分子建模和优化II型E.colil-阿斯巴尔金酶活性通过in silico设计和体外定位导向突变发生
Mahdieh Mahboobi1,2, Ali-Hatef Salmanian3, Hamid Sedighian2
1Department of Systems Biotechnology, Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrake- Pajoohesh Blvd., 15th Km, Tehran-Karaj Highway, P.O. Box 14965-161, Tehran, Iran.
The protein journal
|August 27, 2023
概括
G88Q突变显著增强L-阿斯巴拉基酶 (L-ASNase) 活性,改善其治疗急性淋巴细胞白血病 (ALL) 和工业应用的治疗潜力.
科学领域:
- 生物化学 生物化学
- 酶工程是什么? 酶工程是什么?
- 计算生物学 计算生物学
背景情况:
- L-阿斯巴拉金酶 (L-ASNase) 对于治疗急性淋巴细胞白血病 (ALL) 至关重要,并用于食品加工.
- 改善L-ASNase的活性和动力学对于提高治疗和工业疗效至关重要.
- 针对特定的残留物,如88位的甘氨酸,为酶优化提供了一种策略.
研究的目的:
- 为了识别88甘氨酸突变,增强L-asparaginase活动和运动性质.
- 通过计算和实验验证G88突变对L-ASNase性能的影响.
主要方法:
- 使用了包括分子动力学 (MD) 模拟和分子对接在内的in silico分析.
- 理性设计原则指导了特定氨基酸替代物的选择和实施.
- 实验室实验涉及突变的L-asparaginase变体的动力特征.
主要成果:
- 在in silico选中,G88Q,G88L,G88K和G88A被确定为潜在的有益突变.
- 与野生型大肠杆菌K12 ASNase II相比,G88Q突变体的催化效率 (kcat/Km) 增加了36.32%.
- 动态表征证实了G88Q突变的更高的周转率 (kcat),其中阿斯巴拉金 (ASN) 作为基质.
结论:
- 显然,G88Q突变比其他测试的突变 (G88L,G88K,G88A) 更有效地改善了L-氨酸酶动力学.
- 这种优化的L-ASNase变体对更高效的临床和工业应用具有前景.
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