Engineered small metal-binding protein tag improves the production of recombinant human growth hormone in the periplasm of Escherichia coli.
Bacterial Proteins
/ genetics
Carrier Proteins
/ genetics
Escherichia coli
/ genetics
Human Growth Hormone
/ biosynthesis
Humans
Metabolic Engineering
/ methods
Metalloproteins
/ genetics
Nitrosomonas europaea
/ metabolism
Periplasm
/ metabolism
Polysaccharide-Lyases
/ chemistry
Protein Sorting Signals
Protein Transport
Recombinant Fusion Proteins
/ metabolism
Escherichia coli
PelB-SmbP
SmbP
human growth hormone
periplasm
protein expression and purification
Journal
FEBS open bio
ISSN: 2211-5463
Titre abrégé: FEBS Open Bio
Pays: England
ID NLM: 101580716
Informations de publication
Date de publication:
04 2020
04 2020
Historique:
received:
23
08
2019
revised:
30
01
2020
accepted:
10
02
2020
pubmed:
13
2
2020
medline:
21
7
2021
entrez:
13
2
2020
Statut:
ppublish
Résumé
Fusion proteins play an important role in the production of recombinant proteins in Escherichia coli. They are mostly used for cytoplasmic expression since they can be designed to increase the solubility of the target protein, which then can be easily purified via affinity chromatography. In contrast, fusion proteins are not usually included in construct designs for periplasmic production. Instead, a signal sequence is inserted for protein transport into the periplasm and a C-terminal his-tag added for subsequent purification. Our research group has proposed the small metal-binding protein (SmbP) isolated from the periplasm of Nitrosomonas europaea as a new fusion protein to express recombinant proteins in the cytoplasm or periplasm of E. coli. SmbP also allows purification via immobilized metal affinity chromatography using Ni(II) ions. Recently, we have optimized the periplasmic production of proteins tagged with SmbP by exchanging its native signal peptide with one taken from pectate lyase B (PelB), substantially increasing the amount of protein produced. In this work, we have expressed and purified soluble bioactive human growth hormone (hGH) tagged with PelB-SmbP and obtained the highest periplasmic production reported for this protein so far. Its activity, tested on Nb2-11 cells, was equivalent to commercial growth hormone at 50 ng·mL
Identifiants
pubmed: 32049439
doi: 10.1002/2211-5463.12808
pmc: PMC7137794
doi:
Substances chimiques
Bacterial Proteins
0
Carrier Proteins
0
Metalloproteins
0
Protein Sorting Signals
0
Recombinant Fusion Proteins
0
Human Growth Hormone
12629-01-5
Polysaccharide-Lyases
EC 4.2.2.-
pectate lyase
EC 4.2.2.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
546-551Informations de copyright
© 2020 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.
Références
Data Brief. 2016 Mar 04;7:502-8
pubmed: 27014739
Acta Paediatr Scand Suppl. 1990;366:32-7
pubmed: 2206005
Protein Expr Purif. 2017 Apr;132:44-49
pubmed: 28087367
Protein Expr Purif. 2016 May;121:61-5
pubmed: 26805756
Indian J Pediatr. 2012 Jan;79(1):87-91
pubmed: 22081428
Growth Horm IGF Res. 2015 Oct;25(5):201-6
pubmed: 26187188
Biotechnol Annu Rev. 2005;11:127-52
pubmed: 16216776
Biotechnol J. 2012 May;7(5):620-34
pubmed: 22442034
Protein Expr Purif. 2016 Feb;118:49-54
pubmed: 26494603
Curr Protoc Mol Biol. 2014 Oct 01;108:16.1B.1-16.1B.21
pubmed: 25271713
PLoS One. 2013;8(2):e56168
pubmed: 23409149
Drug Discov Today. 2014 May;19(5):590-601
pubmed: 24246684
J Clin Endocrinol Metab. 1980 Nov;51(5):1058-63
pubmed: 7419681
Protein Expr Purif. 2014 Aug;100:40-7
pubmed: 24859479
Protein Expr Purif. 2015 Apr;108:9-12
pubmed: 25573388
PLoS One. 2014 Mar 10;9(3):e89038
pubmed: 24614134
Biochemistry. 2004 Sep 7;43(35):11206-13
pubmed: 15366930
Braz J Microbiol. 2016 Dec;47 Suppl 1:51-63
pubmed: 27838289
Appl Microbiol Biotechnol. 2006 Sep;72(2):211-22
pubmed: 16791589
Protein Expr Purif. 2013 Feb;87(2):129-35
pubmed: 23168094
Mol Biotechnol. 2019 Jun;61(6):451-460
pubmed: 30997666
Methods Enzymol. 2009;463:29-34
pubmed: 19892164