青砖茶渥堆发酵中嗜热细菌筛选、鉴定及产酶特性研究

朱雯; 吴双; 王文凤; 许文璨; 陈文君; 黄友谊

doi:10.13305/j.cnki.jts.2022.02.001

PDF(782 KB)

茶叶科学 ›› 2022, Vol. 42 ›› Issue (2) : 211-221. DOI: 10.13305/j.cnki.jts.2022.02.001

研究报告

青砖茶渥堆发酵中嗜热细菌筛选、鉴定及产酶特性研究

朱雯, 吴双, 王文凤, 许文璨, 陈文君, 黄友谊^*

作者信息 +

The Screening, Identification and Enzyme Production of Thermophilic Bacteria in Pile-fermentation of Qingzhuan Tea

ZHU Wen, WU Shuang, WANG Wenfeng, XU Wencan, CHEN Wenjun, HUANG Youyi^*

Author information +

文章历史 +

摘要

渥堆发酵是青砖茶独特品质形成的关键技术环节。对来自青砖茶渥堆发酵样品中的细菌进行高温筛选和鉴定。结果表明,通过筛选得到20株能在高温条件下良好生长的细菌,其中15株菌株能够在含茶汤的培养基中生长。3株菌株的最适生长温度为55℃,为嗜热细菌。结合嗜热菌株的形态特征和16 S rDNA基因序列分析,确定1株为枯草芽孢杆菌（Bacillus subtilis）,2株为地衣芽孢杆菌（Bacillus licheniformis）。经嗜热细菌发酵晒青毛茶的产酶试验可知,嗜热菌株在发酵茶叶时能够产生纤维素酶、淀粉酶和单宁酶,其酶活力分别可达215.69、259.28、4.85 U。

Abstract

Pile-fermentation (PF) is a key step for the formation of the unique quality of Qingzhuan tea. In this study, high-temperature screening and identification of bacteria from PF samples of Qingzhuan tea were carried out. The results show that 20 strains of bacteria that could grow under high temperature conditions were obtained through screening, of which 15 strains could grow on a tea-containing medium. The optimum temperature of 3 thermophilic bacteria strains was 55℃. Combined with the morphological characteristics and the 16 S rDNA gene sequence analysis, it was found that one strain was Bacillus subtilis and the other two were Bacillus licheniformis. The enzyme production test on raw tea by thermophilic bacteria shows that three thermophilic bacteria could produce cellulase, amylase and tannin during tea fermentation, and the enzyme activities reached 215.69 U, 259.28 U, and 4.85 U, respectively.

导出引用

朱雯, 吴双, 王文凤, 许文璨, 陈文君, 黄友谊. 青砖茶渥堆发酵中嗜热细菌筛选、鉴定及产酶特性研究[J]. 茶叶科学. 2022, 42(2): 211-221 https://doi.org/10.13305/j.cnki.jts.2022.02.001

ZHU Wen, WU Shuang, WANG Wenfeng, XU Wencan, CHEN Wenjun, HUANG Youyi. The Screening, Identification and Enzyme Production of Thermophilic Bacteria in Pile-fermentation of Qingzhuan Tea[J]. Journal of Tea Science. 2022, 42(2): 211-221 https://doi.org/10.13305/j.cnki.jts.2022.02.001

中图分类号： S571.1

参考文献

[1] 郑鹏程, 谭荣荣, 刘盼盼, 等. 青砖茶渥堆过程中真菌种类及品质变化研究[J]. 食品科技, 2017, 42(11): 22-26.
Zheng P C, Tan R R, Liu P P, et al.Study on the fungi species and quality changes of green brick tea during pile-fermentation process[J]. Food Science and Technology, 2017, 42(11): 22-26.
[2] 张伟. 青砖茶对实验大鼠的减肥和调节血脂作用及其机制研究[D]. 武汉: 华中农业大学, 2009.
Zhang W.Studies on anti-obesity, blood lipid regulation effects and mechanisms of dark brick tea on rats [D]. Wuhan: Huazhong Agricultural University, 2009.
[3] 杨新河, 吕帮玉, 黄明军, 等. 青砖茶清除DPPH·能力及其多酚和黄酮含量研究[J]. 食品研究与开发, 2016, 37(23): 1-5.
Yang X H, Lv B Y, Huang M J, et al.Studies on DPPH free radical scavenging capacity and polyphenols and flavonoids contents of Qingzhuan brick tea[J]. Food Research and Development, 2016, 37(23): 1-5.
[4] 陈玉琼, 张伟, 倪德江, 等. 湖北青砖茶辅助降血脂作用及其抗氧化效果[J]. 茶叶科学, 2010, 30(2): 124-128.
Chen Y Q, Zhang W, Ni D J, et al.Study on the hypolipidemic effect and antioxidative activity of Hubei Qingzhuan tea[J]. Journal of Tea Science, 2010, 30(2): 124-128.
[5] Lin F J, Wei X L, Liu H Y, et al.State-of-the-art review of dark tea: from chemistry to health benefits[J]. Trends in Food Science & Technology, 2021, 109: 126-138.
[6] 苏丹, 唐飞, 张善明, 等. 不同嫩度青砖茶改善小鼠胃肠道功能研究[J]. 湖北农业科学, 2021, 60(14): 103-108.
Su D, Tang F, Zhang S M, et al.Effect of different tenderness of dark brick tea on improving gastrointestinal function in mice[J]. Hubei Agricultural Sciences, 2021, 60(14): 103-108.
[7] Zheng W J, Wan X C, Bao G H.Brick dark tea: a review of the manufacture, chemical constituents and bioconversion of the major chemical components during fermentation[J]. Phytochemistry Reviews, 2015, 14(3): 499-523.
[8] 胡帅. 基于微生物组学及代谢组学技术的青砖茶渥堆过程品质形成机制研究[D]. 武汉: 华中农业大学, 2019.
Hu S.Study on Quality Formation mechanism of Qing Zhuan Tea pile fermentation based on microbiome and metabolomics [D]. Wuhan: Huazhong Agricultural University, 2019.
[9] Cheng L, Wang Y, Zhang J, et al.Dynamic changes of metabolic profile and taste quality during the long-term aging of Qingzhuan tea: the impact of storage age[J]. Food Chemistry, 2021, 359(11): 129953. doi: 10.1016/j.foodchem.2021.129953.
[10] Xu Q, Sun M, Ning J, et al.The Core Role of Bacillus subtilis and Aspergillus fumigatus in Pile-Fermentation Processing of Qingzhuan Brick Tea[J]. Indian Journal of Microbiology, 2019, 59(3): 288-294.
[11] Zhu M Z, Li N, Zhou F, et al.Microbial bioconversion of the chemical components in dark tea[J]. Food Chemistry, 2019, 312: 126043. doi: 10.1016/j.foodchem.2019.126043.
[12] 张丹丹, 王佳佳, 朱雯, 等. 青砖茶渥堆发酵中微生物的变化[J]. 食品科学, 2019, 40(6): 166-172.
Zhang D D, Wang J J, Zhu W, et al.Changes of microbial community during pile fermentation of Qingzhuan tea[J]. Food Science, 2019, 40(6): 166-172.
[13] Hu S, He C, Li Y, et al.Changes of fungal community and non-volatile metabolites during pile-fermentation of dark green tea[J]. Food Research International, 2021, 147(1): 110472. doi: 10.1016/j.foodres.2021.110472.
[14] Zhang W, Yang R, Fang W, et al.Characterization of thermophilic fungal community associated with pile fermentation of Pu-erh tea[J]. International journal of food microbiology, 2016, 227: 29-33.
[15] 东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京: 科学出版社, 2001.
Dong X Z, Cai M Y.Manual of identification of common bacterial systems [M]. Beijing: Science Press, 2001.
[16] 张妮, 张红岩, 杨梦莹, 等. 一株海洋来源高效产角蛋白酶菌株的筛选、鉴定及其酶学性质研究[J]. 食品与发酵工业, 2020, 46(18): 98-104.
Zhang N, Zhang H Y, Yang M Y, et al.Isolation and identification of a highly efficient keratinase producing strain from marine environment and its enzymatic properties[J]. Food and Fermentation Industries, 2020, 46(18): 98-104.
[17] 吴志超. 黑茶发酵优势菌株的胞外酶检测及蛋白组学初探[D]. 福州: 福建师范大学, 2018.
Wu Z C.Detection of extracellular enzymes and initial research on proteomics of the dominant strain for dark tea's fermentation [D]. Fuzhou: Fujian Normal University, 2018.
[18] 中国国家标准化管理委员会. 茶水浸出物测定: GB/T 8305—2013[S]. 北京: 中国标准出版社, 2014: 1-5.
China National Standardization Management Committee. Tea: determination of water extracts content: GB/T 8305—2013 [S]. Beijing: China Standards Press, 2014: 1-5.
[19] 中国国家标准化管理委员会. 茶叶中茶多酚和儿茶素类含量的检测方法: GB/T 8313—2018[S]. 北京: 中国标准出版社, 2018: 1-5.
China National Standardization Management Committee. Determination of total polyphenols and catechins content in tea: GB/T 8313-2018 [S]. Beijing: China Standards Press, 2018: 1-5.
[20] 中国国家标准化管理委员会. 茶游离氨基酸总量的测定: GB/T 8314—2013[S]. 北京: 中国标准出版社, 2013: 1-5.
China National Standardization Management Committee. Tea: determination of water extracts content: GB/T 8305—2013 [S]. Beijing: China Standards Press, 2014: 1-5.
[21] 张志良, 瞿伟菁. 植物生理学实验指导[M]. 北京: 高等教育出版社, 2004.
Zhang Z L, Qu W J.Experimental guidance of plant physiology [M]. Beijing: Higher Education Press, 2004.
[22] 张欢. 茶叶厌氧发酵中高含量没食子酸形成以及茶蛋白降解促进活性肽释放的研究[D]. 武汉: 华中农业大学, 2021.
Zhang H.Research on the formation of high content gallic acid and degradation of tea protein promoting the release of active peptides in tea anaerobic fermentation [D]. Wuhan: Huazhong Agricultural University, 2021.
[23] 姚潇婷. 嗜热地衣芽孢杆菌特性的研究、改良及初步应用研究[D]. 长沙: 中南林业科技大学, 2017.
Yao X T.The research on characteristics, improvement and preliminary application of thermophilic bacillus licheniformis [D]. Changsha: Central South University of Forestry and Technology, 2017.
[24] Zhang W, Cao J, Li Z, et al.HS-SPME and GC/MS volatile component analysis of Yinghong no. 9 dark tea during the pile fermentation process[J]. Food Chemistry, 2021, 357: 129654.
[25] Zhu Y, Luo Y, Wang 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.
[26] Cheng L, Yang Q, Chen Z, et al.Distinct changes of metabolic profile and sensory quality during Qingzhuan tea processing revealed by LC-MS-based metabolomics[J]. Journal of agricultural and food chemistry, 2020, 68(17): 4955-4965.
[27] Guo W, Zhao M, Chen Q, et al.Citrinin produced using strains of Penicillium citrinum from Liupao tea[J]. Food Bioscience, 2019, 28: 183-191.
[28] Chen C, Zhang W, Lv C, et al.Primary study on thermophiles during pile-fermentation of Pu-er tea[J]. IEEE, 2011: 691-695.
[29] Li Q, Huang J, Li Y, et al.Fungal community succession and major components change during manufacturing process of Fu brick tea[J]. Scientific Reports, 2017, 7(1): 6947. doi: 10.1038/s41598-017-07098-8.
[30] Ming Z, Zhang D L, Su X Q, et al.An integrated metagenomics/metaproteomics investigation of the microbial communities and enzymes in solid-state fermentation of Pu-erh tea[J]. Scientific Reports, 2015, 5: 10117. doi: 10.1038/srep10117.