High-level expression, purification, and characterization of recombinant type A botulinum neurotoxin light chain.

Abstract:

:Botulinum neurotoxin light chain (BoNT LC, 50 kDa) is responsible for the zinc endopeptidase activity specific for proteins of neuroexocytosis apparatus. We describe the expression of recombinant type A BoNT LC in Escherichia coli as well as the purification and characterization of the recombinant protein. A high level of expression of BoNT/A LC was obtained by an extended postinduction time of 15 h at 30 degrees C. Recombinant BoNT/A LC was isolated from an Ni(2+) column. Due to its high pI ( approximately 8.7), purification was achieved by a single step of passing the protein through anion-exchange chromatography at pH 8.0 without the need of elution. The purified recombinant BoNT/A LC retained proteolytic activity and had a secondary structure similar to that of native LC determined by CD measurement.

journal_name

Protein Expr Purif

authors

Li L,Singh BR

doi

10.1006/prep.1999.1138

keywords:

subject

Has Abstract

pub_date

1999-12-01 00:00:00

pages

339-44

issue

3

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(99)91138-5

journal_volume

17

pub_type

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