New bacteriophage-derived lysins, LysJ and LysF, with the potential to control Bacillus anthracis,Applied Microbiology and Biotechnology

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New bacteriophage-derived lysins, LysJ and LysF, with the potential to control Bacillus anthracis
Applied Microbiology and Biotechnology ( IF 3.9 ) Pub Date : 2024-01-09 , DOI: 10.1007/s00253-023-12839-z
Aleksandra Nakonieczna ₁ , Agnieszka Topolska-Woś ₂ , Małgorzata Łobocka ₃

Affiliation

Abstract

Bacillus anthracis is an etiological agent of anthrax, a severe zoonotic disease that can be transmitted to people and cause high mortalities. Bacteriophages and their lytic enzymes, endolysins, have potential therapeutic value in treating infections caused by this bacterium as alternatives or complements to antibiotic therapy. They can also be used to identify and detect B. anthracis. Endolysins of two B. anthracis Wbetavirus phages, J5a and F16Ba which were described by us recently, differ significantly from the best-known B. anthracis phage endolysin PlyG from Wbetavirus genus bacteriophage Gamma and a few other Wbetavirus genus phages. They are larger than PlyG (351 vs. 233 amino acid residues), contain a signal peptide at their N-termini, and, by prediction, have a different fold of cell binding domain suggesting different structural basis of cell epitope recognition. We purified in a soluble form the modified versions of these endolysins, designated by us LysJ and LysF, respectively, and depleted of signal peptides. Both modified endolysins could lyse the B. anthracis cell wall in zymogram assays. Their activity against the living cells of B. anthracis and other species of Bacillus genus was tested by spotting on the layers of bacteria in soft agar and by assessing the reduction of optical density of bacterial suspensions. Both methods proved the effectiveness of LysJ and LysF in killing the anthrax bacilli, although the results obtained by each method differed. Additionally, the lytic efficiency of both proteins was different, which apparently correlates with differences in their amino acid sequence.

Key points

• LysJ and LysF are B. anthracis-targeting lysins differing from lysins studied so far

• LysJ and LysF could be overproduced in E. coli in soluble and active forms

• LysJ and LysF are active in killing cells of B. anthracis virulent strains

中文翻译：

新型噬菌体来源的溶素 LysJ 和 LysF，具有控制炭疽杆菌的潜力

抽象的

炭疽杆菌是炭疽病的病原体，炭疽病是一种严重的人畜共患疾病，可传播给人类并导致高死亡率。噬菌体及其溶解酶、内溶素在治疗由该细菌引起的感染方面具有潜在的治疗价值，作为抗生素治疗的替代或补充。它们还可用于识别和检测炭疽杆菌。我们最近描述的两种炭疽芽孢杆菌Wbetavirus噬菌体J5a和F16Ba的内溶素与来自Wbetavirus属噬菌体Gamma和其他一些Wbetavirus属噬菌体的最著名的炭疽芽孢杆菌噬菌体内溶素PlyG显着不同。它们比 PlyG 大（351 个氨基酸残基与 233 个氨基酸残基），在 N 末端含有信号肽，并且根据预测，具有不同折叠的细胞结合结构域，表明细胞表位识别的不同结构基础。我们以可溶形式纯化了这些细胞内溶素的修饰形式，分别命名为 LysJ 和 LysF，并去除了信号肽。在酶谱分析中，两种修饰的细胞内溶素都可以裂解炭疽芽孢杆菌细胞壁。通过在软琼脂中的细菌层上点样并评估细菌悬浮液光密度的降低来测试它们针对炭疽芽孢杆菌和其他芽孢杆菌属物种的活细胞的活性。两种方法都证明了LysJ和LysF杀灭炭疽杆菌的有效性，尽管每种方法获得的结果不同。此外，两种蛋白质的裂解效率不同，这显然与其氨基酸序列的差异相关。