|
|
|
|
|
|
Development of cationic peptide chimeric lysins based on polyvalent phage lysin and their antibacterial activities |
CONG Yu, LIN Hong, WANG Jingxue* |
(College of Food Science and Engineering, Ocean University of China, Qingdao 266000, China) |
|
|
Abstract Polyvalent phage vB_SEqdws-315 can lyse both Salmonella and Escherichia coli strains, which has the potential to be used as antibacterial agents. In order to further study the antibacterial effect of its endolysin Lysin315, the gene was cloned and expressed in Escherichia coli BL21(DE3) with the induction of IPTG. High-purity recombinant Lysin315 was obtained by affinity chromatography. The molecular weight of Lysin315 was close to 25 kDa. Lysin315 needed EDTA as membrane permeability agents to inhibit the gram-negative bacteria because of the limitation of the cell wall structure of gram-negative bacteria. In order to enhance the direct antibacterial activity, Lysin315 was modified by cationic peptides to obtain chimeric lysins (Lysin315-5aa, Lysin315-10aa and Lysin315-15aa). All of the recombinant lysins were cloned and expressed in E. coli BL21(DE3) induced by IPTG and purified by affinity chromatography to acquire high purity. The results showed that cationic lysins could independently inhibit the hosts with high concentration without any membrane permeabilities. Lysin315-10aa also showed good antibacterial activities against Salmonella (106 CFU/mL) in milk at 4°C and 25°C. The development of cationic peptide chimeric lysins provided a solution for the independent application of gram-negative bacteriophages lysins.
|
Keywords
phage
lysin
cationic peptide
antibacterial activity
gram-negative bacterium
|
Issue Date: 18 November 2022
|
|
|
|
|
URL: |
http://sf1970.cnif.cn/EN/10.13995/j.cnki.11-1802/ts.031476 OR http://sf1970.cnif.cn/EN/Y2022/V48/I20/1 |
[1] |
DEWEY-MATTIA D, MANIKONDA K, HALL A J, et al.Surveillance for Foodborne Disease Outbreaks-United States, 2009—2015[J].Morbidity and Mortality Weekly Report Surveillance Summaries, 2018,67(10):1-11.
|
[2] |
LI W, PIRES S M, LIU Z, et al.Surveillance of foodborne disease outbreaks in China, 2003—2017[J].Food Control, 2020, 118:107359.
|
[3] |
史东灵, 解天慧, 石慧.噬菌体EC-p9和SM-p2内溶酶及穿孔素的特性及联合抑菌作用[J].食品与发酵工业, 2021, 47(24):7-12.SHI D L, XIE T H, SHI H.Characteristics and combined bacteriostasis of endolysin and holin in phage EC-p9 and SM-p2[J].Food and Fermentation Industries, 2021, 47(24):7-12.
|
[4] |
GARCíA P, MARTíNEZ B, OBESO J M, et al.Bacteriophages and their application in food safety[J].Letters in Applied Microbiology, 2008, 47(6):479-485.
|
[5] |
KIM J H, KIM H J, JUNG S J, et al.Characterization of Salmonella spp.-specific bacteriophages and their biocontrol application in chicken breast meat[J].Journal of Food Science, 2020, 85(3):526-534.
|
[6] |
GONDIL V S, HARJAI K, CHHIBBER S.Endolysins as emerging alternative therapeutic agents to counter drug-resistant infections[J].International Journal of Antimicrobial Agents, 2020, 55(2):105844.
|
[7] |
PASTAGIA M, SCHUCH R, FISCHETTI V A, et al.Lysins:The arrival of pathogen-directed anti-infectives[J].Journal of Medical Microbiology, 2013, 62(10):1 506-1 516.
|
[8] |
解天慧, 石慧.大肠杆菌O157∶H7噬菌体EC-p9的内溶酶和穿孔素的特性预测及克隆表达[J].食品与发酵工业, 2021, 47(9):107-113.XIE T H, SHI H.Characteristics prediction and expression of endolysin and holin from E.coli O157∶H7 phage EC-p9[J].Food and Fermentation Industries, 2021, 47(9):107-113.
|
[9] |
LOESSNER M J.Bacteriophage endolysins-current state of research and applications[J].Current Opinion in Microbiology, 2005, 8(4):480-487.
|
[10] |
SCHMELCHER M, LOESSNER M J.Bacteriophage endolysins - extending their application to tissues and the bloodstream[J].Current Opinion in Biotechnology, 2021, 68:51-59.
|
[11] |
SCHMELCHER M, DONOVAN D M, LOESSNER M J.Bacteriophage endolysins as novel antimicrobials[J].Future Microbiol, 2012, 7(10):1 147-1 171.
|
[12] |
YAN G M, YANG R J, FAN K J, et al.External lysis of Escherichia coli by a bacteriophage endolysin modified with hydrophobic amino acids[J].AMB Express, 2019, 9(1):106.
|
[13] |
LOOD R, WINER B Y, PELZEK A J, et al.Novel phage lysin capable of killing the multidrug-resistant gram-negative bacterium Acinetobacter baumannii in a mouse bacteremia model[J].Antimicrobial Agents and Chemotherapy, 2015, 59(4):1 983-1 991.
|
[14] |
BRIERS Y, VOLCKAERT G, CORNELISSEN A, et al.Muralytic activity and modular structure of the endolysins of Pseudomonas aeruginosa bacteriophages phiKZ and EL[J].Molecular Microbiology, 2007, 65(5):1 334-1 344.
|
[15] |
BRIERS Y, LAVIGNE R.Breaking barriers:Expansion of the use of endolysins as novel antibacterials against Gram-negative bacteria[J].Future Microbiology, 2015, 10(3):377-390.
|
[16] |
MA Q, GUO Z M, GAO C C, et al.Enhancement of the direct antimicrobial activity of Lysep3 cagainst Escherichia coli by inserting cationic peptides into its C terminus[J].Antonie Van Leeuwenhoek, 2017, 110(3):347-355.
|
[17] |
BRIERS Y, WALMAGH M, VAN PUYENBROECK V, et al.Engineered endolysin-based "Artilysins" to combat multidrug-resistant gram-negative pathogens[J].mBio, 2014, 5(4):e01379-e01314.
|
[18] |
林洪, 李萌, 王静雪.沙门氏菌噬菌体STP4-a重组内溶素抑菌特性分析[J].食品科学, 2015, 36(5):104-108.LIN H, LI M, WANG J X.Characterization of recombinant endolysin from a Salmonella- infecting bacteriophage STP4-a[J].Food Science, 2015, 36(5):104-108.
|
[19] |
NING H Q, CONG Y, LIN H, et al.Development of cationic peptide chimeric lysins based on phage lysin Lysqdvp001 and their antibacterial effects against Vibrio parahaemolyticus:A preliminary study[J].International Journal of Food Microbiology, 2021, 358:109396.
|
[20] |
LI Y Q, HAN Q, FENG J L, et al.Antibacterial characteristics and mechanisms of ε-poly-lysine against Escherichia coli and Staphylococcus aureus[J].Food Control, 2014, 43:22-27.
|
[21] |
RAHMAN M U, WANG W X, SUN Q Q, et al.Endolysin, a promising solution against antimicrobial resistance[J].Antibiotics (Basel), 2021, 10(11):1 277.
|
[22] |
王伟宇, 林洪, 王静雪.副溶血弧菌噬菌体qdvp001重组内溶素诱导表达以及理化性质初步研究[J].食品工业科技, 2016, 37(16):205-209.WANG W Y, LIN H, WANG J X.Characterization of recombinant endolysin from Vibrio parahaemolytics-infecting bacteriophage qdvp001[J].Science and Technology of Food Industry.2016, 37(16):205-209.
|
[23] |
NING H Q, LIN H, WANG J X.Synergistic effects of endolysin Lysqdvp001 and epsilon-poly-lysine in controlling Vibrio parahaemolyticus and its biofilms[J].International Journal of Food Microbiology, 2021, 343:109112.
|
[24] |
GUENTHER S, HERZIG O, FIESELER L, et al.Biocontrol of Salmonella typhimurium in RTE foods with the virulent bacteriophage FO1-E2[J].International Journal of Food Microbiology, 2012, 154(1-2):66-72.
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|