产奶香枯草芽孢杆菌的筛选及其在雪茄烟叶中的应用
Milky flavor-producing Bacillus subtilis and its application in cigar tobacco leaves
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摘要: 为解决国产雪茄烟叶原料香气量不足、刺激感较强等问题,通过嗅香评价筛选烟草源特色产奶香菌株,利用显微特征和16S rDNA测序进行菌株鉴定,结合机制雪茄加香感官评价得分和菌液OD600确定最优菌株培养条件,利用GC-MS分析确定发酵上清液中的关键香味成分,并进行国产雪茄烟叶原料发酵增香应用研究。结果表明:筛选到1株产奶香细菌(F2A),经鉴定其为枯草芽孢杆菌(Bacillus subtilis);该菌株适宜的培养时间为24 h、培养温度为32℃,培养基中碳源和无机盐为葡萄糖和氯化钙;菌株发酵上清液的主要奶香和甜香成分为2-羟基-3-甲基-2-环戊烯酮,且含量较高(7.58 μg/mL);经枯草芽孢杆菌F2A发酵处理后的楚雪14中,香兰素、香豆素、6-乙基-5,6-二氢吡喃-2-酮、2-羟基-3-甲基-2-环戊烯酮等奶香和甜香成分含量均显著增加,奶香香韵及甜感提升明显,巨豆三烯酮、3-羟基-β-大马士革酮、新植二烯等成分含量增加,清香和花香气息增强;由发酵后的楚雪14制成的雪茄中,奶香和甜香香韵显著提升,刺激感降低。该研究首次发现枯草芽孢杆菌F2A能产生2-羟基-3-甲基-2-环戊烯酮等奶甜香香韵成分,可为国产雪茄烟叶品质提升提供新的途径和理论支持。
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关键词:
- 枯草芽孢杆菌 /
- 奶香香韵 /
- 雪茄烟叶 /
- 2-羟基-3-甲基-2-环戊烯酮
Abstract: In order to solve the problems of poor aroma and strong irritation of Sino-cigar tobacco leaves, milky flavor-producing microbial strain isolated from tobacco leaves was screened through smell evaluation in this study. The bacterium was identified by microscopic characteristics and 16S rDNA sequence analysis. The liquid fermentation conditions were optimized by sensory evaluation of flavoring in machine-made cigar and OD600 of bacterial turbidity. The key aroma components of the fermentation broth were determined by GC-MS analysis. Flavoring effects of the Bacillus strain on the sensory quality of cigar tobacco leaves were also studied. The results showed that one bacterium strain F2A was screened out, which was identified as B.subtilis. The preferred conditions for liquid fermentation were temperature of 32 ℃, time of 24 h, carbon sources of glucose, and inorganic salts of CaCl2. The main milky and sweet aroma component in the fermentation broth was identified as 2-hydroxy-3-methyl-2-cyclopentenone with a high content of 7.58 μg/mL. After fermentation by B.subtilis F2A, the contents of milky and sweet flavor components such as vanillin, coumarin, 6-ethyl-5,6-dihydropyran-2-one, and 2-hydroxy-3-methyl-2-cyclopentenone in fermented Chexue 14 cigar tobacco leaves were significantly increased, which significantly enhanced milky and sweet aromas. And the enhancement of tabanone, 3-hydroxy-β-damascone, and neophytadiene could improve freshness and floral smell. After fermentation, the milky and sweet aromas of cigars produced by Chexue 14 tobacco leaves were significantly improved, and the irritation was reduced. This study suggested B.subtilis F2A could produce 2-hydroxy-3-methyl-2-cyclopentenone and other flavor compounds, which provides a new way and theoretical support for the quality improvement of cigar tobacco leaves. -
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