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Shuang Wang,Jingmin Gu,Meng Lv,Zhimin Guo,Guangmou Yan,Ling Yu,Chongtao Du,Xin Feng,Wenyu Han,Changjiang Sun,Liancheng Lei 한국미생물학회 2017 The journal of microbiology Vol.55 No.5
Bacteriophage endolysin is one of the most promising antibioticsubstitutes, but in Gram-negative bacteria, the outermembrane prevents the lysin from hydrolyzing peptidoglycansand blocks the development of lysin applications. Theprime strategy for new antibiotic substitutes is allowing lysinto access the peptidoglycan from outside of the bacteria byreformation of the lysin. In this study, the novel Escherichiacoli (E. coli) phage lyase lysep3, which lacks outside-in catalyticability, was fused with the N-terminal region of theBacillus amyloliquefaciens lysin including its cell wall bindingdomain D8 through the best manner of protein fusionbased on the predicted tertiary structure of lysep3-D8 to obtainan engineered lysin that can lyse bacteria from the outside. Our results showed that lysep3-D8 could lyse both Gramnegativeand Gram-positive bacteria, whereas lysep3 and D8have no impact on bacterial growth. The MIC of lysep3-D8on E. coli CVCC1418 is 60 μg/ml; lysep3-D8 can inhibit thegrowth of bacteria up to 12 h at this concentration. The bactericidalspectrum of lysep3-D8 is broad, as it can lyse of allof 14 E. coli strains, 3 P. aeruginosa strains, 1 Acinetobacterbaumannii strain, and 1 Streptococcus strain. Lysep3-D8 hassufficient bactericidal effects on the 14 E. coli strains testedat the concentration of 100 μg/ml. The cell wall binding domainof the engineered lysin can destroy the integrity of theouter membrane of bacteria, thus allowing the catalytic domainto reach its target, peptidoglycan, to lyse the bacteria. Lysep3-D8 can be used as a preservative in fodder to benefitthe health of animals. The method we used here proved to bea successful exploration of the reformation of phage lysin.
( Hongtao Liu ),( Seng Zhu ),( Yingying Sun ),( Na Li ),( Jingmin Gu ),( Changjiang Sun ),( Xin Feng ),( Wenyu Han ),( Jianxia Jiang ),( Liancheng Lei ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.1
Meningitis caused by Streptococcus suis serotype 2 (S. suis 2) is a great threat to the pig industry and human health. Virulence factors associated with the pathogenesis of meningitis have yet to be clearly defined, even though many potential S. suis 2 virulence factors have been identified. This greatly hinders the progress of S. suis 2 meningitis pathogenesis research. In this study, a co-culture blood-brain barrier (BBB) model was established using primary porcine brain microvascular endothelial cells and astrocytes, and the whole genome library of S. suis 2 was constructed using phage display technology. Finally, a total of 14 potential virulence factors contributing to S. suis 2 adherence to and invasion of the BBB were selected by analyzing the interactions between the phage library and the co-culture model. Twelve of these factors have not been previously reported in meningitis-related research. The data provide valuable insight into the pathogenesis of S. suis 2 meningitis and potential targets for the development of drug therapies.
Wu Yaran,Cai Lei,Xie Xingzi,Yang Shuying,Shi Qing,Jia Hongzhe,Gu Xuqiang,Deng Jingmin,Shi Mingzhao,Chen Qiuping,Cao Shaoqian,Cai Shuangfeng 한국응용생명화학회 2022 Applied Biological Chemistry (Appl Biol Chem) Vol.65 No.2
Resistant starch could be degraded by the fermentation of colonic microorganisms in the large intestine of mammals, but not in the small intestine. In this study, we established a novel strategy by using resistant starch as an endogenous marker to determine the glucose absorption of the small intestine of laboratory animals. By optimization of the classical enzymatic method of starch measurement, the demand for the sample weight was reduced by 90%. Moreover, the amount of resistant starch in normal feed was detectable without any extra addition. The value of small intestine glucose absorption of mice was similar when using resistant starch and titanium dioxide as inert markers. The fermentation of resistant starch by intestinal microorganisms in the small intestine was demonstrated not disturbing the detection of glucose absorption significantly. Artificial sweeteners exposed ICR mice showed different glucose absorption which indicated, first, resistant starch can be used as a novel endogenous marker in the small intestine of small animals; second, although glucose tolerance did not change in mice after short-term exposure to artificial sweeteners, there were significant changes in glucose absorption associated with it; third, the short-term exposure resulted in no significant change in glucose tolerance.