Metabolic Engineering of Rice Cells with Vanillin Synthase Gene (VpVAN) to Produce Vanillin.

Autor: Arya SS; TERI-Deakin Nanobiotechnology Centre, The Energy and Resources Institute, Gurugram, Haryana, 122001, India.; School of Life and Environmental Sciences, Deakin University, Waurn Ponds Campus, Geelong, VIC, 3216, Australia., Mahto BK; TERI-Deakin Nanobiotechnology Centre, The Energy and Resources Institute, Gurugram, Haryana, 122001, India.; University Department of Botany, Ranchi University, Ranchi, Jharkhand, 834008, India., Sengar MS; TERI-Deakin Nanobiotechnology Centre, The Energy and Resources Institute, Gurugram, Haryana, 122001, India., Rookes JE; School of Life and Environmental Sciences, Deakin University, Waurn Ponds Campus, Geelong, VIC, 3216, Australia., Cahill DM; School of Life and Environmental Sciences, Deakin University, Waurn Ponds Campus, Geelong, VIC, 3216, Australia., Lenka SK; TERI-Deakin Nanobiotechnology Centre, The Energy and Resources Institute, Gurugram, Haryana, 122001, India. keshari2u@gmail.com.
Jazyk: angličtina
Zdroj: Molecular biotechnology [Mol Biotechnol] 2022 Aug; Vol. 64 (8), pp. 861-872. Date of Electronic Publication: 2022 Feb 22.
DOI: 10.1007/s12033-022-00470-8
Abstrakt: Vanillin production by metabolic engineering of proprietary microbial strains has gained impetus due to increasing consumer demand for naturally derived products. Here, we demonstrate the use of rice cell cultures metabolically engineered with vanillin synthase gene (VpVAN) as a plant-based alternative to microbial vanillin production systems. VpVAN catalyzes the signature step to convert ferulic acid into vanillin in Vanilla planifolia. As ferulic acid is a phenylpropanoid pathway intermediate in plant cells, rice calli cells are ideal platform for in vivo vanillin synthesis due to the availability of its precursor. In this study, rice calli derived from embryonic rice cells were metabolically engineered with a codon-optimized VpVAN gene using Agrobacterium-mediated transformation. The putative transformants were selected based on their proliferation on herbicide-supplemented N6D medium. Expression of the transgenes were confirmed through a β-glucuronidase (GUS) reporter assay and polymerase chain reaction (PCR) analysis provided evidence of genetic transformation. The semiquantitative RT-PCR and real-time (RT)-qPCR revealed expression of VpVAN in six transgenic calli lines. High-performance liquid chromatography identified the biosynthesis of vanillin in transgenic calli lines, with the highest yielding line producing 544.72 (± 102.50) μg of vanillin -g fresh calli. This work serves as a proof-of-concept to produce vanillin using metabolically engineered rice cell cultures.
(© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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