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Updated: Jul 8, 2025

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一个可扩展的过度表达一个温稳重组合聚胺标签碳酸乙酶在兰巴促进体下的可扩展的过度表达:净化,特性和蛋白质建模

Nadia A Soliman¹, Safaa M Ali², Mahmoud E A Duab³

¹Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City of Scientific Research and Technological Applications (SRTA-City), Universities and Research Institutes Zone, New Borg El-Arab City, Alexandria, 21934, Egypt. nadiastuttgart@yahoo.com.

Journal, genetic engineering & biotechnology

|December 12, 2023

概括

一种来自Geobacillus sp.的新型重组性碳酸乙酶 (EstB). 被成功地表达和净化. 这种酶具有出色的pH值和温度稳定性,使其成为各种工业应用的有希望的生物催化剂.

科学领域:

生物技术是生物技术.
酶学是一种酶学.
蛋白质工程是指蛋白质的工程.

背景情况:

雌激酶是白色生物技术中的关键酶,广泛用于各种工业应用.
它们的应用范围包括制药,化品,食品加工,洗剂和环境修复等.

研究的目的:

为了设计和表达来自Geobacillus sp.的复合性碳酸乙酶 (EstB). 为了提高工业实用性.
为了促进净化和表征复合性EstB的生物化学特性.

主要方法:

在Geobacillus sp.的C端添加了一个聚胺标签. 雌激酶基因 (EstB) 是一种基因.
再组合的EstB在Lambda促进体下表达在大肠杆菌 (E. coli) 中,并在7-L生物反应器中扩大规模.
通过固定金属亲和色谱 (IMAC) 实现净化,随后进行生物化学表征.

主要成果:

再组合的EstB是一种499氨基酸多,与已知的基酶具有很高的同质性.
该酶在pH值8.0时表现出最佳活性,在pH值8-10的范围内稳定.
最佳温度为65°C,在高温下显著保持活动 (在60°C下1小时保持70%).
确定了纯化的酶的动力参数 (Km,Vmax,Kcat,Kcat/Km).

结论:

成功创建和过度表达了从热爱性Geobacillus sp.中标记的复合6x-His标记的EstB. 已经实现了.
通过生物化学表征证实了该酶强大的pH值和温度稳定性.
具有特征的EstB在需要稳定的生物催化剂的各种工业领域具有很大的应用潜力.

关键词:

亲和色谱法是一种亲和色谱法.生物反应器是一种生物反应器.希斯蒂丁的亲和度标签扩大规模扩大规模亚克隆子克隆可以实现.

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