Separation and identification of thermoduric bacteria strains in medium/high temperature Daqu and the analysis of the flavor metabolites
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摘要: 目的:以培菌完成的中高温大曲为样品,分离鉴定耐热细菌并对其风味代谢产物进行分析。方法:通过富集培养和稀释涂布分离、形态学观察及16S r DNA序列分析、生长特性分析,并采用顶空固相微萃取-气相色谱-质谱联用技术对其固态和液态发酵产物进行分析。结果:筛选出一株耐热能力较好的细菌NR2,鉴定为地衣芽孢杆菌(Bacillus licheniformis);该细菌培养第4 h开始进入对数生长期,培养24 h达稳定期;液态发酵可以产2,3-丁二酮、3-羟基-2-丁酮、2,3-丁二醇等风味物质,其中3-羟基-2-丁酮为主要代谢产物;固态发酵代谢产物种类要高于液态发酵,主要是2,3,5-三甲基吡嗪、2,3,5,6-四甲基吡嗪等吡嗪类化合物含量增加。结论:曲香的形成及浓香型白酒风味物质的形成与耐热细菌NR2的代谢特性有关。Abstract: Objective: With the medium / high temperature Daqu bacteria through cultivation as examples,separation of thermoduric bacteria and the analysis of the flavor metabolites. Method: Isolated via enrichment culture and dilution coating,the morphological observation and 16 S r DNA sequence analysis,growth characteristics analysis,the solid and liquid fermentation products were analyzed through headspace solid- phase microextraction and gas chromatography- mass spectrometry technology. Result: The thermoduric bacteria strain NR2 was isolated and it was Bacillus licheniformis,the bacteria culture began to enter the logarithmic phase from the 4 h of cultivation and reached the stable phase after 24 h of cultivation,the liquid state fermentation flavor substances,such as 2,3-diacetyl,3- hydroxy-2- butanone and 2,3- butanediol,with 3- hydroxy-2- butanone being the main metabolites. In addition,there were more species of solid state fermentation metabolites that liquid ones.The contents of pyrazine compounds,such as 2,3,5- methyl pyrazine and 2,3,5,6- tetramethyl pyrazine increased.Conclusion: The formation of liquor and highly- flavored liquor flavor substances was closely related to the metabolic characteristics of the thermoduric bacteria.
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Keywords:
- Daqu /
- thermoduric bacteria /
- separation /
- identification /
- metabolites /
- Bacillus licheniformis
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