Extraction of Ascorbate Oxidase from Cucurbita maxima by Continuous Process in Perforated Rotating Disc Contactor Using Aqueous Two-Phase Systems

被引：14

作者：

Porto, T. S. ^{[1
,2
]}

Marques, P. P. ^{[1
]}

Porto, C. S. ^{[1
]}

Moreira, K. A. ^{[1
,3
]}

Lima-Filho, J. L. ^{[1
]}

Converti, A. ^{[4
]}

Pessoa, A. ^{[2
]}

Porto, A. L. F. ^{[1
,5
]}

机构：

[1] Univ Fed Pernambuco, LIKA, BR-50670901 Recife, PE, Brazil

[2] Univ Sao Paulo, Dept Biochem & Pharmaceut Technol, Sao Paulo, Brazil

[3] Univ Fed Rural Pernambuco, Acad Unit Garanhuns, Recife, PE, Brazil

[4] Univ Genoa, Dept Chem & Proc Engn, Genoa, Italy

[5] Univ Fed Rural Pernambuco, Dept Anim Morphol & Physiol, Recife, PE, Brazil

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 2010年 / 160卷 / 04期

基金：

巴西圣保罗研究基金会;

关键词：

Ascorbate oxidase; ATPS; Continuous extraction; Cucurbita maxima; PRDC; MASS-TRANSFER; PROTEIN EXTRACTION; FERMENTED BROTH; CRUDE EXTRACT; RECOVERY; HYDRODYNAMICS; PERFORMANCE; PREDICTION; COLUMN;

D O I：

10.1007/s12010-009-8631-y

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The ascorbate oxidase is the enzyme used to determine the content of ascorbic acid in the pharmaceutical and food industries and clinics analyses. The techniques currently used for the purification of this enzyme raise its production cost. Thus, the development of alternative processes and with the potential to reduce costs is interesting. The application of aqueous two-phase system is proposed as an alternative to purification because it enables good separation of biomolecules. The objective of this study was to determine the conditions to continuously pre-purify the enzyme ascorbate oxidase by an aqueous two-phase system (PEG/citrate) using rotating column provided with perforated discs. Under the best conditions (20,000 g/mol PEG molar mass, 10% PEG concentration, and 25% citrate concentration), the system showed satisfactory results (partition coefficient, 3.35; separation efficiency, 54.98%; and purification factor, 1.46) and proved suitable for the pre-purification of ascorbate oxidase in continuous process.

引用

页码：1057 / 1064

页数：8