Expression and purification of recombinant TAT-BoNT/A(1-448) under denaturing and native conditions.

Abstract:

:Botulinum toxin type A can temporarily inhibit muscle contraction. Currently, physicians administer this toxin as a bio-drug in treatment of some muscle contraction disorders. TAT-BoNT/A(1-448) is a functional recombinant protein derived from botulinum toxin light chain. Unlike the full length botulinum toxin, TAT-BoNT/A(1-448) is a self-permeable molecule which can pass through bio-surfaces so can be used as a topical therapeutic agent without injection. To maintain the functionality of TAT-BoNT/A(1-448), it is necessary to restore its normal folding upon expression and purification. In this study, we have investigated and optimized expression conditions for this novel recombinant protein. Under denaturing condition (1 mM IPTG, at 37°C), the chimeric protein was produced as inclusion body and required to be purified using denaturing agents (e.g. urea). Yet, lower incubation temperature (18°C) and less IPTG concentration (0.5 mM) induce a protein under native condition. In such condition, about 60% of the chimeric protein was expressed in soluble form.

journal_name

Bioengineered

journal_title

Bioengineered

authors

Saffarian P,Peerayeh SN,Amani J,Ebrahimi F,Sedighianrad H,Halabian R,Imani Fooladi AA

doi

10.1080/21655979.2016.1201252

subject

Has Abstract

pub_date

2016-11-01 00:00:00

pages

478-483

issue

6

eissn

2165-5979

issn

2165-5987

journal_volume

7

pub_type

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