To elucidate the aroma profile of Lushan Yunwu Tea, a Box-Behnken experimental design combined with response surface methodology was applied to optimize the headspace solid-phase microextraction (HS-SPME) conditions, followed by gas chromatography-mass spectrometry (GC-MS) analysis. Additionally, olfactory receptors corresponding to the odorants were inferred from molecular structural features. Ligand-binding sites were predicted using machine-learning methods, and semi-flexible molecular docking was finally conducted for the odorant-olfactory receptor complexes. The results indicate that the optimal extraction conditions obtained by response surface optimization were as follows: sample weight 1.03 g, extraction time 43 min, and extraction temperature 73 ℃, with a validation error of 4.13%. A total of 87 volatile compounds belonging to ten chemical classes, including alcohols, aldehydes and ketones, were identified in Lushan Yunwu Tea. The key odorants with odor activity values (OAVs)≥1 were linalool, β-cyclocitral, geraniol, α-copaene, α-ionone and trans-β-ionone. Molecular docking reveals that all odorant-receptor pairs exhibited binding energies ≤﹣4.0 kcal·mol-1. This study not only optimized the analytical method for profiling tea aroma, but also innovatively employed molecular docking to validate the key aroma-active constituents. The findings provided a foundation for elucidating the formation mechanisms of Lushan Yunwu Tea aroma and offered references for its processing optimization and quality assessment.
[1] 邢湘臣. 五老峰与庐山云雾茶的传说[J]. 农业考古, 1988(1): 323.
Xing X C.The legend of Wulao Peak and Lushan Yunwu tea[J]. Agricultural Archaeology, 1988(1): 323.
[2] Yang Z, Baldermann S, Watanabe N.Recent studies of the volatile compounds in tea[J]. Food Research International, 2013, 53(2): 585-599.
[3] Zhai X, Zhang L, Granvogl M, et al.Flavor of tea (Camellia sinensis): a review on odorants and analytical techniques[J]. Comprehensive Reviews in Food Science and Food Safety, 2022, 21(5): 3867-3909.
[4] Reyes-Garcés N, Gionfriddo E, Gómez-Ríos G, et al.Advances in solid phase microextraction and perspective on future directions[J]. Analytical Chemistry, 2018, 90(1): 302-360.
[5] Risticevic S, Niri V H, Vuckovic D, et al.Recent developments in solid-phase micro extraction[J]. Analytical and Bioanalytical Chemistry, 2009, 393(3): 781-795.
[6] Bezerra M A, Santelli R E, Oliveira E P, et al.Response surface methodology (RSM) as a tool for optimization in analytical chemistry[J]. Talanta, 2008, 76(5): 965-977.
[7] Cao Q Q, Wang J Q, Chen J X, et al.Optimization of brewing conditions for Tieguanyin oolong tea by quadratic orthogonal regression design[J]. NPJ Science of Food, 2022, 6(1): 25. doi: 10.1038/s41538-022-00141-7.
[8] Januszkiewicz J, Sabik H, Azarnia S, et al.Optimization of headspace solid-phase microextraction for the analysis of specific flavors in enzyme modified and natural Cheddar cheese using factorial design and response surface methodology[J]. Journal of Chromatography A, 2008, 1195: 16-24. doi: 10.1016/j.chroma.2008.04.067.
[9] Lancet D, Pace U.The molecular basis of odor recognition[J]. Trends in Biochemical Sciences, 1987, 12: 63-66. doi: 10.1016/0968-0004(87)90032-6.
[10] Fleischer J, Breer H, Strotmann J.Mammalian olfactory receptors[J]. Frontiers in Cellular Neuroscience, 2009, 3: 9. doi: 10.3389/neuro.03.009.2009.
[11] Buck L, Axel R.A novel multigene family may encode odorant receptors: a molecular basis for odor recognition[J]. Cell, 1991, 65(1): 175-187.
[12] Zhang M, Hiki Y, Funahashi A, et al.A deep position-encoding model for predicting olfactory perception from molecular structures and electrostatics[J]. NPJ Systems Biology and Applications, 2024, 10(1): 76. doi: 10.1038/s41540-024-00401-0.
[13] Billesbølle C B, de March C A, van der Velden W J C, et al. Structural basis of odorant recognition by a human odorant receptor[J]. Nature, 2023, 615(7953): 742-749.
[14] Trott O, Olson A J.AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading[J]. Journal of Computational Chemistry, 2010, 31(2): 455-461.
[15] Arya N, Kaur A.Molecular docking: a review paper[J]. International Journal of Innovative Research in Engineering & Management, 2022: 140-146. doi: 10.55524/ijirem.2022.9.1.25.
[16] Xiao Z, Shen T, Niu Y, et al.Unraveling the characteristic aroma compounds in Longjing tea and their interaction mechanisms with S-curve and broad-spectrum olfactory receptors using molecular docking[J]. Food Bioscience, 2024, 60: 104423. doi: 10.1016/j.fbio.2024.104423.
[17] Hill W J, Hunter W G.A review of response surface methodology: a literature survey[J]. Technometrics, 1966, 8(4): 571-590.
[18] Stein S E, Gates R.Mass spectral reference libraries: an ever-expanding resource for chemical identification[J]. Analytical Chemistry, 2012, 84(17): 7274-7282.
[19] Wei X, Koo I, Kim S, et al.Compound identification in GC-MS by simultaneously evaluating mass spectrum and retention index[J]. Analyst, 2014, 139(10): 2507-2514.
[20] Ollitrault G, Achebouche R, Dreux A, et al.Pred-O3, a web server to predict molecules, olfactory receptors and odor relationships[J]. Nucleic Acids Research, 2024, 52(W1): W507-W512.
[21] Krivák R, Hoksza D.P2Rank: machine learning based tool for rapid and accurate prediction of ligand binding sites from protein structure[J]. Journal of Cheminformatics, 2018, 10: 39. doi: 10.1186/s13321-018-0285-8.
[22] Jendele L, Krivak R, Skoda P, et al.PrankWeb: a web server for ligand binding site prediction and visualization[J]. Nucleic Acids Research, 2019, 47(W1): W345-W349.
[23] Jakubec D, Skoda P, Krivak R, et al.PrankWeb 3: accelerated ligand-binding site predictions for experimental and modelled protein structures[J]. Nucleic Acids Research, 2022, 50(1): W593-W597.
[24] Polák L, Škoda P, Riedlová K, et al.PrankWeb 4: a modular web server for protein-ligand binding site prediction and downstream analysis[J]. Nucleic Acids Research, 2025, 53(1): W466-W471.
[25] Bezerra T K A, Araújo A R R, Arcanjo N M O, et al. Optimization of the HS-SPME-GC/MS technique for the analysis of volatile compounds in caprine Coalho cheese using response surface methodology[J]. Food Science and Technology, 2016, 36(1): 103-110.
[26] Ho C T, Zheng X, Li S.Tea aroma formation[J]. Food Science and Human Wellness, 2015, 4(1): 9-27.
[27] 张铭铭, 尹洪旭, 邓余良, 等. 基于HS-SPME/GC×GC-TOFMS/OAV不同栗香特征绿茶关键香气组分分析[J]. 食品科学, 2020, 41(2): 244-252.
Zhang M M, Yin H X, Deng Y L, et al.Analysis of key odorants responsible for different chestnut-like aromas of green teas based on headspace solid-phase microextraction coupled with comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry and odor activity value[J]. Food Science, 2020, 41(2): 244-252.
[28] Van Gemert L J. Compilations of odour threshold values in air, water and other media[M]. 2nd ed. Amsterdam: Oliemans Punter & Partners BV, 2011.
[29] Meng X Y, Zhang H X, Mezei M, et al.Molecular docking: a powerful approach for structure-based drug discovery[J]. Current Computer-Aided Drug Design, 2011, 7(2): 146-157.
[30] Sharma A, Kumar R, Aier I, et al.Sense of smell: structural, functional, mechanistic advancements and challenges in human olfactory research[J]. Current Neuropharmacology, 2019, 17(9): 891-911.
[31] Huang H, Chen X, Wang Y, et al.Characteristic volatile compounds of white tea with different storage times using E-nose, HS-GC-IMS, and HS-SPME-GC-MS[J]. Journal of Food Science, 2024, 89(9): 9137-9153.
[32] Jin G, Bi C, Ji A, et al.Volatile profiling of Tongcheng Xiaohua tea from different geographical origins: a multimethod investigation using sensory analysis, E-nose, HS-SPME-GC-MS, and chemometrics[J]. Foods, 2025, 14(11): 1996. doi: 10.3390/foods14111996.
浙公网安备 33019902000101号