Biofilm-disrupting effects of phage endolysins LysAm24, LysAp22, LysECD7, and LysSi3: breakdown the matrix

Anastasiya M. Lendel, Nataliia P. Antonova, Igor V. Grigoriev, Evgeny V. Usachev, Vladimir A. Gushchin, Daria V. Vasina
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Abstract

The ability of most opportunistic bacteria to form biofilms, coupled with antimicrobial resistance, hinder the efforts to control widespread infections, resulting in high risks of negative outcomes and economic costs. Endolysins are promising compounds that efficiently combat bacteria, including multidrug-resistant strains and biofilms, without a low probability of subsequent emergence of stable endolysin-resistant phenotypes. However, the details of antibiofilm effects of these enzymes are poorly understood. To elucidate the interactions of bacteriophage endolysins LysAm24, LysAp22, LysECD7, and LysSi3 with bacterial films formed by Gram-negative species, we estimated their composition and assessed the endolysins’ effects on the most abundant exopolymers in vitro. The obtained data suggests a pronounced efficiency of these lysins against biofilms with high (Klebsiella pneumoniae) and low (Acinetobacter baumannii) matrix contents, or dual-species biofilms, resulting in at least a twofold loss of the biomass. These peptidoglycan hydrolases interacted diversely with protective compounds of biofilms such as extracellular DNA and polyanionic carbohydrates, indicating a spectrum of biofilm-disrupting effects for bacteriolytic phage enzymes. Specifically, we detected disruption of acid exopolysaccharides by LysAp22, strong DNA-binding capacity of LysAm24, both of these interactions for LysECD7, and neither of them for LysSi3.

Graphical abstract

Abstract Image

噬菌体内溶素 LysAm24、LysAp22、LysECD7 和 LysSi3 的生物膜破坏作用:分解基质
大多数机会性细菌都有形成生物膜的能力,再加上抗菌药耐药性,阻碍了控制大面积感染的努力,导致负面结果和经济成本的高风险。内溶素是一种很有前途的化合物,它能有效对付细菌,包括耐多药菌株和生物膜,而且随后出现稳定的内溶素耐药表型的可能性很低。然而,人们对这些酶的抗生物膜效应的细节了解甚少。为了阐明噬菌体内溶素 LysAm24、LysAp22、LysECD7 和 LysSi3 与革兰氏阴性菌形成的细菌薄膜之间的相互作用,我们估算了它们的组成,并在体外评估了内溶素对最丰富的外聚物的影响。获得的数据表明,这些溶菌素对基质含量高(肺炎克雷伯菌)和基质含量低(鲍曼不动杆菌)的生物膜或双菌种生物膜有明显的作用,导致生物量至少减少两倍。这些肽聚糖水解酶与细胞外 DNA 和聚阴离子碳水化合物等生物膜保护性化合物的相互作用多种多样,表明细菌溶解噬菌体酶具有多种生物膜破坏作用。具体来说,我们发现 LysAp22 能破坏酸性外多糖,LysAm24 有很强的 DNA 结合能力,LysECD7 有这两种相互作用,而 LysSi3 则没有。
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