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

Ma, Yi1,2; Cui, Liu1; Wang, Meng1; Sun, Qiuli1; Liu, Kaisheng3,4; Wang, Jufang1

2021-06-01

10.3390/toxins13060420

TOXINS   影响因子和分区

2072-6651

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 (E-W) from phage Phi X174 and the screened mutant protein E (E-M) in the Escherichia coli BL21(DE3) strain. The results show that the lysis efficiency mediated by protein E-M 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 E-M 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 E-M 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.

bacterial ghosts Phi X174 lysis efficiency pLysS

SCI

英语

通讯

Natural Science Foundation of Guangdong Province[2019A1515011251] ; Fundamental Research Funds for the Central Universities[2018MS54] ; China Scholarship Council[201706155087] ; Science and Technology Foundation of Shenzhen[JCYJ20200109144410181]

Food Science & Technology ; Toxicology

Food Science & Technology ; Toxicology

WOS:000666014300001

MDPI

Web of Science

1.South China Univ Technol, Sch Biol & Biol Engn, Guangzhou 510006, Peoples R China
2.South China Univ Technol, Guangdong Prov Key Lab Fermentat & Enzyme Engn, Guangzhou 510006, Peoples R China
3.Jinan Univ, Clin Med Coll 2, Shenzhen Peoples Hosp, Shenzhen 518020, Peoples R China
4.Southern Univ Sci & Technol, Affiliated Hosp 1, Shenzhen 518020, Peoples R China

Ma, Yi,Cui, Liu,Wang, Meng,et al. A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts[J]. TOXINS,2021,13(6).

Ma, Yi,Cui, Liu,Wang, Meng,Sun, Qiuli,Liu, Kaisheng,&Wang, Jufang.(2021).A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts.TOXINS,13(6).

Ma, Yi,et al."A Novel and Efficient High-Yield Method for Preparing Bacterial Ghosts".TOXINS 13.6(2021).