Expression and Purification of Recombinant Peanut Allergen Ara h 1

As the dominant allergen in peanut, Ara h 1 has a relatively high allergenicity among peanut allergens. According to the previous researches, methods used for Ara h 1 purification usually involve with two or three steps of chromatography, which is tedious and costly. In this study, pET-32a-Ara h 1 plasmid was constructed and Escherichia coli BL21(DE3) pLysS was utilized for recombinant Ara h 1 expression. Ni-NTA affinity column and gradient elution were used for purification. MALDI-TOF MS and western-blot were used to identify recombinant Ara h 1 and analysis its sensitivity respectively. According to the results, the sequence of Ara h 1 in the plasmid was consistent with the gene data of Ara h 1 in the NCBI database. When 300 mmol/L IPTG(isopropyl-β-D-thiogalactopyranoside) was added and cells were cultured at 22℃ for 22 h, the yield of recombinant protein is at a relatively high level. After adding 15 mmol/L sodium laurylsulfonate to the lysis buffer, the recombinant protein could be effectively released to the supernatant. The elution buffer containing 50 mmol/L and 100 mmol/L imidazole were used in the elution process. Recombinant Ara h 1 with high purity and immunoreactivity could be obtained by using the method above. © 2020, Editorial Office of Journal of CIFST. All right reserved.