Studies on Novel Syntheses of Useful Carbohydrates by Enzyme Hybrid

杂化酶新型合成有用碳水化合物的研究

• 批准号: 63560124
• 负责人: USUI Taichi
• 金额: $ 1.28万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63560124/
• 关键词: Lysozyme; Chitinase; Enzyme Hybrid; Transglycosylation; Synthesis Of Useful Oligosaccharide; 糖転移反応体; リゾチーム; 有用オリゴ糖; 糖転位反応

This project is concerned with the enzymatic syntheses of useful chitinoligo- saccharides by chitinolytic enzymes p-Nitrophenyl penta-N-acetyl-beta-chitopentaoside, which is available as a substrate for a lysozyme assay, was efficiently synthesized by transfer of the tetra-N-acetyl-chitotetraosyl residue to the 4-position of p- nitrophenyl 2-acetamido-2-deoxy-beta-D-glucopyranoside using lysozyme in an aqueous solution containing DMSO in a high concentration. Also, a chitinase from Trichoderma reesei catalyzed a transglycosylation reaction from tetra-N-acetyl-chitotetraose as a substrate to hexa-N-acetyl-chitohexaose. The transglycosylation by lysozyme furthermore afforded a more convenient way of preparing hexa-N-acetyl-chitohexaose and hepta-N-acetyl-chitoheptaose from di-N-acetyl-chitobiose than the chitinase reaction.Enzyme hybrid was prepared by shiff-amino reduction of hen egg-white lysozyme with lactose. The modified lysozyme gained more temperature stability and resistance to organic solvent than the native one.

本项目涉及通过几丁质分解酶p-Nitrophenyl penta-N-acetyl-beta-chitopentaoside酶促合成有用的几丁质寡聚体，其可用作溶菌酶测定的底物，通过将四-N-乙酰基-壳聚糖四糖残基转移到对硝基苯基2-乙酰氨基-2-脱氧-β-D-在含有高浓度DMSO的水溶液中使用溶菌酶制备吡喃葡萄糖苷。此外，里氏木霉几丁质酶催化转糖基化反应，从四-N-乙酰基-壳四糖作为底物的六-N-乙酰基-壳六糖。溶菌酶转糖基化反应比几丁质酶反应更方便地从二-N-乙酰基-壳二糖制备六-N-乙酰基-壳六糖和七-N-乙酰基-壳七糖。改性后的溶菌酶比原溶菌酶具有更好的温度稳定性和耐有机溶剂性。

项目成果

碓氷泰市: "Oligosaccharides-Production,Properties and Applications “Chitooligosaccharides"" Gordon and Breach Science Publishers,

Yasushi Usui：“低聚糖 - 生产、特性和应用“壳寡糖”” Gordon and Breach Science Publishers，

碓氷泰市: "Enzymatic Synthesis of Useful Chitin-oligosaccharides by Utilizing Transglycosylation Reaction of Chitinolytic Enzymes in Buffer System Containing Ammonium Sulfate" Carbohydr.Res.

Yasushi Usui：“利用几丁质酶在含有硫酸铵的缓冲系统中的转糖基反应，酶促合成有用的几丁质低聚糖”CarboHydr.Res。

Taichi Usui: "Lysozyme-mediated p-Nitrophenyl Penta-N-acetyl-beta-chitopentaoside Production in Aqueousdimethylsulfoxide Solvent System, as a Substrate for lysozyme assay" Agric. Biol. Chem., 53, 383-388 (1989).

Taichi Usui：“溶菌酶介导的对硝基苯基五-N-乙酰基-β-壳五糖苷在水性二甲基亚砜溶剂系统中的生产，作为溶菌酶测定的底物”Agric。

碓氷泰市: Agric.Biol.Chem.53 (2). 383-388 (1989)

臼井靖：农业生物化学53 (2) (1989)。

碓氷泰市: Euro.J.Biochem.

臼井靖：Euro.J.Biochem。