刘姝琪,郑浩楠,辛园,夏陈,赵江林,郑晓琴,邹亮,向达兵,姜良珍

• 1:成都大学食品与生物工程学院
• 2:四川省农业科学院农产品加工研究所
• 3:农业农村部杂粮加工重点实验室
• 4:成都市农业科技职业学院

摘要(Abstract)：

从8种市售黑茶中分离和鉴定冠突散囊菌，并固态发酵苦荞籽粒。测定发酵前后的营养物质总淀粉、可溶性蛋白、脂肪、还原糖，以及功能成分总黄酮和总多酚的含量，并分析两种重要生物转化酶（苯丙氨酸解氨酶和多酚氧化酶）的活性以及与营养功能成分间的关系，以比较不同冠突散囊菌对苦荞营养功能成分的转化作用。结果表明，分离菌中7株为冠突散囊菌，1株为谢瓦氏曲霉；经冠突散囊菌发酵后，苦荞籽粒营养物质中的总淀粉、可溶性蛋白和脂肪含量皆显著低于发酵前（P<0.05），而还原糖以及功能成分总黄酮和总多酚的含量多数显著高于发酵前（P<0.05）；总黄酮含量较高的为LB和YZ菌株，总多酚含量较高的为JW和YZ菌株。相关性分析显示，发酵苦荞的总黄酮、总多酚分别与苯丙氨酸解氨酶、多酚氧化酶活性呈极显著正相关（P<0.001），而还原糖和总淀粉呈极显著负相关（P<0.001），提示发酵苦荞营养物质的变化与冠突散囊菌的生物转化过程密切相关。本研究为基于益生菌发酵工艺开发新型苦荞产品提供了理论依据。

关键词(KeyWords)： 黑茶;分离鉴定;固态发酵;苦荞;冠突散囊菌;营养成分;功能成分

Abstract:

Keywords:

基金项目(Foundation): 四川省科技厅项目（No.2022NSFSC1706）;; 国家自然科学基金项目（No.32302083）;; 2024年中央“三区”科技人才支持计划（川财教[2024]70号）;; 成都大学教改项目（No.cdsyjg2022047）

作者(Author): 刘姝琪,郑浩楠,辛园,夏陈,赵江林,郑晓琴,邹亮,向达兵,姜良珍

DOI: 10.13386/j.issn1002-0306.2024050324

参考文献(References)：

• [1]温琼英.茯砖茶中优势菌的种名鉴定[J].中国茶叶，1990(6):2-3.[WEN Q Y.Identification of dominant bacterial species in Fuzhuan tea[J].Chinese Tea,1990(6):2-3.]
• [2]HUKBA V,KOLARIK M,KUBATOVA A,et al.Taxonomic revision of Eurotium and transfer of species to Aspergillus[J].Mycologia,2013,105(4):912-937.
• [3]GE Y Y,WANG Y C,LIU Y X,et al.Comparative genomic and transcriptomic analyses of the Fuzhuan brick tea-fermentation fungus Aspergillus cristatus[J].BMC Genomics,2016,17(1):428.
• [4]刘玉倩,杨家干.茯砖茶发花过程菌落及主要内含物质变化[J].蚕桑茶叶通讯，2017(5):30-33.[LIU Y Q,YANG J G.Changes in bacterial colonies and main contents during the flowering process of Fuzhuan tea[J].Sericulture and Tea Communication,2017(5):30-33.]
• [5]温琼英.茯砖茶中主要微生物的研究[J].茶叶通讯，1986(4):19-21.[WEN Q Y.A study on the main microorganisms in Fuzhuan tea[J].Tea Communication,1986(4):19-21.]
• [6]ZHU M Z,LI N,ZHOU F,et al.Microbial bioconversion of the chemical components in dark tea[J].Food Chemistry,2020,312:126043.
• [7]蔡正安,刘素纯,刘仲华,等.茯砖茶中冠突散囊菌纤维素酶的酶学性质研究[J].茶叶科学，2010,30(1):57-62.[CAI Z A,LIU S C,LIU Z H,et al.Study on the enzymatic properties of cellulase from Fusarium oxysporum in Fuzhuan tea[J].Tea Science,2010,30(1):57-62.]
• [8]DU Y,YANG W R,YANG C C,et al.A comprehensive review on microbiome,aromas and flavors,chemical composition,nutrition and future prospects of Fuzhuan brick tea[J].Trends in Food Science&Technology,2022,119:452-466.
• [9]LI Q,JIN Y L,JIANG R G,et al.Dynamic changes in the metabolite profile and taste characteristics of Fu brick tea during the manufacturing process[J].Food Chemistry,2021,344:128576.
• [10]ZHU Y F,CHEN J J,JI X M,et al.Changes of major tea polyphenols and production of four new B-ring fission metabolites of catechins from post-fermented Jing-Wei Fu brick tea[J].Food Chemistry,2015,170:110-117.
• [11]陈然,郝彬秀,田海霞,等.六堡茶真菌分布浅析及金花菌筛选鉴定[J].食品科技，2016,41(4):19-23.[CHEN R,HAO B X,TIAN H X,et al.Analysis of fungal distribution in Liubao tea and screening and identification of golden flower fungi[J].Food Technology,2016,41(4):19-23.]
• [12]JIANG C K,ZENG Z,HUANG Y H,et al.Chemical compositions of Pu'er tea fermented by Eurotium cristatum and their lipidlowering activity[J].LWT-Food Science and Technology,2018,98:204-211.
• [13]LI Q,LIU Z H,HUANG J A,et al.Anti-obesity and hypolipidemic effects of Fuzhuan brick tea water extract in high-fat diet-induced obese rats[J].Journal of the Science of Food and Agriculture,2013,93(6):1310-1316.
• [14]KANG D,SU M,DUAN Y,et al.Eurotium cristatum,a potential probiotic fungus from Fuzhuan brick tea,alleviated obesity in mice by modulating gut microbiota[J].Food Function,2019,10(8):5032-5045.
• [15]KELLER A C,WEIR T L,BROECKLING C D,et al.Antibacterial activity and phytochemical profile of fermented Camellia sinensis (Fuzhuan tea)[J].Food Research International,2013,53(2):945-949.
• [16]LI M Y,WU Y P,ZHONG K,et al.Structural characteristics and hypolipidemic activity of theabrownins from dark tea fermented by single species Eurotium cristatum PW-1[J].Biomolecules,2020,10:204.
• [17]WU X,YAO X S,CHEN Y L,et al.Metabolite profiling,antioxidant and α-glucosidase inhibitory activities of buckwheat processed by solid-state fermentation with Eurotium cristatum YL-1[J].Food Research International,2021,143:110262.
• [18]CHEN Y,WANG Y,CHEN J,et al.Bioprocessing of soybeans (Glycine max L.) by solid-state fermentation with Eurotium cristatum YL-1 improves total phenolic content,isoflavone aglycones,and antioxidant activity[J].RSC Advances,2020,10(29):16928-16941.
• [19]唐梓尧,刘素纯,林辉,等.冠突散囊菌发酵制备燕麦面酱工艺研究[J].中国酿造，2019,38(2):117-121.[TANG Z Y,LIU SC,LIN H,et al.Preparation technology of oat flour paste fermented by Eurotium cristatum[J].China Brewing,2019,38(2):117-121.]
• [20]李粟琳,张翔宇,王洋,等.可发酵三七等中药材的食用菌种筛选和皂苷生物转化产物的分析[J].食品与发酵工业，2017,43(12):164-168.[LI S L,ZHANG X Y,WANG Y,et al.Screening of edible fungi for fermentation of Panax notoginseng Chinese medicinal herbs and analysis of biotransformation products of saponins[J].Food and Fermentation Industries,2017,43(12):164-168.]
• [21]ZOU L,WU D,REN G,et al.Bioactive compounds,health benefits,and industrial applications of tartary buckwheat (Fagopyrum tataricum)[J].Critical Reviews in Food Science and Nutrition,2023,63(5):657-673.
• [22]LUTHAR Z,GOLOB A,GERM M,et al.Tartary buckwheat in human nutrition[J].Plants (Basel,Switzerland),2021,10(4):700.
• [23]石桃雄,郑俊青,黎瑞源,等.苦荞种质资源种子蛋白质及组分含量的变异分析[J].河南农业科学，2021,50(9):25-35.[SHIT X,ZHENG J Q,LI R Y,et al.Analysis of variation in seed protein and component content of tartary buckwheat germplasm resources[J].Henan Agricultural Science,2021,50(9):25-35.]
• [24]姜惠萍,刘万顺,宋福来,等.改良考马斯亮蓝试剂法测定壳聚糖及其衍生物中蛋白质含量[J].现代食品，2022,28(17):173-178.[JIANG H P,LIU W S,SONG F L,et al.Improved Coomassie brilliant blue reagent method for determining protein content in chitosan and its derivatives[J].Modern Food,2022,28(17):173-178.]
• [25]索化夷,赵欣,骞宇,等.永川豆豉发酵过程中总糖和氨基酸变化与滋味的形成[J].食品科学，2015,36(21):100-104.[SUOH Y,ZHAO X,QIAN Y,et al.The changes in total sugar and amino acids during the fermentation process of Yongchuan fermented soybean and the formation of taste[J].Food Science,2015,36(21):100-104.]
• [26]叶兴乾,沈淑妤,黄睿,等.谷物食品总黄酮比色法定量的问题及选用原则[J].中国食品学报，2018,18(2):1-14.[YE X Q,SHEN S Y,HUANG R,et al.The problems and selection principles of colorimetric quantification of total flavonoids in cereal food[J].Chinese Journal of Food Science,2018,18(2):1-14.]
• [27]洪佳敏,林宝妹,张树河,等.4种藜麦抗氧化活性比较研究[J].粮食与油脂，2020,33(8):51-53.[HONG J M,LIN B M,ZHANG S H,et al.Comparative study on antioxidant activity of four types of quinoa[J].Grain and Oil,2020,33(8):51-53.]
• [28]MEHDI R,AHMAD A,BEHNAZ T.Essential oil composition,total phenolic,flavonoid contents,and antioxidant activity of thymus species collected from different regions of Iran[J].Food Chemistry,2017,220:153-161.
• [29]齐祖同.中国真菌志第五卷曲霉属及其相关有性型[M].北京：科学出版社,1997.[QI Z T.The Aspergillus genus and its related sexual types in Volume 5 of the Chinese Fungal Flora[M].Beijing:Science Press,1997.]
• [30]RAJA H A,MILLER A N,PEARCE C J,et al.Fungal identification using molecular tools:A primer for the natural products research community[J].Journal of Natural Products,2017,80(3):756-770.
• [31]PETERSON S W.Aspergillus and Penicillium identification using DNA sequences:Barcode or MLST?[J].Applied Microbiology&Biotechnology,2012,95(2):339-344.
• [32]余烨颖,赵以桥,曾昱龙,等.β-微管蛋白联合ITS序列在金花菌鉴定中的应用[J].茶叶通讯，2020,47(2):255-261.[YU YY,ZHAO Y Q,ZENG Y L,et al.Application of β-tubulin combined with ITS sequence in the identification of golden flower fungus[J].Tea Communication,2020,47(2):255-261.]
• [33]赵仁亮,吴丹,姜依何,等.不同产区加工的茯砖茶中“金花”菌的分离及分子鉴定[J].湖南农业大学学报（自然科学版），2016,42(6):592-600.[ZHAO R L,WU D,JIANG Y K,et al.Isolation and molecular identification of"golden flower"bacteria in Fuzhuan tea processed from different production areas[J].Journal of Hunan Agricultural University (Natural Science Edition),2016,42(6):592-600.]
• [34]WANG Z,JIN Q,LI Q,et al.Multiplex PCR identification of Aspergillus cristatus and Aspergillus chevalieri in Liupao tea based on orphan genes[J].Foods (Basel,Switzerland),2022,11(15):2217.
• [35]陈小红,王靖芳,郑欣萍,等.一株冠突散囊菌的分离鉴定、基因组成及其枇杷花发酵特性分析[J].微生物学通报，2023,50(12):5376-5391.[CHEN X H,WANG J F,ZHENG X P,et al.Isolation and identification,genetic composition,and fermentation characteristics analysis of an Eurotium cristatum in loquat flowers[J].Microbiology Bulletin,2023,50(12):5376-5391.]
• [36]CHEN M X,ZU Z Q,CHEN Q,et al.Untargeted and targeted metabolomics reveal changes in the chemical constituents of instant dark tea during liquid-state fermentation by Eurotium cristatum[J].Food Research International,2021,148:110623.
• [37]ZHU Y C,LUO Y H,WANG P P,et al.Simultaneous determination of free amino acids in Pu-erh tea and their changes during fermentation[J].Food Chemistry,2016,194:643-649.
• [38]姜良珍,王罗,杨涛,等.冠突散囊菌及其发酵应用研究进展[J].食品工业科技，2022,43(4):454-462.[JIANG L Z,WANGL,YANG T,et al.Research progress on the application and fermentation of Eurotium cristatum[J].Food Industry Technology,2022,43(4):454-462.]
• [39]ZHAO Y,LIU D,ZHANG J,et al.Improving soluble phenolic profile and antioxidant activity of grape pomace seeds through fungal solid-state fermentation[J].Foods (Basel,Switzerland),2024,13(8):1158.
• [40]HUANG Y X,CHEN Y L,FAN Z Y,et al.Effects of solidstate fermentation with Eurotium cristatum YL-1 on the nutritional value,total phenolics,isoflavones,antioxidant activity,and volatile organic compounds of black soybeans[J].Agronomy,2021,11:1029.
• [41]ACOSTA-ESTRADA B A,GUTIERREZ-URIBE J A,SER-NA-SALDIVAR S O.Bound phenolics in foods,a review[J].Food Chemistry,2014,152:46-55.
• [42]陈如扬,樊子依,潘一玲,等.冠突散囊菌对植物酚类物质的生物转化及生物活性的影响[J].食品研究与开发，2022,43(8):193-201.[CHEN R Y,FAN Z Y,PAN Y L,et al.The effect of Eurotium cristatum on the biotransformation and biological activities of plant phenolic substances[J].Food Research and Development,2022,43(8):193-201.]
• [43]BEI Q,CHEN G,LU F,et al.Enzymatic action mechanism of phenolic mobilization in oats (Avena sativa L.) during solid-state fermentation with Monascus anka[J].Food Chemistry,2018,245:297-304.
• [44]崔建东,李艳,牟德华.苯丙氨酸解氨酶（PAL）的研究进展[J].食品工业科技，2008,29(7):306-308.[CUI J D,LI Y,MOUD H.Research progress of phenylalanine ammonia lyase[J].Science&Technology of Food Industry,2008,29(7):306-308.]
• [45]LIU R R,XU S H,LI J L,et al.Expression profile of a PALgene from Astragalus membranaceus var.Mongholicus and its crucial role in flux into flavonoid biosynthesis[J].Plant Cell Reports,2006,25(7):705-710.
• [46]黄群,李彦坡,陈林杰,等.冠突散囊菌液态发酵过程中黑茶活性成分变化研究[J].食品科学，2007,28(12):231-234.[HUANG Q,LI Y P,CHEN L J,et al.Study on changes of active components in dark tea during liquid fermentation by Eurotium cristatum[J].Food Science,2007,28(12):231-234.]
• [47]DEY T B,CHAKRABORTY S,JAIN K K,et al.Antioxidant phenolics and their microbial production by submerged and solid-state fermentation process:A review[J].Trends in Food Science&Technology,2016,53:60-74.
• [48]MATINS S,MUSSATTO S I,MARTINES-AVILA G,et al.Bioactive phenolic compounds:Production and extraction by solidstate fermentation.A review[J].Biotechnology Advances,2011,29(3):365-373.
• [49]BHANJA T,KUMARI A,BANERJEE R.Enrichment of phenolics and free radical scavenging property of wheat koji prepared with two filamentous fungi[J].Bioresource Technology,2009,100(11):2861-2866.
• [50]SALAR R K,PUREWAL S S,SANDHU K S.Fermented pearl millet (Pennisetum glaucum) with in vitro DNA damage protection activity,bioactive compounds and antioxidant potential[J].Food Research International,2017,100:204-210.
• [51]XIAO Y,WU X,YAO X,et al.Metabolite profiling,antioxidant and α-glucosidase inhibitory activities of buckwheat processed by solid-state fermentation with Eurotium cristatum YL-1[J].Food Research International (Ottawa,Ont.),2021,143:110262.