Production of novel sugars by microbial fermentation

微生物发酵生产新型糖

• 批准号: 11555219
• 负责人: IIJIMA Shinji
• 金额: $ 8.83万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11555219/
• 关键词: Sugar technology; Microbial polysaccharide; Sugar transferase; Gene Engineering; Gene expression; Design; ガン転移

We are studying biosynthesis of the capsular polysaccharide of Streptococcus agalactiae, which has sialyl lactosamine polymer. Sialyl Lewis sugar was known as a tumor specific antigen. We are trying to produce these kinds of useful sugar by recombinant DNA technology.For this purpose, we tried to increase the production level of the sugar. We have already isolated a gene cluster, which contain multigenes those encoding sugar synthesis genes. Among them, cpsIaR gene was introduced in Escherichia coli with other sugar synthesis genes. This recombinant E.coli showed higher enzyme activity. Under high osmorality with 25% sucrose, both CpsIaR and other sugar biosynthesis enzymes level also increased. Together, these results suggest that CpsIaR positively regulates sugar biosynthesis.We also analyzed the substrate specificity of CpsIaJ which was identified as a β-1-4-galactosyl transferase. We found that the enzyme could transfer galactosyl moiety to both artificial acceptor and GlcNAc-Gal-Glc trisaccharide.

我们正在研究无乳链球菌荚膜多糖的生物合成，它具有唾液酸乳糖胺聚合物。唾液酸刘易斯糖是一种肿瘤特异性抗原。我们正试图通过重组DNA技术生产这些有用的糖，为此，我们试图提高糖的生产水平。我们已经分离出了一个基因簇，它包含了编码糖合成基因的多个基因。其中，cpsIaR基因与其他糖合成基因一起导入大肠杆菌。重组大肠杆菌具有较高的酶活。在25%蔗糖的高渗条件下，CpsIaR和其他糖合成酶的水平也有所提高。我们还分析了CpsIaJ的底物特异性，CpsIaJ被鉴定为β-1-4-半乳糖基转移酶。我们发现该酶可以将半乳糖基转移到人工受体和GlcNAc Gal Glc三糖上。

项目成果

Shinji Mizuarai, Masamichi Kamihira, Ken-Ichi Nishijima and Shinji Iijima: "Integrase-mediated nonviral gene transfection with enhanced integration efficiency"Journal of Bioscience and Bioengineering. 88[5]. 461-467 (1999)

Shinji Mizuarai、Masamichi Kamihira、Ken-Ichi Nishijima 和 Shinji Iijima：“整合酶介导的非病毒基因转染，具有增强的整合效率”生物科学与生物工程杂志。

Yuichi Machida: "Structure and function of a novel coliphage-associated sialidase"FEMS Microbiology Letters. Vol.102. 333-337 (2000)

Yuichi Machida：“新型大肠杆菌噬菌体相关唾液酸酶的结构和功能”FEMS 微生物学快报。

Yuichi Machida,Katsuhide Miyake,Kouji Hattori,Shin Yamamoto,Mitsuo Kawase and Shinji Iijima: "Structure and function of a novel coliphage-associated sialidase"FEMS Microbiology Letters. 182. 333-337 (2000)

Yuichi Machida、Katsuhide Miyake、Kouji Hattori、Shin Yamamoto、Mitsuo Kawase 和 Shinji Iijima：“新型大肠杆菌噬菌体相关唾液酸酶的结构和功能”FEMS 微生物学快报。