Supplementary Table

This study investigated the impact of Bifidobacterium animalis subsp. lactis (B. lactis) IU100 or/and 1.5% resistant starch type III (RS3) on fermented milk during storage. The co-supplementation with enhanced texture, increasing hardness from 10.52 g (control) to 14.88 g and springiness from 1.18 mm to 3.03 mm, and promoted a denser gel network. Volatile profiling combined with OAV analysis revealed that the addition of B. lactis IU100 significantly increased the total content of alcohols (from 1231.77 μg/L to 2841.43 μg/L), particularly promoting the accumulation of compounds such as n-butanol and 1-octen-3-ol are known to contribute fruity and mushroom-like notes in dairy systems. The individual supplementation of 1.5% RS3 markedly elevated the total aldehyde content (from 3761.05 μg/L to 7026.82 μg/L), with compounds such as 2-octenal, (2e)- is associated with distinct fatty and nutty aromas in model systems. When B. lactis IU100 was combined with RS3, the level of 1-hexanol was further elevated, enhancing a fresh green note. Untargeted metabolomics further indicated that 300 significantly differential metabolites were identified in the co-supplemented group, among which key intermediates such as dephospho-CoA and adenosine diphosphate ribose were notably upregulated. These metabolites were mainly mapped to cofactor biosynthesis, purine metabolism, and pyrimidine metabolism, suggesting coordinated roles in the formation and interconversion of flavor precursors. In summary, the combined supplementation of B. lactis IU100 and RS3 effectively enhanced the overall quality and flavor complexity of fermented milk by modulating the volatile composition and core metabolic network.本研究探究了动物双歧杆菌乳酸亚种（Bifidobacterium animalis subsp. lactis，简称B. lactis）IU100单独或联合1.5% III型抗性淀粉（resistant starch type III，RS3）对发酵乳贮藏过程的影响。联合添加该两种成分可改善发酵乳质地：将硬度从对照组的10.52 g提升至14.88 g，弹性从1.18 mm提升至3.03 mm，并形成更为致密的凝胶网络结构。通过挥发性成分谱分析结合香气活度值（odor activity value, OAV）分析发现，添加B. lactis IU100可显著提升醇类总含量（从1231.77 μg/L升至2841.43 μg/L），尤其促进了正丁醇、1-辛烯-3-醇等化合物的积累——此类化合物在乳制品体系中可赋予果香与蘑菇风味。单独添加1.5% RS3则可显著提升醛类总含量（从3761.05 μg/L升至7026.82 μg/L），其中反式-2-辛烯醛（(2E)-2-octenal）等化合物在模型体系中可赋予典型脂肪与坚果香气。当B. lactis IU100与RS3联合添加时，1-己醇含量进一步升高，强化了新鲜青草风味。非靶向代谢组学分析进一步显示，联合添加组共鉴定出300种显著差异代谢物，其中脱磷辅酶A（dephospho-CoA）、二磷酸腺苷核糖（adenosine diphosphate ribose）等关键中间体的表达量显著上调。上述差异代谢物主要富集于辅因子生物合成、嘌呤代谢及嘧啶代谢通路，提示其在风味前体物质的形成与相互转化中发挥协同调控作用。综上，联合添加B. lactis IU100与RS3可通过调控挥发性成分组成与核心代谢网络，有效提升发酵乳的整体品质与风味复杂度。