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Article

A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts

by 1,2, 1, 1, 1, 3,* and 1,*
1
School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China
2
Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou 510006, China
3
Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China
*
Authors to whom correspondence should be addressed.
Received: 25 May 2021 / Accepted: 9 June 2021 / Published: 13 June 2021
(This article belongs to the Special Issue Bacterial Pore-Forming Toxin)
Bacterial ghosts (BGs) are empty cell envelopes possessing native extracellular structures without a cytoplasm and genetic materials. BGs are proposed to have significant prospects in biomedical research as vaccines or delivery carriers. The applications of BGs are often limited by inefficient bacterial lysis and a low yield. To solve these problems, we compared the lysis efficiency of the wild-type protein E (EW) from phage ΦX174 and the screened mutant protein E (EM) in the Escherichia coli BL21(DE3) strain. The results show that the lysis efficiency mediated by protein EM was improved. The implementation of the pLysS plasmid allowed nearly 100% lysis efficiency, with a high initial cell density as high as OD600 = 2.0, which was higher compared to the commonly used BG preparation method. The results of Western blot analysis and immunofluorescence indicate that the expression level of protein EM was significantly higher than that of the non-pLysS plasmid. High-quality BGs were observed by SEM and TEM. To verify the applicability of this method in other bacteria, the T7 RNA polymerase expression system was successfully constructed in Salmonella enterica (S. Enterica, SE). A pET vector containing EM and pLysS were introduced to obtain high-quality SE ghosts which could provide efficient protection for humans and animals. This paper describes a novel and commonly used method to produce high-quality BGs on a large scale for the first time. View Full-Text
Keywords: bacterial ghosts; ΦX174; lysis efficiency; pLysS bacterial ghosts; ΦX174; lysis efficiency; pLysS
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MDPI and ACS Style

Ma, Y.; Cui, L.; Wang, M.; Sun, Q.; Liu, K.; Wang, J. A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts. Toxins 2021, 13, 420. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins13060420

AMA Style

Ma Y, Cui L, Wang M, Sun Q, Liu K, Wang J. A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts. Toxins. 2021; 13(6):420. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins13060420

Chicago/Turabian Style

Ma, Yi, Liu Cui, Meng Wang, Qiuli Sun, Kaisheng Liu, and Jufang Wang. 2021. "A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts" Toxins 13, no. 6: 420. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins13060420

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