Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein

Anh Ngoc Nguyen, Jung A. Song, Minh Tan Nguyen, Bich Hang Do, Grace G. Kwon, Sang Su Park, Jiwon Yoo, Jaepyeong Jang, Jonghwa Jin, Mark J. Osborn, Yeon Jin Jang, Thu Trang Thi Vu, Heung Bum Oh, Han Choe

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Human fibroblast growth factor 21 (hFGF21) has been characterized as an important regulator of glucose and lipid metabolism homeostasis. Here, to produce hFGF21 efficiently in Escherichia coli, the expression and solubility of hFGF21 were tested and optimised by fusing the protein with one of eight tags: hexahistidine (His6), thioredoxin (Trx), small ubiquitin-related modifier (Sumo), glutathione S-transferase (GST), maltose-binding protein (MBP), N-utilisation substance protein A (NusA), human protein disulphide isomerase (PDI), and the b′a′ domain of PDI (PDIb′a′). Each tag increased solubility of the protein when the expression temperature was 18°C. Unlike many other tags that were tested, MBP significantly enhanced the solubility of the protein also in the culture condition at 37°C. Thus, the MBP-hFGF21 construct was further pursued for optimisation of affinity chromatography purification. After tag removal, 8.1 mg of pure hFGF21 was obtained as a final product from 500 mL of starting culture. The protein was then characterised by mass spectroscopy and an in vitro functional assay using NIH-3T3 cells transfected with a β-klotho reporter gene. These characteristics are similar to those of commercial hFGF21. Thus, the MBP tag is useful for efficient prokaryotic production and purification of bioactive hFGF21.

Original languageEnglish (US)
Article number16139
JournalScientific reports
Volume7
Issue number1
DOIs
StatePublished - Dec 1 2017

Bibliographical note

Funding Information:
This work was supported by research funds from the National Research Foundation of Korea (NRF-2015K1A4A3046807, 2017M3A9F8031039) and by a grant (2015-219) from the Asan Institute for Life Sciences, Seoul, Korea.

Publisher Copyright:
© 2017 The Author(s).

Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.

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