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 Purifjournal_title
Protein expression and purificationauthors
Li L,Singh BRdoi
10.1006/prep.1999.1138keywords:
subject
Has Abstractpub_date
1999-12-01 00:00:00pages
339-44issue
3eissn
1046-5928issn
1096-0279pii
S1046-5928(99)91138-5journal_volume
17pub_type
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