Genetic and proteomic analysis of novel antimicrobial proteins from rice and their application to harmful microorganisms

水稻新型抗菌蛋白的遗传和蛋白质组学分析及其在有害微生物中的应用

• 批准号: 18560749
• 负责人: TANIGUCHI Masayuki
• 金额: $ 2.55万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18560749/
• 关键词: Antimicrobial protein; Rice protein; Antimicrobial activity; Protein purification; Periodontal disease; システインプロテアーゼ; 歯周病菌; プロテアーゼ阻害剤

In this study, the preparation of cysteine protease inhibitors (oryzacystatins: Ocs) as a recombinant protein and their properties were investigated. The screening of antimicrobial proteins from rice and rice bran against Porphyromonas gingivalis was also studied. The results obtained in this study were summarized as follows.1. Five types of recombinant-OCs were expressed in Escherichia coli BL21 as the GST or His-tag fusion protein and purified by the conventional method. The inhibitory activities of OCs for several cysteine proteases were estimated.2. To measure sensitively antimicrobial activity, turbidity assay, ATP-chemical luminescence assay and fluorescence assay using DNA-binding reagent were developed and their performances were compared.3. The proteins inhibitory to proteases (Arg- and Lys-gingipains) from P. gingivalis were detected in the extracts of rice and rice bran.4. The proteins of MW 1-16 kDa obtained from the extract of rice bran have antimicrobial activity against P. gingivalis. The chemical structure of the proteins purified by two-dimensional electrohoresis was analyzed by MALDI-TOF-MS. The several proteins in the purified samples were identified as candidates of antimicrobial proteins.

本研究以水稻半胱氨酸蛋白酶抑制剂（Oryzacystatins：OCs）为重组蛋白，对其制备及性质进行了研究。本文还对大米和米糠中抗菌蛋白的筛选进行了初步研究。本研究的主要结果如下：1.在大肠杆菌BL 21中表达5种类型的重组OC作为GST或His标签融合蛋白，并通过常规方法纯化。测定了OCs对几种半胱氨酸蛋白酶的抑制活性.为了更灵敏地测定抗菌活性，建立了浊度法、ATP化学发光法和DNA结合剂荧光法，并比较了它们的性能.在大米和米糠提取物中检测到牙龈卟啉单胞菌蛋白酶（Arg-和Lys-gingipains）的抑制蛋白.从米糠提取物中获得的分子量为1-16 kDa的蛋白质对牙龈卟啉单胞菌具有抗微生物活性。通过二维电泳纯化的蛋白质的化学结构进行了分析，MALDI-TOF-MS的纯化样品中的几个蛋白被确定为抗菌蛋白的候选者。

项目成果

Oryzacystatin-II, a cystatin from rice (Oryza sativa L. japonica), is adimeric protein: the possible involvement of the interconversion between dimer and monomer in the regulation of the reactivity of oryzacystatin-II.

Oryzacystatin-II 是一种来自水稻 (Oryza sativa L. japonica) 的半胱氨酸蛋白酶抑制剂，是二聚体蛋白：二聚体和单体之间的相互转化可能参与 oryzacystatin-II 反应性的调节。

• 发表时间: 2007
• 期刊: J. Agric. Food Chem. 55
• 作者: S.;Ohtsubo;M.;Taiyoji;T.;Kawase;M.;Taniguchi;E.;Saitoh;寺嶋 正明;Sadami Ohtsubo
• 通讯作者: Sadami Ohtsubo

プロテアーゼ阻害剤ならびに抗菌剤

蛋白酶抑制剂和抗菌剂

• 发表时间: 2007

Oryzacystatin-II, a Cystatin from Rice(Oryza sativa L. japonica), is a Dimeric Protein: the Possible Involvement of the Interconversion between Dimer and Monomer in the Regulation of the Activity of Oryzacystatin-II

Oryzacystatin-II 是一种来自水稻 (Oryza sativa L. japonica) 的半胱氨酸蛋白酶抑制剂，是一种二聚体蛋白：二聚体和单体之间的相互转化可能参与 Oryzacystatin-II 活性的调节

• 发表时间: 2007
• 期刊: J.Agric.Food Chem Vol.55, No.5
• 作者: S.;Ohtsubo;M.;Taiyoji;T.;Kawase;M.;Taniguchi;E.;Saitoh
• 通讯作者: Saitoh

Screening of antimicrobial proteins from rice against Porphyromonas gingivalis and their properties

大米中抗牙龈卟啉单胞菌抗菌蛋白的筛选及其特性

• 发表时间: 2007
• 作者: Yuki;Niwa;Akira;Ikarashi;Yasuyuki;Shitomi;Tomokazu;Kawase;Takaaki;Tanaka;Masayuki;Taniguchi
• 通讯作者: Taniguchi

Inhibition of the growth and proteolytic activity of Porphyromonas gingivalis by recombinant oryzacystatins.

重组 oryzacystatins 抑制牙龈卟啉单胞菌的生长和蛋白水解活性。

• 发表时间: 2007
• 作者: S.;Ohtsubo;M.;Taiyoji;T.;Kawase;M.;Taniguchi;E.;Saitoh;寺嶋 正明;Sadami Ohtsubo;M. Terashima;丹羽 祐基;Yasuyuki Shitomi
• 通讯作者: Yasuyuki Shitomi