稻花香馫香型白酒酒曲酶活测定与高通量测序分析

Enzyme Activity Analysis and High-Throughput Sequencing of Daqu of Daohuaxiang Xinxiang Baijiu

以稻花香馫香型白酒生产中所用到的中高温大曲(ZGW),高温大曲(GW)和根霉曲(GM)为例,测定了它们的糖化酶、液化酶、酯化酶和酸性蛋白酶活力;检测了可培养微生物种类及数量(通过稀释平板涂布法);分析了微生物群落结构组成(通过高通量测序)。酶活测定结果表明:中高温大曲(ZGW)的糖化力最高,为189.5 U/g;根霉曲(GM)的液化力、酯化力和酸性蛋白酶活力最高,分别为16.36 U/g、164.69 mg/g和189.32 U/g。传统稀释涂平板结果发现,霉菌、酵母菌、总细菌和芽孢杆菌均以中高温大曲为最高,分别达到9.07×10^(2)cfu/g、6.02×10^(4)cfu/g、1.90×10^(7)cfu/g和1.74×10^(7)cfu/g。高通量测序结果表明,中高温大曲细菌和真菌的物种多样性最高,而根霉曲的最低。在细菌属水平上,高温大曲的优势菌种有Bacillus(芽孢杆菌属),Kroppenstedtia,Thermoactinomyces(高温放线菌属),Saccharopolyspora(糖多孢菌属),Norank_f_Pseudonocardiaceae,Planifilum,Staphylococcus(葡萄球菌属)和Pediococcus(片球菌属);中高温大曲的优势菌种有Bacillus(芽孢杆菌属),Kroppenstedtia,Thermoactinomyces(高温放线菌属),Weissella(魏斯氏菌属),Lactobacillus(乳杆菌属),Pediococcus(片球菌属),Planifilum和Staphylococcus(葡萄球菌属),其中Bacillus(芽孢杆菌属)丰度最高。在真菌属水平上,高温大曲的优势菌种有Thermoascus(嗜热子囊菌属),Thermomyces(嗜热真菌属)和Aspergillus(曲霉菌属),其中Thermomyces(嗜热真菌属)丰度最高;中高温大曲的优势菌种有Thermoascus(嗜热子囊菌属),Thermomyces(嗜热真菌属),Cutaneotrichosporon,Candida(假丝酵母属),Rhizopus(根霉菌属),Aspergillus(曲霉菌属),Hyphopichia,Trichomonascus和Xeromyces(耐干霉菌属),其中丰度较高的是Thermoascus(嗜热子囊菌属)。而Rhizopus(根霉菌属)作为根霉曲中唯一的优势菌种,丰度可达99.99%。

Taking the medium-temperature Daqu,high-temperature Daqu and Rhizopus Xiaoqu used in the production of Daohuaxiang Xinxiang Baijiu as the research objects,we determined the activities of the saccharifying enzyme,liquefying enzyme,esterifying enzyme and acid protease,detected the species and quantity of cultivable microorganisms by dilution-spread plate method,and ana-lyzed the composition of microbial communities by HTS.The results showed that the saccharifying power of the medium-temperature Daqu was the highest(189.5 U/g);the liquefying power,esterifying power and acid protease activity of the Rhizopus Xiaoqu were the highest(16.36 U/g,164.69 mg/g and 189.32 U/g);the number of mould,yeast,bacteria and bacillus strains in the medium-temperature Daqu was the highest(9.07×10^(2)cfu/g,6.02×10^(4)cfu/g,1.90×10^(7)cfu/g and 1.74×10^(7)cfu/g);the species diversity of bacteria and fungi of the medium-temperature Daqu was the highest,while that of the Rhizopus Xiaoqu was the lowest.The dominant species of the high-temperature Daqu were Bacillus,Kroppenstedtia,Thermoactinomyces,Saccharopolyspora,Norank_f_Pseudonocardiaceae,Planifilum,Staphylococcus,Pediococcus,Thermoascus,Thermomyces and Aspergillus,among which Thermomyces was the most abundant.The dominant species of the medium-temperature Daqu were Bacillus,Kroppenstedtia,Thermoactinomyces,Weissella,Lactobacillus,Pediococcus,Planifilum,Staphylococcus,Thermoascus,Thermomyces,Curaneotrichosporon,Candida,Rhizopus,Aspergillus,Hyphopichia,Trichomonascus and Xeromyces,among which Bacillus and Thermoascus were the most abundant.The only dominant species of the Rhizopus Xiaoqu was Rhizopus,the abundance of which reached 99.99%.