Department of Bio and Nano Technology, Guru Jambheshwar University of Science and Technology, Hisar, Haryana, 125001, India.
Plant Tissue Culture and Genetic Engineering Lab, Department of Biotechnology, Ministry of Science and Technology (Government of India), National Agri-Food Biotechnology Institute (NABI), Sector 81, Knowledge City, S.A.S. Nagar, Mohali, Punjab, 140306, India.
Planta. 2023 Oct 28;258(6):107. doi: 10.1007/s00425-023-04256-3.
The present investigation profoundly asserted the catalytic potential of plant-based aldo-ketoreductase, postulating its role in polyketide biosynthesis and providing new insights for tailored biosynthesis of vital plant polyketides for therapeutics. Plants hold great potential as a future source of innovative biocatalysts, expanding the possibilities within chemical reactions and generating a variety of benefits. The aldo-keto reductase (AKR) superfamily includes a huge collection of NAD(P)H-dependent oxidoreductases that carry out a variety of redox reactions essential for biosynthesis, detoxification, and intermediary metabolism. The present study involved the isolation, cloning, and purification of a novel aldo-ketoreductase (AvAKR) from the leaves of Aloe vera (Aloe barbadensis Miller) by heterologous gene expression in Escherichia coli based on the unigene sequences of putative ketoreductase and cDNA library screening by oligonucleotide hybridization. The in-silico structural analysis, phylogenetic relationship, and molecular modeling were outranged to approach the novelty of the sequence. Additionally, agroinfiltration of the candidate gene tagged with a green fluorescent protein (GFP) was employed for transient expression in the Nicotiana benthamiana to evaluate the sub-cellular localization of the candidate gene. The AvAKR preferred cytoplasmic localization and shared similarities with the known plant AKRs, keeping the majority of the conserved active-site residues in the AKR superfamily enzymes. The enzyme facilitated the NADPH-dependent reduction of various carbonyl substrates, including benzaldehyde and sugars, proclaiming a broad spectrum range. Our study successfully isolated and characterized a novel aldo-ketoreductase (AvAKR) from Aloe vera, highlighting its versatile NADPH-dependent carbonyl reduction proficiency therewith showcasing its potential as a versatile biocatalyst in diverse redox reactions.
本研究深入探讨了植物来源的醛酮还原酶的催化潜力，提出其在聚酮生物合成中的作用，并为定制生物合成具有治疗价值的重要植物聚酮提供了新的见解。植物作为未来创新生物催化剂的来源具有巨大的潜力，扩展了化学反应的可能性，并产生了多种益处。醛酮还原酶（AKR）超家族包含大量的 NAD(P)H 依赖性氧化还原酶，它们执行各种氧化还原反应，这些反应对于生物合成、解毒和中间代谢至关重要。本研究通过基于假定的酮还原酶的基因和 cDNA 文库筛选的寡核苷酸杂交的同源基因表达，从库拉索芦荟（Aloe barbadensis Miller）的叶子中分离、克隆和纯化了一种新型的醛酮还原酶（AvAKR）。通过计算机结构分析、系统发育关系和分子建模对序列的新颖性进行了研究。此外，通过带有绿色荧光蛋白（GFP）的候选基因的农杆菌浸润进行瞬时表达，在 Nicotiana benthamiana 中评估候选基因的亚细胞定位。AvAKR 优先定位于细胞质，与已知的植物 AKR 具有相似性，保留了 AKR 超家族酶中的大多数保守活性位点残基。该酶促进各种羰基底物的 NADPH 依赖性还原，包括苯甲醛和糖，表现出广泛的谱范围。我们的研究成功地从库拉索芦荟中分离和鉴定了一种新型的醛酮还原酶（AvAKR），突出了其在多种羰基还原中的多功能 NADPH 依赖性碳还原能力，展示了其作为各种氧化还原反应中多功能生物催化剂的潜力。
