Lantibiotic engineering: peptide engineering approach using the modification enzymes

羊毛硫抗生素工程：使用修饰酶的肽工程方法

• 批准号: 23380050
• 负责人: SONOMOTO Kenji
• 金额: $ 12.06万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23380050/
• 关键词: ランチビオティック; 異常アミノ酸; nukacin ISK-1; ペプチドデザイン; 翻訳後修飾; 脱水・環化; 基質認識ドメイン; リーダーペプチド; 異常アミノ酸形成酵素NukM; 結晶化; ペプチドの機能改変; プロタミン; 表面プラズモン共鳴

Nukacin ISK-1 is a lantibiotic produced by Staphylococcus warneri ISK-1. Ribosomally synthesized precursor peptide, NukA undergoes introduction of unusual amino acids by a single modification enzyme, NukM, and the leader sequence is removed and the mature peptide is secreted by ABC transporter NukT. NukM was divided into two domains, NukMN and NukMC, based on a prediction that each domain is responsible for dehydration (phosphorylation and further phosphate elimination) and cyclization, respectively. The in vitro reconstitution of NukM, NukMN, and NukMC revealed that NukM can fully modify NukA, and NukMN could partially phosphorylate and dehydrate it. A series of NukT mutants were constructed and their transport activity in vivo and peptidase activity in vitro were investigated. It was suggested that peptidase activity of NukT depends on ATP hydrolysis, and the N-terminal peptidase domain of NukT may cooperatively function with the C-terminal ATP-binding domain.

努卡星ISK-1是由华纳葡萄球菌ISK-1产生的一种抗生素。核糖体合成的前体多肽Nuka通过一个单一的修饰酶NukM引入不寻常的氨基酸，并去除前导序列，由ABC转运蛋白NukT分泌成熟多肽。根据一个预测，NukM被分为两个结构域，NukMN和NukMC，每个结构域分别负责脱水(磷酸化和进一步的磷酸消除)和环化。NukM、NukMN和NukMC的体外重组表明，NukM可以完全修饰Nuka，NukMN可以部分磷酸化和脱水。构建了一系列NukT突变体，并对其体内转运活性和体外肽酶活性进行了研究。提示NukT的多肽酶活性依赖于ATP的水解，N端的多肽酶结构域可能与C端的三磷酸腺苷结合区协同作用。

项目成果

Enhanced production of nukacin D13E in Lactococcus lactis NZ9000 by the additional expression of immunity genes

通过免疫基因的额外表达增强乳酸乳球菌 NZ9000 中 nukacin D13E 的产量

• DOI: 10.1007/s00253-011-3563-1
• 发表时间: 2012
• 期刊: Applied Microbiology and Biotechnoly
• 作者: Tijo Varghese Puramattathu;et al
• 通讯作者: et al

New Era of Lactic Acid Bacteria to Open the Future

乳酸菌新时代，开启未来

• 发表时间: 2012
• 作者: S.Koyama;et al;石橋直樹，善藤威史，園元謙二;Kenji Sonomoto
• 通讯作者: Kenji Sonomoto

Methodologies and strategies for the bioengineering of lantibiotics

羊毛硫抗生素生物工程的方法和策略

• DOI: 10.2174/138920111796117274
• 发表时间: 2011
• 期刊: Current Pharmaceutical Biotechnology
• 影响因子: 2.8
• 作者: Jun-ichi Nagao;Mami Nishie & Kenji Sonomoto
• 通讯作者: Mami Nishie & Kenji Sonomoto

九州大学大学院農学研究院生命機能科学部門分子微生物学・バイオマス資源化学講座微生物工学分野

九州大学农学研究生院微生物工学系、分子微生物与生物质资源化学系、生物功能科学系

New Insight of Lantibiotic Engineering : Characterization of the enzymes, design and mode of action of lantibiotics, and immunity proteins

羊毛硫抗生素工程的新见解：酶的表征、羊毛硫抗生素的设计和作用方式以及免疫蛋白

• 发表时间: 2011
• 作者: Kenji Sonomoto;Mami Nishie;Mohammad R.Islam;Jun-ichi Nagao;Takeshi Zendo;Jiro Nakayama
• 通讯作者: Jiro Nakayama