NT*-HRV3CP: An optimized construct of human rhinovirus 14 3C protease for high-yield expression and fast affinity-tag cleavage

Elwy H. Abdelkader; Gottfried Otting

doi:10.1016/j.jbiotec.2020.11.005

NT*-HRV3CP: An optimized construct of human rhinovirus 14 3C protease for high-yield expression and fast affinity-tag cleavage

Elwy H. Abdelkader, Gottfried Otting^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

20 Citations (Scopus)

Abstract

The human rhinovirus 14 3C protease (HRV3C protease), in fusion with glutathione S-transferase also referred to as PreScission™ protease, is a cysteine protease of particular interest for affinity tag removal from fusion proteins due to its stringent recognition sequence specificity (LEVLFQ/GX) and superior activity at low temperature. Here we report the expression, purification and use of a fusion construct of HRV3C protease, NT*-HRV3CP, that affords high expression yield in E. coli (over 300 mg/L cell culture), facile single-step purification, high solubility (>10 mg/mL) and excellent storage properties. NT*-HRV3CP cleaves affinity tags at 4 °C in minutes, making it an attractive tool for the production of recombinant proteins for biotechnological, industrial and pharmaceutical applications.

Original language	English
Pages (from-to)	145-151
Number of pages	7
Journal	Journal of Biotechnology
Volume	325
DOIs	https://doi.org/10.1016/j.jbiotec.2020.11.005
Publication status	Published - 10 Jan 2021

Access to Document

10.1016/j.jbiotec.2020.11.005

Cite this

@article{acf10c054fd44768b8aa37cc58b33517,

title = "NT*-HRV3CP: An optimized construct of human rhinovirus 14 3C protease for high-yield expression and fast affinity-tag cleavage",

abstract = "The human rhinovirus 14 3C protease (HRV3C protease), in fusion with glutathione S-transferase also referred to as PreScission{\texttrademark} protease, is a cysteine protease of particular interest for affinity tag removal from fusion proteins due to its stringent recognition sequence specificity (LEVLFQ/GX) and superior activity at low temperature. Here we report the expression, purification and use of a fusion construct of HRV3C protease, NT*-HRV3CP, that affords high expression yield in E. coli (over 300 mg/L cell culture), facile single-step purification, high solubility (>10 mg/mL) and excellent storage properties. NT*-HRV3CP cleaves affinity tags at 4 °C in minutes, making it an attractive tool for the production of recombinant proteins for biotechnological, industrial and pharmaceutical applications.",

keywords = "Fusion tag cleavage, Human rhinovirus 14 3C protease, NT* fusion protein, PreScission protease",

author = "Abdelkader, {Elwy H.} and Gottfried Otting",

note = "Publisher Copyright: {\textcopyright} 2020 Elsevier B.V.",

year = "2021",

month = jan,

day = "10",

doi = "10.1016/j.jbiotec.2020.11.005",

language = "English",

volume = "325",

pages = "145--151",

journal = "Journal of Biotechnology",

issn = "0168-1656",

publisher = "Elsevier B.V.",

}

TY - JOUR

T1 - NT*-HRV3CP

T2 - An optimized construct of human rhinovirus 14 3C protease for high-yield expression and fast affinity-tag cleavage

AU - Abdelkader, Elwy H.

AU - Otting, Gottfried

PY - 2021/1/10

Y1 - 2021/1/10

N2 - The human rhinovirus 14 3C protease (HRV3C protease), in fusion with glutathione S-transferase also referred to as PreScission™ protease, is a cysteine protease of particular interest for affinity tag removal from fusion proteins due to its stringent recognition sequence specificity (LEVLFQ/GX) and superior activity at low temperature. Here we report the expression, purification and use of a fusion construct of HRV3C protease, NT*-HRV3CP, that affords high expression yield in E. coli (over 300 mg/L cell culture), facile single-step purification, high solubility (>10 mg/mL) and excellent storage properties. NT*-HRV3CP cleaves affinity tags at 4 °C in minutes, making it an attractive tool for the production of recombinant proteins for biotechnological, industrial and pharmaceutical applications.

AB - The human rhinovirus 14 3C protease (HRV3C protease), in fusion with glutathione S-transferase also referred to as PreScission™ protease, is a cysteine protease of particular interest for affinity tag removal from fusion proteins due to its stringent recognition sequence specificity (LEVLFQ/GX) and superior activity at low temperature. Here we report the expression, purification and use of a fusion construct of HRV3C protease, NT*-HRV3CP, that affords high expression yield in E. coli (over 300 mg/L cell culture), facile single-step purification, high solubility (>10 mg/mL) and excellent storage properties. NT*-HRV3CP cleaves affinity tags at 4 °C in minutes, making it an attractive tool for the production of recombinant proteins for biotechnological, industrial and pharmaceutical applications.

KW - Fusion tag cleavage

KW - Human rhinovirus 14 3C protease

KW - NT fusion protein

KW - PreScission protease

UR - http://www.scopus.com/inward/record.url?scp=85096134762&partnerID=8YFLogxK

U2 - 10.1016/j.jbiotec.2020.11.005

DO - 10.1016/j.jbiotec.2020.11.005

M3 - Article

SN - 0168-1656

VL - 325

SP - 145

EP - 151

JO - Journal of Biotechnology

JF - Journal of Biotechnology

ER -

NT*-HRV3CP: An optimized construct of human rhinovirus 14 3C protease for high-yield expression and fast affinity-tag cleavage

Abstract

Access to Document

Other files and links

Fingerprint

Cite this