Corcoran B M, Stanton C, Fitzgerald G F, Ross R P
Teagasc, Dairy Products Research Centre, Moorepark, Fermoy, Co. Cork, Ireland.
Appl Environ Microbiol. 2005 Jun;71(6):3060-7. doi: 10.1128/AEM.71.6.3060-3067.2005.
Lactobacillus rhamnosus GG is an industrially significant probiotic strain with proven health benefits. In this study, the effect of glucose on L. rhamnosus GG survival was analyzed in simulated gastric juice at pH 2.0. It was found that the presence of 19.4 mM glucose resulted in up to 6-log10-enhanced survival following 90 min of exposure. Further work with dilute HCl confirmed that glucose was the sole component responsible. Comparative analysis with other Lactobacillus strains revealed that enhanced survival was apparent in all strains, but at different pH values. The presence of glucose at concentrations from 1 to 19.4 mM enhanced L. rhamnosus GG survival from 6.4 to 8 log10 CFU ml(-1) in simulated gastric juice. The mechanisms behind the protective effect of glucose were investigated. Addition of N',N'-dicyclohexylcarbodiimide to simulated gastric juice caused survival to collapse, which was indicative of a prominent role in inhibition of F0F1-ATPase. Further work with neomycin-resistant mutants that exhibited 38% to 48% of the F0F1-ATPase activity of the parent confirmed this, as the survival in the presence of glucose of these mutants decreased 3 x 10(6)-fold compared with the survival of the wild type (which had a viability of 8.02 log10 CFU ml(-1)). L. rhamnosus GG survival in acidic conditions occurred only in the presence of sugars that it could metabolize efficiently. To confirm the involvement of glycolysis in the glucose effect, iodoacetic acid was used to inhibit glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity. The reduction in GAPDH activity caused survival to decrease by 8.30 log10 CFU ml(-1) in the presence of glucose. The data indicate that glucose provides ATP to F0F1-ATPase via glycolysis, enabling proton exclusion and thereby enhancing survival during gastric transit.
鼠李糖乳杆菌GG是一种具有重要工业意义的益生菌菌株,已证实对健康有益。在本研究中,分析了葡萄糖对鼠李糖乳杆菌GG在pH 2.0的模拟胃液中存活的影响。结果发现,在暴露90分钟后,19.4 mM葡萄糖的存在使存活率提高了6个对数级。用稀盐酸进一步研究证实葡萄糖是唯一起作用的成分。与其他乳酸杆菌菌株的比较分析表明,所有菌株在不同pH值下存活率均有提高。在模拟胃液中,1至19.4 mM浓度的葡萄糖使鼠李糖乳杆菌GG的存活率从6.4 log10 CFU/ml提高到8 log10 CFU/ml。研究了葡萄糖保护作用背后的机制。向模拟胃液中添加N',N'-二环己基碳二亚胺导致存活率下降,这表明其在抑制F0F1-ATP酶方面起重要作用。对新霉素抗性突变体的进一步研究证实了这一点,这些突变体的F0F1-ATP酶活性为亲本的38%至48%,与野生型(存活率为8.02 log10 CFU/ml)相比,这些突变体在葡萄糖存在下的存活率降低了3×10(6)倍。鼠李糖乳杆菌GG仅在存在其能有效代谢的糖类时,才能在酸性条件下存活。为了证实糖酵解参与葡萄糖效应,使用碘乙酸抑制甘油醛-3-磷酸脱氢酶(GAPDH)活性。在葡萄糖存在下,GAPDH活性的降低导致存活率下降8.30 log10 CFU/ml。数据表明,葡萄糖通过糖酵解为F0F1-ATP酶提供ATP,实现质子排出,从而提高在胃转运过程中的存活率。