传统鱼露发酵过程中细菌群落演替及对其挥发性风味形成的影响分析
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摘要
采用16S rRNA高通量测序技术研究传统鱼露发酵过程细菌群落演替规律。结果显示,鱼露细菌种类随着发酵时间的延长逐渐上升,在发酵时间6个月达到最大(312种),随着发酵时间延长至12个月,菌群多样性有所下降但菌群间差异性变小。厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)是鱼露整个发酵过程中最主要的门。盐厌氧菌属(Haloanaerobium)是鱼露发酵过程中最主要的属,随着发酵时间的延长其相对丰度先上升而后下降。与发酵初期(0个月)相比,发酵中后期发光杆菌属(Photobacterium)、四联球菌属(Tetragenococcus)和盐单胞菌属(Halomonas)等微生物菌群的相对丰度增加最为显著;发酵末期(12个月),Halomonas成为优势菌群。采用气相色谱-质谱联用技术研究鱼露发酵过程中的挥发性风味成分,共检测出醛、醇、酮、酯、烃、酸、含氮化合物7大类共54种挥发性化合物,并根据气味活性值(≥1)筛选到9种主体呈香风味化合物。Pearson相关性分析显示Halanaerobium和Halomonas与多种主体挥发性风味化合物的变化呈现一定的相关性。结果表明,细菌群落结构对鱼露特殊的风味形成具有重要影响。
In this study, 16 S rRNA high-throughput sequencing was used to study the bacterial community succession in fish sauce during its fermentation process. The results showed that number of bacterial species in fish sauce increased first, peaking(312 species) at month 6, and then decreased with prolonged fermentation time. Less abundant bacterial diversity and a smaller difference among the bacterial communities were observed from month 6 to 12 of fermentation. Firmicutes and Proteobacteria were the most dominant phyla during the whole fermentation process. Haloanaerobium was the most predominant genus, and its relative abundance increased first and then decreased with increasing fermentation time. The relative abundances of Photobacterium, Tetragenococcus and Halomonas were significantly increased during the mid to late fermentation stages compared with those at month 0. At the end of the 12-month fermentation period, Halomonas became the dominant genus. Gas chromatography-mass spectrometry(GC-MS) was used to study the volatile flavor components in fish sauce during the fermentation process. A total of 54 volatile flavor components including aldehydes, alcohols, ketones, esters, hydrocarbons, acids and nitrogen compounds were detected, and 9 key volatile flavor compounds were identified according to their odour activity value(OAV)( ≥ 1). Pearson's correlation analysis showed that Halanaerobium and Halomonas had a significant correlation with the changes in major volatile compounds. These results suggest that the bacterial community structure has an important influence on the formation of volatile flavor components in Chinese fish sauce.
In this study, 16 S rRNA high-throughput sequencing was used to study the bacterial community succession in fish sauce during its fermentation process. The results showed that number of bacterial species in fish sauce increased first, peaking(312 species) at month 6, and then decreased with prolonged fermentation time. Less abundant bacterial diversity and a smaller difference among the bacterial communities were observed from month 6 to 12 of fermentation. Firmicutes and Proteobacteria were the most dominant phyla during the whole fermentation process. Haloanaerobium was the most predominant genus, and its relative abundance increased first and then decreased with increasing fermentation time. The relative abundances of Photobacterium, Tetragenococcus and Halomonas were significantly increased during the mid to late fermentation stages compared with those at month 0. At the end of the 12-month fermentation period, Halomonas became the dominant genus. Gas chromatography-mass spectrometry(GC-MS) was used to study the volatile flavor components in fish sauce during the fermentation process. A total of 54 volatile flavor components including aldehydes, alcohols, ketones, esters, hydrocarbons, acids and nitrogen compounds were detected, and 9 key volatile flavor compounds were identified according to their odour activity value(OAV)( ≥ 1). Pearson's correlation analysis showed that Halanaerobium and Halomonas had a significant correlation with the changes in major volatile compounds. These results suggest that the bacterial community structure has an important influence on the formation of volatile flavor components in Chinese fish sauce.
引文
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[22]PEREZ-PALACIOS T,PETISCA C,MELO A,et al.Quantification of furanic compounds in coated deep-fried products simulating normal preparation and consumption:optimisation of HS-SPME analytical conditions by response surface methodology[J].Food Chemistry,2012,135(3):1337-1343.DOI:10.1016/j.foodchem.2012.05.100.
[23]SIDIRA M,KANDYLIS P,KANELLAKI M,et al.Effect of immobilized Lactobacillus casei on the evolution of flavor compounds in probiotic dry-fermented sausages during ripening[J].Meat Science,2015,100:41-51.DOI:10.1016/j.meatsci.2014.09.011.
[24]IVANOVA-PETROPULOS V,MITREV S,STAFILOV T,et al.Characterisation of traditional Macedonian edible oils by their fatty acid composition and their volatile compounds[J].Food Research International,2015,77(3):506-514.DOI:10.1016/j.foodres.2015.08.014.
[25]冯云子.高盐稀态酱油关键香气物质的变化规律及形成机理的研究[D].广州:华南理工大学,2015:131-132.
[26]SUN S Y,JIANG W G,ZHAO Y P.Profile of volatile compounds in 12 Chinese soy sauces produced by a high-salt-diluted state fermentation[J].Journal of the Institute of Brewing,2010,116(3):316-328.DOI:10.1002/j.2050-0416.2010.tb00437.x.
[27]高先楚,王锡昌,顾赛麒,等.中华绒螯蟹性腺加热熟制前后挥发性成分和脂肪酸组成分析[J].现代食品科技,2014,30(9):265-274.DOI:10.13982/j.mfst.1673-9078.2014.09.044.
[28]顾赛麒,王锡昌,陶宁萍,等.顶空固相微萃取-气质联用及电子鼻技术检测中华绒螯蟹不同可食部位中的香气成分[J].食品科学,2013,34(18):239-244.DOI:10.7506/spkx1002-6630-201318049.
[29]FUKAMI K,ISHIYAMA S,YAGURAMAKI H,et al.Identification of distinctive volatile compounds in fish sauce[J].Journal of Agricultural and Food Chemistry,2002,50(19):5412-5416.DOI:10.1021/jf020405y.
[30]STEINHAUS P,SCHIEBERLE P.Characterization of the key aroma compounds in soy sauce using approaches of molecular sensory science[J].Journal of Agricultural and Food Chemistry,2007,55(15):6262-6269.DOI:10.1021/jf0709092.
[31]LEE S,AHN B.Comparison of volatile components in fermented soybean pastes using simultaneous distillation and extraction(SDE)with sensory characterisation[J].Food Chemistry,2009,114(2):600-609.DOI:10.1016/j.foodchem.2008.09.091.
[2]肖宏艳,曾庆孝.潮汕鱼露发酵过程中挥发性风味成分分析[J].中国调味品,2010,35(2):92-96.
[3]SINSUWAN S,RODTONG S,YONGSAWATDIGUL J.Production and characterization of NaCl-activated proteinases from Virgibacillus sp.SK33 isolated from fish sauce fermentation[J].Process Biochemistry,2008,43(2):185-192.DOI:10.1016/j.procbio.2007.11.015.
[4]UCHIDA H,KONDO D,YAMASHITA S,et al.Purification and properties of a protease produced by Bacillus subtilis CN2isolated from a Vietnamese fish sauce[J].World Journal of Microbiology&Biotechnology,2004,20(6):579-582.DOI:10.1023/B:WIBI.0000043168.15067.44.
[5]MONTRIWONG A,RODTONG S,YONGSAWATDIGUL J.Detergent-stable salt-activated proteinases from Virgibacillus halodenitrificans SK1-3-7 isolated from fish sauce fermentation[J].Applied Biochemistry and Biotechnology,2015,176(2):505-517.DOI:10.1007/s12010-015-1591-5.
[6]JIANG W,LI C,XU B,et al.Halomonas shantousis sp nov.,a novel biogenic amines degrading bacterium isolated from Chinese fermented fish sauce[J].Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology,2014,106(6):1073-1080.DOI:10.1007/s10482-014-0275-4.
[7]ZAMAN M Z,BAKAR F A,SELAMAT J,et al.Degradation of histamine by the halotolerant Staphylococcus carnosus FS19isolate obtained from fish sauce[J].Food Control,2014,40:58-63.DOI:10.1016/j.foodcont.2013.11.031.
[8]KIM M J,KWAK H S,JUNG H Y,et al.Microbial communities related to sensory attributes in Korean fermented soy bean paste(doenjang)[J].Food Research International,2016,89:724-732.DOI:10.1016/j.foodres.2016.09.032.
[9]DUAN S,HU X,LI M,et al.Composition and metabolic activities of the bacterial community in shrimp sauce at the flavor-forming stage of fermentation as revealed by metatranscriptome and 16S rRNA gene sequencings[J].Journal of Agricultural and Food Chemistry,2016,64(12):2591-2603.DOI:10.1021/acs.jafc.5b05826.
[10]LV X G,JIA R B,LI Y,et al.Characterization of the dominant bacterial communities of traditional fermentation starters for Hong Qu glutinous rice wine by means of MALDI-TOF mass spectrometry fingerprinting,16S rRNA gene sequencing and speciesspecific PCRs[J].Food Control,2016,67:292-302.DOI:10.1016/j.foodcont.2016.03.005.
[11]焦晶凯,莫蓓红.Illumina MiSeq平台高覆盖率测定干酪中的细菌微生物多样性[J].中国酿造,2014,33(5):34-38.DOI:10.11882/j.issn.0254-5071.2014.05.009.
[12]吴燕燕,钱茜茜,李来好,等.基于Illumina MiSeq技术分析腌干鱼加工过程中微生物群落多样性[J].食品科学,2017,38(12):1-8.DOI:10.7506/spkx1002-6630-201712001.
[13]吴燕燕,王悦齐,李来好,等.基于电子鼻与HS-SPME-GC-MS技术分析不同处理方式腌干带鱼挥发性风味成分[J].水产学报,2016,40(12):1931-1940.DOI:10.11964/jfc.20160410346.
[14]SCHLOSS P D,WESTCOTT S L,RYABIN T,et al.Introducing mothur:open-source,platform-independent,community-supported software for describing and comparing microbial communities[J].Applied and Environmental Microbiology,2009,75(23):7537-7541.DOI:10.1128/AEM.01541-09.
[15]LEE S H,JUNG J Y,JEON C O.Microbial successions and metabolite changes during fermentation of salted shrimp(saeu-jeot)with different salt concentrations[J].PLoS ONE,2014,9(2):e90115.DOI:10.1371/journal.pone.0090115.
[16]KOBAYASHI T,KIMURA B,FUJII T.Strictly anaerobic halophiles isolated from canned Swedish fermented herrings(Surstromming)[J].International Journal of Food Microbiology,2000,54(1/2):81-89.DOI:10.1016/S0168-1605(99)00172-5.
[17]LEE H W,CHOI Y J,HWANG I M.Relationship between chemical characteristics and bacterial community of a Korean salted-fermented anchovy sauce,Myeolchi-Aekjeot[J].LWT-Food Science and Technology,2016,73:251-258.DOI:10.1016/j.lwt.2016.06.007.
[18]NAM Y D,LEE S Y,LIM S I.Microbial community analysis of Korean soybean pastes by next-generation sequencing[J].International Journal of Food Microbiology,2012,155(1/2):36-42.DOI:10.1016/j.ijfoodmicro.2012.01.013.
[19]UDOMSIL N,CHEN S,RODTONG S,et al.Quantification of viable bacterial starter cultures of Virgibacillus sp.and Tetragenococcus halophilus in fish sauce fermentation by real-time quantitative PCR[J].Food Microbiology,2016,57:54-62.DOI:10.1016/j.fm.2016.01.004.
[20]WANG Y,JIANG Y T,CAO J X,et al.Study on lipolysis-oxidation and volatile flavour compounds of dry-cured goose with different curing salt content during production[J].Food Chemistry,2016,190:33-40.DOI:10.1016/j.foodchem.2015.05.048.
[21]WAH T T,WALAISRI S,ASSAVANIG A,et al.Co-culturing of Pichia guilliermondii enhanced volatile flavor compound formation by Zygosaccharomyces rouxii in the model system of Thai soy sauce fermentation[J].International Journal of Food Microbiology,2013,160(3):282-289.DOI:10.1016/j.ijfoodmicro.2012.10.022.
[22]PEREZ-PALACIOS T,PETISCA C,MELO A,et al.Quantification of furanic compounds in coated deep-fried products simulating normal preparation and consumption:optimisation of HS-SPME analytical conditions by response surface methodology[J].Food Chemistry,2012,135(3):1337-1343.DOI:10.1016/j.foodchem.2012.05.100.
[23]SIDIRA M,KANDYLIS P,KANELLAKI M,et al.Effect of immobilized Lactobacillus casei on the evolution of flavor compounds in probiotic dry-fermented sausages during ripening[J].Meat Science,2015,100:41-51.DOI:10.1016/j.meatsci.2014.09.011.
[24]IVANOVA-PETROPULOS V,MITREV S,STAFILOV T,et al.Characterisation of traditional Macedonian edible oils by their fatty acid composition and their volatile compounds[J].Food Research International,2015,77(3):506-514.DOI:10.1016/j.foodres.2015.08.014.
[25]冯云子.高盐稀态酱油关键香气物质的变化规律及形成机理的研究[D].广州:华南理工大学,2015:131-132.
[26]SUN S Y,JIANG W G,ZHAO Y P.Profile of volatile compounds in 12 Chinese soy sauces produced by a high-salt-diluted state fermentation[J].Journal of the Institute of Brewing,2010,116(3):316-328.DOI:10.1002/j.2050-0416.2010.tb00437.x.
[27]高先楚,王锡昌,顾赛麒,等.中华绒螯蟹性腺加热熟制前后挥发性成分和脂肪酸组成分析[J].现代食品科技,2014,30(9):265-274.DOI:10.13982/j.mfst.1673-9078.2014.09.044.
[28]顾赛麒,王锡昌,陶宁萍,等.顶空固相微萃取-气质联用及电子鼻技术检测中华绒螯蟹不同可食部位中的香气成分[J].食品科学,2013,34(18):239-244.DOI:10.7506/spkx1002-6630-201318049.
[29]FUKAMI K,ISHIYAMA S,YAGURAMAKI H,et al.Identification of distinctive volatile compounds in fish sauce[J].Journal of Agricultural and Food Chemistry,2002,50(19):5412-5416.DOI:10.1021/jf020405y.
[30]STEINHAUS P,SCHIEBERLE P.Characterization of the key aroma compounds in soy sauce using approaches of molecular sensory science[J].Journal of Agricultural and Food Chemistry,2007,55(15):6262-6269.DOI:10.1021/jf0709092.
[31]LEE S,AHN B.Comparison of volatile components in fermented soybean pastes using simultaneous distillation and extraction(SDE)with sensory characterisation[J].Food Chemistry,2009,114(2):600-609.DOI:10.1016/j.foodchem.2008.09.091.