人类3'-adenosine-5'-phosphosulfate合成酶1的结构引导工程,以增强PAPS的生物合成
Ruixue Zhang1, Jiajun Huang1, Jiali Gu2
1National Key Laboratory for Development and Utilization of Forest Food Resources, Zhejiang A & F University, Hangzhou, Zhejiang 311300, China.
Journal of agricultural and food chemistry
|November 6, 2025
概括
研究人员使用半比率设计将3'-adenosine 5'-phosphosulfate synthase 1 (hPAPSS1) 活性提高了2.22倍. 这改善了PAPS的生物合成,PAPS是硫化修饰的关键分子.
科学领域:
- 生物化学 生物化学
- 酵素工程是什么意思 酵素工程
- 分子生物学分子生物学
背景情况:
- 3 - adenosine 5 -phosphosulfate (PAPS) 对于生物硫化至关重要.
- 人类双功能PAPS合成酶1 (hPAPSS1) 有效地合成PAPS.
- 目前hPAPSS1的局限性包括ATP亲和力和催化活性较差.
研究的目的:
- 为了增强hPAPSS1.1.的活性和ATP结合亲和力.
- 克服PAPS生物合成中对硫烯化合物的局限性.
- 为了阐明hPAPSS1催化增强的机制.
主要方法:
- 半合理的酶设计策略.
- 位点定向的突变发生 (C207G,F560W).
- 酶活性测定和分子动力学模拟.
主要成果:
- 一种组合变种 (hPAPSS1-C207G/F560W) 显示活动增加了2.22倍.
- 实现了34.45%的PAPS转换率.
- C207G突变提高了C环的灵活性和周转率;F560W优化了ATP结合.
结论:
- 确定了hPAPSS1酶活性中的关键瓶.
- 证明了增强催化性能的机制.
- 启用了有效的PAPS合成用于生物技术应用.
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