Isolation，Identification，and Functional Analysis of Culturable Microorganisms in Cigar Tobacco Leaves from Various Regions

doi:10.15933/j.cnki.1004-3268.2024.09.004

Abstract

Abstract: To clarify the species and functionality of culturable microorganisms in domestic cigar tobacco leaves,cured cigar tobacco leaves from Wuzhishan,Hainan Province(variety Haiyan 103)and and from Dazhou，Sichuan Province(variety Chuanxue No.1) were used as experimental materials.The culture‑dependent method were employed to isolate microorganisms,which were subsequently identified through molecular sequencing.The enzyme and aromatic substances production abilities of the isolated microorganisms were investigated.The results showed the isolation of 52 different bacterial strains from Haiyan 103 and Chuanxue No.1 cigar leaves,belonging to five different families:Bacillaceae,Planococcaceae,Erwiniaceae,Enterobacteriales,and Moraxellaceae.Bacillus subtilis E‑11,isolated from Hainan tobacco,exhibited the highest protease and pectinase activities of 85.02 U/mL and 256.71 U/mL,respectively.Additionally,four yeast strains were isolated from Hainan tobacco leaves,namely one strain from Pichia,one strain from Rhodotorula,and two strains from Debaryomyces.These yeast strains did not produce extracellular protease and pectinase,and Pichia sp.J‑2 demonstrated the ability to metabolize 2‑phenylethanol.The highest yield of 2‑phenylethanol,333.81 mg/L,was observed after 72 h of fermentation.In conclusion,cigar tobacco contains a rich variety of culturable microbial resources.Bacillus subtilis E‑11 and Pichia sp.J‑2,isolated from Haiyan 103,show promising potential for degrading macromolecular substances and enhancing the aroma content in tobacco leaves.

Key words: Cigar tobacco leaf, Bacteria, Yeast, Isolation, Identification, Functional analysis

摘要： 为了明确我国雪茄烟叶叶际可培养微生物种类及功能，以海南省五指山市海烟103和四川省达州市川雪一号经晾制后的雪茄烟叶为试验材料，通过传统的分离纯化方法对雪茄烟叶叶际可培养微生物进行分离，并进行分子鉴定；同时，对所分离微生物的产酶和产香能力进行研究。结果表明，从海烟103和川雪一号雪茄烟叶中共分离到52株不同的细菌，这些细菌来自5个不同的科，分别是芽孢杆菌科（Bacillaceae）、动球菌科（Planococcaceae）、欧文菌科（Erwiniaceae）、肠杆菌科（Enterobacteriaceae）和莫拉菌科（Moraxellaceae）。两地雪茄烟叶中的优势菌均为芽孢杆菌科细菌，其中，从海南雪茄烟叶中分离的枯草芽孢杆菌E-11的蛋白酶和果胶酶活性最高，分别为85.02 U/mL和256.71 U/mL。此外，还分离到4株酵母菌，均来自海南烟叶，包括1株毕赤酵母属（Pichia）、1株胶红酵母属（Rhodotorula）、2株德巴利酵母属（Debaryomyces），这些酵母菌不具备产生胞外蛋白酶、果胶酶能力，分离到的毕赤酵母J-2可以代谢产生具有玫瑰花香的2-苯乙醇，在发酵72 h时2-苯乙醇产量最高，为333.81 mg/L。综上，雪茄烟叶中含有丰富的可培养微生物资源，从海烟103中分离的枯草芽孢杆菌E-11和毕赤酵母J-2在降解烟叶大分子物质、增加烟叶香气物质含量方面具有较好的应用前景。

关键词: 雪茄烟叶, 细菌, 酵母菌, 分离, 鉴定, 功能分析

CLC Number:

S572

SI Hongyang, YANG Weili, CUI Bing, LAI Miao, ZHAO Mingqin. Isolation，Identification，and Functional Analysis of Culturable Microorganisms in Cigar Tobacco Leaves from Various Regions[J]. Journal of Henan Agricultural Sciences, 2024, 53(9): 37-45.

司洪阳, 阳苇丽, 崔冰, 来苗, 赵铭钦. 不同产地雪茄烟叶叶际可培养微生物分离、鉴定及功能研究[J]. 河南农业科学, 2024, 53(9): 37-45.

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