The present study aimed to determine microbial community, short-chain fatty acids (SCFAs), and volatilome of Bulang pickled tea during fermentation. Sequencing of 16S rRNA and ITS revealed that Bualng pickled tea was dominated by Lactobacillus plantarum, unclassified Enterobacteriaceae, unclassified Debaryomyces, Candida metapsilosis, Cladosporium sphaerospermum, and unclassified Aspergillus. The overall contents of SCFAs increased, with acetic acid showing the highest content. A total of 398 differential volatile metabolites were detected using differential metabolomics analysis. Out of these different volatile compounds, ten key volatile compounds including (Z)-4-heptenal, 1-(2-thienyl)-ethanone, 5-methyl-(E)-2-hepten-4-one, 2-ethoxy-3-methylpyrazine, p-cresol, 2-methoxy-phenol, ethy-4-methylvalerate, 3-ethyl-phenol, p-menthene-8-thiol, and 2-s-butyl-3-methoxypyrazinewere were screened based on odor activity value (OAV). The Spearman correlation analysis showed a high correlation of SCFAs and volatile compounds with microorganisms, especially L. plantarum and C. sphaerospermum. This study provided a theoretical basis for elucidating the flavor quality formation mechanism of Bulang pickled tea.
A mimicked biosynthetic pathway of catechin metabolite genes from C. sinensis, consisting of flavanone 3 hydroxylase (F3H), dihydroflavonol reductase (DFR), and leucoanthocyanidin reductase (LCR), was designed and arranged in two sets of constructs: (a) single promoter in front of F3H and ribosome-binding sequences both in front of DFR and LCR; (b) three different promoters with each in the front of the three genes and ribosome-binding sequences at appropriate positions. Recombinant E. coli BL (DE3) harbouring the constructs were cultivated for 65 h at 26 °C in M9 medium consisting of 40 g/L glucose, 1 mM IPTG, and 3 mM eriodictyol. Compounds produced were extracted in ethyl acetate in alkaline conditions after 1 h at room temperature and identified by HPLC. Two of the four major catechins, namely, (-)-epicatechin (0.01) and (-)-epicatechin gallate (0.36 mg/L), and two other types ((+)-catechin hydrate (0.13 mg/L) and (-)-catechin gallate (0.04 mg/L)) were successfully produced.