Ksenia Lvova, Manuel Marcos, Benita Pérez-Cid, Xanel Vecino, José Manuel Cruz, Ana Belén Moldes
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引用次数: 0
Abstract
In the existing literature, there is ample information available concerning the analogs constituting linear Gramicidin (Gramicidin A). However, the literature lacks information regarding the microbial production of cyclic Gramicidin S (GR-S) and the study of its analogs' amino acid composition. Thus, in this study, GR-S was produced by Aneurinibacillus aneurinilyticus, isolated from corn steep liquor (CSL) and grown in synthetic or CSL-based medium. The extract was obtained from microbial biomass and underwent analysis with ultra performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) to ascertain the presence and abundance of GR-S clusters, identifying five GR S analogs. Hence, when using a synthetic medium, the dominant cluster compromised the sequence Val-Orn-Leu-Phe-Pro-Val-Orn-Leu-Phe-Pro. Contrarily, when using CSL-based medium, the main analog was composed of Val-Orn-Leu-Phe-Pro-Val-Lys-Leu-Phe-Pro. In the remaining analogs, one or both Ornithine (Orn) residues were replaced by Lysine (Lys) or diaminobutyric acid (D-Dab). Microscopy images revealed that the use of CSL-based medium resulted in a lower yield of GR-S and a higher accumulation of vacuoles compatible with the presence of intracellular polyhydroxyalkanoates (PHA) when comparing with TSB medium. The antifungal assay revealed a superior conidial inhibition activity of GR-S extract under evaluation against Aspergillus brasiliensis in comparison to the commercial GR-S. Consequently, it can be concluded that this novel microbial GR-S extract exhibits favorable antifungal activity, positioning it as a promising candidate for antimicrobial treatments. However, further research is necessary to address the global challenge of antibiotic-resistant pathogens.
在现有文献中,关于构成线性Gramicidin (Gramicidin A)的类似物有充足的信息。然而,文献缺乏关于微生物生产环Gramicidin S (GR-S)及其类似物氨基酸组成的研究。因此,在本研究中,GR-S是由从玉米浆(CSL)中分离出来的动脉瘤杆菌产生的,并在合成或基于CSL的培养基中生长。通过超高效液相色谱-电喷雾串联质谱(UPLC-ESI-MS/MS)分析,确定了GR-S簇的存在和丰度,鉴定出5种GR-S类似物。因此,当使用合成培养基时,优势簇破坏了序列Val-Orn-Leu-Phe-Pro-Val-Orn-Leu-Phe-Pro。相反,当使用基于csl的介质时,主要模拟物由Val-Orn-Leu-Phe-Pro-Val-Lys-Leu-Phe-Pro组成。在剩下的类似物中,一个或两个鸟氨酸(Orn)残基被赖氨酸(Lys)或二氨基丁酸(D-Dab)取代。显微镜图像显示,与TSB培养基相比,使用基于csl的培养基导致GR-S的产量较低,并且与细胞内聚羟基烷酸酯(PHA)的存在相兼容的液泡积累较多。抗真菌实验表明,与市售GR-S相比,GR-S提取物对巴西曲霉的分生孢子抑制活性更高。因此,可以得出结论,这种新型微生物GR-S提取物具有良好的抗真菌活性,将其定位为有希望的抗菌治疗候选者。然而,需要进一步的研究来应对抗生素耐药病原体的全球挑战。
期刊介绍:
Microbial Cell Factories is an open access peer-reviewed journal that covers any topic related to the development, use and investigation of microbial cells as producers of recombinant proteins and natural products, or as catalyzers of biological transformations of industrial interest. Microbial Cell Factories is the world leading, primary research journal fully focusing on Applied Microbiology.
The journal is divided into the following editorial sections:
-Metabolic engineering
-Synthetic biology
-Whole-cell biocatalysis
-Microbial regulations
-Recombinant protein production/bioprocessing
-Production of natural compounds
-Systems biology of cell factories
-Microbial production processes
-Cell-free systems