A LfcinB-derived peptide LfcinB-W4,10 was designed based on the structure-function relationship of antimicrobial peptides based on antimicrobial peptide database and bioinformatics analysis. In order to verify the rationality of the designed derivative peptide and the functional correctness of the expression product, the optimized gene sequence was digested with restriction endonucleases and ligated into the secretory expression vector pPIC9K with T4 DNA ligase. The recombinant expression vector pPIC9K-LFcinB-W4,10 was linearized by Sac I and transformed into Pichia pastoris GS115 by electroporation, and G418 was used to obtain high copy transformants. The results showed the target gene was stably integrated with the yeast genome. The expression of positive transformants were induced by 2.5% (v/v) methanol, and the expressed product was LfcinB-W4,10 with a relative molecular weight of 3.2 kDa detected by Tricine-SDS-PAGE. Moreover, the antibacterial activity of the fermentation supernatant was measured every 24 h. The results showed the derived peptide LfcinB-W4,10 had good antibacterial activity against Staphylococcus aureus, and the bacteriostatic ring diameter was the largest after 96 h induction. In conclusion, this study lays a foundation to further explore the relationship between function and structure of bovine lactoferrin peptides.
LYU Zili
,
ZHANG Enpeng
,
GUO Aizhen
,
LUO Bin
,
LIANG Xin
,
SHAN Xudong
,
ZHUANG Jing
,
ZHANG Xia
,
WANG Liang
. Design of a recombinant bovine lactoferricin peptide and its expression in Pichia pastoris[J]. Food and Fermentation Industries, 2019
, 45(15)
: 24
-29
.
DOI: 10.13995/j.cnki.11-1802/ts.020507
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