Growth characteristics of Lactobacillus brevis KB290 in the presence of bile.
Live Lactobacillus brevis KB290 have several probiotic activities, including immune stimulation and modulation of intestinal microbial balance. We investigated the adaptation of L. brevis KB290 to bile as a mechanism of intestinal survival. Strain KB290 was grown for 5 days at 37 °C in tryptone-yeast extract-glucose (TYG) broth supplemented with 0.5% sodium acetate (TYGA) containing 0.15%, 0.3%, or 0.5% bile. Growth was determined by absorbance at 620 nm or by dry weight. Growth was enhanced as the broth's bile concentration increased. Bile-enhanced growth was not observed in TYG broth or with xylose or fructose as the carbon source, although strain KB290 could assimilate these sugars. Compared with cells grown without bile, cells grown with bile had twice the cell yield (dry weight) and higher hydrophobicity, which may improve epithelial adhesion. Metabolite analysis revealed that bile induced more lactate production by glycolysis, thus enhancing growth efficiency. Scanning electron microscopy revealed that cells cultured without bile for 5 days in TYGA broth had a shortened rod shape and showed lysis and aggregation, unlike cells cultured for 1 day; cells grown with bile for 5 days had an intact rod shape and rarely appeared damaged. Cellular material leakage through autolysis was lower in the presence of bile than in its absence. Thus lysis of strain KB290 cells cultured for extended periods was suppressed in the presence of bile. This study provides new role of bile and sodium acetate for retaining an intact cell shape and enhancing cell yield, which are beneficial for intestinal survival.
NARO Institute of Livestock and Grassland Science, Tsukuba, Ibaraki, Japan. Electronic address: anne@affrc.go.jp.
Hiromi
H
Kimoto-Nira
Nature-Wellness Research Department, Kagome Co., Ltd., Nasushiobara, Tochigi, Japan.
Shigenori
S
Suzuki
Nature-Wellness Research Department, Kagome Co., Ltd., Nasushiobara, Tochigi, Japan.
Hiroyuki
H
Suganuma
NARO Institute of Livestock and Grassland Science, Tsukuba, Ibaraki, Japan.
Naoko
N
Moriya
NARO Institute of Livestock and Grassland Science, Tsukuba, Ibaraki, Japan.
Chise
C
Suzuki
D001422:Bacterial Adhesion; D001433:Bacteriolysis; D001646:Bile / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D006019:Glycolysis; D006801:Humans; D019344:Lactic Acid / Q000378:metabolism; D052196:Levilactobacillus brevis / Q000254:growth & development* / Q000378:metabolism* / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D019346:Sodium Acetate / Q000378:metabolism; D013696:Temperature; D013997:Time Factors
D003470:Culture Media; D019344:Lactic Acid; D019346:Sodium Acetate
Bile resistance; Cell lysis; Hydrophobicity; Lactobacillus brevis