粗神经孢子虫醛脱氢酶的工程研究

IF 6.2 1区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY
Zhengzhuang Chen, Chang Wang, Ruijun Wang, Li Li* and Jikun Du*, 
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引用次数: 0

摘要

微生物合成是一种很有前途的番石榴素生产策略,但产量低。醛脱氢酶(ALDH)的催化效率有限,使其成为微生物合成藏红花素的关键靶点。本研究对粗神经孢子虫NcALDH进行了生物化学表征,并通过结构引导酶工程技术显著提高了NcALDH的催化活性。NcALDH在30℃、pH 8.0条件下对西红花素二醛的活性最佳。然后,通过重新设计NcALDH的底物结合袋,获得了催化活性增强的G118W突变体。此外,通过分子动力学模拟阐明了G118W突变体活性增强的机制。最后,利用含G118W的重组菌株作为全细胞生物催化剂,获得了77.86 μM crocetin,比对照菌株的滴度提高了113%。本研究为微生物高效合成西红花苷奠定了坚实的基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Engineering of an Aldehyde Dehydrogenase from Neurospora crassa for Crocetin Production

Engineering of an Aldehyde Dehydrogenase from Neurospora crassa for Crocetin Production

Microbial synthesis represents a promising strategy for crocetin production but suffers from a low yield. The limited catalytic efficiency of aldehyde dehydrogenase (ALDH) highlighted it as a critical target for the microbial synthesis of crocetin. In this study, the NcALDH from Neurospora crassa was biochemically characterized and its catalytic activity was significantly improved through structure-guided enzyme engineering. NcALDH showed optimal activity toward crocetin dialdehyde at 30 °C and pH 8.0. Then, the G118W mutant with enhanced catalytic activity was obtained by redesigning the substrate-binding pocket of NcALDH. Furthermore, the mechanism behind the enhanced activity of the G118W mutant was elucidated through molecular dynamics simulations. Finally, 77.86 μM crocetin was achieved by using the recombinant strain harboring G118W as a whole-cell biocatalyst, marking a 113% increase compared with the titer achieved by the control strain. This study establishes a strong foundation for high-efficiency microbial crocetin synthesis.

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来源期刊
Journal of Agricultural and Food Chemistry
Journal of Agricultural and Food Chemistry 农林科学-农业综合
CiteScore
9.90
自引率
8.20%
发文量
1375
审稿时长
2.3 months
期刊介绍: The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.
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