菌酶协同发酵前后低次烟叶香气差异性分析
Analysis of aroma difference of low-grade tobacco leaves prior to and after treatment by bacterial-enzyme co-fermentation
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摘要: 为探究菌酶协同处理前后的烟叶香气成分的差异,使用从云烟87烟叶表面筛选到的一株贝莱斯芽孢杆菌(Bacillus velezensis) YUNM-4和3种生物酶(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,分别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。结果表明: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,糖碱比显著增加。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53.7%,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,可以明显提升卷烟的感官品质。Abstract: In order to explore the difference of aroma components of tobacco leaves before and after the synergistic treatment of bacteria and enzymes, a strain of Bacillus velezensis YUNM-4 screened from the surface of Yunyan 87 tobacco leaves and three kinds of biological enzymes (cellulase, lipase and neutral protease) were used to make a compound biological enzyme preparation, and the upper low-grade tobacco leaves were subjected to synergistic fermentation treatment. The conventional chemical components and aroma components of the treated tobacco leaves were determined by continuous flow analyzer and gas chromatography-mass spectrometry (GC-MS), respectively. Orthogonal partial least squares discriminant analysis (OPLS-DA) and independent t test (P<0. 05) were used to screen out the components with significant differences in the content of tobacco leaves before and after fermentation, and the relative odor activity value (ROAV) was used to analyze the difference in the contribution of aroma components in tobacco leaves before and after fermentation. The results showed that: 1) Compared with the control group, the total potassium, nicotine, total sugar, reducing sugar and potassium chloride ratio of the conventional chemical components of the fermented tobacco leaves reduced significantly, while the sugar-nicotine ratio increased significantly. 2) The total content of volatile aroma components after fermentation tobacco leaves significantly increased, reaching 498. 09 μg/g, an increase of 53. 7%, mainly alcohols, ketones, hydrocarbons, organic acids, furans and phenols; The OPLS-DA model had good experimental repeatability without over-fitting phenomenon. Through this model, 32 components with significant differences in content were screened out can better distinguish the differences in aroma components of tobacco leaves before and after fermentation. 3) The ROAV of 12 volatile aroma components in the fermented tobacco leaves was higher than that in the pre-fermented tobacco leaves. 4) The ROAV values of 11 components such as benzyl alcohol, phenylethyl alcohol, damascenone, β-ionone, acetophenone and dihydroactinidiolide were greater than 1, which were the key aroma components of fermented tobacco leaves, and could significantly improve the sensory quality of cigarettes.
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