Expression of nisin in Scenedesmus acutus and evaluation of its antimicrobial activity.

IF 2.1 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-06-12 DOI:10.1007/s10529-025-03603-6

Julieta Ponce-Alonso, Luzmila Martínez-González, Ángel Gabriel Alpuche-Solís, Fidel Martinez-Gutierrez, Ana Cecilia Lorenzo-Leal, Horacio Bach, Ruth Elena Soria-Guerra

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Abstract

Objectives: 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).

Results: 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.

Conclusions: 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.

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nisin在针叶草中的表达及抑菌活性评价。

目的：在绿色微藻（Scenedesmus acutus） UTEX 72的细胞核中表达人工合成的nisin基因，并评价其体外对金黄色葡萄球菌（ATCC 29213）、粪肠球菌（ATCC 29212）和单核增生李斯特菌（ATCC 19118）的抑菌活性。结果：通过与农杆菌共培养，实现了尖锐葡萄球菌的核转化。PCR分析证实转化系中存在Nis基因，ELISA测定总蛋白含量。结果表明，nisin主要在总可溶性蛋白部分表达，产率最高可达2.24µg/µl。抗菌试验表明，所有三种检测的革兰氏阳性细菌都有生长抑制作用。14.3µg/ml转基因nisin对金黄色葡萄球菌和单核增生乳杆菌的抑制作用为60 min，对粪肠球菌的抑制作用为55µg/ml，作用时间为180 min。野生型尖锐葡萄球菌总可溶性蛋白无抑菌活性。结论：Nisin在尖锐葡萄球菌细胞核中成功表达，对金黄色葡萄球菌、单核细胞增生乳杆菌和粪肠球菌具有抗菌活性。这些发现突出了尖锐葡萄球菌作为生产转基因抗菌肽的一个有希望的平台，为对抗致病菌提供了潜在的替代方案。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.