Abstract
Microbes are thought to have key roles in the development of the special properties of post-fermented pu-erh tea (pu-erh shucha), a well-known traditional Chinese tea; however, little is known about the bacteria during the fermentation. In this work, the structure and dynamics of the bacterial community involved in the production of pu-erh shucha were investigated using 16S rRNA gene clone libraries constructed from samples collected on days zero (LD-0), 5 (LD-5), 10 (LD-10), 15 (LD-15) and 20 (LD-20) of the fermentation. A total of 747 sequences with individual clone library containing 115–174 sequences and 4–20 unique operational taxonomic units (OTUs) were obtained. These OTUs were grouped into four phyla (Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria) and further identified as members of 10 families, such as Alcaligenaceae, Bacillaceae, Enterobacteriaceae, etc. The dominant bacteria were Enterobacteriaceae in the raw material (LD-0) and in the initial stages of fermentation (LD-5 and LD-10), which changed to Bacillaceae at the last stages of fermentation (LD-15 and LD-20) at a temperature of 40–60 °C. It is interesting that the dominant OTUs in libraries LD-15 and LD-20 were very closely related to Bacillus coagulans, which is a safe thermoduric probiotic. Together the bacterial diversity and dynamics during a fermentation of pu-erh shucha were demonstrated, and a worthy clue for artificial inoculation of B. coagulans to improve the health benefits of pu-erh shucha or produce probiotic pu-erh tea were provided.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- GRAS:
-
Generally recognized as safe
- OTUs:
-
Operational taxonomic units
- QPS:
-
Qualified presumption of safety
- PCR-DGGE:
-
Polymerase chain reaction denaturing gradient gel electrophoresis
- SSF:
-
Solid-state fermentation
References
Abe M, Takaoka N, Idemoto Y, Takagi C, Imai T, Nakasaki K (2008) Characteristic fungi observed in the fermentation process for puer tea. Int J Food Microbiol 124:199–203
Amann R, Ludwig W (2000) Ribosomal RNA-targeted nucleic acid probes for studies in microbial ecology. FEMS Microbiol Rev 24:555–565
Chen H, Jen-Kun S-Y, Lin-Shiau (2008) Pu-erh tea its manufacturing and health benefits. In: Ho CT, Lin JK, Shahidi F (eds) Tea and tea products: chemistry and health-promoting properties. CRC Press, Boca Raton, pp 9–16
Chen Y, Liu B, Chang Y (2010) Bioactivities and sensory evaluation of Pu-erh teas made from three tea leaves in an improved pile fermentation process. J Biosci Bioeng 109:557–563
Colwell RK, Chao A, Gotelli NJ, Lin SY, Mao CX, Chazdon RL, Longino JT (2012) Models and estimators linking individual-based and sample-based rarefaction, extrapolation and comparison of assemblages. J Plant Ecol 5:3–21
Denis C, Cadot P, Leguerinel I, Thuault D, Sohier D (2006) Heat resistance of coliform species isolated from cooked ham, snail flesh, and ‘bouchees a lareine’. Lett Appl Microbiol 42:160–164
Endres JR, Clewell A, Jade KA, Farber T, Hauswirth J, Schauss AG (2009) Safety assessment of a proprietary preparation of a novel Probiotic, Bacillus coagulans, as a food ingredient. Food Chem Toxicol 47:1231–1238
Endres JR, Qureshi I, Farber T, Hauswirth J, Hirka G, Pasics I, Schauss AG (2011) One-year chronic oral toxicity with combined reproduction toxicity study of a novel probiotic, Bacillus coagulans, as a food ingredient. Food Chem Toxicol 49:1174–1182
European Food Safety Authority (2008) Scientific Opinion of the Panel on Biological Hazards on a request from EFSA on the maintenance of the QPS list of microorganisms intentionally added to food or feed. EFSA J: 923:1–48. http://www.efsa.europa.eu/en/efsajournal/doc/923.pdf
Ganeden Biotech I (2011) Notice to US Food and Drug Administration that Bacillus coagulans GBI-30, 6086, a Novel Probiotic, is Generally Recognized as Safe for use in Foods. http://www.accessdata.fda.gov/scripts/fcn/gras_notices/grn000399.pdf
Gong S, Chengyin L, Xu L, Yuxiang Z, Hong S, Yalin G, Jihong W, Lei Z, Zilei G (2009) Methodology of sensory evaluation of tea. GB/T 23776-2009
Good IJ (1953) The population frequencies of species and the estimation of population parameters. Biometrika 40:237–264
Ho CT, Lin JK, Shahidi F (2008) Tea and tea products: chemistry and health-promoting properties. CRC Press, Boca Raton
Jeng KC, Chen CS, Fang YP, Hou RC, Chen YS (2007) Effect of microbial fermentation on content of statin, GABA, and polyphenols in Pu-Erh tea. J Agric Food Chem 55:8787–8792
Kim BC, Jeong WJ, Kim DY, Oh HW, Kim H, Park DS, Park HM, Bae KS (2009) Paenibacillus pueri sp. nov., isolated from Pu’er tea. Int J Syst Evol Microbiol 59:1002–1006
Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, New York
Li XR, Ma EB, Yan LZ, Meng H, Du XW, Zhang SW, Quan ZX (2011) Bacterial and fungal diversity in the traditional Chinese liquor fermentation process. Int J Food Microbiol 146:31–37
Oh HW, Kim BC, Lee KH, Kim DY, Park DS, Park HM, Bae KS (2008) Paenibacillus camelliae sp. nov., isolated from fermented leaves of Camellia sinensis. The J Microbiol 46:530–534
Peng X, Yu M (2011) Isolation and identification of culturable microorganisms in a 10-year-old fermented Pu-erh Tea. Food Science 196–199:1049–1054 (in Chinese with English abstract)
Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microb 75:7537–7541
Sun T (2011) Fungal communities during pu-erh tea fermentation revealed by ITS library. Thesis of Yunnan Agricultural University (in Chinese with English abstract)
Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267
Williams KP, Gillespie JJ, Sobral BW, Nordberg EK, Snyder EE, Shallom JM, Dickerman AW (2010) Phylogeny of gammaproteobacteria. J Bacteriol 192:2305–2314
Wright ES, Yilmaz LS, Noguera DR (2012) DECIPHER, a search-based approach to chimera identification for 16S rRNA Sequences. Appl Environ Microb 78:717–725
Xu A, Wang Y, Wen J, Liu P, Liu Z, Li Z (2011) Fungal community associated with fermentation and storage of Fuzhuan brick-tea. Int J Food Microbiol 146:14–22
Zhao ZJ, Zhou L, Wang EX, Liu QJ (2010) Fungal colonization of pu-erh tea in Yunnan. J Food Safety 30:769–784
Acknowledgments
This work was supported by grants from The National Natural Science Foundation of China (grant no. 31160174), the Key Program for Applied Basic Researches of Yunnan Province, China (grant no. 2009CC005) and the Natural Science Foundation of Yunnan Province, China (grant no. 2010ZC084).
Author information
Authors and Affiliations
Corresponding authors
Additional information
Ming Zhao and Wei Xiao have contributed equally to this work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Zhao, M., Xiao, W., Ma, Y. et al. Structure and dynamics of the bacterial communities in fermentation of the traditional Chinese post-fermented pu-erh tea revealed by 16S rRNA gene clone library. World J Microbiol Biotechnol 29, 1877–1884 (2013). https://doi.org/10.1007/s11274-013-1351-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11274-013-1351-z