类胡萝卜素降解菌株的原位筛选及其在雪茄提质增香中的应用

doi:10.13560/j.cnki.biotech.bull.1985.2023-0258

生物技术通报 ›› 2023, Vol. 39 ›› Issue (9): 192-201.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0258

类胡萝卜素降解菌株的原位筛选及其在雪茄提质增香中的应用

吴巧茵¹^,²^,³(), 施友志⁴, 李林林⁴, 彭政¹^,³, 谭再钰⁴, 刘利平⁴, 张娟¹^,³(), 潘勇⁴()

1.江南大学工业生物技术教育部重点实验室，无锡 214122
2.江南大学生物工程学院，无锡 214122
3.江南大学未来食品科学中心，无锡 214122
4.湖北中烟工业有限责任公司，武汉 430040

收稿日期:2023-03-21 出版日期:2023-09-26 发布日期:2023-10-24
通讯作者: 张娟，女，博士，教授，研究方向：基于系统生物学的微生物群落分析与代谢工程改造、新型食品酶制剂的开发；E-mail: zhangj@jiangnan.edu.cn；
潘勇，男，工程师，研究方向：雪茄原料、工艺加工技术研究与评价；E-mail: sx_panyong@sx.hbtobacoo.cn
作者简介:吴巧茵，女，硕士研究生，研究方向：雪茄微生物与发酵研究；E-mail: 6200201045@stu.jiangnan.edu.cn
基金资助:
国家重点研发计划(2019YFC1605800)

In Situ Screening of Carotenoid Degrading Strains and the Application in Improving Quality and Aroma of Cigar

WU Qiao-yin¹^,²^,³(), SHI You-zhi⁴, LI Lin-lin⁴, PENG Zheng¹^,³, TAN Zai-yu⁴, LIU Li-ping⁴, ZHANG Juan¹^,³(), PAN Yong⁴()

1. Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122
2. School of Biotechnology, Jiangnan University, Wuxi 214122
3. Science Center for Future Foods, Jiangnan University, Wuxi 214122
4. China Tobacco Hubei Industry Co., Ltd., Wuhan 430040

Received:2023-03-21 Published:2023-09-26 Online:2023-10-24

摘要/Abstract

摘要：

通过筛选可降解类胡萝卜素的烟草内生菌，并应用于国产雪茄烟叶发酵从而实现烟叶香韵和感官质量提升。首先，原位培养基中添加烟叶粉末模拟原位生长环境以富集烟叶内生菌，结合流式分选技术，从优质雪茄烟叶内生菌中筛选具有β-胡萝卜素降解能力的菌株；将菌株接种至国产雪茄烟叶发酵，检测发酵烟叶类胡萝卜素降解产物含量并进行感官评价。结果显示，共筛选获得21株内生菌对β-胡萝卜素的降解率超过50%，主要为农杆菌属、根瘤菌属、鞘氨醇杆菌属、肠杆菌属等。菌株C31、C11、H4能够增加烟叶香韵类型，明显改善烟叶香气质、香气量、杂气、刺激性等指标，发酵烟叶类胡萝卜素降解产物含量分别是对照的1.38、1.28、1.43倍，其中法尼基丙酮、香叶基丙酮、巨豆三烯酮的增加强化了烟叶焦甜香、花香、木香香韵。综上，原位筛选获得的3株类胡萝卜素降解内生菌C31、C11、H4能够较好地改善国产雪茄烟叶品质，可作为新型雪茄工业微生物制剂和特征香韵雪茄微生物制剂进一步开发应用。

关键词: 类胡萝卜素, 降解, 内生菌, 原位筛选, 雪茄烟叶, 发酵, 提质, 增香

Abstract:

Endophyte degrading carotenoids were screened and applied to domestic cigar leaves to increase domestic cigar tobacco leaves（CTLs）aroma and improve sensory quality. CTLs powder was added into in situ medium to simulate the growth environment of tobacco endophytes, and enrichment culture combined with flow cytometry technology was used to screen strains with β-carotene-degrading ability from high-quality cigar tobacco endophytes. The strains inoculated into domestic CTLs, and the contents of carotenoid degradation products were detected and the sensory quality of CTLs was analyzed. Total 21 endophytes had β-carotene degradation rate over 50%, mainly including Agrobacterium, Rhizobium, Sphingobacterium, Enterobacter, etc. These strains made different effects on CTLs sensory quality, among which strains C31, C11 and H4 increased the aroma type, significantly improved the aroma quality, aroma amount, miscellaneous gas, irritation and other CTLs sensory index. The carotenoid-degrading product content from strain C31, C11 and H4 CTLs was 1.38, 1.28 and 1.43 times compared with CK, respectively. The farnesyl acetone, geranyl acetone and megastigmatrienone strengthened the burnt sweet, flowery and woody aroma of CTLs. The three endophytes C31, C11 and H4 obtained by in situ screening can better improve the domestic CTLs sensory quality, and can be further developed and applied as microbial preparations of new cigar industry and characteristic aromatic cigar.

Key words: carotenoids, degradation, endophyte, situ screening, cigar tobacco leaves, fermentation, quality improvement, aroma enhancement

吴巧茵, 施友志, 李林林, 彭政, 谭再钰, 刘利平, 张娟, 潘勇. 类胡萝卜素降解菌株的原位筛选及其在雪茄提质增香中的应用[J]. 生物技术通报, 2023, 39(9): 192-201.

WU Qiao-yin, SHI You-zhi, LI Lin-lin, PENG Zheng, TAN Zai-yu, LIU Li-ping, ZHANG Juan, PAN Yong. In Situ Screening of Carotenoid Degrading Strains and the Application in Improving Quality and Aroma of Cigar[J]. Biotechnology Bulletin, 2023, 39(9): 192-201.

图/表 7

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菌株 Strain	香气质 Aroma quality	香气量 Aroma amount	杂气 Miscellaneous gas	刺激性Thrillness	余味 Remaining taste	甜润感Sweetness	燃烧性Flammability	灰色Gray	质量得分 Quality score
CK	3.10	3.10	2.90	2.90	3.10	2.90	4.10	4.00	60.47
C31	3.50	3.30	3.00	3.30	3.30	3.50	4.50	4.00	67.07
C11	3.30	3.50	3.30	3.50	3.30	3.30	4.50	3.50	66.93
H4	3.00	3.00	3.00	3.30	3.30	2.80	4.00	3.80	63.00
A72	3.30	3.30	3.50	3.30	3.00	2.80	4.50	4.00	62.73
F5	3.00	3.30	3.00	2.80	3.30	2.80	4.00	3.50	61.07
1D3	3.50	3.00	3.50	3.00	3.00	2.50	4.00	4.00	61.00
A5	3.00	2.80	2.80	3.00	2.50	2.30	4.00	3.00	60.93
H92	2.90	3.10	3.10	3.20	2.70	2.80	3.80	3.70	60.83
2F2	3.00	3.00	3.00	3.50	3.00	2.50	3.80	3.50	59.87
2B1	2.80	2.80	2.80	3.20	3.10	2.80	3.30	3.20	59.75
C7	3.30	3.30	3.00	2.50	3.30	2.80	3.00	3.50	59.73
A71	3.30	2.50	3.00	2.50	2.80	2.80	4.00	4.00	59.73
F92	3.00	2.80	3.00	2.80	2.80	2.50	4.00	3.50	59.73
2B2	2.50	2.80	2.80	2.80	2.80	3.00	4.00	4.00	59.33
1F9	2.50	2.50	2.50	2.50	2.50	2.50	4.00	4.00	58.80
2E5	3.00	2.50	3.00	3.00	3.00	3.50	4.00	4.00	57.00
2C11	3.00	3.50	3.00	3.00	3.00	2.50	3.00	3.00	57.30
1C1	2.50	2.50	3.00	3.00	3.00	2.50	4.00	3.00	56.70
B11	2.80	2.80	2.50	3.00	2.80	1.50	4.00	3.30	56.00
F91	3.00	2.80	2.50	2.50	3.00	1.50	3.50	3.00	55.47
C42	3.00	2.80	2.50	2.80	2.80	2.80	3.30	3.30	55.07

菌株 Strain	香气质 Aroma quality	香气量 Aroma amount	杂气 Miscellaneous gas	刺激性Thrillness	余味 Remaining taste	甜润感Sweetness	燃烧性Flammability	灰色Gray	质量得分 Quality score
CK	3.10	3.10	2.90	2.90	3.10	2.90	4.10	4.00	60.47
C31	3.50	3.30	3.00	3.30	3.30	3.50	4.50	4.00	67.07
C11	3.30	3.50	3.30	3.50	3.30	3.30	4.50	3.50	66.93
H4	3.00	3.00	3.00	3.30	3.30	2.80	4.00	3.80	63.00
A72	3.30	3.30	3.50	3.30	3.00	2.80	4.50	4.00	62.73
F5	3.00	3.30	3.00	2.80	3.30	2.80	4.00	3.50	61.07
1D3	3.50	3.00	3.50	3.00	3.00	2.50	4.00	4.00	61.00
A5	3.00	2.80	2.80	3.00	2.50	2.30	4.00	3.00	60.93
H92	2.90	3.10	3.10	3.20	2.70	2.80	3.80	3.70	60.83
2F2	3.00	3.00	3.00	3.50	3.00	2.50	3.80	3.50	59.87
2B1	2.80	2.80	2.80	3.20	3.10	2.80	3.30	3.20	59.75
C7	3.30	3.30	3.00	2.50	3.30	2.80	3.00	3.50	59.73
A71	3.30	2.50	3.00	2.50	2.80	2.80	4.00	4.00	59.73
F92	3.00	2.80	3.00	2.80	2.80	2.50	4.00	3.50	59.73
2B2	2.50	2.80	2.80	2.80	2.80	3.00	4.00	4.00	59.33
1F9	2.50	2.50	2.50	2.50	2.50	2.50	4.00	4.00	58.80
2E5	3.00	2.50	3.00	3.00	3.00	3.50	4.00	4.00	57.00
2C11	3.00	3.50	3.00	3.00	3.00	2.50	3.00	3.00	57.30
1C1	2.50	2.50	3.00	3.00	3.00	2.50	4.00	3.00	56.70
B11	2.80	2.80	2.50	3.00	2.80	1.50	4.00	3.30	56.00
F91	3.00	2.80	2.50	2.50	3.00	1.50	3.50	3.00	55.47
C42	3.00	2.80	2.50	2.80	2.80	2.80	3.30	3.30	55.07

类胡萝卜素降解产物含量Content of carotenoid degradation product/（μg·g^-1）	对照CK	C11	C31	H4
柑橘酮3,7-Nonadien-2-one, 4,8-dimethyl	-	-	-	0.05
香叶基丙酮Geranyl acetone	0.60	1.04	0.97	0.96
氧化紫罗兰醇（+）-3-Oxo-alpha-ionol	-	0.14	-	0.08
巨豆三烯酮Megastigmatrienone	0.70	0.91	0.27	0.50
橙化基丙酮Neryl acetone	0.60	-	0.79	0.96
六氢假紫罗兰酮2-Undecanone, 6,10-dimethyl	0.06	0.09	0.13	0.09
二氢猕猴桃内酯Dihydroactinidiolide	0.45	0.66	0.64	0.53
4-氧化异佛尔酮4-Oxoisophorone	0.01	0.03	-	0.04
法尼基丙酮Farnesyl acetone	0.38	0.83	0.98	0.68
甲基庚烯酮5-Hepten-2-one, 6-methyl	0.03	0.09	0.18	0.19
β-二氢紫罗兰酮Dihydro-beta-ionone	0.06	-	-	0.05
总量Total	2.88	3.70	3.96	4.13

类胡萝卜素降解产物含量Content of carotenoid degradation product/（μg·g^-1）	对照CK	C11	C31	H4
柑橘酮3,7-Nonadien-2-one, 4,8-dimethyl	-	-	-	0.05
香叶基丙酮Geranyl acetone	0.60	1.04	0.97	0.96
氧化紫罗兰醇（+）-3-Oxo-alpha-ionol	-	0.14	-	0.08
巨豆三烯酮Megastigmatrienone	0.70	0.91	0.27	0.50
橙化基丙酮Neryl acetone	0.60	-	0.79	0.96
六氢假紫罗兰酮2-Undecanone, 6,10-dimethyl	0.06	0.09	0.13	0.09
二氢猕猴桃内酯Dihydroactinidiolide	0.45	0.66	0.64	0.53
4-氧化异佛尔酮4-Oxoisophorone	0.01	0.03	-	0.04
法尼基丙酮Farnesyl acetone	0.38	0.83	0.98	0.68
甲基庚烯酮5-Hepten-2-one, 6-methyl	0.03	0.09	0.18	0.19
β-二氢紫罗兰酮Dihydro-beta-ionone	0.06	-	-	0.05
总量Total	2.88	3.70	3.96	4.13

类胡萝卜素降解菌株的原位筛选及其在雪茄提质增香中的应用

In Situ Screening of Carotenoid Degrading Strains and the Application in Improving Quality and Aroma of Cigar

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