Camellia sinensis polyphenol oxdiase (PPO) plays essential roles in the quality formation of tea processing. Despite that it always is a hotspot, there is no report on expressing C. sinensis PPO in vitro. Based on purifying the active Yihongzao PPO with transit-peptide expressed in Escherichia coli, the active mature form was also expressed in E. coli and purified from the inclusion bodies. The specific activity of the mature form was higher about three times than that of PPO with transit-peptide, which suggests that the transit-peptide should restrain the activity of PPO in vivo and in vitro. It is helpful to yield and apply for the C. sinensis engineering PPO with high activity in a large-scale.
LIU Jing-wei
,
HUANG YOU-yi
,
DING Jian
,
LIU Cong
,
XIAO xiu-dan
,
NI De-jiang
. Prokaryotic Expression of Polyphenol Oxidase Mature Protein from Camellia sinensis[J]. Journal of Tea Science, 2010
, 30(2)
: 136
-140
.
DOI: 10.13305/j.cnki.jts.2010.02.010
[1] Yoruk R, Marshall M R.Physicochemical properties and function of plant polyphenol oxidase: A review[J]. J Food Biochem, 2003, 27(5): 361-422.
[2] Dai Li, Gong Changrong, Shi Lin, et al. Polyphenol Oxidase in Plants[J]. Chinese Agric Science Bulletin, 2007, 23(6): 313-315.
[3] Coetzer C, Corsini D, Love S, et al. Control of enzymatic browning in potato (Solanum tuberosum L.) by sense and antisense RNA from tomato polyphenol oxidase[J]. J Agric Food Chem, 2001, 49(2): 652-657.
[4] 曹艳红, 章镇, 姚泉洪, 等. 苹果多酚氧化酶双链RNA干扰(RNAi)研究[J]. 实验生物学报, 2004, 37(6): 487-493.
[5] Thanaraj S N Stephen, Ramachandran Seshadri. Influence of polyphenol oxidase activity and polyphenol content of tea shoot on quality of black tea[J]. J Sci Food Agri, 2006, 51(1): 57-69.
[6] Jain J C, Takeo T.A review: the enzyme of tea and their role in tea making[J]. J Food Biochem, 2007, 8(4): 243-279.
[7] 赵淑娟, 王坤波, 傅冬和, 等. 茶多酚氧化酶酶学性质研究[J]. 湖南农业大学学报(自然科学版), 2008, 34(1): 84-86.
[8] 赵东, 刘祖生, 奚彪. 茶树多酚氧化酶基因的克隆及其序列比较[J]. 茶叶科学, 2001, 21(2): 94-98.
[9] 陈忠正, 李斌, 黎秋华, 等. 茶叶多酚氧化酶的基因克隆与序列分析[J]. 食品工业科技, 2006(10): 109-111,117.
[10] 黄建安, 黄意欢, 罗军武, 等. 茶树多酚氧化酶基因的SNP分析[J]. 湖南农业大学学报(自然科学版), 2007, 33(4): 454-458, 485.
[11] 黄建安, 黄意欢, 李家贤, 等. 茶树多酚氧化酶基因的PCR-RFLP多态性分析[J]. 茶叶科学, 2008, 28(5):370-378.
[12] Tu You-Ying, Xu Xin-Qing, Xia Hui-Long, et al. Optimization of theaflavin biosynthesis from tea polyphenols using an immobilized enzyme system and response surface methodology[J]. Biotech Letters, 2005, 27(4): 269-274.
[13] 曾盔, 刘仲华. 用茶叶多酚氧化酶生物传感器测定茶多酚[J]. 食品科学, 2006, 27(7): 95-98.
[14] 谷记平, 刘仲华, 黄建安, 等. 利用外源多酚氧化酶酶促氧化制备茶黄素的研究[J]. 天然产物研究与开发, 2007, 19(B08): 299-302, 317.
[15] 黄意欢. 茶学实验技术 [M]. 北京: 中国农业出版社, 1997.
[16] Sommer A, Ne'eman E, Steffens J C, et al. Import, targeting, and processing of a plant polyphenol oxidase[J]. Plant Physiol, 1994, 105(4): 1301-1311.
[17] Nakayama T, Yonekura-Sakakibara K, Sato T, et al. Aureusidin Synthase: A Polyphenol Oxidase Homolog Responsible for Flower Coloration[J]. Science, 2000, 290(5494): 1163-1166.
[18] Hunt M D, Eannetta N T, Yu H, et al. cDNA cloning and expression of potato polyphenol oxidase[J]. Plant Mol Biol, 1993, 21(1): 59-68.
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