Novel modification of growth medium enables efficient E. coli expression and simple purification of an endotoxin-free recombinant murine Hsp70 protein

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15310%2F09%3A00010292" target="_blank" >RIV/61989592:15310/09:00010292 - isvavai.cz</a>

Alternative codes found

RIV/61989592:15110/09:00009520

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Novel modification of growth medium enables efficient E. coli expression and simple purification of an endotoxin-free recombinant murine Hsp70 protein

Original language description

We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-D-fructose-6-phosphate aminotransferase (GFAT), was identified. N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies.

Czech name

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Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

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Project

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Continuities

Z - Vyzkumny zamer (s odkazem do CEZ)<br>S - Specificky vyzkum na vysokych skolach

Publication year

2009

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Microbiology and Biotechnology

ISSN

1017-7825

e-ISSN

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Volume of the periodical

19

Issue of the periodical within the volume

7

Country of publishing house

KR - KOREA, REPUBLIC OF

Number of pages

7

Pages from-to

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UT code for WoS article

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EID of the result in the Scopus database

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