Optimization of Fermentation Process for Yunnan Produced Cigar Tobacco Leaves Using Response Surface Methodology

doi:10.11924/j.issn.1000-6850.casb2024-0476

Abstract

Abstract:

The paper aims to study the influence of process parameter changes during the bioenzyme-assisted fermentation of cigar tobacco leaves produced in Yunnan on their sensory quality and neutral aroma components. The ‘Yunxue 36’ cigar tobacco leaves were selected as the research object. According to the Box-Behnken Design principle, with different enzyme combinations, fermentation time, and fermentation temperature as influencing factors and sensory quality as the response value, the response surface methodology was used to optimize the fermentation process conditions of cigar tobacco leaves produced in Yunnan, and the corresponding prediction mathematical model was established. The simultaneous distillation extraction (SDE) method was used to extract volatile components, and the gas chromatography-mass spectrometry (GC-MS) method was used to analyze their component composition. The results showed that the regression equation model fitted well with the experiment, and the fermentation time had a greater impact on the sensory quality of tobacco leaves. Combining model optimization with practical operation, when the enzyme ratio was cellulase 0.7 mg/g + neutral protease 0.3 mg/g, the fermentation time was 15 days, and the fermentation temperature was 35℃, the sensory score of tobacco leaves was 84.333, which was relatively close to the sample of the best sensory quality test treatment. A total of 37 neutral aroma components were detected in 17 treated samples. Except for neophytadiene and other categories, the content of neutral aroma components in all treated samples was: carotenoid degradation products> phenylalanine conversion products> cembrane degradation products> non-enzymatic browning reaction products. The best test treatment for tobacco leaf sensory quality was an enzyme ratio of cellulase 0.7 mg/g+ neutral protease 0.3 mg/g, a fermentation time of 15 days, and a fermentation temperature of 40℃, which was mainly manifested in the improvement of the texture of the smoke, the enhancement of the richness of the aroma, and the improvement of the off-flavor, irritation, and aftertaste. The contents of the turkeyenone and the chlorophyll degradation product neophytadiene were the highest. This study used bioenzyme preparations to assist in exploring the fermentation process of cigar tobacco leaves produced in Yunnan, and found the optimal fermentation conditions of a specific enzyme preparation composition, which could provide a theoretical basis for the production practice of cigar tobacco.

Key words: biological enzyme, assisted fermentation, response surface methodology, cigar tobacco leaves, Yunnan produced cigars, fermentation process, sensory quality, neutral aroma components

ZHOU Xiao, XU Zhengrong, LU Xin, YANG Gang, DONG Junzhong, WANG Yue, WANG Bingbing, LI Tian, LIU Yan. Optimization of Fermentation Process for Yunnan Produced Cigar Tobacco Leaves Using Response Surface Methodology[J]. Chinese Agricultural Science Bulletin, 2025, 41(1): 141-147.

Figures/Tables 6

References 25

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水平	因素
水平	A.纤维素酶+中性蛋白酶/(mg/g)	B.发酵时间/d	C.发酵温度/℃
-1	0.3+0.7	5	35
0	0.5+0.5	10	40
1	0.7+0.3	15	45

水平	因素
水平	A.纤维素酶+中性蛋白酶/(mg/g)	B.发酵时间/d	C.发酵温度/℃
-1	0.3+0.7	5	35
0	0.5+0.5	10	40
1	0.7+0.3	15	45

来源	平方和	自由度	均方	F值	P值	显著性
模型	162.230	9	18.026	8.200	0.006	**
A	9.031	1	9.031	4.108	0.082
B	34.031	1	34.031	15.481	0.006	**
C	15.125	1	15.125	6.881	0.034	*
AB	10.563	1	10.563	4.805	0.064
AC	16.000	1	16.000	7.279	0.031	*
BC	1.000	1	1.000	0.455	0.522
A²	44.816	1	44.816	20.388	0.003	**
B²	26.580	1	26.580	12.091	0.010	*
C²	3.132	1	3.132	1.425	0.271
残差	15.388	7	2.198
失拟项	11.688	3	3.896	4.212	0.099	不显著
纯误差	3.700	4	0.925
总和	177.618	16

来源	平方和	自由度	均方	F值	P值	显著性
模型	162.230	9	18.026	8.200	0.006	**
A	9.031	1	9.031	4.108	0.082
B	34.031	1	34.031	15.481	0.006	**
C	15.125	1	15.125	6.881	0.034	*
AB	10.563	1	10.563	4.805	0.064
AC	16.000	1	16.000	7.279	0.031	*
BC	1.000	1	1.000	0.455	0.522
A²	44.816	1	44.816	20.388	0.003	**
B²	26.580	1	26.580	12.091	0.010	*
C²	3.132	1	3.132	1.425	0.271
残差	15.388	7	2.198
失拟项	11.688	3	3.896	4.212	0.099	不显著
纯误差	3.700	4	0.925
总和	177.618	16

类别	致香物质	B1	B2	B3	B4	B5	B6	B7	B8	B9	B10	B11	B12	B13	B14	B15	B16	B17
苯丙氨酸转化产物	苯甲醛	—	1.34	0.89	1.29	1.08	0.68	0.81	1.29	0.90	0.86	0.84	0.77	0.75	0.69	0.44	0.70	0.66
	苯甲醇	3.01	3.94	0.88	0.40	1.31	1.39	3.46	1.14	4.06	0.78	0.81	1.09	0.77	0.93	1.10	2.16	2.69
	苯乙醛	9.07	20.22	13.48	15.32	16.38	12.21	15.98	18.26	16.90	14.80	11.75	11.63	10.90	15.04	10.87	16.13	14.87
	苯乙醇	13.88	14.90	5.61	5.85	5.70	8.14	16.90	8.84	18.31	3.51	5.44	5.63	4.01	4.79	5.93	8.69	7.71
	小计	25.96	40.40	20.86	22.87	24.48	22.42	37.15	29.53	40.17	19.95	18.84	19.12	16.43	21.44	18.34	27.68	25.94
非酶棕色化反应产物	糠醛	—	1.61	—	—	—	—	—	—	—	—	—	—	—	—	—	—	—
	糠醇	—	3.10	0.34	0.37	0.49	0.52	1.59	1.25	1.38	—	0.35	—	—	—	—	0.93	1.13
	小计	—	4.71	0.34	0.37	0.49	0.52	1.59	1.25	1.38	—	0.35	—	—	—	—	0.93	1.13
西柏烷类降解产物	茄酮	11.30	15.76	11.70	13.69	12.53	11.14	5.03	4.79	4.17	13.81	11.29	11.39	11.60	9.74	11.48	9.82	10.13
类胡萝卜素降解产物	芳樟醇	—	0.47	0.33	—	0.38	0.35	—	—	0.44	0.26	0.24	0.23	0.28	0.29	0.29	0.33	0.30
	茶香酮	0.34	—	0.32	0.50	0.33	0.38	0.43	0.52	0.53	0.36	0.30	0.29	0.35	0.25	0.36	0.34	0.41
	β-环柠檬醛	0.20	0.25	0.20	0.31	0.29	0.17	—	—	0.17	0.24	0.22	0.21	0.24	0.21	0.18	0.16	0.11
	突厥烯酮	—	4.01	—	—	—	3.57	—	3.84	—	4.13	5.22	5.03	—	—	3.89	—	—
	大马士酮	7.68	5.90	3.50	3.80	5.08	—	3.17	—	4.15	0.98	1.44	1.01	3.32	3.77	2.79	3.99	4.00
	香叶基丙酮	—	—	1.00	—	0.96	1.45	—	—	—	1.14	0.94	1.03	1.18	0.83	1.33	1.81	2.17
	β-紫罗兰酮	0.82	0.38	—	0.27	0.78	0.40	—	—	—	0.61	0.54	0.97	0.89	0.63	0.96	0.54	0.60
	二氢猕猴桃内酯	9.67	9.02	3.30	3.84	3.07	4.88	12.25	14.48	9.15	2.74	1.90	2.85	2.05	1.47	3.18	3.88	4.02
	巨豆三烯酮A	4.71	3.25	1.80	2.23	2.33	2.33	2.81	3.27	2.72	2.23	1.91	2.78	2.02	1.85	2.29	2.16	2.36
	巨豆三烯酮B	14.30	8.35	9.98	12.44	11.96	11.44	16.01	16.07	14.52	11.72	9.32	9.31	10.11	9.04	10.31	10.00	11.01
	4-羟基-β-二氢大马酮	—	—	—	—	—	—	0.57	0.89	—	—	—	—	—	—	—	—	—
	巨豆三烯酮C	4.44	2.72	1.82	2.17	1.81	2.44	4.69	1.19	3.25	1.91	1.65	1.58	1.59	1.31	1.66	1.67	2.01
	巨豆三烯酮D	8.47	6.37	7.38	8.19	8.49	8.23	9.90	11.22	10.52	8.16	6.81	6.78	7.13	6.47	7.79	7.43	6.94
	法尼基丙酮	1.73	0.54	0.30	0.40	—	0.47	—	—	—	0.52	0.62	0.77	0.57	0.24	—	—	—
	小计	52.36	41.27	29.93	34.14	35.50	36.10	49.83	51.49	45.44	34.99	31.12	32.85	29.72	26.36	35.04	32.31	33.95
叶绿素降解产物	新植二烯	31.57	23.26	23.05	31.62	19.03	26.64	10.27	12.69	8.90	32.49	37.18	36.01	29.44	20.50	20.64	17.65	18.22
其他		17.37	19.81	14.24	14.74	18.05	30.26	32.58	14.29	8.77	26.80	24.02	32.43	17.29	19.65	18.92	22.41	19.77
合计		138.56	145.21	100.12	117.44	110.06	127.07	136.46	114.02	108.84	128.04	122.81	131.80	104.48	97.70	104.41	110.81	109.15