高温大曲中霉菌的筛选及其挥发性产物分析
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摘要
本研究以本地酒厂高温大曲为研究对象,从中分离得到1株功能性霉菌,通过形态学、生理生化及分子生物学鉴定,确定为黑曲霉(Aspergillus niger)。同时对该株霉菌进行固态发酵试验,并利用固相微萃取和气质联用对该菌株发酵产物的挥发性物质进行分析。结果表明,该菌对固态发酵产物贡献香味成分最多的物质是酚类、醇类和呋喃类,其中2-甲氧基苯酚、4-乙烯基愈创木酚、仲丁醇的含量最高,依次为17.21%、14.88%、5.63%。另外,对该菌的产酶特性进行了研究,发现其产蛋白酶的活力最高,是高温大曲微生物组成的重要成员之一。
In this study, one functional mold strain was isolated from local high-temperature Daqu. The strain was identified as Aspergillus niger through morphological, physiological, physiochemical, and molecular biological analysis. Meanwhile, the strain was used in solid-state fermentation experiments. And the volatile fermenting products were analyzed by solid-phase microextraction coupled with GC-MS. The results showed that phenolic substances, alcohols and furans were the most important volatile fermenting products,among which the content of 2-methoxyphenol, 2-methoxy-4-vinylphenol and 2-butanol was the highest, accounting for 17.21 %,14.88 %, and 5.63 % respectively. In addition, the enzyme-producing properties of the strain were studied. It was found that the strain had strong protease-producing capabilities and the strain was one of the most important microbial strains in high-temperature Daqu.
In this study, one functional mold strain was isolated from local high-temperature Daqu. The strain was identified as Aspergillus niger through morphological, physiological, physiochemical, and molecular biological analysis. Meanwhile, the strain was used in solid-state fermentation experiments. And the volatile fermenting products were analyzed by solid-phase microextraction coupled with GC-MS. The results showed that phenolic substances, alcohols and furans were the most important volatile fermenting products,among which the content of 2-methoxyphenol, 2-methoxy-4-vinylphenol and 2-butanol was the highest, accounting for 17.21 %,14.88 %, and 5.63 % respectively. In addition, the enzyme-producing properties of the strain were studied. It was found that the strain had strong protease-producing capabilities and the strain was one of the most important microbial strains in high-temperature Daqu.
引文
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[16]莫新良.黄酒储存期间4-乙烯基愈创木酚和香草醛的变化及影响因素[J].食品与发酵工业,2016,42(2):29-33.
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[18]范文来.白酒79个风味化合物嗅觉阈值测定[J].酿酒,2011,38(4):80-84
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[20]候兆乾.不同原料组成对甜高粱秸秆白酒风味物质种类及相对含量的影响[D].呼和浩特:内蒙古农业大学,2017.
[21]FAN W L,XU Y,QIAN M C.Identification of aroma compounds in Chinese“Moutai”and“Langjiu”liquors by normal phase liquid chromatography fractionation followed by gas chromatography/olfactometry[M]//QIAN M C,SHELLHAMMER T H.Flavor chemistry of wine and other alcoholic beverages.Washington,DC:American Chemical Society,2012:303-338.
[22]徐丹,孙秀兰,李永仙,等.黑曲霉对黄曲霉生长、产毒及黄曲霉毒素B1的影响[J].中国微生态学杂志,2011,23(6):490-499.
[23]沈怡方.白酒生产技术全书[M].北京:中国轻工业出版社,2013:105-121.
[24]陈笔.酱香型白酒酿造过程中霉菌群落结构以及霉菌与酵母相互作用的研究[D].无锡:江南大学,2014.
[25]SHI S,ZHANG L,WU Z Y,et al.Analysis of the fungi community in multiple-and single-grains Zaopei from a Luzhou-flavor liquor distillery in western China[J].World j microbiol biotechnol,2011,27:1869-1874.
[26]徐佳.酱香型白酒酿造过程中霉菌的功能性研究[J].酿酒,2015,42(5):32-37.
[27]常吕珍,杨云娟,张润美,等.黑曲霉F5产高温α-糖化酶酶学性质及产酒精能力研究[J].酿酒科技,2016(10):30-33.
[28]龙茜萍,王晓丹,谭静,等.一株高产糖化酶菌株的筛选与鉴定[J].酿酒科技,2013(8):7-9.
[29]黄永光.酱香型白酒酿造中Aspergillus hennebergii及其分泌酸性蛋白酶的研究[D].无锡:江南大学,2014.
[2]王晓丹,徐佳.酱香型大曲中分离到的阿姆斯特丹散囊菌产酶产香特性[J].食品科学,2016,37(11):154-159.
[3]张荣.产酱香功能细菌的筛选及其特征风味化合物的研究[D].无锡:江南大学,2009.
[4]班世栋,王晓丹,陈孟强,等.酱香型大曲中具产酶功能霉菌的分离筛选[J].酿酒,2014,41(4):31-35.
[5]张文学,乔宗伟,胡承,等.浓香型白酒糟醅中真菌菌群的多样性分析[J].四川大学学报:工程科学版,2006,38(5):97-101.
[6]蒋英丽,卓毓崇,聂正东,等.高温大曲中产果香霉菌的分离及研究[J].酿酒科技,2015(2):21-24.
[7]沈玉洁.高温大曲中嗜热功能菌的筛选及应用性能研究[D].武汉:湖北工业大学,2011.
[8]中国科学院微生物研究所.常见与常用真菌[M].北京:科学出版社,1973.
[9]邓皖玉,王会,赵荣寿,等.高温大曲中产酱味霉菌的分离及研究[J].酿酒科技,2015(3):9-11.
[10]赵建新,顾小红,刘杨岷,等.传统豆酱挥发性风味化合物的研究[J].食品科学,2006,27(12):684-687.
[11]黄红霞,孟鸳,康旭,等.顶空固相微萃取-气质联用技术分析传统豆豉中的挥发性成分[J].农产品加工(学刊),2010(9):25-29.
[12]宋昊,郑玉芝.黄豆豆渣发酵产物中挥发性风味化合物成分分析[J].食品科学,2016,37(10):176-182.
[13]班世栋.酱香大曲中霉菌类群和酶系研究[D].贵阳:贵州大学,2015.
[14]WANG C,SHI D,GONG G.Microorganisms in Daqu:a starter culture of Chinese Maotai-flavor liquor[J].World journal of microbiology and biotechnology,2008,24(10):2183-2190.
[15]黄蕴利,黄永光,郭旭.白酒中的主要生物活性功能成分研究进展[J].食品工业科技,2016,37(15):375-379.
[16]莫新良.黄酒储存期间4-乙烯基愈创木酚和香草醛的变化及影响因素[J].食品与发酵工业,2016,42(2):29-33.
[17]杨玉波.高粱蒸煮风味物质香气活力研究[J].食品与发酵工业,2018,44(5):222-226.
[18]范文来.白酒79个风味化合物嗅觉阈值测定[J].酿酒,2011,38(4):80-84
[19]许佩勤.白酒中吡嗪、呋喃类化合物的分析方法研究[J].食品与发酵科技,2017,53(2):85-93.
[20]候兆乾.不同原料组成对甜高粱秸秆白酒风味物质种类及相对含量的影响[D].呼和浩特:内蒙古农业大学,2017.
[21]FAN W L,XU Y,QIAN M C.Identification of aroma compounds in Chinese“Moutai”and“Langjiu”liquors by normal phase liquid chromatography fractionation followed by gas chromatography/olfactometry[M]//QIAN M C,SHELLHAMMER T H.Flavor chemistry of wine and other alcoholic beverages.Washington,DC:American Chemical Society,2012:303-338.
[22]徐丹,孙秀兰,李永仙,等.黑曲霉对黄曲霉生长、产毒及黄曲霉毒素B1的影响[J].中国微生态学杂志,2011,23(6):490-499.
[23]沈怡方.白酒生产技术全书[M].北京:中国轻工业出版社,2013:105-121.
[24]陈笔.酱香型白酒酿造过程中霉菌群落结构以及霉菌与酵母相互作用的研究[D].无锡:江南大学,2014.
[25]SHI S,ZHANG L,WU Z Y,et al.Analysis of the fungi community in multiple-and single-grains Zaopei from a Luzhou-flavor liquor distillery in western China[J].World j microbiol biotechnol,2011,27:1869-1874.
[26]徐佳.酱香型白酒酿造过程中霉菌的功能性研究[J].酿酒,2015,42(5):32-37.
[27]常吕珍,杨云娟,张润美,等.黑曲霉F5产高温α-糖化酶酶学性质及产酒精能力研究[J].酿酒科技,2016(10):30-33.
[28]龙茜萍,王晓丹,谭静,等.一株高产糖化酶菌株的筛选与鉴定[J].酿酒科技,2013(8):7-9.
[29]黄永光.酱香型白酒酿造中Aspergillus hennebergii及其分泌酸性蛋白酶的研究[D].无锡:江南大学,2014.