Metabolic-Engineering Approach to Enhance Vanillin and Phenolic Compounds in Ocimum Sanctum (CIM-Angana) via VpVAN Overexpression.

IF 5.4 2区生物学 Q1 PLANT SCIENCES

Physiologia plantarum Pub Date : 2024-11-01 DOI:10.1111/ppl.70005

Zakir Husain, Sana Khan, Aqib Sarfraz, Zafar Iqbal, Ashish Chandran, Kahkashan Khatoon, Gazala Parween, Farah Deeba, Shama Afroz, Feroz Khan, Ratnasekhar Ch, Laiq Ur Rahman

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引用次数: 0

Abstract

Transgenic Ocimum sanctum plants were engineered to produce vanillin by overexpressing the VpVAN gene using Agrobacterium-mediated transformation. Positive transformants developed shoots within 4-5 weeks and were transferred to a root induction medium and four independent transformants with no observed adverse effects were kept for anlysis. Quantitative RT-PCR indicated significantly higher VpVAN expression in transgenic lines AG_3 and AG_1, impacting the phenylpropanoid pathway and phenolic compound accumulation. Molecular docking studies indicated ferulic acid's higher binding affinity to vanillin synthase than eugenol. LC-MS/MS analysis revealed a marked increase in vanillin production in transgenic lines compared to wild type, with AG_3 exhibiting the highest vanillin content (1.98 ± 0.0047 mg/g extract) and AG_1 following (1.49 ± 0.0047 mg/g extract). AG_3 also showed elevated levels of benzoic acid, 4-hydroxy benzyl alcohol, and ferulic acid. This study highlights the potential of metabolic engineering in O. sanctum for enhanced vanillin production, suggesting pathways for large-scale production of natural vanillin and other valuable compounds in transgenic plants.

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利用农杆菌介导的转化法，通过过表达 VpVAN 基因，培育出可生产香兰素的转基因欧琴圣草植株。阳性转化体在 4-5 周内长出新芽，然后转移到根诱导培养基上，保留四个未观察到不良影响的独立转化体进行分析。定量 RT-PCR 结果表明，转基因品系 AG_3 和 AG_1 的 VpVAN 表达量明显增加，影响了苯丙氨酯途径和酚类化合物的积累。分子对接研究表明阿魏酸与香兰素合成酶的结合亲和力高于丁香酚。LC-MS/MS 分析显示，与野生型相比，转基因品系的香兰素产量明显增加，其中 AG_3 的香兰素含量最高（1.98 ± 0.0047 mg/g），AG_1次之（1.49 ± 0.0047 mg/g）。AG_3 的苯甲酸、4-羟基苯甲醇和阿魏酸含量也有所提高。这项研究强调了代谢工程在提高香兰素产量方面的潜力，为转基因植物大规模生产天然香兰素和其他有价值的化合物提供了途径。

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来源期刊

Physiologia plantarum 生物-植物科学

CiteScore

11.00

自引率

3.10%

发文量

224

审稿时长

3.9 months

期刊介绍： Physiologia Plantarum is an international journal committed to publishing the best full-length original research papers that advance our understanding of primary mechanisms of plant development, growth and productivity as well as plant interactions with the biotic and abiotic environment. All organisational levels of experimental plant biology – from molecular and cell biology, biochemistry and biophysics to ecophysiology and global change biology – fall within the scope of the journal. The content is distributed between 5 main subject areas supervised by Subject Editors specialised in the respective domain: (1) biochemistry and metabolism, (2) ecophysiology, stress and adaptation, (3) uptake, transport and assimilation, (4) development, growth and differentiation, (5) photobiology and photosynthesis.