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Synthesis of 1,2-Azaborines and the Preparation of Their Protein Complexes with T4 Lysozyme Mutants
Published on: March 25, 2017
Construction and evaluation of an inside-out lysis system based on T4 lysozyme
Wenfeng Ni1, Mengya Qin1, Xiaoge Yang1
1Anhui Provincial Key Laboratory of Biodiversity Conservation and Characteristic Resource Utilization in Southwest Anhui, School of Life Sciences and Food Engineering, Anqing Normal University, Anqing, Anhui, China.
None:
Lysozyme can effectively lyse Escherichia coli (E. coli), providing a method for the release of intracellular substances. In this study, an inside-out lysis system based on T4 lysozyme was constructed. Three secretion signal peptides (PelB, NapA, and HybA) were fused to T4 lysozyme to create fusion proteins, among which, NapA-T4 lysozyme (N-T4L) exhibited the highest level of intracellular protein leakage. Microscopic analysis revealed significant cell wall damage in N-T4L expressing cells, confirming the initiation of cell lysis. The influence of chemical factors on cell lysis was also evaluated. Expression of N-T4L reduced cell viability by 2.09 log units following Triton X-100 treatment and pH adjustment. Antibacterial assays demonstrated that N-T4L in the fermentation supernatant maintained substantial inhibitory activity against bacteria. These findings provide a strong foundation for synthetic biology applications involving programmed cell destruction in E. coli.
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