黄水芽胞杆菌Bacillus aquiflavi 3H-10分离自浓香型白酒黄水样品,是我国四川省宜宾地区首株酿酒微生物新种菌株。为了深入挖掘其功能特性,利用高通量测序技术测定菌株的全基因组序列,并通过数据库比对、平板透明圈法、福林酚法和GC-MS技术解析该菌株的功能基因、产生物酶和发酵风味物质的能力。研究结果表明,菌株B.aquiflavi 3H-10基因组大小3.62 Mb,G+C含量35.40 mol%,其中预测到3 224个蛋白质编码基因。通过与KEGG、NR和CAZy数据库比对发现,基因组中含有编码22种蛋白酶和多种碳水化合物酶类的基因;B.aquiflavi 3H-10在蛋白酶培养基平板上透明圈较明显,蛋白酶活力可达85 U/mL;在大豆酪蛋白液体培养基37 ℃培养72 h的菌体发酵液中,可检测到56种代谢风味化合物,包括酯类、醇类、芳香族、羰基化合物、杂环化合物等,其中,相对含量较高的物质包括苯甲醛(杏仁香,坚果香)、2,5-二甲基吡嗪(炒芝麻香)、正丁醇(水果香)、3-甲基己醇以及浓香型大曲酒的主体香味物质己酸乙酯(甜香,水果香,窖香,青瓜香)。研究表明,黄水芽胞杆菌B.aquiflavi 3H-10具有显著的产蛋白酶能力,可发酵产生多种风味物质,促进浓香型白酒风味的形成,推测其在白酒发酵过程中具有潜在应用价值。
Bacillus aquiflavi 3H-10 is the first strain of brewing novel species, isolated from yellow water of strong-flavor Chinese Baijiu in Yibin, Sichuan province. In order to explore its function characteristics, the whole genome sequence of B. aquiflavi 3H-10 was determined by high-throughput sequencing. And potential functional genes, biological enzymes and fermentation flavor substances were analyzed with BLAST against sequence database, plate transparent circle method, Folin-Ciocalteu method and GC-MS. The results showed that the genome size of B. aquiflavi 3H-10 was 3.62 MB and the G+C content was 35.40 mol% with 3 224 protein-coding genes predicted. Analysis with KEGG, NR and CAZy databases showed that the genome harbors genes responsible for 22 proteases and various carbohydrate enzymes. Clear transparent circle was observed for B. aquiflavi 3H-10 on the protease medium plate and the protease enzyme activity was 85 U/mL. After cultivation in TSB at 37 ℃ for 72 h, the strain could produce 56 metabolic flavor compounds, including esters, alcohols, aromatics, carbonyl compounds and heterocyclic compounds, etc. Among them, the substances with relatively high contents include benzaldehyde (almond flavor, nut flavor), 2,5-dimethylpyrazine (fried sesame flavor), n-butanol (fruit flavor), 3- methylhexanol and ethyl hexanoate (sweet, fruity, cellar and cucumber), the main flavoring substrate of strong-flavor Chinese Baijiu. Our study demonstrated that B. aquiflavi 3H-10 had significant protease-producing ability and could produce various flavor substances by fermentation, which promoted the formation of flavors in strong-flavor Chinese Baijiu. It was speculated that B. aquiflavi 3H-10 had potential application value in Chinese Baijiu fermentation.
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