High-level expression of the Endo-beta-N-acetylglucosaminidase F2 gene in E.coli: one step purification to homogeneity.

Abstract:

:The Endo F2gene was overexpressed in E.coli as a fusion protein joined to the maltose-binding protein. MBP-Endo F2was found in a highly enriched state as insoluble, inactive inclusion bodies. Extraction of the inclusion bodies with 20% acetic acid followed by exhaustive dialysis rendered the fusion protein active and soluble. MBP-Endo F2was digested with Factor Xaand purified on Q-Sepharose. The enzyme was homogeneous by SDS-PAGE, and appeared as a single symmetrical peak on HPLC. Analysis of the amino-terminus demonstrated conclusively that recombinant Endo F2was homogeneous and identical to the native enzyme.

journal_name

Glycobiology

journal_title

Glycobiology

authors

Reddy A,Grimwood BG,Plummer TH,Tarentino AL

doi

10.1093/glycob/8.6.633

subject

Has Abstract

pub_date

1998-06-01 00:00:00

pages

633-6

issue

6

eissn

0959-6658

issn

1460-2423

pii

cwb068

journal_volume

8

pub_type

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