Ali Sahd, Karaynir Abdulkerim, Salih Doğan Hanife, Nachimuthu Ramesh, Badu Kingsley, Bozdoğan Bülent
Recombinant DNA and Protein Research Center (REDPROM), Aydin Adnan Menderes University, Aydin, Türkiye.
Antibiotic Resistance and Phage Therapy Laboratory, SBST, Vellore Institute of Technology, Vellore, India.
Phage (New Rochelle). 2025 Mar 17;6(1):20-31. doi: 10.1089/phage.2024.0029. eCollection 2025 Mar.
Biofilm is a major challenge across several sectors and contributes to serious risks to public health. This study aimed to evaluate the antibiofilm efficacy of disintegrated phages, whose lytic activities have been eliminated, against bacterial biofilms.
A total of seven lytic phages were disintegrated by sonication and confirmed to have completely lost their lytic activities by the spot test. Subsequently, both the disintegrated and intact phages were tested on the biofilms produced by five different biofilm-producing bacteria. The effects of heat and proteinase K on the ability of disintegrated phages to disrupt biofilms were determined. Moreover, the structural proteins released after the disintegration of phages were screened for their presence of lipase, amylase, protease, and DNase activities. Genome analysis of all the seven phages were screened for the presence of genes encoding proteins with enzymatic activities.
The disintegrated phages showed more effectiveness in degrading the bacterial biofilm when compared with intact phages. The amylase test results were positive for all the seven disintegrated phages tested, confirming the presence of starch-degrading enzymes. Genomic analysis of five phages revealed the presence of genes encoding transglycosylases, amidases, and glycosaminidases, which could contribute to biofilm degradation. However, only four of these proteins were also structural proteins of phages.
Our study demonstrated that disintegrated phages without lytic effects can still possess biofilm degrading ability, probably associated with the enzymatic activities of their structural proteins. This study showed that phages may have activities beyond lytic phage activities exhibited by their structural enzymes.
生物膜是多个领域面临的重大挑战，对公众健康构成严重风险。本研究旨在评估已消除裂解活性的裂解性噬菌体对细菌生物膜的抗生物膜功效。
通过超声处理使总共七种裂解性噬菌体解体，并通过点滴试验确认其完全丧失了裂解活性。随后，对由五种不同的生物膜形成细菌产生的生物膜测试了解体后的噬菌体和完整噬菌体。测定了加热和蛋白酶K对解体后的噬菌体破坏生物膜能力的影响。此外，筛选了噬菌体解体后释放的结构蛋白是否存在脂肪酶、淀粉酶、蛋白酶和脱氧核糖核酸酶活性。对所有七种噬菌体进行基因组分析，筛选是否存在编码具有酶活性蛋白质的基因。
与完整噬菌体相比，解体后的噬菌体在降解细菌生物膜方面表现出更高的有效性。对所有七种测试的解体后噬菌体的淀粉酶测试结果均为阳性，证实存在淀粉降解酶。对五种噬菌体的基因组分析显示存在编码转糖基酶、酰胺酶和糖胺酶的基因，这些基因可能有助于生物膜降解。然而，这些蛋白质中只有四种也是噬菌体的结构蛋白。
我们的研究表明，没有裂解作用的解体后噬菌体仍然可能具有生物膜降解能力，这可能与其结构蛋白的酶活性有关。本研究表明，噬菌体可能具有超出其结构酶所表现出的裂解噬菌体活性的其他活性。
