Optimization of enzymatic parameters for the production of formononetin from red clover ( Trifolium pratense L.) through a response surface methodology

Trifolium pratense, is a forage found worldwide, but it is negatively impacted by the clover root borer, Hylastinus obscurus. Methanolic extraction has been reported for isolating formononetin from vegetal tissues, with an antifeeding effect on H. obscurus. However, this methodology is time-consumin...

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Bibliographic Details
Published in:Natural product research 2022-09, Vol.36 (18), p.4713-4718
Main Authors: Muñoz, María Teresa, Maldonado, Vanessa, Herrera, Wence, Mutis, Ana, Bardehle, Leonardo, Medina, Cristian, Hormazábal, Emilio, Ortega, Fernando, Quiroz, Andrés
Format: Article
Language:English
Subjects:
Borers
Clover
Cultivars
Hylastinus obscurus
Metabolites
Optimization
Parameters
pH effects
Red clover
Response surface methodology
Secondary metabolites
Trifolium pratense
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Summary:Trifolium pratense, is a forage found worldwide, but it is negatively impacted by the clover root borer, Hylastinus obscurus. Methanolic extraction has been reported for isolating formononetin from vegetal tissues, with an antifeeding effect on H. obscurus. However, this methodology is time-consuming and also extracts other secondary metabolites, whereas enzymatic assays can provide higher specificity. Hence, the objective of this work was to determine the optimal conditions in pH, temperature, and incubation time for the activity of isoflavone synthase via a response surface model. Once these parameters were optimized, the concentration of formononetin in cultivars and experimental lines of T. pratense was evaluated enzymatically. The results showed that the best condition for developing the enzymatic assay was pH 9.1 with an incubation at 34.5 °C for 155 min. The formononetin content fluctuated between 0.74 and 1.96 mg/g of fresh weight, where Precoz-3, Precoz-1, and Superqueli-INIA presented the highest production.
ISSN:1478-6419
1478-6427
DOI:10.1080/14786419.2021.2002324