UDP-N-acetylglucosamine:N-acetylmuramoyl-(pentapeptide) pyrophosphoryl undecaprenol N-acetylglucosamine transferase from Escherichia coli: overproduction, solubilization, and purification.

Abstract:

:Plasmids for the high-level overproduction of wild-type, and C- and N-terminal His-tagged MurG N-acetylglucosaminyl transferase from Escherichia coli were constructed. In complementation tests the three forms were active in vivo. After IPTG induction, growth, spheroplast formation and lysis, overproduced MurG proteins were mainly present (90%) in the particulate fraction. Readily solubilized by CHAPS, they were purified without any detergent to over 80% purity for both His-tagged forms but only up to 20% for the wild-type form. The enzymatic activity of each purified MurG protein was determined and found to be inhibited to the same extent by ramoplanin.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Crouvoisier M,Mengin-Lecreulx D,van Heijenoort J

doi

10.1016/s0014-5793(99)00412-3

keywords:

subject

Has Abstract

pub_date

1999-04-23 00:00:00

pages

289-92

issue

2-3

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(99)00412-3

journal_volume

449

pub_type

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