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野生型高产苯乙醇生产菌株的筛选及发酵参数优化

Screening of High-Yield 2-Phenylethanol Producing Strain from Wild-Type and Optimization of Fermentation Parameters.

作者信息

Zhang Chenshuo, Fan Tingwen, Wang Zhichun, Yu Jiamu, Guo Xiaoming, Jiang Wei, Miao Lili, Yang Huaiyi

机构信息

State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.

State Key Laboratory of Animal Biotech Breeding, College of Biological Sciences, China Agricultural University, Beijing 100193, China.

出版信息

Foods. 2025 Jul 11;14(14):2444. doi: 10.3390/foods14142444.

DOI:10.3390/foods14142444
PMID:40724265
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12294199/
Abstract

2-Phenylethanol (2-PE), an aromatic alcohol with a rose-like fragrance, is widely used in the food, pharmaceutical, and high-end cosmetic industries. In this study, a high-yield 2-PE-producing strain was isolated and identified as based on morphological characterization and taxonomic identification. Fermentation medium components (carbon and nitrogen sources) were optimized through single-factor experiments in shaking flasks, and fermentation medium with 40 g/L glucose, 5 g/L malt extract, 1.75 g/L corn steep liquor, 2.5 g/L yeast extract, 5 g/L malt extract, 1.75 g/L corn steep liquor was considered suitable for 2-PE production. RT-qPCR results indicated that corn steep liquor activates expression of genes related to the shikimate pathway and Ehrlich pathway (, , , and ), thereby promoting the synthesis of 2-PE through these pathways. Excess yeast extract inhibited the expression of and , while enhancing the expression of and , thus promoting the de novo synthesis of 2-PE. Furthermore, fermentation in a 5 L bioreactor was applied to investigate the effects of feeding strategies, inoculum proportion, and pH on 2-PE production. With a pH of 5.5 and10% inoculum proportion, the supplementation of the substrate L-Phe led to a 2-PE production of 4.81 g/L after 24 h of fermentation. Finally, in situ product recovery (ISPR) techniques was applied to alleviate 2-PE cytotoxicity, achieving a production of 6.41 g/L. This process offers a promising strategy for producing 2-PE efficiently and naturally, paving the way for further industrial applications in food, pharmaceutical, and cosmetic sectors.

摘要

2-苯乙醇(2-PE)是一种具有玫瑰香味的芳香醇,广泛应用于食品、制药和高端化妆品行业。在本研究中,通过形态学特征和分类学鉴定,分离并鉴定出一株高产2-PE的菌株。通过摇瓶单因素实验优化了发酵培养基成分(碳源和氮源),含有40 g/L葡萄糖、5 g/L麦芽提取物、1.75 g/L玉米浆、2.5 g/L酵母提取物、5 g/L麦芽提取物、1.75 g/L玉米浆的发酵培养基被认为适合2-PE生产。RT-qPCR结果表明,玉米浆激活了与莽草酸途径和埃利希途径相关基因(、、、和)的表达,从而通过这些途径促进2-PE的合成。过量的酵母提取物抑制了和的表达,同时增强了和的表达,从而促进了2-PE的从头合成。此外,在5 L生物反应器中进行发酵,以研究补料策略、接种比例和pH对2-PE生产的影响。在pH为5.5和接种比例为10%的情况下,添加底物L-苯丙氨酸,发酵24 h后2-PE产量达到4.81 g/L。最后,应用原位产物回收(ISPR)技术减轻2-PE的细胞毒性,产量达到6.41 g/L。该工艺为高效、天然地生产2-PE提供了一种有前景的策略,为在食品、制药和化妆品领域的进一步工业应用铺平了道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/3b49f87c4c20/foods-14-02444-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/914ed5792ddf/foods-14-02444-g0A1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/7e27d71b3676/foods-14-02444-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/2cff61a11c2d/foods-14-02444-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/d7c19290e0cb/foods-14-02444-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/f13057f8259e/foods-14-02444-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/27915de8503a/foods-14-02444-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/641849f529d7/foods-14-02444-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/3b49f87c4c20/foods-14-02444-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/914ed5792ddf/foods-14-02444-g0A1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/7e27d71b3676/foods-14-02444-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/2cff61a11c2d/foods-14-02444-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/d7c19290e0cb/foods-14-02444-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/f13057f8259e/foods-14-02444-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/27915de8503a/foods-14-02444-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/641849f529d7/foods-14-02444-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6ce6/12294199/3b49f87c4c20/foods-14-02444-g007.jpg

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本文引用的文献

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Highly efficient production of 2-phenylethanol by wild-type Saccharomyces bayanus strain.野生型巴氏酵母菌株高效生产 2-苯乙醇。
Bioresour Technol. 2024 Jul;403:130867. doi: 10.1016/j.biortech.2024.130867. Epub 2024 May 20.
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Metabolic engineering strategies for de novo biosynthesis of sterols and steroids in yeast.酵母中甾醇和类固醇从头生物合成的代谢工程策略。
Bioresour Bioprocess. 2021 Nov 5;8(1):110. doi: 10.1186/s40643-021-00460-9.
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RT-qPCR based quantitative analysis of ARO and ADH genes in Saccharomyces cerevisiae and Metschnikowia pulcherrima strains growth white grape juice.基于 RT-qPCR 的酿酒酵母和毕赤酵母菌株在生长白葡萄汁中 ARO 和 ADH 基因的定量分析。
Mol Biol Rep. 2024 Apr 20;51(1):547. doi: 10.1007/s11033-024-09444-2.
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Investigation on screening, identification, and fermentation characteristics of Yunnan olive in the fermented liquid utilizing five strains of Saccharomyces cerevisiae.利用五株酿酒酵母对滇橄榄发酵液进行筛选、鉴定及发酵特性研究。
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