Recombinant expression of antimicrobial peptides in Pichia pastoris: A strategy to inhibit the Penicillium expansum in pears
Yining Huang, Liguang Gao, Ming Lin and Ting Yu
Postharvest Biology and Technology, Volume 171, January 2021, 111298
2021
บทคัดย่อ
The alteration of safety-secured yeast is a crucial step before scale applications. Based on previous studies, Pichia pastoris showed great potential and value to improve its biocontrol ability. The original sequences of antimicrobial peptide Ac-AMP2 and MiAMP1 were optimized according to the preference of Pichia pastoris and ligated into pPICZαA plasmid which is emerged as a high-performance vector for transformation and expression. The results of RT-qPCR and Western blotting could imply that pPICZαA/Ac-AMP2 and pPICZαA/MiAMP1 were successfully overexpressed in Pichia pastoris GS115. The peptide concentration of GS115/Ac-AMP2 reached a maximum value of 210 mg L−1 at 60 h while GS115/MiAMP1 was 220 mg L−1 at 96 h. The biocontrol experiment indicated that the recombinant strain GS115/Ac-AMP2 and GS115/MiAMP1 could highly suppress the pathogen Penicillium expansum in vivo, which was respectively 42% (GS115/Ac-AMP2) and 29.2% (GS115/MiAMP1) of incidence disease lower than the sterile distilled water treatment. In the case of the experimental results considered, the modified GS115/Ac-AMP2 and GS115/MiAMP1 might be promising biological agent in postharvest applications.