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工程双功能银河酶/基转移酶奇米拉用于增强的UDP-d-基生产

Jin-Da Zhuang¹, Jin-Min Shi¹, Chen-Cheng Hong¹

¹Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology Nanjing Agricultural University, 1 Weigang, 210095 Nanjing, China.

|July 26, 2024

概括

研究人员设计了新的双功能酶,以高效地生物技术生产尿素二酸盐-d-xylose (UDP-d-xylose),这是生物制药的关键前体,如肝素. 这一突破简化了必不可少的葡萄糖氨基酸甘的合成.

科学领域:

生物技术和生物化学
酵素工程是什么意思酵素工程
葡萄糖生物学葡萄糖生物学

背景情况:

尿素二酸-d-xylose (UDP-d-xylose) 是合成生物制药,如氨酸和糖氨酸甘油的关键的糖氨酸供体.
目前用于UDP-d-xylose生产的方法在效率和可扩展性方面面临挑战.
糖基工程的进步需要改善基本前体的生物催化路径.

研究的目的:

设计新的双功能化学生物催化剂,用于直接将d-西洛斯转化为UDP-d-西洛斯.
为了研究域定向和链接长度对酶功能的影响.
为了优化大规模UDP-d-西洛斯生产的反应条件.

主要方法:

从 *Solitalea canadensis* galactokinase/uridyltransferase 中设计了八种融合蛋白质结构.
在大肠杆菌中融合蛋白的功能表达和特征.
优化pH值,温度,酶度和基质比率,用于催化转化.

主要成果:

首个双功能酶的成功合成,使得直接的d-西洛斯转化为UDP-d-西洛斯.
具有NH₂-GSGGGSGHM-COOH链接器的融合结构表现出优越的表达和催化活性.
确定了最佳条件:pH值7.0,30°C,酶度3.3 mg/mL,未化酶的比例为1:1.
通过优化聚变酶,观察到催化效率 (K_cat/K_m) 提高了10%.

结论:

工程双功能酶为UDP-d-西洛斯合成提供了高效和方便的生物催化途径.
这一进步支持复杂碳水化合物和生物制药的生物技术生产.
这项研究为进一步发展糖基工程和制药应用提供了基础.

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