Husain Zakir, Khan Sana, Sarfraz Aqib, Iqbal Zafar, Chandran Ashish, Khatoon Kahkashan, Parween Gazala, Deeba Farah, Afroz Shama, Khan Feroz, Ch Ratnasekhar, Rahman Laiq Ur
Plant Biotechnology Division, CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, Utter Pradesh, India.
Technology Dissemination and Computational Biology Division, CSIR-Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, Utter Pradesh, India.
Physiol Plant. 2024 Nov-Dec;176(6):e70005. doi: 10.1111/ppl.70005.
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.
通过农杆菌介导的转化过表达VpVAN基因，对转基因罗勒植株进行工程改造以生产香草醛。阳性转化体在4-5周内长出芽，然后转移到生根诱导培养基上，并保留4个未观察到不良影响的独立转化体用于分析。定量RT-PCR表明，转基因株系AG_3和AG_1中VpVAN的表达显著更高，影响了苯丙烷途径和酚类化合物的积累。分子对接研究表明，阿魏酸对香草醛合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，转基因株系中香草醛的产量显著增加，AG_3的香草醛含量最高（1.98±0.0047mg/g提取物），AG_1次之（1.49±0.0047mg/g提取物）。AG_3中苯甲酸、4-羟基苄醇和阿魏酸的含量也有所升高。这项研究突出了罗勒代谢工程在提高香草醛产量方面的潜力，为在转基因植物中大规模生产天然香草醛和其他有价值的化合物提供了途径。
