Kovács Renátó, Erdélyi Lóránd, Fenyvesi Ferenc, Balla Noémi, Kovács Fruzsina, Vámosi György, Klusóczki Ágnes, Gyöngyösi Alexandra, Bácskay Ildikó, Vecsernyés Miklós, Váradi Judit
Department of Medical Microbiology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary.
Faculty of Pharmacy, University of Debrecen, 4032 Debrecen, Hungary.
Pharmaceutics. 2022 Dec 21;15(1):18. doi: 10.3390/pharmaceutics15010018.
The antimicrobial effect of chitosan and synthetic chitosan derivatives has been confirmed on many Gram-positive and Gram-negative bacteria and fungi. The tests were carried out on pathogenic microorganisms, so the mechanism and concentration dependence of the inhibitory effect of chitosan were revealed. We conducted our tests on a probiotic strain, . Commercially available chitosan derivatives of different molecular weights were added to suspension in increasing concentrations. The minimum inhibitory concentration (MIC) value of chitosan was determined and confirmed the viability decreasing effect at concentrations above the MIC with a time-kill assay. The release of bacterium cell content was measured at 260 nm after treatment with 0.001-0.1% concentration chitosan solution. An increase in the permeability of the cell membrane was observed only with the 0.1% treatment. The interaction was also investigated by zeta potential measurement, and the irreversible interaction and concentration dependence were established in all concentrations. The interaction of fluorescein isothiocyanate (FITC) labeled low molecular weight chitosan and bacterial cells labeled with membrane dye (FM 4-64) was confirmed by confocal microscopy. In conclusion, the inhibitory effect of chitosan was verified on a probiotic strain, which is an undesirable effect in probiotic preparations containing chitosan additives, while the inhibitory effect experienced with pathogenic strains is beneficial.
壳聚糖及合成壳聚糖衍生物对多种革兰氏阳性菌、革兰氏阴性菌和真菌的抗菌作用已得到证实。这些试验是针对致病微生物进行的,因此揭示了壳聚糖抑制作用的机制及浓度依赖性。我们对一种益生菌菌株进行了试验。将不同分子量的市售壳聚糖衍生物以递增浓度添加到悬浮液中。测定了壳聚糖的最低抑菌浓度(MIC)值,并通过时间杀菌试验证实了在高于MIC的浓度下其对活力的降低作用。用0.001 - 0.1%浓度的壳聚糖溶液处理后,在260 nm处测量细菌细胞内容物的释放。仅在0.1%处理时观察到细胞膜通透性增加。还通过zeta电位测量研究了相互作用,并确定了所有浓度下的不可逆相互作用及浓度依赖性。通过共聚焦显微镜证实了异硫氰酸荧光素(FITC)标记的低分子量壳聚糖与用膜染料(FM 4 - 64)标记的细菌细胞之间的相互作用。总之,证实了壳聚糖对一种益生菌菌株具有抑制作用,这在含有壳聚糖添加剂的益生菌制剂中是一种不良作用,而对致病菌株的抑制作用则是有益的。