为了筛选具有协同作用的乳酸菌并从代谢产物角度探究乳酸菌协同作用的机理,通过测定乳酸菌单独及混合培养的生长情况、乳酸和游离氨基氮代谢物的产量、乳酸脱氢酶(lactic dehydrogenase,LDH)关键酶活性及转录水平等指标评价乳酸菌的相互作用关系,进一步利用非靶向代谢组学技术探究乳酸菌之间的协同作用机理。结果表明,植物乳植杆菌HC 1775(Lactiplantibacillus plantarum HC 1775,Lpl)和鼠李糖乳酪杆菌GG HC 1509(Lacticaseibacillus rhamnosus GG HC 1509,LGG)混合培养后细菌数量明显增加,乳酸含量达到71.20 μg/mL,较Lpl、LGG单独培养增加了23.52%、38.68%,氨基氮含量为64.68 μg/mL;添加无细胞上清液后Lpl活菌数增加了23.70%,LGG增加了32.83%;ldh基因对Lpl+LGG混合菌调控明显,ldh表达量为单菌的2.15倍,Lpl和LGG之间具有相互促进生长的协同作用;非靶向代谢组学分析表明,Lpl+LGG混合培养代谢特征谱与单菌间存在显著差异,代谢物丰度较高,其中天冬氨酸、苏氨酸等氨基酸类物质为关键差异代谢物。Lpl和LGG间存在协同作用,混合培养体系中氨基酸类物质显著增加影响了乳酸菌间的代谢,乳酸菌协同培养研究为乳酸菌的应用提供了坚实的理论基础。
To screen for lactic acid bacteria (LAB) with synergistic effects and explore the mechanisms of their synergistic interactions from a metabolic perspective, the growth of LAB, lactic acid production and free amino nitrogen metabolites, lactate dehydrogenase (LDH) activity, and transcription regulation were measured in both monocultures and co-cultures.These indicators were used to evaluate the interactions between LAB, and non-targeted metabolomics further elucidated metabolic mechanisms underlying their synergy.The results displayed that the number of bacteria in the mixed culture of Lactiplantibacillus plantarum HC 1775 (Lpl) and Lacticaseibacillus rhamnosus GG HC 1509 (LGG) was significantly enhanced, with total lactic acid production reaching 71.20 μg/mL, which was 23.52% and 38.68% higher than Lpl and LGG monocultures, respectively.The free amino nitrogen content was 64.68 μg/mL.When cell-free supernatant was added, the viable counts of Lpl increased by 23.70% and those of LGG by 32.83%.The ldh gene was significantly regulated in Lpl+LGG co-culture, with an expression level 2.15-fold higher than in single cultures.The results displayed that Lpl and LGG exhibited a synergistic relationship that promoted each other’s growth.Metabolomic profiling revealed distinct metabolic signatures in co-culture, characterized by elevated amino acid biosynthesis (notably aspartate and threonine) and heightened metabolic flux.Lpl and LGG exhibit synergistic interactions, with amino acids significantly increasing in the co-culture system, thereby influencing the metabolism of LAB.This mechanistic understanding of LAB interactions advances foundational knowledge for optimizing microbial consortia in industrial and probiotic applications.
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