Production of an antioxidative peptide from hairy basil seed waste by a recombinant Escherichia coli.
OBJECTIVE
To circumvent the time-consuming and costly problems associated with natural product extraction, a potential antioxidative peptide selected from hairy basil waste after oil extraction was produced by recombinant DNA technology.


RESULTS
Because the target peptide is short, the recombinant peptide containing seven repeats of the target sequence, QTFQYSRGWTN, and the DNA fragment coding this sequence was cloned into the pQE-30 Xa expression vector and transformed into Escherichia coli. After 6 h of recombinant peptide expression in E. coli, the target peptide was purified by Ni(2+) affinity chromatography and gel extraction. The expected 15 kDa recombinant target peptide construct was verified by modified dot blot analysis. Compared with the chemically synthesized peptide, the recombinant peptide revealed significantly higher antioxidant activities (p < 0.05), as determined by DPPH and ABTS radical scavenging assays, and in vitro DNA damage induced by hydroxyl radicals.


CONCLUSIONS
This approach provides an alternative to produce an antioxidative peptide that provides a potential scaffold for the further development of antioxidative peptides for industrial applications.
Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
Apapun
A
Kongcharoen
Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
Wannapun
W
Poolex
Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok, Thailand.
Thanaporn
T
Wichai
Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok, Thailand. ruethairat.b@chula.ac.th.
Ruethairat
R
Boonsombat
D000975:Antioxidants / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D018649:Ocimum basilicum / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D012639:Seeds / Q000737:chemistry / Q000378:metabolism*
D000975:Antioxidants; D010455:Peptides
Antioxidant; Escherichia coli; Hairy basil; Ocimum basilicum; Peptide; Recombinant protein