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Microbial enzymes during solid-state fermentation (SSF), which play important roles in the food, chemical, pharmaceutical and environmental fields, remain relatively unknown. In this work, the microbial communities and enzymes in SSF of Pu-erh tea, a well-known traditional Chinese tea, were investigated by integrated metagenomics/metaproteomics approach. The dominant bacteria and fungi were identified as Proteobacteria (48.42%) and Aspergillus (94.98%), through pyrosequencing-based analyses of the bacterial 16S and fungal 18S rRNA genes, respectively. In total, 335 proteins with at least two unique peptides were identified and classified into 28 Biological Processes and 35 Molecular Function categories using a metaproteomics analysis. The integration of metagenomics and metaproteomics data demonstrated that Aspergillus was dominant fungus and major host of identified proteins (50.45%). Enzymes involved in the degradation of the plant cell wall were identified and associated with the soft-rotting of tea leaves. Peroxiredoxins, catalase and peroxidases were associated with the oxidation of catechins. In conclusion, this work greatly advances our understanding of the SSF of Pu-erh tea and provides a powerful tool for studying SSF mechanisms, especially in relation to the microbial communities present.

An in-depth knowledge of the microbiota and metabolites in the solid-state fermentation (SSF) of Post-fermented Pu-erh tea (Pu-erh Shucha, PFPT), a Chinese traditional tea with various health benefits, is essential to develop modern fermentation technology. In this work, the microbial diversity and succession in two laboratory-developed SSF protocols for PFPT were investigated using pyrosequencing analyses of the bacterial 16S rRNA and fungal 18S rRNA genes. The active bacteria in the initial stages of SSF (seven days) were from the raw materials and environment, with a dominance of Proteobacteria in both the raw materials and SSF after seven days. The environmental bacteria were inoculated into the tea mass throughout the fermentation process and multiplied, with a dominance of Firmicutes at day 14 and 21, and then Firmicutes and Actinobacteria at the last stages of fermentation (day 28 and 35). The dominant fungi came from the raw material and were identified at the genus level as Aspergillus throughout the SSF process. The contents of tea polyphenols, free amino acids, gallic acid, theaflavin, thearubigin, and catechins decreased significantly (p < 0.05), while the level of theabrownin increased significantly (p < 0.05). The caffeine content showed no significant change (p > 0.05). In total, 30 bacterial and three fungal genera showed significant correlations to 1-8 and 3-4 identified tea compounds, respectively (p < 0.05). The dynamics of the microbiota and chemical compounds, and correlations between their changes in the SSF of PFPT were revealed, and present a foundation for further studies on the microbial effects on chemical compounds.

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