Purification and characterization of theobromine synthase in a Theobromine-Enriched wild tea plant (Camellia gymnogyna Chang) from Dayao Mountain, China. (1st May 2020)

• Record Type: Journal Article
• Authors: Teng, Jie; Yan, Changyu; Zeng, Wen; Zhang, Yuqian; Zeng, Zhen; Huang, Yahui
• Is Part Of: Food chemistry. Volume 311(2020)
• Publication Date: 2020-05-01
• Subjects: CgC Camellia gymnogyna Chang -- TS theobromine synthase -- GFP green fluorescent protein -- NMT N-methyltransferase -- XMT xanthosine methy transferase -- MXMT 7-methy xanthine methyl transferase -- DXMT 3, 7-dimethyl xanthine methyl transferase -- SAM S-adenosyl-l-methionine -- TCS tea caffeine synthase -- SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis -- HPLC high performance liquid chromatography -- qRT-PCR quantitative real-time PCR -- CNS central nervous system; Caffeine (PubChem CID: 2519) -- Theobromine (PubChem CID: 5429) -- Theophylline (PubChem CID: 2153) -- Theacrine (PubChem CID: 75324) -- 7-Methylxanthine (PubChem CID: 68374) -- Paraxanthine (PubChem CID: 5687) -- Xanthine (PubChem CID: 1188) -- Xanthosine (PubChem CID: 64959); Purine alkaloid -- Theobromine synthase -- Purification -- Wild tea -- Dayao Mountain; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2019.125875
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: Camellia gymnogyna Chang tea plant, found in Guangxi Province contains three different purine alkaloids. Theobromine synthase was from Camellia gymnogyna Chang, purified, and characterized. 7-Methylxanthine was a favorable substrate for theobromine synthase. Full length cloning, subcellular localization, and polygenetic tree analysis of the theobromine synthase gene. Abstract: Camellia gymnogyna Chang ( Cg C), a wild tea plant, was discovered on Dayao Mountain, China. However, research regarding this tea plant is limited. Our study found that Cg C contains theobromine, caffeine, and theacrine, among which theobromine content was the highest (14.37–39.72 mg/g). In addition, theobromine synthase (TS) was partially purified from Cg C leaves, up to 35.87-fold, with consecutive chromatography, and its molecular weight was found to be approximately 62 kDa. The optimum reaction time, pH, and temperature for theobromine synthase from 7-methylxanthine was found to be 6 h, 4, and 45 °C, respectively. TS expression at both mRNA and protein stages was higher in the first than in the fourth leaf ( P < 0.05). Subcellular localization of TS indicated that it was localized in the nucleus. These results indicate that Cg C can be of scientific value and could lead to efficient utilization of this rare wild tea germplasm.