Ponce-Alonso Julieta, Martínez-González Luzmila, Alpuche-Solís Ángel Gabriel, Martinez-Gutierrez Fidel, Lorenzo-Leal Ana Cecilia, Bach Horacio, Soria-Guerra Ruth Elena
Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava No.6, Zona Universitaria, C.P. 78210, San Luis Potosí, México.
Instituto Potosino de Investigación Ciencia y Tecnología, A.C. Camino a La Presa San José 2055. Col. Lomas 4 Sección, C.P. 78216, San Luis Potosí, México.
Biotechnol Lett. 2025 Jun 12;47(3):60. doi: 10.1007/s10529-025-03603-6.
The study aimed to express a synthetic nisin gene in the nucleus of the green microalga Scenedesmus acutus UTEX 72 and evaluate its antimicrobial activity in vitro against Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), and Listeria monocytogenes (ATCC 19118).
Nuclear transformation of S. acutus was achieved through co-culture with Agrobacterium tumefaciens. PCR analysis confirmed the presence of the Nis gene in the transformant lines, while ELISA quantified the total protein content. Results showed that nisin was predominantly expressed in the total soluble protein fraction, with the highest yield reaching 2.24 µg/µl. Antimicrobial assays demonstrated growth inhibition in all three tested Gram-positive bacteria. S. aureus and L. monocytogenes were inhibited after 60 min of exposure to 14.3 µg/ml of transgenic nisin, whereas E. faecalis needed 55 µg/ml and 180 min for inhibition. No antimicrobial activity was observed with wild-type S. acutus total soluble protein.
Nisin was successfully expressed in the nucleus of S. acutus, exhibiting antimicrobial activity against S. aureus, L. monocytogenes, and E. faecalis. These findings highlight S. acutus as a promising platform for the production of transgenic antimicrobial peptides, offering a potential alternative for combating pathogenic bacteria.
本研究旨在将合成的乳链菌肽基因在绿色微藻尖锐栅藻(Scenedesmus acutus UTEX 72)的细胞核中表达,并在体外评估其对金黄色葡萄球菌(ATCC 29213)、粪肠球菌(ATCC 29212)和单核细胞增生李斯特菌(ATCC 19118)的抗菌活性。
通过与根癌农杆菌共培养实现了尖锐栅藻的核转化。PCR分析证实了转化株系中存在Nis基因,而ELISA定量了总蛋白含量。结果表明,乳链菌肽主要在总可溶性蛋白组分中表达,最高产量达到2.24微克/微升。抗菌试验表明,对所有三种测试的革兰氏阳性菌均有生长抑制作用。金黄色葡萄球菌和单核细胞增生李斯特菌在暴露于14.3微克/毫升的转基因乳链菌肽60分钟后受到抑制,而粪肠球菌需要55微克/毫升和180分钟才能被抑制。野生型尖锐栅藻总可溶性蛋白未观察到抗菌活性。
乳链菌肽在尖锐栅藻的细胞核中成功表达,对金黄色葡萄球菌、单核细胞增生李斯特菌和粪肠球菌具有抗菌活性。这些发现突出了尖锐栅藻作为生产转基因抗菌肽的有前景的平台,为对抗病原菌提供了一种潜在的替代方法。