查佩罗宁的共同表达和化学修饰使得从大肠杆菌细胞质中产生活性微生物转谷氨酶的产生成为可能
Zitao Li1, Shuang Xing1, Jing Liu2
1National Glycoengineering Research Center, NMPA Key Laboratory for Quality Research and Evaluation of Carbohydrate-Based Medicine, and Shandong Key Laboratory of Carbohydrate Chemistry and Glycobiology, Shandong University, Qingdao 266237, Shandong, China.
International journal of biological macromolecules
|October 14, 2023
概括
这项研究提出了一种新的方法,用于生产高纯度的微生物转质氨酶 (MTG),使用分子沙佩罗宁策略和化学修饰. 优化过程产生了活性MTG,适用于生物宏分子修饰应用.
科学领域:
- 生物化学 生化学
- 蛋白质工程是指蛋白质工程.
- 酶学 是一种酶学.
背景情况:
- 微生物转胺酶 (MTG) 是一种用于修改生物大分子的有价值酶.
- 有效生产和净化活跃的MTG仍然具有挑战性.
研究的目的:
- 开发一种高产量的表达和微生物转谷氨酶 (MTG) 在大肠杆菌中的净化策略.
- 为了实现生物技术应用的MTG的高纯度和特异性活性.
主要方法:
- 在大肠杆菌细胞质中用于高水平MTG表达的分子Chaperonin策略.
- 利用小分子介导的化学修饰 (乙化) 在净化过程中有效地去除 propeptide chaperonin.
- 使用质谱法 (MS) 鉴定了纯化的乙化MTG.
主要成果:
- 从养批次培养中获得高MTG产量,高达300mg/L.
- 在化学脱后获得具有高特异性活性 (49.6 U/mg) 的净化MTG.
- 通过MS分析确认了MTG的氨基基组的乙化,表明了掩盖的活性位点.
结论:
- 开发的方法使高纯度,高活性MTG的高效生产成为可能.
- 乙化MTG通过防止自我聚合和基质交叉链接提供优势.
- 这一战略增强了MTG在蛋白质聚合和PEGylation应用中的实用性.
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