细胞色素P450和ALDH在脂质耶氏菌D21173转化柑橘香精（+）价酮中的关键作用

IF 3.4 2区农林科学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Food and Bioproducts Processing Pub Date : 2025-09-15 DOI:10.1016/j.fbp.2025.09.010

Xiao Li , Gang Fan , Yanli Xie , Jinchu Yang , Yingjie Feng , Siyi Pan , Linhua Huang , Guijie Li , Yujiao Cheng

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引用次数: 0

摘要

（+）-诺卡酮是一种重要的倍半萜化合物，用于食品香精、香料、农业和医药。本研究采用基因敲除技术，研究了脂质Y. lipolytica D21173基因2911（CYP450）和基因0658（ALDH）对柑橘风味(+)-价烯生物转化为(+)-努卡酮的影响。结果表明，CYP450基因共含有513个氨基酸，其中α-螺旋44.44 %，随机盘绕38.60 %，延伸链13.06 %，β-转角3.90 %。ALDH包含572个氨基酸，α-螺旋占44.23 %，延伸链占12.06 %，β-转角占5.42 %，随机卷曲占38.29 %。此外，CYP450和ALDH参与调节Y. lipolytica D21173的生长，并参与其对各种环境胁迫的响应，包括不同的离子和氧化应激。当Fe2 +浓度为15 mmol/L时，野生型菌株的相对抑制率为68.61 %，溶脂耶氏菌-Δ0658-NTCR菌株的相对抑制率为74.87 %，溶脂耶氏菌-Δ0658-NTCR菌株的相对抑制率为81.93 %。当H₂O₂浓度为30 mmol/L时，3株菌株对野生型的相对抑制值分别为21.91 %、37.13 %和37.67 %。聚脂耶氏菌-Δ0658-NTCR和聚脂耶氏菌-Δ2911-NTCR转化的(+)-努卡酮含量分别为203.89 mg/L （P <； 0.05）和20.51 mg/L (P <； 0.001)，说明基因2911（CYP450）和基因0658（ALDH）参与了(+)-价位生物转化为(+)-努卡酮的调控。这是首次利用基因缺失（NTC作为耐药基因）来研究细胞色素P450和ALDH在脂质体Y. lipolytica D21173中(+)-价键生物转化为(+)-nootkatone的重要功能。为今后构建柑橘香精(+)-诺卡酮生物制造平台的细胞工厂奠定基础。

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The key role of Cytochrome P450 and ALDH in citrus flavor (+)-valencene biotransformation to (+)-nootkatone with Yarrowia lipolytica D21173

(+)-Nootkatone is an important sesquiterpene compound that is used in food flavor, fragrances, agriculture, and medicine. In this study, gene knockout technology was used to investigate the effect of gene2911(CYP450) and gene0658(ALDH) of Y. lipolytica D21173 on citrus flavor (+)-valencene biotransformation to (+)-nootkatone. The results showed that the prediction of CYP450 contained 513 amino acids with 44.44 % α-helix, 38.60 % random coiling, 13.06 % extended chain, and 3.90 % β-turn angle. The ALDH included 572 amino acids with 44.23 % of α-helices, 12.06 % of extended strands, 5.42 % of β-turn angles and 38.29 % of random coiling. Besides, CYP450 and ALDH were involved in regulating the growth of Y. lipolytica D21173 and contributed to its response to various environmental stresses, including different ionic and oxidative stresses. The highest relative inhibition observed was 68.61 % for the wild-type strain, 74.87 % for the Yarrowia lipolytica-Δ0658-NTCR strain, and 81.93 % for the Yarrowia lipolytica-Δ0658-NTCR strain when the concentration of Fe^2 + was 15 mmol/L. When exposed to a H₂O₂ concentration of 30 mmol/L, the relative inhibition values of the three strains were 21.91 % for the wild-type, 37.13 % for Yarrowia lipolytica-Δ0658-NTCR, and 37.67 % for Yarrowia lipolytica-Δ0658-NTCR. The content of (+)-nootkatone transformed by Yarrowia lipolytica-Δ0658-NTCR and Yarrowia lipolytica-Δ2911-NTCR were 203.89 mg/L (P < 0.05) and 20.51 mg/L (P < 0.001), which indicated that gene2911(CYP450) and gene0658(ALDH) were involved in the regulation of (+)-valencene biotransformation to (+)-nootkatone. This is the first time that gene deletion (NTC as resistance gene) is used to investigate the vital function of cytochrome P450 and ALDH in Y. lipolytica D21173 of (+)-valencene biotransformation to (+)-nootkatone. It will provide a solid foundation on constructing cell factories of the citrus flavor (+)-nootkatone biomanufacturing platform in the future.

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来源期刊

Food and Bioproducts Processing 工程技术-工程：化工

CiteScore

9.70

自引率

4.30%

发文量

115

审稿时长

24 days

期刊介绍： Official Journal of the European Federation of Chemical Engineering: Part C FBP aims to be the principal international journal for publication of high quality, original papers in the branches of engineering and science dedicated to the safe processing of biological products. It is the only journal to exploit the synergy between biotechnology, bioprocessing and food engineering. Papers showing how research results can be used in engineering design, and accounts of experimental or theoretical research work bringing new perspectives to established principles, highlighting unsolved problems or indicating directions for future research, are particularly welcome. Contributions that deal with new developments in equipment or processes and that can be given quantitative expression are encouraged. The journal is especially interested in papers that extend the boundaries of food and bioproducts processing. The journal has a strong emphasis on the interface between engineering and food or bioproducts. Papers that are not likely to be published are those: • Primarily concerned with food formulation • That use experimental design techniques to obtain response surfaces but gain little insight from them • That are empirical and ignore established mechanistic models, e.g., empirical drying curves • That are primarily concerned about sensory evaluation and colour • Concern the extraction, encapsulation and/or antioxidant activity of a specific biological material without providing insight that could be applied to a similar but different material, • Containing only chemical analyses of biological materials.