Molecular design of lysozyme for switching the antimicrobial action

用于切换抗菌作用的溶菌酶的分子设计

• 批准号: 12660115
• 负责人: KATO Akio
• 金额: $ 2.3万
• 依托单位: Yamaguchi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660115/
• 关键词: Lysozyme; Bactericidal lysozyme; Lipophilized lysozyme; Hyrdophobic peptide-fused lysozyme; ソポフィル化リゾチ-ム; 疎水ペプチド結合リゾチ-ム; 抗菌性スイッチング; Pichia pastoris

In order to expand the antimicrobial action to Gram-negative and Gram-positive, the molecular design of hen egg white lysozyme was attempted by genetic modification. The penta hydrophobic peptide (Phe-Phe-Val-Ala-Pro) was fused to the C terminus of lysozyme. The resulting petapeptide fused lysozyme (H5-Lz) showed strong bactericidal action against Gram-negative and -positive bacteria. The secretion amount of H5-Lz greatly increased in Pichia pastoris expression system, although it was very poor in Saccharomyces . cerevisiae. The conformation of H5-Lz was kept in a folded form, but the stability was lightly decreased, reflecting the fusion of hydrophobic peptide. The H5-Lz can be useful for industrial applications. As a typical example, the H5-Lz was introduce into tobacco leaf through agrobacterium infection. The transgenic tobacco showed strong resistance to gray mold and powdery mildew infection.

为了扩大对革兰氏阴性菌和革兰氏阳性菌的抗菌作用，尝试通过基因改造对鸡蛋白色溶菌酶进行分子设计。将五肽（Phe-Phe-Val-Ala-Pro）与溶菌酶的C端融合。合成的肽融合溶菌酶（H5-Lz）对革兰氏阴性菌和革兰氏阳性菌均有较强的杀菌作用。H5-Lz在酵母中的分泌量很低，但在毕赤酵母表达系统中的分泌量大大增加。啤酒。H5-Lz的构象保持在折叠状态，但稳定性略有下降，反映了疏水肽的融合。H5-Lz可用于工业应用。作为一个典型的例子，通过农杆菌感染将H5-Lz导入烟草叶片。转基因烟草对灰霉病和白粉病表现出较强的抗性。

项目成果

KATO, A., LIU, S., KATO, M., AZAKAMI, H.: "Moleuclar designs of hen lysozyme by genetic modification"Industrial Proteins. 9. 12-14 (2001)

KATO, A.、LIU, S.、KATO, M.、AZAKAMI, H.：“通过基因修饰进行母鸡溶菌酶的分子设计”工业蛋白质。

A.Kato, S.Liu, M.Kato, H.Azakami: "Molecular designs of hen lysozyme by genetic modification."Industrial Proteins. Vol9, No2. 12-14 (2001)

A.Kato、S.Liu、M.Kato、H.Azakami：“通过基因修饰进行母鸡溶菌酶的分子设计。”工业蛋白质。

LIU S.T, SUGIMOTO T, AZAKAMI H, KATO A: "Lipophilization of lysome by short and middle chain fatty acids"Journal of Agricultural and Food Chemistry. 48. 265-269 (2000)

LIU S.T、SUGIMOTO T、AZAKAMI H、KATO A：“短链和中链脂肪酸对溶酶体的亲脂化”农业与食品化学杂志。

S.T.Liu, T.Sugimoto, H.Azakami, A.Kato: "Lipophilization of lysozyme by short and middle chain fatty acids."J. Agric. Food Chem.. 48. 265-269 (2000)

S.T.Liu、T.Sugimoto、H.Azakami、A.Kato：“短链和中链脂肪酸对溶菌酶的亲脂化”。

LIU, S., AZAKAMI, H., KATO, A.: "Improvement in the yield of lipophilized lysozyme by the combination with Maillard-type glycosylation"Nahruna-Food. 44. 407-410 (2000)

LIU, S.、AZAKAMI, H.、KATO, A.：“通过与美拉德型糖基化组合提高亲脂溶菌酶的产量”Nahruna-Food。