乳酸菌接种方式对模拟葡萄汁发酵特性和香气物质的影响

    Effects of lactic acid bacteria inoculation methods on fermentation characteristics and aroma compounds of simulated grape juice

    • 摘要: 为探究酿酒酵母、酒酒球菌和植物乳杆菌的菌株组合与接种方式对模拟葡萄汁发酵动力学及香气品质的影响,试验采用酿酒酵母ES 488启动酒精发酵后,将本土酒酒球菌(ZX-1和GF-2)与植物乳杆菌(GF-26和GF-20)两两组合,分别在酒精发酵期间(同时接种)和结束后(顺序接种)启动苹果酸-乳酸发酵(MLF),动态测定发酵体系中还原糖和L-苹果酸含量,以及酿酒酵母、酒酒球菌和植物乳杆菌活菌数;借助顶空固相微萃取-气相色谱-质谱联用技术(HS-SPME-GC-MS)测定酒样挥发性香气化合物。结果表明,相较于顺序接种MLF,同时接种处理组的总发酵时间缩减了7~19 d。其中酿酒酵母、酒酒球菌(ZX-1)和植物乳杆菌(GF-26)同时接种发酵酒样的总发酵时间为14 d。与接种单一乳酸菌进行MLF相比,接入2种乳酸菌的MLF发酵时间缩短了4~6 d。酒精发酵结束时,同时接种处理组的乙醇体积分数相对较低,而乳酸菌增殖速率则高于相应的顺序接种处理组。值得注意的是,在酒精发酵启动阶段同时接种ZX-1和GF-26进行MLF处理组的降酸效果最佳,酒样中高级醇含量是顺序接种ZX-1和GF-26菌株酒样的1.91倍,酯类化合物也增加了33.97%,关键香气物质辛酸乙酯、壬酸乙酯和己酸乙酯增强了酒样的花香味、果香味和甜香味。综合分析,ES 488、ZX-1和GF-26三元菌株同时接种发酵具有提高MLF效率,改善葡萄酒香气品质的生产应用潜力。

       

      Abstract: This study systematically investigated the effects of inoculation timing and specific strain combinations involving Saccharomyces cerevisiae, Oenococcus oeni and Lactobacillus plantarum on fermentation kinetics and the final aromatic profile in a simulated grape juice medium. The primary objective was to evaluate the potential of simultaneous inoculation as a strategy to enhance both winemaking efficiency and sensory quality, thereby providing a scientific basis for optimizing traditional fermentation protocols. Alcoholic fermentation was initiated using the yeast strain S.cerevisiae ES 488. Two indigenous lactic acid bacteria (LAB) consortia were tested: one combining O.oeni ZX-1 and L.plantarum GF-26, and the other pairing O.oeni GF-2 with L.plantarum GF-20. The key experimental variable was the timing of LAB inoculation: simultaneous inoculation introduced LAB at the start of alcoholic fermentation alongside yeast, whereas sequential inoculation did so only after alcoholic fermentation was complete. Throughout the process, critical biochemical parameters including reducing sugar and L-malic acid degradation, as well as the microbial viability of all three strains were dynamically monitored at regular intervals to capture the real-time metabolic progression. After stabilization, volatile aroma compounds in the final wines were analyzed using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS), enabling precise identification and quantification of key odor-active molecules. Results showed that simultaneous inoculation significantly accelerated overall fermentation, reducing the total duration of alcoholic and malolactic fermentation by 7 to 19 days compared to the sequential method. Notably, the consortium of ES 488, ZX-1, and GF-26 achieved full microbial stability within 14 days, indicating high process efficiency and potential for rapid turnaround in commercial settings. Moreover, the dual-species LAB inoculum shortened the malolactic fermentation phase by an additional 4 to 6 days compared to single-strain inoculations, suggesting synergistic metabolic interactions that enhance substrate utilization and stress tolerance. Although ethanol levels immediately following alcoholic fermentation were slightly lower in simultaneous treatments, likely due to early competition for nutrients, the resulting low-ethanol environment promoted significantly higher proliferation rates of the LAB, which otherwise face inhibitory conditions in the high-ethanol post-fermentation environment of sequential trials. This improved microbial activity facilitated faster malic acid consumption and greater metabolic diversity. The simultaneous treatment with the ZX-1 and GF-26 consortium stood out not only for its rapid malic acid degradation but also for producing a sensorially superior wine. Chemical analysis revealed 1.91 times higher concentrations of higher alcohols and 33.97% more total esters compared to the sequential control using the same strains. Consequently, this wine exhibited significantly enhanced intensities of floral, fruity, and sweet aromatic notes, contributing to a more balanced and appealing sensory perception. Statistical analysis confirmed a strong positive correlation between these sensory attributes and elevated levels of key esters (ethyl octanoate, ethyl nonanoate, diethyl succinate, and ethyl caproate), which are known to contribute to fruity aroma complexity. These findings underscore the role of microbial synergy in shaping wine aroma. In conclusion, simultaneous inoculation using the tailored three-strain consortium of S.cerevisiae ES 488, O.oeni ZX-1, and L.plantarum GF-26 represents a promising enological approach. The integrated strategy offers dual benefits: it substantially improves production efficiency by shortening fermentation time, increasing tank turnover, and reducing operational costs, while simultaneously enhancing wine sensory quality through a more complex and desirable ester-rich aroma profile. These outcomes support the tailored inoculation strategy’s practical application for producing wines with greater aromatic intensity, typicity, and consistency in a time-efficient and predictable manner, effectively addressing key challenges in modern winemaking and offering a reliable method to improve both economic viability and sensory appeal. The present study provides novel insights into microbial co-cultivation dynamics and offers actionable strategies for advancing fermentation technologies in the wine industry.

       

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