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作物学报 ›› 2010, Vol. 36 ›› Issue (07): 1169-1175.doi: 10.3724/SP.J.1006.2010.01169

• 耕作栽培·生理生化 • 上一篇    下一篇

转小麦铁蛋白基因酵母的抗氧化活性

赵永亮1,2,陈静1,王丹1,王卫国1,李云辉1   

  1. 1河南工业大学生物工程学院,河南郑州450001;2中国农业科学院作物科学研究所农业部品种资源重点实验室/农作大科学工程,北京100081
  • 收稿日期:2009-12-17 修回日期:2010-03-19 出版日期:2010-07-12 网络出版日期:2010-04-28
  • 基金资助:

    本研究由国家重点基础研究发展计划(973计划)项目(2004CB117200)和河南工业大学博士基金项目(150216)资助。

Antioxidative Activities of Transgenic Yeast with Ferritin Gene from Wheat

ZHAO Yong-Liang1,2,CHEN Jing1,WANG Dan1,WANG Wei-Guo1, LI Yun-Hui1   

  1. 1Department of Biology Engineering,Henan University of Technology,Zhengzhou,45001,China;2The National Key Facility for Crop Gene Resources and Genetic Improvement,Key Laboratory of Crop Germplasm and Biotechnology,Ministry of Agriculture,Institute of Crop Sciences,Chinese Academy of Agricultural Sciencses,Beijing 100081.China
  • Received:2009-12-17 Revised:2010-03-19 Published:2010-07-12 Published online:2010-04-28

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摘要： 铁蛋白是广泛存在于动、植物和微生物中的储铁关键蛋白，具有调节铁代谢平衡、消除亚铁离子引起的氧化毒性等作用。本研究克隆了小麦铁蛋白基因，构建其真核表达载体，经筛选、检测得到具有形成活性小麦铁蛋白多聚体能力的转基因酵母。转基因酵母与转空载体的对照酵母相比不仅可有效清除过氧化氢(H2O2)，而且还具清除羟自由基(.OH)和超氧阴离子(O2?)的能力；×OH、O2?和H2O2对转基因酵母的半致死浓度(LD50)分别为0.40、1.00和36.91 mmol L-1，比对照酵母分别提高了37.93%、47.06%和77.03%。不同浓度转基因酵母对×OH、O2?和H2O2清除能力的回归方程分别为一元三次多项式、对数方程和负指数方程。说明小麦铁蛋白基因的真核表达产物可以有效清除3种活性氧，而且具有较为复杂的互不相同的作用机制。

关键词: 小麦, 铁蛋白, 转基因酵母, 抗氧化活性

Abstract: Ferritin, a key protein to store iron, is widely distributed in animal, plant, and microbe. Ferritin can regulate the balance in iron metabolism and decrease the oxidative damage caused by ferrous ion. It is generally recognized that the antioxidant activity of ferritin is mainly caused by integrating ferrous ion among cells, and thereby decreasing the generation of Fenton reaction. In addition to the antioxidant pathway against hydrogen peroxide (H2O2), less information is available on other antioxidative pathways and mechanism of ferritin. In this study, a ferritin gene was cloned from wheat (Triticum aestivum L.), and its expression vehicle of eukaryotic cell was constructed and transformed into yeast (Saccharomyces cerevisiae). Compared with the control yeasts, the transgenic yeasts were able to scavenge H2O2,O2?and ·OH effectively. The half lethal density (LD50) of transgenic yeasts induced by the three types of active oxygen (·OH, O2?and H2O2) was 0.40, 1.00, and 36.91 mmol L-1, which increasedby37.93%, 47.06%, and 77.03% compared with the control respectively. The abilities for eliminating ·OH, O2?, and H2O2 of the trsnsgenic yeast per unit concentiation were quantitated with cubic polynomial, logarithmic, and negative exponential equations based on yeast concentration respectively. These results demonstrated that expressed products of wheat ferritin gene in eukaryotic orgamism can effectivly in eliminate various types of active oxygen with different and complex mechanisms.

Key words: Wheat, Ferritin, Trangenic yeast, Antioxdative activity

[1]    Zheng Y-L(郑荣梁), Huang Z-Y(黄中洋). Free Radical Biology (自由基生物学), 3rd edn. Beijing: Higher Education Press, 2007. pp 1–25 (in Chinese)
[2]    Blokhina O, Virolatinen E, Kurt V F. Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Bot, 2003, 91: 179–194
[3]    Theil E C, Matzapetakis M, Liu X. Ferritins: Iron/oxygen biominerals in protein nanocages. J Biol Inorg Chem, 2006, 11, 803–810
[4]    Harrison P M, Arosio P. Ferritins: molecular properties, iron storage function and cellular regulation.  Biochim Biophys Acta,1996, 1275: 161–203
[5]    Theil, E C. Ferritin: structure, gene regulation, and cellular function in animals, plants, and microorganisms. Annu Rev Biochem, 1987, 56: 289–315
[6]    Wade V J, Treffry A, Laulhere J P, Bauminger E R, Cleton M I, Mann S, Briat J F, Harrison P M. Structure and composition of ferritin cores from pea seed (Pisum sativum). Biochim Biophys Acta, 1993, 1161: 91–96
[7]    Goto F, Yoshihara T, Saiki H, Takaiwa F. Iron accumulation and enhanced growth in transgenic lettuce plants expressing the iron-binding protein ferritin. Theor Appl Genet,2000, 100: 658–664
[8]    Ye X(叶霞), Huang X-D(黄晓德张振).Agrobacterium-mediated transformation of tomato with PvFer. Acta Hort Sin (园艺学报), 2007, 34(2): 489–492 (in Chinese with English abstract)), Yao H-Q(姚泉洪), Zhang Z(
[9]    Ye H X, Li M, Guo Z J. Evaluation and application of two high-Iron transgenic rice lines expressing a pea ferritin gene. Rice Sci, 2008, 15: 51–56
[10] Deak M, Horvath G V, Davletova S, Torok K, Sass L, Vass I, Barna B, Kiraly Z, Dudits D. Plants ectopically expressing the iron-binding protein, ferritin, are tolerant to oxidative damage and pathogens. Nat Biotechnol, 1999, 17: 192–196
[11] Hegedus A, Janda T, Horváth G VDudits D. Accumulation of overproduced ferritin in the chloroplast provides protection against photoinhibition induced by low temperature in tobacco plants. J Plant Physiol, 2008, 165: 1647–1651,
[12] Halliwell B, Gutteridge J M. Protection against oxidants in biological systems: the superoxide theory of oxygen toxicity. In: Free Radicals in Biology and Medicine. New York: Oxford University Press, 1989. pp 86–179
[13] Balla G, Jacob H S, Balla J, Rosenberg M, Nath K, Apple F, Eaton J W, Vercellotti G M. Ferritin: a cytoprotective antioxidant strategem of endothelium. J Biol Chem, 1992, 267: 18148–18153
[14] Baldi A, Lombardi D, Russo P ,Palescandolo Ferritin contributes to melanoma progression by modulating cell growth and sensitivity to oxidative stress., 2005, 11: 3175–3183 E, Luca A D, Santini D, Baldi F, Rossiello L, Dellanna M L, Mastrofrancesco A, Maresca V, Flori E, Natali P G, Picardo Mauro, Pagg M G.Clin Cancer Res
[15] Pham C G Papa S, Jones J, Alvarez K, Jayawardena S, Smaele D E, Cong R, Beaumont C. Ferritin heavy chain upregulation by NF-kappaB inhibits TNF alpha-induced apoptosis by suppressing reactive oxygen species. , 2004, 119: 529, Bubici C, Zazzeroni F,Cell–542
[16] Ravet K, Touraine B, Boucherez J, Briat J F, Gaymard F, Cellier F. Ferritins control interaction between iron homeostasis and oxidative stress in Arabidopsis. Plant J, 2009, 57: 400–412
[17] Kim H J, Kim H M, Kim J H, Ryu K S, Park S M, Jahng K Y, Yang M K, Kim D H. Expression of heteropolymeric ferritin improves iron storage in Saccharomyces cerevisiae. Appl Environ Microbiol, 2003, 69: 1999–2005
[18] Shin Y M, Kwon T H, Kim K S, Chae K S, Kim D H, Kim J H, Yang M S.Enhanced iron uptake of Saccharomyces cerevisiae by heterologous expression of a tadpole ferritin gene. Appl Environ Microbiol, 2001, 67: 1280–1283
[19] Campanella A, Isaya G, O’Neill H A, Santambrogio P, Cozzi A, Arosio P, Levi Sonia. The expression of human mitochondrial ferritin rescues respiratory function in frataxin-deficient yeast. Human Mol Genet, 2004, 13: 2279–2288
[20] Zhao Y-L(赵永亮), Zhou R-H(周荣华), Jia J-Z(贾继增). The homologous cloning of the ferritin gene from wheat. J Triticeae Crops (麦类作物学报), 2008, 28(1): 25–30 (in Chinese with English abstract)
[21] Schiestl R H, Gietz R D. High efficiency transformation of intact yeast cells using single stranded DNA as a carrier. Currt Genet, 1989, 16: 339–346
[22] Laemmli U K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 1970, 227: 680–685
[23] Barcelo F, Arean C O, Moore G R. Isolation and preliminary characterization of ferritin from clover seeds. BioMetals, 1995, 8: 47–52
[24] Bradford M M. Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Anal Biochem, 1976, 72: 248–254
[25] Jin M(金鸣), Cai Y-X(蔡亚欣), Li J-R(李金荣), Zhao H(赵辉). 1,10-phenanthroline-Fe(2+) oxidative assay of hydroxyl radical produced by H2O2/Fe(2+). Prog Biochem Biophys (生物化学与生物物理进展), 1996, 23(6): 553–555 (in Chinese with English abstract)
[26] Marlund S, Marlund G. Involvement of superoxide anion radicals in the autoxidation of pyrogallol and a convininent assay for superoxide dismutase. Eur J Biochem,1974, 47: 469–471
[27] Zhang H-R(张海容). Study of the scavenging radical activities of polysaccharide of hippophae rhamnosides fruit peel. Bull Bot (植物学通报), 2005, 22(6): 703–707 (in Chinese with English abstract)
[28] Cai H-J(蔡华静), Xiong Z-Q(熊智强), Tu G-Q(涂国全). A preliminary study on the antibacterial action of the bioactive compound produced by streptomyces 702. Acta Agric Univ Jiangxiensis (江西农业大学学报), 2005, 27(2): 274–278 (in Chinese with English abstract)
[29] Wang Y-H(王义华), Xu M-Z(徐梅珍), Jiang P(江萍), He Z-F(何照范), Xiong L-Y(熊绿芸).Purification, chemical component and antioxidative analysis of polysaccharide from Saccharomyces cerevisiae.Microbiology (微生物学通报), 2003, 30(4): 51–53 (in Chinese with English abstract)
[30] Sheng W(盛玮).Study on glutathione peroxidese activity in yeast.J Huaibei Ind Teachers Coll (淮北煤炭师范学院学报), 2003, 24(1): 40–43 (in Chinese with English abstract)
[31] Lin Z-L(林稚兰), Hao F-Y(郝福英), Wang L(王龙), Jia L(贾乐), Tian F(田枫), Li Q(李奇). Study on antioxidation of Saccharomyces cerevisiae Cu-metallothionein. Mycosystema (菌物学报), 1997, 15(4): 291–296 (in Chinese with English abstract) Zhang K-S(张坤生), Zhang L(张亮), Lü X-L(吕晓玲), Chen X-J(陈晓娟). The study on antioxidant activity of Rhodotorular glutinis pigment. Food Sci(食品科学), 2004, 25(3): 61–63 (in Chinese with English abstract)
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