Lactobacillus brevis is an important strain used in food fermentation. Bacteriophage contamination is one of the main factors resulting in the failure of fermentation, which has caused great losses to food fermentation production. A virulent bacteriophage of L. brevis was isolated and characterized from fermented pickle water. Double-layer plate method was used to isolate and purify phage. The isolated phage was identified via the characteristics in negative staining observed with transmission electron microscopy (TEM), and its nucleic acid type. The phage genome was digested with various nuclease, and analyzed with electrophoresis subsequently to determine the type of nucleic acid, genome size and packaging mechanism. The phage titer was determined by double plate method. Characterization of the phage was investigated by determination of the optimal multiplicity of infection, one-step growth curve, and the effect of temperature, pH, other physicochemical factors and common disinfectants on their survival. A virulent phage named φB23 was isolated and purified. It consisted of a head with a polyhedral structure and a long-curved tail. The genome was double-stranded DNA, about 52.4 kb in size, and packaged with sticky terminal ligation in mechanism. The optimal multiplicity of infection was 0.001. One-step growth curve showed that the latent time and burst time were 30 min and 90 min, respectively, and the burst size was (103.2±9.0) pfu/cell. φB23 could be effectively inactivated by pasteurization under temperature of 63 ℃ and 72 ℃, treated for 10 min and 5 min, respectively. It was stable in the range of pH 3.0-8.0. In terms of disinfection effect of biocidal, φB23 was inactivated by 75% ethanol treatment for 10 min, and completely inactivated by 500 mg/L sodium hypochlorite treatment for 5 min. φB23 was identified as a phage belongs to the family Siphoviridae in the order Caudovirales, and its biochemical properties were characterized and analyzed. The results provide a basis for further elucidating the phage infection mechanism against L. brevis and developing effective phage control methods.
LIN Zeyong
,
LIANG Qianwen
,
SU Sui
,
GU Caiting
,
GUO Jieer
,
LUO Yuyu
,
LI Yun
. Isolation, identification and characterization of a virulent phage of Lactobacillus brevis[J]. Food and Fermentation Industries, 2022
, 48(12)
: 226
-232
.
DOI: 10.13995/j.cnki.11-1802/ts.028461
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