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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[1]
邵灯寅,李雨芮,邹恩凯,等.黑曲霉和酒糟二次发酵提升低次烟叶品质的研究[J].安徽农业科学,2023,51(16):178-181.
SHAO D Y,LI Y R,ZOU E K,et al.Study on the quality improvement of low-grade tobacco leaves by the secondary fermentation of Aspergillus niger and vinasse[J].Journal of Anhui Agricultural Sciences,2023,51(16):178-181. -
[2]
郝捷,季嫱,李力群,等.生物酶和微生物技术改善烟叶香气的研究进展[J].生物技术进展,2022,12(6):817-824.
HAO J,JI Q,LI L Q,et al.Research progress on improving aroma of tobacco leaf with bio-enzymes and microorganism technology[J].Current Biotechnology,2022,12(6):817-824. -
[3]
刘洪坤,毛文龙,游敏,等.改善低次烟叶品质的生物发酵工艺优化[J].中国烟草科学,2024,45(2):99-107.
LIU H K,MAO W L,YOU M,et al.Optimization of biological fermentation for improving the quality of discarded tobacco leaves[J].Chinese Tobacco Science,2024,45(2):99-107. -
[4]
马林.利用生物技术改变烟叶化学组分提高其吸食品质和安全性的研究[J].郑州工程学院学报,2001(3):40-42,45.
MA L.Study on changing tobacco chemical components for improving its smoking quality and safty by bio-technology[J].Journal of Zhengzhou Institute of Technology, 2001(3):40-42,45. -
[5]
赵铭钦,岳雪梅,邱立友.微生物发酵增质剂对卷烟酸性组分含量及品质效应的影响[J].中国烟草科学,2000,21(1):11. ZHAO M Q,YUE X M,QIU L Y.Effect of microbial fermentation quality enhancer on acid component content and quality effect of cigarette[J].Chinese Tobacco Science,2000
,21(1):11. -
[6]
黄申,芦尧,刘强,等.生物酶在烟草工业中的应用研究进展[J].轻工学报,2023,38(5):112-118.
HUANG S,LU Y,LIU Q,et al.Review on application of biological enzymes in tobacco industry[J].Journal of Light Industry,2023,38(5):112-118. -
[7]
代丽,黄永成,宫长荣,等.密集式烘烤条件下不同变黄温湿度对烤后烟叶致香物质的影响[J].华北农学报,2008,23(6):148-152.
DAI L,HUANG Y C,GONG C R,et al.Effects of different temperature and humidity yellowing conditions on aroma constituents of tobacco leaves during bulk curing[J].Acta Agriculturae Boreali-Sinica,2008,23(6):148-152. -
[8]
-
[9]
国家烟草专卖局.烟草及烟草制品 总植物碱的测定 连续流动法:YC/T 160—2002[S].北京:中国标准出版社,2004. State Tobacco Monopoly Administration.Tobacco and tobacco products—Determination of total alkaloids—Continuous flow method:YC/T 160—2002[S].Beijing:Standards Press of China,2004.
-
[10]
国家烟草专卖局.烟草及烟草制品 氯的测定 连续流动法:YC/T 162—2011[S].北京:中国标准出版社,2011. State Tobacco Monopoly Administration.Tobacco and tobacco products—Determination of chloride—Continuous flow method:YC/T 162—2011[S].Beijing:Standards Press of China,2011.
-
[11]
国家烟草专卖局.烟草及烟草制品 钾的测定 连续流动法:YC/T 217—2007[S].北京:中国标准出版社,2007. State Tobacco Monopoly Administration.Tobacco and tobacco products—Determination of Potassium—Continuous flow method:YC/T 217—2007[S].Beijing:Standards Press of China,2007.
-
[12]
国家烟草专卖局.烟草及烟草制品 水溶性糖的测定 连续流动法:YC/T 159—2019[S].北京:中国标准出版社,2019. State Tobacco Monopoly Administration.Tobacco and tobacco products—Determination of water soluble sugars—Continuous flow method:YC/T 159—2019[S].Beijing:Standards Press of China,2019.
-
[13]
国家烟草专卖局.烟草及烟草制品 总氮的测定 连续流动法:YC/T 161—2002[S].北京:中国标准出版社,2004. State Tobacco Monopoly Administration.Tobacco and tobacco products—Determination of total nitrogen—Continuous flow method:YC/T 161—2002[S].Beijing:Standards Press of China,2004.
-
[14]
国家市场监督管理总局,国家标准化管理委员会.感官分析 方法学 采用三点强迫选择法(3-AFC)测定嗅觉、味觉和风味觉察阈值的一般导则:GB/T 22366—2022[S].北京:中国标准出版社,2022. State Administration for Market Regulation,National Standardization Administration.Sensory analysis—Methodology—General guidance for measuring odour,flavour and taste detection thresholds by a three-alternative forced-choice (3-AFC)procedure:GB/T 2236—2022
[S].Beijing:Standards Press of China,2022. -
[15]
方嘉炜,严锦申,钟帅,等.毕节烟区不同产地烟叶的感官质量特征及关键香气成分[J].烟草科技,2024,57(8):21-31.
FANG J W,YAN J S,ZHONG S,et al.Sensory quality characteristics and key aroma components of tobacco leaves from different tobacco planting areas in Bijie[J].Tobacco Science & Technology,2024,57(8):21-31. -
[16]
LIH H,QIN D,WU Z Y,et al.Characterization of key aroma compounds in Chinese Guojing sesame-flavor Baijiu by means of molecular sensory science[J].Food Chemistry,2019,284:100-107.
-
[17]
李登科,田楠,田志章,等.应用顶空气相-离子迁移谱与相对气味活度值法分析卷烟特征香气物质[J].分析测试学报,2021,40(8):1164-1170.
LI D K,TIAN N,TIAN Z Z,et al.Analysis of typical aromatic matters in cigarettes using headspace-gas chromatography-ion mobility spectroscopy combined with relative odor activity value[J].Journal of Instrumental Analysis,2021,40(8):1164-1170. -
[18]
倪超,宋小宁,张松杰,等.高碳基土壤修复肥对舞阳烟叶化学成分和致香物质的影响[J].江西农业学报,2017,29(12):63-67.
NI C,SONG X N,ZHANG S J,et al.Effects of high-carbon-based soil remediation fertilizer on chemical constituents and aromatic substances of tobacco leaves in Wuyang area[J].Acta Agriculturae Jiangxi,2017,29(12):63-67. -
[19]
ELMENHORST H.The identification of maltol in cigarette smoke/nachweis von maltol in cigarettenrauch[J].Contributions to Tobacco Research,1971,6(2):70-73.
-
[20]
史清照,李河霖,王超,等.卷烟烟气内源性花香成分分析及花香特征调控[J].烟草科技,2023,56(5):56-67.
SHI Q Z,LI H L,WANG C,et al.Analysis of endogenous floral aroma components and regulation of floral aroma characteristics of mainstream cigarette smoke[J].Tobacco Science & Technology,2023,56(5):56-67. -
[21]
刘俊辉,杨春强,范武,等.卷烟烟气中7种香韵特征赋予组群的筛查[J].烟草科技,2019,52(4):44-50.
LIU J H,YANG C Q,FAN W,et al.Screening of seven note character impact groups in cigarette smoke[J].Tobacco Science & Technology,2019,52(4):44-50. -
[22]
吴彦辉,薛立新,许自成,等.烤烟巨豆三烯酮研究现状与展望[J].中国农业科技导报,2013,15(3):150-156.
WU Y H,XUE L X,XU Z C,et al.Research status and prospects of megastigmatrienone in flue-cured tobacco[J].Journal of Agricultural Science and Technology,2013,15(3):150-156. -
[23]
MOOKHERJEE B D,WILSON R A.Tobacco constituents-their importance in flaor and fragrance chemistry[J].Perfumer and Flavorist,1990,15:27-49.
-
[24]
ZHAN J,ZHOU F F,ZHU H B,et al.Classification and distribution of common aroma components in tobacco leaf based on their physical and chemical properties[J].Agricultural Science & Technology,2013,14(9):1358-1364.
-
[25]
张小慧,连梦瑶,李欢,等.不同品种辣椒挥发性香味物质的对比[J].食品工业,2023,44(12):119-125.
ZHANG X H,LIAN M Y,LI H,et al.Comparison of volatile aroma substances in different varieties of pepper[J].The Food Industry,2023,44(12):119-125. -
[26]
李萌,王旭东,罗昭标,等.混菌固态发酵低次烟叶工艺优化及挥发性致香成分分析[J].河南农业科学,2022,51(9):171-180.
LI M,WANG X D,LUO Z B,et al.Optimization of mixed microbial solid-state fermentation and analysis of volatile aroma components in low-grade tobacco leaves[J].Journal of Henan Agricultural Sciences,2022,51(9):171-180. -
[27]
杜康,张尉,顾丽莉,等.初烤烟叶中烟碱和新植二烯的提取工艺研究[J].核农学报,2021,35(6):1394-1401.
DU K,ZHANG W,GU L L,et al.Study on extraction technology of nicotine and neophytadiene from flue-cured tobacco leaves[J].Journal of Nuclear Agricultural Sciences,2021,35(6):1394-1401. -
[28]
胡安福,范武,夏倩,等.卷烟主流烟气焦甜、奶香和豆香特征成分组群的分布特征和感官贡献[J].烟草科技,2020,53(12):27-36.
HU A F,FAN W,XIA Q,et al.Distribution characteristics in mainstream cigarette smoke and sensory contributions of burnt-sweet,creamy and bean aroma components[J].Tobacco Science & Technology,2020,53(12):27-36. -
[29]
卢绍浩,谢永恒,许利平,等.基于OPLS-DA模型的雪茄烟叶香气差异分析[J].中国农业科技导报,2024,26(12):176-186.
LU S H,XIE Y H,XU L P,et al.Differential analysis of aroma components of cigar tobacco leaves based on OPLS-DA model[J].Journal of Agricultural Science and Technology,2024,26(12):176-186. -
[30]
邵淑贤,徐梦婷,林燕萍,等.基于电子鼻与HS-SPME-GC-MS技术对不同产地黄观音乌龙茶香气差异分析[J].食品科学,2023,44(4):232-239.
SHAO S X,XU M T,LIN Y P,et al.Differential analysis of aroma components of Huangguanyin oolong tea from different geographical origins using electronic nose and headspace solid-phase microextraction-gas chromatography-mass spectrometry[J].Food Science,2023,44(4):232-239. -
[31]
孙凤林,郝志峰.OPLS在非线性偏最小二乘回归模型的应用[J].计算机工程与设计,2010,31(12):2826-2829.
SUN F L,HAO Z F.Application of OPLS in non-linear partial least squares regression model[J].Computer Engineering and Design,2010,31(12):2826-2829. -
[32]
杨志晓,夏海乾,蔡凯,等.特色烟草新品种贵烟28的选育及其特征特性[J].中国烟草科学,2024,45(5):1-7.
YANG Z X,XIA H Q,CAI K,et al.Breeding and characteristics of a new special tobacco variety Guiyan28[J].Chinese Tobacco Science,2024,45(5):1-7. -
[33]
贾学伟,代玉祥,崔春,等.甘草固体颗粒香料的改良及卷烟加香应用[J].中国烟草学报,2025,31(1):30-39.
JIA X W,DAI Y X,CUI C,et al.Improvement of licorice solid granular spice and its application in cigarette flavoring[J].Acta Tabacaria Sinica,2025,31(1):30-39.
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