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Molecular Biology Reports

, Volume 46, Issue 3, pp 2599–2608 | Cite as

Recombinant production of a bioactive peptide from spotless smooth-hound (Mustelus griseus) muscle and characterization of its antioxidant activity

  • Fahimeh Ahmadi-Vavsari
  • Jamshid FarmaniEmail author
  • Ali Dehestani
Original Article
  • 152 Downloads

Abstract

Bioactive peptides are short amino acid sequences with desirable health effects which are derived from animals, plants, and marine sources. In this study, recombinant production of a bioactive peptide (GIISHR) from spotless smooth-hound (Mustelus griseus) muscle and its antioxidant properties is discussed. A gene composed of 12 tandem copies of the peptide sequence was cloned in pET-28a and expressed as a His-tagged polypeptide in Escherichia coli. The recombinant polypeptide was then purified by Ni–NTA affinity chromatography, cleaved by Trypsin and purified by ultrafiltration. DPPH (1,1-diphenyl-2-picrylhydrazyl), ABTS (2,2′-azinobis-3-ethylbenzotiazoline-6-sulfonic acid) and hydroxyl radical scavenging activity assays, ferric reducing antioxidant power (FRAP) assay and β-carotene bleaching test were used to characterize the antioxidant activity of the GIISHR. Liquid chromatography–mass spectrometry analysis revealed 60% purity for released bioactive peptide. Production yield was estimated as 60–80 mg GIISHR active peptide per 1 L bacterial culture. Antioxidant activity assays indicated that the antioxidant activity was increased with increase in peptide concentration. Though the DPPH radical scavenging activity, FRAP and β-carotene bleaching power of the peptide were lower than those of the synthetic antioxidant tert-butylhydroquinone (TBHQ), the ABTS and hydroxyl radical scavenging activities of the peptide (at a concentration of 20 mg/mL) were similar to those of TBHQ (at a concentration of 0.1 mg/mL). The findings of the present study may be helpful in development of a process for production of the bioactive antioxidant peptides and its application in food industry.

Keywords

GIISHR Bioactive peptide Antioxidant activity Recombinant expression Mustelus griseus 

Abbreviations

GIISHR

Gly-Ile-Ile-Ser-His-Arg

DPPH

1,1-Diphenyl-2-picrylhydrazyl

ABTS

2,2′-Azinobis-3-ethylbenzotiazoline-6-sulfonic acid

FRAP

Ferric reducing antioxidant power

TBHQ

Tert-butylhydroquinone

PCR

Polymerase chain reaction

BSA

Bovine serum albumin

TBST

Tris-buffed saline with Tween

PBS

Phosphate buffer solution

LC–MS

Liquid chromatography–mass spectrometry

Prx

Peroxiredoxin

EM

Electron migration

HAT

Hydrogen atom transfer

DNA

Deoxyribonucleic acid

DAB

3,3′-Diaminobenzidine

IPTG

Isopropyl β-d-1-thiogalactopyranoside

Notes

Acknowledgements

This research was funded by of the Biotechnology Development Council of the Islamic Republic of Iran (Grant No: 950901) and conducted at Genetics and Agricultural Biotechnology Institute of Tabarestan.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Food Science and Technology, Faculty of Agricultural EngineeringSari Agricultural Sciences and Natural Resources UniversitySariIran
  2. 2.Genetics and Agricultural Biotechnology Institute of TabarestanSari Agricultural Sciences and Natural Resources UniversitySariIran

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