FOOD SCIENCE ›› 2023, Vol. 44 ›› Issue (10): 224-230.doi: 10.7506/spkx1002-6630-20220428-375
• Bioengineering • Previous Articles Next Articles
DI Nana, JIANG Bo, ZHANG Ran, CAO Hongzhen, CHEN Jingjing, ZHANG Tao
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Published:
Abstract: In this study, the antihypertensive peptide WQVLPNAVPAK was selected for the synthesis of the gene encoding the hexameric peptide angiotensin I-converting enzyme inhibitory peptide (ACEIP) based on the codon preference of Escherichia coli. The synthetic gene was cloned into the expression vector pET30a to construct recombinant plasmid pET30a-ACEIP. The recombinant plasmid was identified by double digestion with restriction endonucleases NdeI and HindIII and polymerase chain reaction (PCR), and transformed into competent E. coli BL21 (DE3) cells to construct the expression system E. coli BL21(DE3)/pET30a-ACEIP, which was induced with isopropyl-β-D-thiogalactopyranoside (IPTG) at a final concentration of 0.8 mmol/L. A clear band of 8.7 kDa was seen on Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The hexameric peptide ACEIP was purified by Ni2+ affinity chromatography and the expression of the fusion protein was 61.30 mg/L. The expression of ACEIP was 57.84 mg/L. The half-maximal inhibitory concentration (IC50) of the hexameric peptide was 6.39 mg/mL after trypsin digestion, whose ACE inhibitory activity was higher than that of the monomeric peptide WQVLPNAVPAK (IC50 of 12.34 mg/mL).
Key words: antihypertensive peptide; biosynthesis; peptide preparation; hypotensive activity; hypertension
CLC Number:
Q78
DI Nana, JIANG Bo, ZHANG Ran, CAO Hongzhen, CHEN Jingjing, ZHANG Tao. Expression and Activity Identification of Recombinant Antihypertensive Peptide (Angiotensin I-Converting Enzyme Inhibitory Peptide)[J]. FOOD SCIENCE, 2023, 44(10): 224-230.
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URL: https://www.spkx.net.cn/EN/10.7506/spkx1002-6630-20220428-375
https://www.spkx.net.cn/EN/Y2023/V44/I10/224