一个序列基因使得在自然酶中广泛使用非正规的氧化辅因子成为可能
Samer Saleh1,2, Ning-Hsiang Hsu3,2, Emma Luu4
1Department of Chemical and Biomolecular Engineering, University of California, Irvine, Irvine, CA, USA.
bioRxiv : the preprint server for biology
|August 6, 2025
概括
研究人员在化 aldehyde脱酶 (ALDH) 中发现了一个保存的 RH/QxxR 基因,该基因能够有效地使用非正规的氧化还原因子 (NRC),如尼古丁胺胺 mononucleotide (NMN+). 这一发现开启了生物制造的新可能性,通过扩大酶兼容性以更便宜,可控制的辅助因子.
科学领域:
- 生物化学 生物化学
- 合成生物学 合成生物学
- 酶工程是什么? 酶工程是什么?
背景情况:
- 非正规的氧化还原共因子 (NRCs) 在生物制造中比尼古丁胺胺氨基二核酸 (酸盐) (NAD(P) +) 具有优势,包括更低的成本和精确的电子转移控制.
- 由于缺乏兼容的酶,NRCs的广泛采用受到阻碍,这对生物技术应用构成了重大挑战.
研究的目的:
- 为了识别具有广泛NRC活动的天然酶.
- 阐明NRC识别和酶内活性的分子基础.
- 为生物制造设计具有增强NRC活性的酶.
主要方法:
- 选阿尔代脱酶 (ALDH) 蛋白家族的NRC活动.
- 保存的RH/QxxR序列图案的识别和表征.
- 对ALDH活跃地点的结构和动态分析.
- 通过将RH/QxxR基因引入到多种ALDH支架中来进行酶工程.
主要成果:
- 确定了一个保存的RH/QxxR动机,使得NRC在自然ALDHs中的广泛活性成为可能.
- 博斯塔鲁斯ALDH3a1和Pseudanabaena bicepsALDH表现出高的转换率与尼古丁胺胺单核酸 (NMN+),相当于或超过NAD+.
- 在工程ALDH中,RH/QxxR动机增强了NMN+活性高达60倍,并支持了与其他合成NRC,如1 - - 2 - 碳甲基尼古丁胺 (AmNA+) 的活性.
结论:
- RH/QxxR动机是ALDH中NRC活动的关键决定因素,促进了辅因子结合和活性位点预组织.
- 这个图案代表了大自然设计NRC兼容酶的战略,为未来的生物催化剂开发提供了宝贵的蓝图.
- 这些发现为利用酶中潜伏的进化可塑性提供了一条途径,为生物制造NRC活性生物催化剂的工程创造了优越的起点.
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