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Separation of recombinant β-glucuronidase from transgenic tobacco by aqueous two-phase extraction

Paper ID Volume ID Publish Year Pages File Format Full-Text
3978 202 2010 8 PDF Available
Title
Separation of recombinant β-glucuronidase from transgenic tobacco by aqueous two-phase extraction
Abstract

Transgenic plants hold many promises as viable production hosts for therapeutic recombinant proteins. Many efforts have been devoted to increase the expression level of the proteins, but the efforts for developing economic processes to purify those proteins are lacking. In this report, aqueous two-phase extraction (ATPE) was investigated as an alternative for the separation of an acidic recombinant protein, β-glucuronidase (rGUS), from transgenic tobacco. Screening experiments by fractional factorial designs showed that PEG concentration and ionic strength of the system significantly affected the partitioning of native tobacco proteins and GUS. Response surface methodology was used to determine an optimized aqueous two-phase system for the purification of rGUS from transgenic tobacco. In a 13.4% (w/w) PEG 3400/18% (w/w) potassium phosphate system, 74% of the rGUS was recovered in the top PEG-rich phase while more than 90% of the native tobacco proteins were removed in the interphase and the bottom phase. A purification factor of about 20 was achieved in this process. The most important impurity from tobacco, Rubisco, was largely removed from the rGUS in the recovered phase.

Keywords
Transgenic tobacco; Recombinant protein; Downstream processing; Aqueous two-phase system; Aqueous two-phase extraction; Recombinant glucuronidase
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Separation of recombinant β-glucuronidase from transgenic tobacco by aqueous two-phase extraction
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 49, Issue 3, 15 May 2010, Pages 343–350
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us