering
2014;31(1):136-141
- CountryChina
- Language:Chinese
-
Abstract:
Alpha-glycerophosphate oxidase (alpha-GPO) from Enterococcus casseli flavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
