本文選題：人銅鋅超氧化物歧化酶 + 釀酒酵母�。� 參考：《鄭州大學(xué)》2005年碩士論文

【摘要】：背景和目的 超氧化物歧化酶(SOD)是一種廣泛存在于動(dòng)物、植物、微生物中的金屬酶,按其結(jié)合的金屬離子可分為Cu/Zn-SOD、Mn-SOD、Fe-SOD三種,它們主要催化超氧化物陰離子自由基發(fā)生歧化反應(yīng),生成氧和過(guò)氧化氫,因而具有防御氧毒性,增強(qiáng)機(jī)體抗輻射損傷能力,還可預(yù)防衰老,在一些疾病如腫瘤、炎癥及自身免疫疾病等的治療中有良好的療效。通過(guò)研究寄生蟲(chóng)及宿主體內(nèi)SOD的含量、結(jié)構(gòu)及功能有助于確立寄生蟲(chóng)免疫診斷指標(biāo)、制備疫苗和尋找特異性抑制蟲(chóng)體相關(guān)酶的藥物。其中,Cu/Zn-SOD(SOD1)分布最廣,占總SOD的90%以上。它是一種普遍存在的酶,在家族性肌萎縮性側(cè)索硬化癥(FALS)、帕金森病(PD)、登革熱、癌癥、Down’s綜合癥、白內(nèi)障和多種神經(jīng)紊亂綜合癥中具有重要的生理學(xué)意義和治療潛力。 天然SOD來(lái)源有限,且具有異體蛋白免疫原性,外源SOD不易被人體接受等,使之在應(yīng)用方面受到很大限制,SOD基因工程是廣開(kāi)酶源、降低成本和獲得無(wú)抗原性的人源SOD的有效途徑。目前國(guó)外對(duì)SOD進(jìn)行了一系列的基礎(chǔ)研究、應(yīng)用開(kāi)發(fā)和臨床試驗(yàn)。國(guó)內(nèi)主要集中在與應(yīng)用有關(guān)的基礎(chǔ)理論的研究上,如分離純化及其性質(zhì)等方面的研究,SOD基因克隆和表達(dá)方面的研究報(bào)道較少。利用大腸桿菌表達(dá)系統(tǒng)來(lái)生產(chǎn)SOD的生物技術(shù)研究已比較成熟,并已達(dá)到醫(yī)學(xué)臨床試驗(yàn)階段,而用釀酒酵母等真核生物表達(dá)系統(tǒng)的研究和開(kāi)發(fā)則較少。本文首次利用釀酒酵母交配因子(MF)α的分泌信號(hào)肽序列與hCu/Zn-SOD cDNA融合,構(gòu)建了分泌表達(dá)載體,實(shí)現(xiàn)其在釀酒酵母中的克隆和分泌表達(dá),其表達(dá)產(chǎn)物有可能不經(jīng)過(guò)傳統(tǒng)的純化工藝而直接應(yīng)用于化妝品、醫(yī)療保健食品等領(lǐng)域,發(fā)揮其抗氧化,抗衰老等功能。該酵母工程菌株的成功構(gòu)建為開(kāi)發(fā)hCu/Zn-SOD的基因工程產(chǎn)品奠定了良好的基礎(chǔ)。
[Abstract]:Background and purpose Superoxide dismutase (SOD) is a kind of metallozyme widely found in animals, plants and microbes. According to the metal ions combined with it, it can be divided into three kinds: Cu- / Zn-SOD- Mn-SOD- Fe-SOD. They mainly catalyze the disproportionation of superoxide anion free radicals. Oxygen and hydrogen peroxide can be used to protect against oxygen toxicity, enhance the ability of body to resist radiation damage, and prevent aging. It has a good effect in the treatment of some diseases such as tumor, inflammation and autoimmune diseases. By studying the content, structure and function of SOD in parasite and host, it is helpful to establish the diagnostic index of parasite immunity, to prepare vaccine and to search for drugs that can specifically inhibit the enzyme related to parasite. Among them, CuZn-SOD1) is the most widely distributed, accounting for more than 90% of the total SOD. It is a ubiquitous enzyme that has important physiological and therapeutic potential in familial ALS, PD, dengue, Downs syndrome, cataract and multiple neurological disorders. The source of natural SOD is limited, and it has the immunogenicity of alloprotein, and the exogenous SOD is not easily accepted by human body, so its application is restricted greatly. It is an effective way to open the enzyme source, reduce the cost and obtain the human SOD without antigenicity. At present, a series of basic research, application development and clinical trials on SOD have been carried out abroad. There are few reports on the cloning and expression of superoxide dismutase (SOD) gene in China, such as isolation, purification and properties of superoxide dismutase (SOD) gene. The biotechnological research on the production of SOD by E. coli expression system has been mature and has reached the stage of medical clinical trial, but the research and development of eukaryotic expression system using Saccharomyces cerevisiae and other eukaryotes is less. In this paper, the secretory expression vector of Saccharomyces cerevisiae was constructed by fusion of the secreting signal peptide sequence of yeast mating factor MF- 偽 with hCu/Zn-SOD cDNA for the first time, and its cloning and secretory expression in Saccharomyces cerevisiae were realized. The expressed products may be directly used in cosmetics, health food and other fields without the traditional purification process, and play their functions of anti-oxidation and anti-aging. The successful construction of the yeast engineering strain laid a good foundation for the development of genetic engineering products of hCu/Zn-SOD.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類(lèi)號(hào)】：R341

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 馮可;王丕武;付永平;張卓;;重組大豆錳超氧化物歧化酶酵母工程菌發(fā)酵條件研究[J];農(nóng)業(yè)機(jī)械;2011年26期

相關(guān)碩士學(xué)位論文 前1條

1 馮可;利用酵母工程菌生產(chǎn)大豆MnSOD發(fā)酵條件的優(yōu)化[D];吉林農(nóng)業(yè)大學(xué);2011年

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本文編號(hào)：1950868