基于HS-SPME-GC-MS的浓香型白酒窖泥中可培养Clostridium spp.挥发性代谢物成分分析
Component analysis of volatile metabolites of culturable Clostridium spp. isolated from pit muds of Luzhou-flavor liquor based on HS-SPME-GC-MS
-
摘要: 对某酒企窖泥进行分离纯化培养，得到88株细菌纯培养物，通过16S rRNA基因序列比对和系统发育树分析，其中75株细菌被分为8个系统发育簇（cluster Ⅰ—Ⅷ），其最高相似菌分别为C. tyrobutyricum，C. butyricum，C. indolis，C. acetobutylicum，C. bifermentans，C. sporogenes，C. cadaveris和C. kluyveri.采用顶空固相微萃取-气相色谱-质谱联用法（HS-SPME-GC-MS）对8个代表性梭菌菌株的挥发性代谢物进行检测，共检测出75种挥发性物质，其中醇类23种、酸类20种、酯类14、醛类2种、酮类2种和其他挥发性物质14种.通过主成分分析发现，菌株RL1（C. tyrobutyricum）和EL7（C. butyricum）主要与乙酸和丁酸等化合物的形成有关；EL46（C. kluyveri）、RL20（C. acetobutylicum）和RL10（C. indolis）主要与己酸和己酸乙酯等化合物的形成有关；EL12（C. bifermentans）和RL23（C. sporogenes）主要与硫代醋酸S-甲酯、1-癸醇和正辛醇有关；而EL43（C. cadaveris）主要与多种酯类化合物的合成有关.
-
关键词:
- 浓香型白酒 /
- 窖泥 /
- 梭菌 /
- 顶空固相微萃取-气质联用 /
- 挥发性代谢产物
Abstract: A total of 88 pure cultures was obtained from the pit muds of one wine corporation following separation and purification culture. The 75 strains of bacteria were assigned into 8 phylogenetic clusters (cluster Ⅰ-Ⅷ) based on the analysis of 16S rRNA gene sequences and phylogenetic tree. In them, the most similar bacteria were C. tyrobutyricum, C. butyricum, C. indolis, C. acetobutylicum, C. bifermentans, C. sporogenes, C. cadaveris and C. kluyveri, respectively. The volatile metabolites of 8 representative Clostridium spp. were analyzed using headspace solid-phase microextraction (HS-SPME) method followed by gas chromatography-mass spectrometry (GC-MS). A total of 75 volatile metabolites was detected, including 23 alcohols,20 acids,14 esters,2 aldehydes,2 ketones and other 14 volatile substances. The principal component analysis revealed that the representative strain RL1 (C. tyrobutyricum) and EL7 (C. butyricum) were mainly related to the generation of acetic acid and butyric acid, and EL46 (C. kluyveri), RL20 (C. acetobutylicum) and RL10 (C. indolis) mainly contributed to the formation of caproic acid and ethyl hexanoate, and EL12 (C. bifermentans) and RL23 (C. sporogenes) were mainly associated with the S-methyl acetate, 1-Decyl alcohol and octanol, and EL43 (C. cadaveris) was mainly correlated to the biosynthesis of various esters.-
Key words:
- Luzhou-flavor liquor /
- pit muds /
- Clostridium spp. /
- HS-SPME-GC-MS /
- volatile metabolite
-
-
[1]
FAN W L,QIAN M C.Characterization of aroma compounds of Chinese "Wuliangye" and "Jiannanchun" liquors by aroma extract dilution analysis[J].Journal of Agricultural and Food Chemistry,2006,54(7):2695.
-
[2] ZHENG J,LIANG R,WU C,et al.Discrimination of different kinds of Luzhou-flavor raw liquors based on their volatile features[J].Food Research International,2013,56:77.
-
[3] FAN W,QIAN M C.Identification of aroma compounds in Chinese "Yanghe Daqu" liquor by normal phase chromatography fractionation followed by gas chromatography/olfactometry[J].Flavour and Fragrance Journal,2006,21(2):333.
-
[4] HU X L,DU H,XU Y.Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production[J].International Journal of Food Microbiology,2015,214:116.
-
[5] ZHENG X W,TABRIZI M R,NOUT M J R,et al.Daqu-A traditional Chinese liquor fermentation starter[J].Journal of the Institute of Brewing,2011,117(1):82.
-
[6] HU X,DU H,REN C,et al.Illuminating anaerobic microbial community and cooccurrence patterns across a quality gradient in Chinese liquor fermentation pit muds[J].Applied and Environmental Microbiology,2016,82(8):2506.
-
[7] TAO Y,LI J,RUI J,et al.Prokaryotic communities in pit mud from different-aged cellars used for the production of Chinese strong-flavored liquor[J].Applied and Environmental Microbiology,2014,80(7):2254.
-
[8] SJÖBLOM M,MATSAKAS L,CHRISTAKOPOULOS P,et al.Production of butyric acid by Clostridium tyrobutyricum,(ATCC25755) using sweet sorghum stalks and beet molasses[J].Industrial Crops and Products,2015,74:535.
-
[9] CAI G,JIN B,SAINT C,et al.Genetic manipulation of butyrate formation pathways in Clostridium butyricum[J].Journal of Biotechnology,2011,155(3):269.
-
[10]
[10] LI H G,LI W,GU Q Y,et al.Acetone,butanol,and ethanol production from cane molasses using Clostridium beijerinckii mutant obtained by combined low-energy ion beam implantation and N-methyl-N-nitro-N-nitrosoguanidine induction[J].Bioresource Technology,2013,137:254.
-
[11]
[11] ZHENG J,LIANG R,ZHANG L,et al.Characterization of microbial communities in strong aromatic liquor fermentation pit muds of different ages assessed by combined DGGE and PLFA analyses[J].Food Research International,2013,54(1):660.
-
[12]
[12] ELSDEN S R,HILTON M G,WALLER J M.The end products of the metabolism of aromatic amino acids by Clostridia[J].Archives of Microbiology,1976,107(3):283.
-
[13]
[15] WANG M Y,ZHANG W X.Analysis of microbial community structure in pit mud from two Chinese Luzhou-flavor liquor producing areas[J].Microbiology China,2014,41(8):1498.
-
[14]
[16] WANG C,CHEN Q,WANG Q,et al.Long-term batch brewing accumulates adaptive microbes,which comprehensively produce more flavorful Chinese liquors[J].Food Research International,2014,62:894.
-
[15]
[17] HU X L,WANG H Y,WU Q,et al.Development,validation and application of specific primers for analyzing the clostridial diversity in dark fermentation pit mud by PCR-DGGE[J].Bioresource Technology,2014,163:40.
-
[16]
[18] STACKEBRANDT E,EBERS J.Taxonomic parameters revisited:tarnished gold standards[J].Microbiology Today,2006,33(4):152.
-
[17]
[19] LIU C,HUANG D,LIU L,et al.Clostridium swellfunianum sp.nov.,a novel anaerobic bacterium isolated from the pit mud of Chinese Luzhou-flavor liquor production[J].Antonie Van Leeuwenhoek,2014,106(4):817.
-
[1]
-
点击查看大图
计量
- PDF下载量: 94
- 文章访问数: 8808
- 引证文献数: 0
下载:
