造纸法再造烟叶浓缩液中抗逆酵母的分离鉴定与代谢活性研究
Isolation, identification and metabolic activity of stress resistant yeast in the concentrated tobacco waste aqueous extract during the paper-making reconstituted tobacco process
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摘要: 采用稀释离心、富集培养、梯度平板筛选等方法，从造纸法再造烟叶生产过程中的烟草废弃物水提液（TWE）浓缩液中分离获得具有正常代谢活动的菌株，对其进行形态、生理生化特性等鉴定，并将其添加到TWE浓缩液中进行发酵，考查其生长代谢特性和对TWE浓缩液中代谢物组成的影响。结果表明：从TWE浓缩液中筛选获得一株高抗逆性的酵母菌株MC-9A1，其属于拜耳接合酵母（Zygosaccharomyces bailii）；该菌株能在TWE浓缩液中生长并进行正常代谢活动，消耗大量的糖分；菌株MC-9A1在TWE浓缩液中消耗了大量强极性物质和少量中极性物质，并生成了大量弱极性或非极性物质，310多种代谢物发生显著上调或下调，如苯乙酰胺、木糖醇等极显著增加，而熊果苷、D-半乳糖醛酸等显著减少；菌株MC-9A1发酵改善了TWE浓缩液感官效果，使其香气质量增加，杂气、残余下降。Abstract: After dilution centrifugation, enrichment culture, and gradient plate screening, a series of strains that can adapt to concentrated tobacco waste water extract (TWE) and exhibit metabolic activity were isolated from concentrated TWE during the paper-making reconstituted tobacco process. Their morphological, physiological and biochemical characteristics were identified. The isolated strains were added into concentrated TWE for fermentation. The growth and metabolism characteristics and their effects on the composition of metabolites in TWE were investigated. The results showed that a yeast strain MC-9A1 resistant to high stress was isolated from concentrated TWE, which belonged to the species Zygosaccharomyces bailii. The strain MC-9A1 could grow in concentrated TWE and carried out normal metabolic activities, consuming a lot of sugar. At the same time, MC-9A1 consumed a large number of strongly polar substances and a small number of moderately polar substances in concentrated TWE, and generated a large number of weakly polar or non-polar substances. More than 310 metabolites were significantly up-regulated or down-regulated. For example, acetamide and xylitol increased significantly, while arbutin and D-galacturonic acid decreased significantly. The sensory evaluation results showed that the sensory effect was improved by fermentation of MC-9A1 in concentrated TWE, which increased aroma quality, while decreased odor and the residual.
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