Overexpression of Ace-AMP1 in Pichia pastoris on enhancing the inhibition of blue mold on pears
Conventional methods of Ace-AMP1 antimicrobial peptide extraction from onion seeds are usually difficult, costly with a low extraction rate. To solve these problems, the original gene of antimicrobial peptide Ace-AMP1 was optimized according to the preference of Pichia pastoris and ligated into pPIC...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2019-02-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2017.10.092 |
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| Summary: | Conventional methods of Ace-AMP1 antimicrobial peptide extraction from onion seeds are usually difficult, costly with a low extraction rate. To solve these problems, the original gene of antimicrobial peptide Ace-AMP1 was optimized according to the preference of Pichia pastoris and ligated into pPICZα A vector, which was designated as pPICZ α A/Ace-AMP1. The recombinant vector was then transformed P. pastoris through electroporation. The results of real-time quantitative polymerase chain reaction (RT-qPCR) and Western-blotting confirmed that the Ace-AMP1 was successfully overexpressed in GS115, and the antibacterial peptide could be stably expressed in yeast cell. The protein expression level showed a significant increase in the supernatant after the induction, reached the maximum concentration (109 μg/mL) at 84 h, while the control group still remained a low level. The biocontrol experiment indicated that the recombinant strain GS115/Ace-AMP1 could highly control the blue mold compared with the controls, which was 34% of disease incidence lower than the sterile distilled water treatment. The same trend was observed in the lesion diameter, which dropped by 65% with the GS115/Ace-AMP1 treatment compared with the control. Therefore, the GS115/Ace-AMP1 can be as an Ace-AMP1 peptide manufacturer, as well as a biocontrol agent. |
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| ISSN: | 1008-9209 2097-5155 |