本研究以EGCG-O-甲基转移酶（EOMT）催化EGCG生成EGCG3"Me的产量为主要指标,探讨了重组大肠杆菌内EGCG-O-甲基转移酶的诱导表达条件。结果表明,当诱导剂IPTG终浓度为0.05mmol/L,诱导时间为20h,培养基初始pH为7.0,以及诱导温度为20℃时,EOMT的表达效果最佳。
EGCG3"Me could be produced from EGCG catalyzed by EGCG-O-Methyltransferase. Taken the yield of EGCG3"Me as main index, the present study focused on the producing conditions of EGCG-O-Methyltransferase induced by IPTG in recombinant E. coli bacteria. Results showed that the optimum producing conditions were as follows: the concentration of IPTG was 0.05mmol/L, the induction time was 20h, the initial pH of medium was 7.0 and the induction temperature was 20℃.
[1] Sano M, Suzuki M, Miyase T, et al. Novel anti-allergic catechin derivatives isolated from oolong tea[J]. Journal of Agriculture and Food Chemistry, 1999, 47(5): 1906-1910.
[2] Maeda-Yamamoto M, Kawahara H, Matsuda N, et al. Effects of tea infusions of various varieties or different manufacturing types on inhibition of mouse mast cell activation[J]. Biosci Biotechnol Biochem, 1998, 62(11): 2277-2279.
[3] Mari Maeda-Yamamoto, Naoki Inagaki, Jiro Kitaura, et al. O-Methylated Catechins from Tea Leaves Inhibit Mutiple Protein Kinases in Mast Cells[J]. The Journal of Immunology, 2004, 172(7): 4486-4492.
[4] Chiu feng-lan, Lin jen-kun. HPLC Analysis of Naturally Occurring Methylated Catechins 3″-and 4″-Methyl-epigallocatechin Gallate, in Various Fresh Tea Leaves and Commercial Teas and Their Potent Inhibitory Effects on Inducible Nitric Oxide Synthase in Macrophages[J]. Journal of Agriculture and Food Chemistry, 2005, 53(8): 7035-7042.
[5] Mari Maeda-Yamamoto, Mitsuaki Sano, Hirofumi Tachiban.Anti-Allergic and Anti-Cancer Metastatic Agents in Green Tea (Camellia sinensis L.)[J]. Kagaku to Seibutsu, 2003, 41(7): 442-447.
[6] 吕海鹏, 谭俊峰, 林智. 茶树种质资源EGCG3"Me含量及其变化规律研究[J]. 茶叶科学, 2006, 26(4): 310-314.
[7] 吕海鹏, 孙业良, 林智, 等. 表没食子儿茶素没食子酸酯的甲基化分子修饰[J]. 食品科学, 2010, 31(15): 139-142.
[8] 伍妍俊, 汪小钢, 宛晓春. 甲基化EGCG的合成及其在人工模拟胃肠液中的稳定性[J]. 安徽农业大学学报, 2010, 37(4): 688-691.
[9] Masanobu Kirita, Daiki Honma, Yoshihisa Tanaka, et al. Cloning of a Novel O-Methyltransferases from Camellia sinensis and Synthesis of O-Methylated EGCG and Evaluation of their Bioactivity[J]. J Agric Food Chem, 2010, 58(12): 7196-7201.
[10] 李波,倪志勇,石庆华,等.棉花咖啡酸-O-甲基转移酶基因的原核表达及蛋白纯化鉴定[J]. 西北植物学报, 2010, 30(9): 1738-1743.
[11] Mohammad-Wadud Bhuiya, Chang-Jun Liu.A cost-effective colorimetric assay for phenolic O-methyltransferases and characterization of caffeate 3-O-methyltransferases from Populus trichocarpa[J]. Analytical Biochemistry, 2009, 384(1):151-158.
[12] 张佳艺, 关怡新, 姚善泾. 小分子伴侣GroEL(191-345)在E. coli中的表达及其培养条件的优化[J]. 浙江大学学报: 农业与生命科学版, 2003, 29(6): 603-608.
[13] 陈卫, 葛佳佳, 张灏, 等. 半乳糖苷酶基因在大肠杆菌中过量表达及IPTG诱导条件[J]. 无锡轻工大学学报, 2002, 21(5): 492-495.
浙公网安备 33019902000101号 
