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Identification and functional characterization of a novel aldo–keto reductase from Aloe vera

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Abstract

Main conclusion

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.

Abstract

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.

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Data availability

The authors declare that all data supporting the findings of this study are available within the article. The nucleotide and deduced amino acid sequence of AvAKR have been submitted to the NCBI database (https://www.ncbi.nlm.nih.gov/) via GenBank accession number BankIt2620826 AvAKR. The original transcriptome data set could be accessed through Sequence Read Archive (NCBI) via accession number SRR5167034 (https://www.ncbi.nlm.nih.gov/sra).

Abbreviations

AKR:

Aldo–keto reductase

GFP:

Green fluorescent protein

LB:

Luria broth

NCBI:

National Center for Biotechnology Information

PKR:

Polyketide reductase

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Acknowledgements

The authors would like to thank Guru Jambheshwar University of Science and Technology (GJUS&T), Hisar, Haryana, National Agri-Food Biotechnology Institute (NABI), Mohali and Government of Uttar Pradesh for providing the necessary facilities and a suitable environment for executing the present study. AJ, SC, SS, and GS acknowledge the Council of Scientific and Industrial Research (CSIR), Department of Biotechnology (DBT), University Grant Commission (UGC) and the Department of Science and Technology (DST), respectively for the fellowship. AJ, SS and GS are thankful to GJUS&T for Ph.D registration.

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Authors and Affiliations

  1. Department of Bio and Nano Technology, Guru Jambheshwar University of Science and Technology, Hisar, Haryana, 125001, India

    Alka Jangra, Sonia Sihag, Garima Sharma & Vinod Chhokar

  2. 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

    Siddhant Chaturvedi & Siddharth Tiwari

  3. Goswami Tulsidas Government Post Graduate College (Bundelkhand University, Jhansi), Karwi, Chitrakoot, Uttar Pradesh, 210205, India

    Siddhant Chaturvedi

Authors
  1. Alka Jangra
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  2. Siddhant Chaturvedi
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  3. Sonia Sihag
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  4. Garima Sharma
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  6. Vinod Chhokar
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Contributions

VC and AJ conceptualized the article and designed the experiments. AJ and SC conducted experiments and prepared the original draft. SS and GS contributed to writing the manuscript and data analysis. AJ, SC, and ST contributed to data interpretation and curation. VC contributed to the proofreading of manuscript. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Vinod Chhokar.

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Communicated by Dorothea Bartels.

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Jangra, A., Chaturvedi, S., Sihag, S. et al. Identification and functional characterization of a novel aldo–keto reductase from Aloe vera. Planta 258, 107 (2023). https://doi.org/10.1007/s00425-023-04256-3

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