菌酶协同发酵前后低次烟叶香气差异性分析
Analysis of aroma differences in upper low-grade tobacco leaves pre- and post- bacterial-enzyme synergistic fermentation
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摘要: 【目的】 探究菌酶协同处理前后烟叶香气成分的差异。【方法】 使用从云烟87烟叶表面筛选到的一株贝莱斯芽孢杆菌YUNM-4(Bacillus velezensisYUNM-4),分别与纤维素酶、脂肪酶和中性蛋白酶制成复合生物酶制剂,对上部低次烟叶进行协同发酵处理,利用连续流动仪和气相色谱-质谱联用(GC-MS)仪测定处理后烟叶的常规化学成分和香气成分,采用正交偏最小二乘法判别分析(OPLS-DA)和独立t检验(P<0.05)筛选出烟叶发酵前后含量具有显著性差异的成分,并通过相对气味活性值(ROAV)分析发酵前后烟叶中香气成分的贡献度差异。【结果】 1)与对照组相比,发酵后烟叶的常规化学成分总钾、烟碱、总糖、还原糖和钾氯比显著降低,糖碱比显著升高。2)发酵后烟叶的挥发性香味成分总含量明显提高,达到498.09 μg/g,提高了53.66%,主要以醇类、酮类、烃类、有机酸类、呋喃类和酚类化合物为主;构建的OPLS-DA模型具有良好的实验重复性且不存在过拟合现象,通过该模型筛选出32种含量具有显著性差异的成分,能较好地区分发酵前后烟叶香味成分的差异;3)发酵后烟叶中有12种挥发性香气成分的ROAV高于发酵前烟叶,其中苯甲醇、苯乙醇、大马士酮、β-紫罗兰酮、苯乙酮、二氢猕猴桃内酯等11种挥发性香气成分的ROAV>1,是发酵后烟叶的关键香气成分。【结论】 利用复合生物制剂对上部低次烟叶进行菌酶协同发酵处理,可以明显提升卷烟的感官品质。Abstract: 【Objective】 To investigate differences in aroma components of tobacco leaves before and after bacterial-enzyme synergistic treatment. 【Methods】 A strain of Bacillus velezensis YUNM-4, isolated from the surface of Yunyan 87 tobacco leaves, was combined with cellulase, lipase, and neutral protease to formulate a composite enzyme preparation. This preparation was applied in synergistic fermentation of upper low-grade flue-cured tobacco leaves. Conventional chemical components and aroma components of fermented tobacco leaves were analyzed using a continuous flow analyzer and gas chromatography-mass spectrometry (GC-MS), respectively. Orthogonal partial least squares discriminant analysis (OPLS-DA) and independent t-tests (P<0.05) were employed to identify components with significant content differences between pre- and post-fermentation samples, while relative odor activity values (ROAV) were applied to analyze differences in aroma component contributions. 【Results】 1) Compared to the control group, fermented tobacco leaves exhibited significant decreases in total potassium, nicotine, total sugar, reducing sugar, and K/Cl ratio, while the sugar/nicotine ratio increased significantly. 2) The total content of volatile aroma components in post-fermentation samples increased significantly to 498.09 μg/g, representing a 53.66% increase, primarily due to alcohols, ketones, hydrocarbons, organic acids, furans, and phenols. The OPLS-DA model demonstrated excellent reproducibility and no overfitting, as confirmed by cross-validation, identifying 32 components with significant content differences that effectively distinguished pre- and post-fermentation aroma profiles. 3) Twelve volatile aroma components in post-fermentation samples showed higher ROAV values than pre-fermentation samples, with 11 components (including benzyl alcohol, phenylethyl alcohol, damascenone, β-ionone, acetophenone, and dihydroactinidiolide) having ROAV>1, indicating their role as key aroma components. 【Conclusion】 Treatment of upper low-grade flue-cured tobacco leaves with bacterial-enzyme synergistic fermentation significantly improves cigarette sensory quality.
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