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产果香微生物筛选及其对雪茄烟叶品质的影响

Screening of fruity flavor-producing bacteria and their effects on quality of cigar tobacco leaves

  • 摘要: 为筛选产果香微生物并提升国产雪茄烟叶品质,通过稀释涂布法进行菌种分离,以产香能力、留香时间和刺激性为指标进行菌种筛选,通过16S rDNA序列测定结合显微特征进行菌种鉴定,以机制雪茄加香感官评价和菌株发酵液的OD600值为指标进行菌株培养条件优化,采用GC-MS法对发酵液香味成分进行分析,并进行国产雪茄烟叶发酵应用研究。结果表明:①从巴西、河南和云南烤烟烟叶表面分离出20株细菌,其中M-8菌株产香效果较好,鉴定为马胃葡萄球菌(Staphylococcus equorum),优选培养条件为温度30 ℃、pH值7.0、培养时间36 h,优选的碳源和无机盐分别为葡萄糖和氯化钙。②从该菌株发酵液中共鉴定出29种挥发性成分,其中3-甲基丁酸、2-甲基丁酸、2-甲基丁酸乙酯、苯甲醇、苯甲酸乙酯和3-甲基-3, 5-二羟基戊酸内酯为关键果香、甜香和花香成分,且3-甲基丁酸和2-甲基丁酸乙酯的OAVsROAVs较大,感官贡献度大。③马胃葡萄球菌发酵楚雪14茄芯烟叶后,烟叶提取物中的3-甲基丁酸、2-甲基丁酸、4-丁基苯甲酸2-苯乙酯、二氢猕猴桃内酯、巨豆三烯酮和6-乙基-5, 6-二氢-2H-吡喃-2-酮等香味成分含量显著增加,雪茄烟叶的果香、蜜甜香和豆香香韵提升,香气质量明显改善,杂气和刺激性降低,余味舒适。马胃葡萄球菌可用于提升国产雪茄烟叶品质。

     

    Abstract: To screen fruity flavor-producing bacteria and improve the quality of domestic cigar tobacco leaves, flavor-producing bacteria were isolated by the dilution coating method. The bacteria were screened for flavor-producing ability, aroma retention time and irritation, and identified by 16S rDNA sequence analysis combined with microscopic characteristics. The sensory evaluation scores of the machine-made cigars and the OD600 values of the fermentation broths of the bacterial strains were used as indexes to optimize the culture conditions. The aroma components in the fermentation broth were analyzed by GC-MS, and the fermentation broth was then used to ferment domestic cigar tobacco leaves. The results showed that: 1) Twenty bacterial strains were isolated from the surface of flue-cured tobacco leaves from Brazil, Henan Province and Yunnan Province, in which M-8 strain had better flavor-producing ability and was identified as Staphylococcus equorum. The optimized culture conditions for M-8 strain were temperature at 30 ℃, pH at 7.0, and culture time at 36 h, and the optimized carbon source and inorganic salt were glucose and calcium chloride, respectively. 2) A total of 29 volatile components were identified in the fermentation broth of strain M-8. Among them, 3-methylbutyric acid, 2-methylbutyric acid, 2-methylbutanoic acid ethyl ester, benzenemethanol, ethyl benzoate and 3, 5-dihydroxy-3-methylpentanoic acid δ-lactone were revealed as the main aroma components with fruity, sweet and floral scents. 3-Methylbutyric acid and 2-methylbutanoic acid ethyl ester contributed more to the sensory quality due to their higher OAVs and ROAVs. 3) After fermentation with S. equorum, the contents of aroma components, such as 3-methylbutyric acid, 2-methylbutyric acid, 4-butylbenzoic acid 2-phenylethyl ester, 5, 6, 7, 7a-tetrahydro-4, 4, 7a-trimethyl-2(4H)-benzofuranone, megastigmatrienone and 6-ethyl-5, 6-dihydro-2H-pyrano-2-one, in Chuxue 14 cigar filler tobacco leaves were significantly increased, the fruity, honeysweet and bean aroma scents were significantly promoted, the offensive odor and irritation were reduced and the aftertaste was more pleasurable. Therefore, S. equorum derived from the fermentation could be used to improve the quality of domestic cigar tobacco leaves.

     

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