Department of Laboratory Medicine, Qingdao Central Hospital, Qingdao, China; College of Marine Life Science, Ocean University of China, Qingdao, China.
Department of Laboratory Medicine, Qingdao Central Hospital, Qingdao, China.
Protein Expr Purif. 2025 Feb;226:106608. doi: 10.1016/j.pep.2024.106608. Epub 2024 Sep 16.
Currently, there is an urgent to develop safe and environmentally friendly alternatives to antibiotics for combating Vibrio parahaemolyticus. Endolysins are considered promising antibacterial agents due to their desirable range of action and ability to deal with antibiotic-resistant bacteria. While numerous Vibrio phages have been identified, the research on their endolysins is still in its infancy. In this study, a novel endolysin called LysVPB was cloned and expressed in Pichia pastoris. Phylogenetic analysis revealed that LysVPB bears little resemblance to other known endolysins, highlighting its unique nature. Homology modeling identified a putative calcium-binding site in LysVPB. The recombinant LysVPB achieved a lytic activity of 64.8 U/mL and had a molecular weight of approximately 17 kDa. LysVPB exhibited enhanced efficacy at pH 9.0, with 60 % of its maximum activity observed within the broad pH range of 6.0-10.0. The catalytic efficiency of LysVPB peaked at 30 °C but significantly declined beyond 50 °C. Ba, Co, and Cu showed inhibitory effects on the activity of LysVPB, while Ca can boost it to 126.8 %. Furthermore, LysVPB exhibited satisfactory efficacy against strains of V. parahaemolyticus. LysVPB is an innovative phage lysin with good characteristics that are specific to certain hosts. The modular nature of LysVPB allows for efficient domain exchange with alternative lysins as antimicrobial components and fusion with antimicrobial peptides. This opens up possibilities for engineering chimeric lysins in a broader range of target hosts with high antimicrobial effectiveness and strong activity under physiological conditions.
目前，迫切需要开发安全且环保的抗生素替代品来对抗副溶血性弧菌。溶菌酶被认为是有前途的抗菌剂，因为它们具有理想的作用范围和应对抗生素耐药菌的能力。虽然已经鉴定出许多弧菌噬菌体，但对其溶菌酶的研究仍处于起步阶段。在这项研究中，一种新型的溶菌酶 LysVPB 在毕赤酵母中被克隆和表达。系统发育分析表明，LysVPB 与其他已知的溶菌酶没有相似之处，突出了其独特的性质。同源建模确定了 LysVPB 中一个假定的钙结合位点。重组 LysVPB 实现了 64.8 U/mL 的溶菌活性，分子量约为 17 kDa。LysVPB 在 pH 9.0 时表现出增强的功效，在 pH 6.0-10.0 的较宽范围内观察到其最大活性的 60%。LysVPB 的催化效率在 30°C 时达到峰值，但超过 50°C 时显著下降。Ba、Co 和 Cu 对 LysVPB 的活性表现出抑制作用，而 Ca 可以将其提高到 126.8%。此外，LysVPB 对副溶血性弧菌菌株表现出令人满意的功效。LysVPB 是一种具有良好特性的创新噬菌体溶菌酶，对特定宿主具有特异性。LysVPB 的模块性质允许与替代溶菌酶进行有效的结构域交换，作为抗菌成分，并与抗菌肽融合。这为在更广泛的目标宿主中工程合成具有高抗菌效果和在生理条件下强活性的嵌合溶菌酶开辟了可能性。
