Fusion Partner Facilitates Expression of Cell-Penetrating Peptide L2 in Pichia pastoris.

IF 4.3 2区医学 Q1 INFECTIOUS DISEASES Antibiotics-Basel Pub Date : 2024-12-11 DOI:10.3390/antibiotics13121207

Xuan Li, Na Yang, Yuxin Fang, Ruoyu Mao, Ya Hao, Da Teng, Na Dong, Anshan Shan, Jianhua Wang

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Abstract

Background: L2 is formed by combining the pheromone of Streptococcus agalactiae (S. agalactiae) and a cell-penetrating peptide (CPP) with cell-penetrating selectivity. L2 has more significant penetration and better specificity for killing S. agalactiae. However, the production of AMPs by chemical synthesis is always a challenge because of the production cost.

Methods: This study was devoted to the heterologous expression of the cell-penetrating peptide L2 in Pichia pastoris using SUMO and a short acidic fusion tag as fusion partners, and the high-density expression of SUMO-L2 was achieved in a 5 L fermenter.

Results: The results showed that SUMO-L2 expression in the 5 L fermenter reached 629 mg/L. The antibacterial activity of recombinant L2 was examined; the minimum inhibitory concentration (MICs) and minimum bactericidal concentration (MBCs) of purified L2 were 4-8 μg/mL and 8-16 μg/mL against S. agalactiae after 84 h of lysis with 50% formic acid.

Conclusions: The findings suggest that SUMO is a suitable fusion tag to express cell-penetrating peptide L2.

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融合伙伴促进细胞穿透肽L2在毕赤酵母中的表达。

背景：L2是由无乳链球菌（S. agalactiae）的信息素与具有细胞穿透选择性的细胞穿透肽（CPP）结合形成的。L2对无乳链球菌具有更显著的穿透性和更好的特异性。然而，由于生产成本的原因，化学合成AMPs一直是一个挑战。方法：本研究以SUMO和短酸性融合标签作为融合伙伴，在毕赤酵母中进行细胞穿透肽L2的异源表达，并在5l发酵罐中实现SUMO-L2的高密度表达。结果：5 L发酵罐中SUMO-L2表达量达到629 mg/L。检测重组蛋白L2的抑菌活性；50%甲酸裂解84 h后，纯化的L2对无乳链球菌的最低抑菌浓度（mic）为4 ~ 8 μg/mL，最低杀菌浓度（MBCs）为8 ~ 16 μg/mL。结论：SUMO是表达细胞穿透肽L2的合适融合标签。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Antibiotics-Basel Pharmacology, Toxicology and Pharmaceutics-General Pharmacology, Toxicology and Pharmaceutics

CiteScore

7.30

自引率

14.60%

发文量

1547

审稿时长

11 weeks

期刊介绍： Antibiotics (ISSN 2079-6382) is an open access, peer reviewed journal on all aspects of antibiotics. Antibiotics is a multi-disciplinary journal encompassing the general fields of biochemistry, chemistry, genetics, microbiology and pharmacology. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. Therefore, there is no restriction on the length of papers.