Joshi Himanshu, Seniya Surya P, Suryanarayanan Venkatesan, Patidar Neelam D, Singh Sanjeev K, Jain Vikas
Microbiology and Molecular Biology Laboratory, Department of Biological Sciences, Indian Institute of Science Education and Research (IISER), Bhopal, India.
Computer Aided Drug Designing and Molecular Modeling Laboratory, Department of Bioinformatics, Alagappa University, Karaikudi, India.
FEBS Lett. 2017 Oct;591(20):3276-3287. doi: 10.1002/1873-3468.12848. Epub 2017 Sep 20.
Most bacteriophages rapidly infect and kill bacteria and, therefore, qualify as the next generation therapeutics for rapidly emerging drug-resistant bacteria such as Mycobacterium tuberculosis. We have previously characterized the mycobacteriophage D29-generated endolysin, Lysin A, for its activity against mycobacteria. Here, we present a detailed characterization of the lysozyme domain (LD) of D29 Lysin A that hydrolyzes peptidoglycan of both gram-positive and gram-negative bacteria with high potency. By characterizing an exhaustive LD protein variant library, we have identified critical residues important for LD activity and stability. We further complement our in vitro experiments with detailed in silico investigations. We present LD as a potent candidate for developing phage-based broad-spectrum therapeutics.
大多数噬菌体能够迅速感染并杀死细菌，因此，它们有望成为治疗诸如结核分枝杆菌这类快速出现的耐药菌的新一代疗法。我们之前已对分枝杆菌噬菌体D29产生的溶菌酶Lysin A针对分枝杆菌的活性进行了表征。在此，我们详细描述了D29 Lysin A的溶菌酶结构域（LD），该结构域能够高效水解革兰氏阳性菌和革兰氏阴性菌的肽聚糖。通过对一个详尽的LD蛋白变体文库进行表征，我们确定了对LD活性和稳定性至关重要的关键残基。我们还通过详细的计算机模拟研究对体外实验进行了补充。我们提出，LD是开发基于噬菌体的广谱疗法的有力候选物。
