Serum metabolomics analysis of rat after intragastric infusion of Pu-erh theabrownin.

BACKGROUND: The aim was to study the effects of Pu-erh theabrownin (TB) (Mw > 50 kDa) on the metabolism of rat serum by nuclear magnetic resonance (NMR)-based metabolomics and identify candidate marker metabolites associated with Pu-erh TB, and thus provide fundamental information for a better understanding of the metabolism of Pu-erh tea in animals. RESULTS: TB infusion induced different changes in endogenous serum metabolites depending on the type of diet. Compared with the control group, the TB infusion group showed significantly reduced serum glycine and choline levels, as well as significantly increased taurine, carnitine and high-density lipoprotein (all P < 0.05). Compared with the high-lipid group, the high-lipid TB infusion group exhibited significantly reduced low-density lipoprotein and acetate levels, as well as significantly increased inositol, carnitine and glycine levels (all P < 0.05). CONCLUSION: Examination of the variations of these differential expressed metabolites and their individual functions revealed that the TB extract accelerated lipid catabolism in rats and might affect glucose metabolism. Of these, carnitine level significantly increased after intragastric infusion of TB regardless of the type of diet, and activities of carnitine palmitoyltransferases I and II changed significantly, suggesting carnitine may be a candidate serum marker for tracking the metabolism of TB in rats. © 2015 Society of Chemical Industry.

[Study on the UV-quantitative analysis of theabrownins in Pu-Erh tea].

An UV-quantitative analysis method for the theabrownin (TB) in Pu-erh tea and its derived products was established in the present study. The results showed that the Pu-erh tea shows characteristic absorption at the wavelength of 270 nm, which can be used as wavelength for the content determination of TB. The preparation methods of standard TB and standard curve were also established. The determination results show that the method is simple, the results have certain credibility, and the established method can be used for the determination of TB in Pu-erh tea and its derived products.

Influence of different fermentation raw materials on pyrolyzates of Pu-erh tea theabrownin by Curie-point pyrolysis-gas chromatography-mass spectroscopy.

SCOPE: In this study, Pu-erh tea was prepared by solid-state fermentation of the sun-dried green tea, short-fermentation black tea, and black tea. Ultraviolet-visible spectroscopy, infrared spectroscopy and Curie-point pyrolysis-gas chromatography-mass spectroscopy (CP-Py-GC/MS) were used to study the characteristics and chemical compositions of the TB formed in these Pu-erh teas. METHODS AND RESULTS: The pyrolysates of the Pu-erh teas' theabrownin (TB1, TB2, TB3) were analyzed at 386°C using a Curie point pyrolysis instrument. TB1, TB2, and TB3 produced 158, 135 and 148 pyrolysis products, respectively. These compounds could be grouped as follows: alkaloids, 25.23% (TB1), 25.67% (TB2), and 21.91% (TB3); phenols, 24.19% (TB1), 23.08% (TB2), and 29.91% (TB3); nitrogen-compounds, 11.79% (TB1), 11.25% (TB2), and 13.39% (TB3); hydrocarbons, 10.96% (TB1), 10.37% (TB2), and 11.37% (TB3); ketones, 10.34% (TB1), 6.58% (TB2), and 8.22% (TB3); furan, 5.40% (TB1), 4.06% (TB2), and 2.33% (TB3); alcohol, 3.30% (TB1), 4.16% (TB2), and 3.34% (TB3); fatty acids, 2.80% (TB1), 4.03% (TB2), and 1.42% (TB3); esters 2.17% (TB1), 0.99% (TB2) and 2.35% (TB3); and amines 1.61% (TB1), 2.81% (TB2), and 0.86% (TB3). CONCLUSION: Theabrownin mainly contains alkaloids (caffeine), tea pigments, polysaccharides, proteins and lipids. Its precursors include various polyphenols, tea pigments, alkaloids, polysaccharides, proteins and lipids. These compounds in the raw materials impact the chemical composition of theabrownin, and black tea is not a suitable raw fermentation material for Pu-erh tea.

[Spectroscopic and structural characteristics of the main components of Theabrownin in Pu-erh tea].

In the present study, we fractionated the main components of TB in Pu-erh tea by dialysis and investigated their compositions, structures and properties. TB in the Pu-erh tea was fractionated by dialyses using films with different pore sizes. The highest TB concentration was obtained in the fraction with molecular weight species > 25 000 Da. The carboxyl and hydroxyl concentrations also increased in fractions where the molecular weight size selection was highest (i. e. , > 25 000 Da). AFM images revealed that TB particles with different molecular weights had different morphologies and the TB particles appeared as scattered islands or aggregates. CP-MAS NMR experiments revealed that the TB fraction with molecular weight species between 3 500 and 25 000 Da in size was characterized by a polymeric material of polyphenols. This polymeric substance contained primarily multiple benzene rings, polysaccharides, protein residue groups and various other functional groups. After acidic hydrolysis, this fraction formed a black precipitate. CP-MAS NMR found that the precipitate was a polymeric material with multiple benzene rings. CP-GC/MS identified 16 candidate compounds from the pyrolysis products of the black precipitate obtained from the main fraction with molecular weight species between 3 500 and 25 000 Da.

Influence of large molecular polymeric pigments isolated from fermented Zijuan tea on the activity of key enzymes involved in lipid metabolism in rat.

Influence of large molecular polymeric pigments (LMPP) isolated from fermented Zijuan tea on the activity and mRNA expression of key enzymes involved in lipid metabolism in rat was explored. The results show that intragastric infusion of high-dose LMPP (1.215 g/kg body weight) effectively suppressed the elevation in TC and LDL-C (p<0.05), and prevented the reduction in HDL-C (p<0.05), compared with the hyperlipidemia model group. LMPP significantly enhanced the activity of HL and HSL, and increased the HSL mRNA expression in the liver tissue and adipose tissue. High-LMPP treatment significantly reduced the HMG-CoA reductase expression by 56.5% in the liver compared with hyperlipidemia model group. In contrast, LDL-R expression was increased by 120% in the presence of high-LMPP treatment. These results suggest that LMPP have the hypolipidemic effect to some extent and significantly enhance HSL mRNA expression in the liver and adipose tissue, thereby increasing HSL activity in rat.

[The molecular composition and spectral properties of polysaccharide isolated from pu-erh tea and its material].

Pu-erh tea, a kind of well-known tea from the ancient time, is originally produced in the Yunnan Lanchan River basin through a special solid state fermentation by fungi. It uses sun-dried green tea as its starting materials. To investigate the variation of composition and spectral properties of polysaccharide during solid state fermentation of pu-erh tea by using Saccharomyces cerevisiae as preponderant starter and using sun-dried green tea as materials in the present study. The results showed that the content of water soluble polysaccharide was increased, and the activity of hydrolase such as cellulase, pectinase and glucomylase were also enhanced. The content of neutral sugar increased with the ferment time increasing and the M(w) of raw polysaccharide showed significant difference during fermentation. The main polysaccharide TPS2 and TPS1 were isolated and purified from pu-erh tea and its materials by DEAE-52 and Sephadex G-150 column chromatography. TPS2 contains the higher content of uronic acid, but TPS1 contains the higher contents of neutral sugar and protein. Monosaccharide analysis by GC-MS revealed that TPS1 and TPS2 were composed of arabinose, galactose, glucose, rhamnose, xylose and mannose with molar ratios of 24.2 : 23.6 : 5.9 : 3.2 : 1.8 : 1.1 and 19.3 : 26.9 : 3.2 : 2.7 : 1.3 : 5.5, respectively. The average molecular weight of TPS1 and TPS2 was 1.68 x 10(4) and 1.21 x 10(4) Daltons, respectively. UV scanning spectrum showed that TPS1 and TPS2 had no characteristic absorption between 200 and 400 nm wavelength, it suggested that they contain trace protein. IR spectrum of TPS1 and TPS2 demonstrated that pyranoid rings were contained in them. As shown in the image of atomic force microscope, the molecular appearance of TPS1 and TPS2 resembled islands and apparently consisted of conglomerations. The height of conglomerations of TPS2 was about 40 nm and the length or width was 0.5-0.8 microm, while the height of conglomerations of TPS1 was about 4nm and the length or width was 0.2-0.4 microm. TPS2 shows sheet conglomerations with rough surface, but TPS1 shows squama conglomerations with smooth surface in the image of scanning electron micrograph. The experimental data suggested that the variation of composition and spectral properties of polysaccharide isolated from pu-erh tea and its materials owed to the action of microorganism and humid and thermal action for long time process.

[Construction and identification of exogenous gene expression system in Lactococcus lactis].

OBJECTIVE: To construct and characterize an exogenous gene expression system in Lactococcus lactis. METHODS: PCR-based gene assembly was used to synthesize the gene sequence containing P59 promoter, USP45 signal peptide, ribosome binding site and multiple cloning site. Plasmid pBS-pu was obtained after ligation of the assembled sequence with pBluescript II SK (+), and the terminator of USP45 protein was added to construct the recombinant plasmid pNBC1000. The gene coding for Streptococcus pyogenes M6 protein was amplified and cloned into pNBC1000 to obtain the plasmid pNBC2000. To characterize the expression system, enhanced green fluorescent protein (EGFP) gene was amplified from the plasmid pEGFP-N1 and cloned into pNBC2000. Laser scanning confocal microscope was used to observe the bacteria containing pNBC2000, pNBC2000-EGFP or pBS-EGFP. RESULTS: Green fluorescence was visualized in the bacteria containing pNBC2000-EGFP, but not in the bacteria containing pNBC2000 or pBS-EGFP. CONCLUSION: The plasmids pNBC1000 and pNBC2000 containing P59 promoter, USP45 signal peptide, ribosome binding site and multiple cloning sites are successfully constructed, which are capable of expressing exogenous gene in Lactococcus lactis.