Expression and purification of antimicrobial peptide AP2 using SUMO fusion partner technology inEscherichia coli

2018 ◽  
Vol 67 (6) ◽  
pp. 606-613 ◽  
Author(s):  
Q. Mo ◽  
A. Fu ◽  
Z. Lin ◽  
W. Wang ◽  
L. Gong ◽  
...  
Keyword(s):  
Antimicrobial Peptide ◽  
Fusion Partner ◽  
Inescherichia Coli ◽  
Expression And Purification ◽  
Sumo Fusion

scholarly journals Heterologous Expression and Purification of the Antimicrobial Peptide Buforin II

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Boda Ferenc-András ◽  
Szabó Zoltán-István ◽  
Szőcs Erika ◽  
Salamon Pál ◽  
Orbán Csongor ◽  
...  
Keyword(s):  
Antimicrobial Peptides ◽  
Heterologous Expression ◽  
Antimicrobial Peptide ◽  
Solid Phase ◽  
Host Cells ◽  
Fusion Partner ◽  
Monitoring And Control ◽  
Expression And Purification ◽  
Recombinant Peptide ◽  
Bacterial Membranes

AbstractAntimicrobial peptides are natural substances that have played a role in the development of the adaptive immune system, and are currently involved in the prevention of infections, through their direct antimicrobial and immunomodulatory properties. While the amino acid composition and spatial structure vary, most antibacterial peptides have a positive surface charge, which allows them to bind to the negative bacterial membranes. Buforin II is a widely studied antimicrobial peptide first obtained through the structural modification of buforin I, a peptide isolated from Bufo gargarizans. The peptide showed significant antibacterial activity against Gram-positive and Gram-negative bacterial strains. The mechanism of action of buforin II differs from that of other antimicrobial peptides, as it binds directly to bacterial DNA and RNA. The aim of our study was to obtain recombinant buforin II with a ubiquitin fusion partner, through heterologous expression in Escherichia coli Rosetta™ (DE3)pLysS cells, using a laboratory scale biore-actor. The incubation of expression host cells in a bioreactor allowed the constant monitoring and control of the process parameters, leading to high biomass levels and an increased production rate of the peptide. The parameters used during incubation were: 37°C, pH=6.9 and dissolved oxygen level above 40%. Purification of the recombinant protein was accomplished by affinity chromatography using a Ni-chelate solid phase to which the 10xHistag of our construct showed affinity. Method optimisation consisted in the use of gradient and linear elution, of which the latter was found to be more effective. Digestion of the fusion partner from the target peptide was performed with ubiquitin carboxyl-terminal hydrolase enzyme. The expression and purification protocols developed in our experiment allow the production of a significant amount of buforin II, allowing its use for further research. Furthermore, the presented methods could be suitable for industrial production of the recombinant peptide..

Cloning, Direct Expression, and Purification of a Snake Venom Cardiotoxin inEscherichia coli

1996 ◽  
Vol 219 (2) ◽  
pp. 450-456 ◽  
Author(s):  
T.K.S. Kumar ◽  
P.W. Yang ◽  
S.H. Lin ◽  
C.Y. Wu ◽  
B. Lei ◽  
...  
Keyword(s):  
Snake Venom ◽  
Inescherichia Coli ◽  
Expression And Purification

scholarly journals Cloning, Expression, and Purification of a Putative Prostaglandin Synthase from Nostoc as a Sumo Fusion Protein

2013 ◽  
Vol 104 (2) ◽  
pp. 562a
Author(s):  
Margaret V. Butchy ◽  
Kevin Backus ◽  
Huasong Tang ◽  
Barry S. Selinsky
Keyword(s):  
Fusion Protein ◽  
Expression And Purification ◽  
Prostaglandin Synthase ◽  
Sumo Fusion

scholarly journals High Level Expression and Purification of the Clinically Active Antimicrobial Peptide P-113 in Escherichia coli

Molecules ◽  
2018 ◽  
Vol 23 (4) ◽  
pp. 800 ◽  
Author(s):  
Kuang-Ting Cheng ◽  
Chih-Lung Wu ◽  
Bak-Sau Yip ◽  
Hui-Yuan Yu ◽  
Hsi-Tsung Cheng ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Peptide ◽  
Expression And Purification ◽  
High Level Expression ◽  
High Level

Functional expression of hepassocin in Escherichia coli using SUMO fusion partner and molecular chaperones

2013 ◽  
Vol 92 (2) ◽  
pp. 135-140 ◽  
Author(s):  
Qi Wang ◽  
Jiaojiao Zhao ◽  
Yan Wang ◽  
Honglou Sun ◽  
Yi Jiang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Molecular Chaperones ◽  
Functional Expression ◽  
Fusion Partner ◽  
Sumo Fusion
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