为提高香兰素、4-羟基苯甲酸和香草醇的产量而对红豆杉毛根培养物进行代谢工程改造。

IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Frontiers in Bioengineering and Biotechnology Pub Date : 2024-10-02 eCollection Date: 2024-01-01 DOI:10.3389/fbioe.2024.1435190
Zakir Husain, Zafar Iqbal Warsi, Sana Khan, Ganesan Mahendran, Shama Afroz, Ashish Chandran, Praveen Kumar Kashyap, Kahkashan Khatoon, Gazala Parween, Sudeep Tandon, Laiq Ur Rahman
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引用次数: 0

摘要

香草的味道由多种复杂的化合物混合而成,其中最突出的是香兰素,此外还有香草醇和 4-羟基苯甲酸。从香草豆中提取的天然香兰素价格昂贵,因此研究人员采用异源合成法,在植物宿主体内生产与天然相同的香兰素。因此,需要采用其他传统种植和采集方法来弥补供需之间的巨大差距。目前的研究成功开发了一种从叶片诱导毛根形成的方法。该研究将香兰素合成酶(VpVAN)基因整合到转基因毛细根系中,合成香兰素相关化合物。通过 PCR 分析证实了转基因根中存在 VpVAN 基因。此外,RT-qPCR 分析也证明了 VpVAN 基因在转基因根系中的表达。转基因毛根克隆 H1、H2 和 H5 的香兰素、香草醇和 4-羟基苯甲酸产量均有所提高。茉莉酸甲酯(MJ)和水杨酸(SA)的激发进一步提高了粗壮毛根中这些化合物的产量。60 天后观察到毛状根生物量最大,毛状根克隆 H5 和 HR2 分别获得了最大的香兰素和 4-羟基苯甲酸合成量。香草醇 HR2 是在培养的第 45 天获得的。伤口相关激素茉莉酸甲酯和水杨酸的激发提高了香兰素、香草醇和 4-羟基苯甲酸的产量,其中香兰素增加了 215 倍,香草醇增加了 13 倍,4-羟基苯甲酸增加了 21 倍。研究结果表明,用 VpVAN 基因建立转基因毛根培养物是提高香草香味化合物(如香兰素、香草醇和 4-羟基苯甲酸)产量的一种很有前途的替代方法。目前已开发出一种具有成本效益的方案,利用毛根培养法大规模生产酚类化合物。这种方法既能满足对这些物质日益增长的需求,又能降低天然香兰素的生产成本,适合工业规模的应用。
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Metabolic engineering of hairy root cultures in Beta vulgaris for enhanced production of vanillin, 4-hydroxybenzoic acid, and vanillyl alcohol.

The flavor of vanilla is a complex blend of compounds, with vanillin as the most prominent, along with vanillyl alcohol and 4-hydroxybenzoic acid. Natural vanillin extracted from vanilla beans is expensive, so researchers use heterologous synthesis to produce nature-identical vanillin in plant hosts. Consequently, alternative traditional farming and gathering methods are required to bridge the significant disparity between supply and demand. The current research successfully developed a method to induce hairy root formation from leaves. It integrated the Vanillin synthase (VpVAN) gene into transgenic hairy root lines of Beta vulgaris, synthesizing vanillin-related compounds. The presence of the VpVAN gene in transgenic roots was confirmed using PCR analysis. Additionally, RT-qPCR analysis demonstrated the expression of the VpVAN gene in the transgenic root lines. The transgenic hairy root clones H1, H2, and H5 showed enhanced vanillin production, vanillyl alcohol, and 4-hydroxybenzoic acid. Elicitation with methyl jasmonate (MJ) and salicylic acid (SA) further improved the production of these compounds in B. vulgaris hairy roots. The maximum hairy root biomass was observed after 60 days, with the maximum synthesis of vanillin and 4-hydroxybenzoic acid obtained from hairy root clones H5 and HR2, respectively. Vanillyl alcohol HR2 was obtained on the 45th day of cultivation. Elicitation with wound-associated hormone methyl jasmonate and salicylic acid enhanced the yield of vanillin, vanillyl alcohol, and 4-hydroxybenzoic acid, with a 215-fold increase in vanillin, a 13-fold increase in vanillyl alcohol, and a 21 fold increase in 4-hydroxybenzoic acid. The study results indicate that establishing transgenic hairy root cultures with the VpVAN gene is a promising alternative method for enhancing the production of vanilla flavor compounds such as vanillin, vanillyl alcohol, and 4-hydroxybenzoic acid. A cost-effective protocol has been developed to mass-produce phenolic compounds using a hairy root culture of B. vulgaris. This approach addresses the increasing demand for these substances while reducing the cost of natural vanillin production, making it suitable for industrial-scale applications.

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来源期刊
Frontiers in Bioengineering and Biotechnology
Frontiers in Bioengineering and Biotechnology Chemical Engineering-Bioengineering
CiteScore
8.30
自引率
5.30%
发文量
2270
审稿时长
12 weeks
期刊介绍: The translation of new discoveries in medicine to clinical routine has never been easy. During the second half of the last century, thanks to the progress in chemistry, biochemistry and pharmacology, we have seen the development and the application of a large number of drugs and devices aimed at the treatment of symptoms, blocking unwanted pathways and, in the case of infectious diseases, fighting the micro-organisms responsible. However, we are facing, today, a dramatic change in the therapeutic approach to pathologies and diseases. Indeed, the challenge of the present and the next decade is to fully restore the physiological status of the diseased organism and to completely regenerate tissue and organs when they are so seriously affected that treatments cannot be limited to the repression of symptoms or to the repair of damage. This is being made possible thanks to the major developments made in basic cell and molecular biology, including stem cell science, growth factor delivery, gene isolation and transfection, the advances in bioengineering and nanotechnology, including development of new biomaterials, biofabrication technologies and use of bioreactors, and the big improvements in diagnostic tools and imaging of cells, tissues and organs. In today`s world, an enhancement of communication between multidisciplinary experts, together with the promotion of joint projects and close collaborations among scientists, engineers, industry people, regulatory agencies and physicians are absolute requirements for the success of any attempt to develop and clinically apply a new biological therapy or an innovative device involving the collective use of biomaterials, cells and/or bioactive molecules. “Frontiers in Bioengineering and Biotechnology” aspires to be a forum for all people involved in the process by bridging the gap too often existing between a discovery in the basic sciences and its clinical application.
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