Kwon Seok-Joon, Kim Domyoung, Lee Inseon, Kim Jungbae, Dordick Jonathan S
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York, 12180.
Department of Chemical and Biological Engineering, Korea University, Seoul, 02841, Republic of Korea.
Biotechnol Bioeng. 2017 Aug;114(8):1648-1657. doi: 10.1002/bit.26300. Epub 2017 May 18.
Targeting infectious bacterial pathogens is important for reducing the evolution of antibiotic-resistant bacteria and preserving the endogenous human microbiome. Cell lytic enzymes including bacteriophage endolysins, bacterial autolysins, and other bacteriolysins are useful antibiotic alternatives due to their exceptional target selectivity, which may be used to lysins rapidly kill target bacteria and their high specificity permit the normal commensal microflora to be left undisturbed. Genetic information of numerous lysins is currently available, but the identification of their antimicrobial function and specificity has been limited because most lysins are often poorly expressed and exhibit low solubilities. Here, we report the development of bacterial cell chip for rapidly accessing the function of diverse genes that are suggestive of encoding lysins. This approach can be used to evaluate rapidly the species-specific antimicrobial activity of diverse lysins synthesized from in vitro transcription and translation (TNT) of plasmid DNA. In addition, new potent lysins can be assessed that are not expressed in hosts and display low solubility. As a result of evaluating the species-specific antimicrobial function of 11 (un)known lysins with an in vitro TNT-coupled bacterial cell chip, a potent recombinant lysin against Staphylococcus strains, SA1, was identified. The SA1 was highly potent against not only S. aureus, but also both lysostaphin-resistant S. simulans and S. epidermidis cells. To this end, the SA1 may be applicable to treat both methicillin-resistant S. aureus (MRSA) and lysostaphin-resistant MRSA mutants. Biotechnol. Bioeng. 2017;114: 1648-1657. © 2017 Wiley Periodicals, Inc.
靶向感染性细菌病原体对于减少抗生素耐药菌的进化和保护人体内生微生物群至关重要。包括噬菌体溶菌酶、细菌自溶素和其他溶菌素在内的细胞裂解酶因其卓越的靶标选择性而成为有用的抗生素替代品，这种选择性可用于使溶菌素迅速杀死靶标细菌，其高特异性使正常共生微生物群不受干扰。目前已有许多溶菌素的遗传信息，但由于大多数溶菌素通常表达不佳且溶解度低，其抗菌功能和特异性的鉴定受到限制。在此，我们报告了一种细菌细胞芯片的开发，用于快速了解提示编码溶菌素的各种基因的功能。这种方法可用于快速评估由质粒DNA的体外转录和翻译（TNT）合成的各种溶菌素的物种特异性抗菌活性。此外，还可以评估在宿主中不表达且溶解度低的新型强效溶菌素。通过使用体外TNT偶联细菌细胞芯片评估11种已知和未知溶菌素的物种特异性抗菌功能，鉴定出一种针对葡萄球菌菌株SA1的强效重组溶菌素。SA1不仅对金黄色葡萄球菌具有高效力，而且对耐溶葡萄球菌素的模仿葡萄球菌和表皮葡萄球菌细胞也具有高效力。为此，SA1可能适用于治疗耐甲氧西林金黄色葡萄球菌（MRSA）和耐溶葡萄球菌素的MRSA突变体。生物技术与生物工程。2017年；114：1648 - 1657。©2017威利期刊公司。
