Processing of soybean (Glycine max) extracts in aqueous two‐phase systems as a first step for the potential recovery of recombinant proteins

Abstract

BACKGROUND: The potential use of plants as production systems to establish bioprocesses has been established over the past decade. However, the lack of efficient initial concentration and separation procedures affect the generic acceptance of plants as economically viable systems. In this context the use of aqueous two-phase systems (ATPS) can provide strategies to facilitate the adoption of plants as a base for bioprocesses. Among the crops, soybeans (Glycine max) represent an attractive alternative since potentially they can produce high levels of recombinant protein. In this paper the processing of fractionated soybean extracts using ATPS is evaluated as a first step to recover recombinant proteins expressed in plants, using β-glucuronidase (GUS; E.C. 3.2.1.31) as a model protein.

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

“…Low PEG concentration associated with high citrate concentration and PEG molar mass favored the transfer of the enzyme to the disperse phase, thus increasing the partition coefficient. Some authors [3][4][5][6] proposed that, when the system has high M PEG , a remarkable volume exclusion effect might have taken place, hence suggesting that there was no space enough for the enzyme in the PEG phase of the system; however, this phenomenon was not observed in the present study. Figure 1 shows the interaction between the three variables.…”

“…Therefore, the establishment of efficient primary recovery procedures for the recovery of recombinant proteins from transgenic crop is needed. In this context, aqueous two-phase systems (ATPS) are an attractive alternative to facilitate the adoption of bioprocesses based on plants as production systems and proved to have great potential for the recovery and purification of biological compounds [4].…”

Aqueous two-phase extraction of 2,3-butanediol from fermentation broths using an ethanol/phosphate system

“…ATPE has been widely applied in the separation of biomacromolecules, such as proteins and nucleic acids because of its mild conditions and high capacity [12][13][14][15]. Up to now, most aqueous twophase systems (ATPS) used for purification were based either on polyethylene glycol (PEG)/salt systems or polymer/polymer (e.g., PEG/dextran) systems.…”

Aqueous Two-Phase System Strategies for the Recovery of Proteins from Plants

“…The methodology described in the next section is to obtain soybean extracts to evaluate ATPS as a fi rst step to recover recombinant proteins expressed in seeds [ 11 ].…”