Recombinant expression of bioactive peptide lunasin in Escherichia coli
Journal
Applied Microbiology and Biotechnology
Journal Volume
88
Journal Issue
1
Pages
177-186
Date Issued
2010
Author(s)
Abstract
Lunasin, a cancer-preventive peptide, was isolated from soybean, barley, and wheat. Previous studies showed that this 43-amino acid peptide has the ability to suppress chemical carcinogen-induced transformation in mammalian cells and skin carcinogenesis in mice. In this study, we attempted to use the Escherichia coli T7 expression system for expression of lunasin. The lunasin gene was synthesized by overlapping extension polymerase chain reaction and expressed in E. coli BL21(DE3) with the use of vector pET29a. The recombinant lunasin containing his-tag at the C-terminus was expressed in soluble form which could be purified by immobilized metal affinity chromatography. After 4 h, the expression level is above 4.73 mg of recombinant his-tagged lunasin/L of Luria-Bertani broth. It does not affect the bacterial growth and expression levels. This is the first study that successfully uses E. coli as a host to produce valuable bioactive lunasin. The result of in vitro bioassay showed that the purified recombinant lunasin can inhibit histone acetylation. Recombinant lunasin also inhibits the release of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-1β, and nitric oxide production). Compared with other research methods on extraction or chemical synthesis to produce lunasin, our method is very efficient in saving time and cost. In the future, it could be applied in medicine and structure-function determination. ? 2010 Springer-Verlag.
Subjects
Core histone acetylation; Escherichia coli; Lunasin; Overlapping extension polymerase chain reaction
SDGs
Other Subjects
Amino acid peptides; Bacterial growth; Bioactive peptides; C-terminus; Chemical carcinogens; Chemical synthesis; E. coli; Expression levels; His-tag; Histone acetylation; Immobilized metal affinity chromatography; In-vitro; Inflammatory cytokines; Interleukin-1; Lunasin; Luria-Bertani broths; Mammalian cells; Overlapping extension polymerase chain reaction; Recombinant expression; research methods; Skin carcinogenesis; T7 expression system; Tumor necrosis factors; Acetylation; Affinity chromatography; Amino acids; Bioassay; Escherichia coli; Mammals; Nitric oxide; Organic acids; Peptides; Polymerization; Polymers; Purification; Synthesis (chemical); Polymerase chain reaction; bacterial protein; interleukin 1beta; lunasin; nitric oxide; polyhistidine tag; recombinant protein; tumor necrosis factor alpha; unclassified drug; amino acid; barley; bioactivity; bioassay; cancer; carcinogen; chromatography; coliform bacterium; immobilization; metal binding; nitric oxide; peptide; polymerase chain reaction; recombination; soybean; transformation; tumor; wheat; acetylation; article; bacterial strain; bioassay; carboxy terminal sequence; controlled study; cytokine production; Escherichia coli; extraction; gene expression system; immobilized metal affinity chromatography; in vitro study; male; nonhuman; polymerase chain reaction; protein expression; protein purification; rat; synthesis; Bacteriophage T7; Culture Media; Cytokines; DNA-Directed RNA Polymerases; Escherichia coli; Gene Expression; Histone Acetyltransferases; Hordeum; Immunologic Factors; Plant Proteins; Promoter Regions, Genetic; Recombinant Fusion Proteins; Soybeans; Triticum; Viral Proteins; Bacteria (microorganisms); Escherichia coli; Glycine max; Hordeum; Luria; Mammalia; Mus; Triticum aestivum
Type
journal article