Trends in the biotechnological production of isoflavonoids in plant cell suspension cultures

Rani, Dolly; Vimolmangkang, Sornkanok

doi:10.1007/s11101-022-09811-6

Cited by 11 publications

(3 citation statements)

References 122 publications

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“…With recent developments in synthetic biology and metabolic engineering, there is a drive for the biotechnology industry to enhance isoflavonoid production in-planta and ex-planta [ 48 , 49 ]. The potential to introduce exogenous isoflavonoid biosynthesis into cereal crops is an exciting avenue, but one in which the outcomes may be limited due to the negative public sentiment and regulatory barriers surrounding genetically modified crops [ 31 ].…”

Section: Discussionmentioning

confidence: 99%

Phylogenetic Analysis and Protein Modelling of Isoflavonoid Synthase Highlights Key Catalytic Sites towards Realising New Bioengineering Endeavours

2022

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Isoflavonoid synthase (IFS) is a critical enzyme for the biosynthesis of over 2400 isoflavonoids. Isoflavonoids are an important class of plant secondary metabolites that have a range of pharmaceutical and nutraceutical properties. With growing interest in isoflavonoids from both research and industrial perspectives, efforts are being forwarded to enhance isoflavonoid production in-planta and ex-planta; therefore, in-silico analysis and characterisation of available IFS protein sequences are needed. The present study is the first-ever attempt toward phylogenetic analysis and protein modelling of available IFS protein sequences. Phylogenetic analysis has shown that IFS amino acid sequences have 86.4% pairwise identity and 26.5% identical sites, and the sequences were grouped into six different clades. The presence of a β-hairpin and extra loop at catalytic sites of Trifolium pratense, Beta vulgaris and Medicago truncatula, respectively, compared with Glycyrrhiza echinata are critical structural differences that may affect catalytic function. Protein docking highlighted the preference of selected IFS for liquiritigenin compared with naringenin and has listed T. pratense as the most efficient candidate for heterologous biosynthesis of isoflavonoids. The in-silico characterisation of IFS represented in this study is vital in realising the new bioengineering endeavours and will help in the characterisation and selection of IFS candidate enzymes for heterologous biosynthesis of isoflavonoids.

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Section: Discussionmentioning

confidence: 99%

Phylogenetic Analysis and Protein Modelling of Isoflavonoid Synthase Highlights Key Catalytic Sites towards Realising New Bioengineering Endeavours

2022

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“…For the past few years, the genistein biosynthetic pathway has been well-characterized in plants [ 6 , 7 ]. It starts from the single-step conversion of tyrosine to p -coumaric acid (CA) by tyrosine ammonia lyase (TAL), or the two-step conversion of phenylalanine to CA by phenylalanine ammonia lyase (PAL) and cinnamate 4-hydroxylase (C4H).…”

Section: Introductionmentioning

confidence: 99%

Modular Engineering of Saccharomyces cerevisiae for De Novo Biosynthesis of Genistein

Meng

Zhang

et al. 2022

Microorganisms

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Genistein, a nutraceutical isoflavone, has various pharmaceutical and biological activities which benefit human health via soy-containing food intake. This study aimed to construct Saccharomyces cerevisiae to produce genistein from sugar via a modular engineering strategy. In the midstream module, various sources of chalcone synthases and chalcone isomerase-like proteins were tested which enhanced the naringenin production from p-coumaric acid by decreasing the formation of the byproduct. The upstream module was reshaped to enhance the metabolic flux to p-coumaric acid from glucose by overexpressing the genes in the tyrosine biosynthetic pathway and deleting the competing genes. The downstream module was rebuilt to produce genistein from naringenin by pairing various isoflavone synthases and cytochrome P450 reductases. The optimal pair was used for the de novo biosynthesis of genistein with a titer of 31.02 mg/L from sucrose at 25 °C. This is the first report on the de novo biosynthesis of genistein in engineered S. cerevisiae to date. This work shows promising potential for producing flavonoids and isoflavonoids by modular metabolic engineering.

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“…Otherwise, an equal concentration of NAA and 2,4-D (0.5 mg L − 1 each) was optimal for the initiation and production of Sageretia thea cell cultures(Kim et al, 2023). Moreover, the optimization of the PGRs concentration of 2,4-D, NAA, and kinetin to 1.33, 1.76, and 0.15 mg L − 1 , respectively, resulted in higher cell biomass of 70.62 g L − 1 DW of soybean cell cultures(Rani et al, 2022). Consequently, the PGRs concentration as well as the mineral content ensure the reliability, e ciency, and regeneration of the cell suspension initiation protocol(Rani et al., 2022; Schmitz et al, 2016).…”

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confidence: 96%

Production of phenolic compounds, flavonoids, and lupeol by Lycium schweinfurthii suspension cultures in shake flasks and a rocking-motion bioreactor

Mamdouh,

Nebauer,

Junne

et al. 2024

Preprint

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Lycium schweinfurthii, a wild shrub of the Solanaceae family, has received increasing attention in the last decade for its therapeutic potential in traditional medicine due to its diverse array of secondary metabolites, including phenolic substances and terpenoids. The aim of this study was to investigate the accumulation of phenolics, flavonoids, and the terpenoid lupeol in L. schweinfurthii cell suspension cultures cultivated in flasks and in a single-use 2-dimensional rocking motion bioreactor. 3 different media formulations were compared for in vitro cell cultures. Various parameters, such as biomass accumulation, settled cell volume, cell viability (assessed via a 2,3,5-triphenyl tetrazolium chloride assay), and sucrose consumption were recorded as indicators of cell activity and growth. Total phenolic and flavonoid contents were estimated spectrophotometrically and lupeol was quantified via High-Performance Thin Layer Chromatography (HPTLC). Although a higher fresh biomass concentration of 464 g L− 1 was obtained in MS medium supplemented with a combination of each, 1 mg L− 1 of 2,4-Dichlorophenoxyacetic acid (2,4-D) and 1-Naphthaleneacetic acid (NAA), the rocking-motion bioreactor cultivation was performed with 2 mg L− 1 NAA due to its superior reproducibility in viability, productivity, and content of bioactive compounds. A final fresh biomass concentration of 185 g L− 1 was achieved in a 16 L cultivation scale with a notable increase in the concentration of phenolics (1.4-fold) and flavonoids (1.7-fold). Most importantly, the concentration of lupeol, a pentacyclic triterpenoid known for its anti-inflammatory, antibacterial, and anti-atherogenic properties, exhibited a remarkable 5.5-fold increase in the bioreactor cultivation (585 µg g− 1) compared to shake flask cultivations (106 µg g− 1). The current study demonstrated the profound impact of media composition and especially of controlled cultivation conditions in a rocking-motion bioreactor on the accumulation of bioactive compounds. The findings are also relevant for other plant cell cultures.

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Trends in the biotechnological production of isoflavonoids in plant cell suspension cultures

Cited by 11 publications

References 122 publications

Phylogenetic Analysis and Protein Modelling of Isoflavonoid Synthase Highlights Key Catalytic Sites towards Realising New Bioengineering Endeavours

Phylogenetic Analysis and Protein Modelling of Isoflavonoid Synthase Highlights Key Catalytic Sites towards Realising New Bioengineering Endeavours

Modular Engineering of Saccharomyces cerevisiae for De Novo Biosynthesis of Genistein

Production of phenolic compounds, flavonoids, and lupeol by Lycium schweinfurthii suspension cultures in shake flasks and a rocking-motion bioreactor

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