Molecular Modification of Kex2 P1' Site Enhances Expression and Druggability of Fungal Defensin

文献类型： 外文期刊

第一作者： Jin, Yanjie

作者： Jin, Yanjie;Yang, Na;Teng, Da;Hao, Ya;Mao, Ruoyu;Wang, Jianhua;Jin, Yanjie;Yang, Na;Teng, Da;Hao, Ya;Mao, Ruoyu;Wang, Jianhua;Jin, Yanjie;Yang, Na;Teng, Da;Hao, Ya;Mao, Ruoyu;Wang, Jianhua

作者机构：

关键词： fungal defensin; Pichia pastoris; P1' site of Kex2; expression; druggability

期刊名称：ANTIBIOTICS-BASEL （ 影响因子：4.8； 五年影响因子：4.9 ）

ISSN： 2079-6382

年卷期： 2023 年 12 卷 4 期

页码：

收录情况： SCI

摘要： Pichia pastoris is the widely used expression system for producing recombinant secretory proteins. It is known that Kex2 protease plays a vital role in the process of protein secretion, in which the P1' site affects its cleavage efficiency. To enhance the expression level of fungal defensin-derived peptide NZ2114, this work attempts to optimize the P1' site of Kex2 by replacing it with 20 amino acids in turn. The results showed that when the amino acid of the P1' site was changed to Phe (F), the yield of target peptide significantly increased from 2.39 g/L to 4.81 g/L. Additionally, the novel peptide F-NZ2114 (short for FNZ) showed strong antimicrobial activity against Gram-positive (G(+)) bacteria, especially for Staphylococcus aureus and Streptococcus agalactiae (MIC: 4-8 mu g/mL). The FNZ was very stable and retained high activity in various conditions; in addition, a low cytotoxicity and no hemolysis were observed even at a high concentration of 128 mu g/mL, and a longer postantibiotic effect was reached. The above results indicate that this engineering strategy provided a feasible optimization scheme for enhancing the expression level and druggability of this antimicrobial peptide from fungal defensin and other similar targets by this updated recombinant yeast.

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