Plant Milking Technology—An Innovative and Sustainable Process to Produce Highly Active Extracts from Plant Roots

Abstract

We have used an original technology (Plant Milking Technology) based on aeroponic cultivation of plants associated with the gentle recovery of active ingredients from roots. Extraction of bioactive molecules was achieved by soaking the roots, still attached to the living plants, into a nontoxic solvent for a 2 h period. This nondestructive recovery process allows using the same root biomass for successive harvesting dates, in a recyclable way. We have applied this technology to Morus alba L. (mulberry tree), an emblematic tree of the Traditional Chinese Medicine (TCM). Trees were aeroponically grown in large-scale devices (100 m2) and were submitted to nitrogen deprivation to increase the content in active molecules (prenylated flavonoids). The Plant Milking technology applied to Morus alba L. allowed to produce an extract enriched in prenylated compounds (18-fold increase when compared to commercial root extract). Prenylated flavonoids (moracenin A and B, kuwanon C, wittiorumin F, morusin) presented a high affinity for the aged-associated collagenase enzyme, which was confirmed by activity inhibition. In accordance, M. alba extract presents efficient properties to regulate the skin matrisome, which is critical during skin aging. The benefits have been especially confirmed in vivo on wrinkle reduction, in a clinical study that involved aged women. Plant Milking technology is an optimal solution to produce active ingredients from plant roots, including trees, that meet both customer expectations around sustainability, as well as the need for an efficient production system for biotechnologists.

Collagenase and Tyrosinase Inhibitory Effect of Isolated Constituents from the Moss Polytrichum formosum

“…The inhibitory potential of the phytochemical constituents from the 70% ethanol and methanol extracts, against collagenase and tyrosinase, respectively, were investigated by the Target Binding ® approach [ 19 ]. Briefly, Target Binding ® is based on the interactions of a given protein target with a whole plant extract.…”

“…4006713.0025) to FAL (N-(3[2-Furyl]acryloyl)-Leu) + Gly-Pro-Ala (GPA). The collagenase activity from Clostridium histolyticum (type IA, specific activity ≥ 125 CDU/mg solid) from Sigma-Aldrich was determined by the procedure previously described by Chajra et al [ 19 ]. Ethylenediaminetetraacetic acid (EDTA) disodium salt dihydrate (purity ≥ 99%) purchased from Alfa Aesar was used as a control.…”

“…The affinity of the constituents from P. formosum to collagenase and tyrosinase was investigated by the Target Binding ® technology [ 30 ], a method for pre-selection of the inhibitor candidates in complex extracts, which is described in detail by Chajra et al [ 19 ] with few modifications. Collagenase from Clostridium histolyticum (type IA) was prepared at 1.5 mg/mL in 50 mM phosphate buffer (pH 7.5).…”

“…The points in the linear range of the absorbance versus time plots were applied to calculate the slopes, directly proportional to tyrosinase activity. Then, the values of tyrosinase inhibition, expressed as the percent of the activity of the test samples versus the control experiment (pure solvent), were calculated for all samples according to the following equation: The affinity of the constituents from P. formosum to collagenase and tyrosinase was investigated by the Target Binding ® technology [30], a method for pre-selection of the inhibitor candidates in complex extracts, which is described in detail by Chajra et al [19] with few modifications. Collagenase from Clostridium histolyticum (type IA) was prepared at 1.5 mg/mL in 50 mm phosphate buffer (pH 7.5).…”

“…In the present study, P. formosum extracts and isolated constituents were investigated as a new source of collagenase and tyrosinase inhibitors. A specific ligand–protein approach, Target Binding ® [ 19 ], was used to retrieve candidate molecules for both collagenase and tyrosinase inhibition activities. Subsequent preparative chromatography purification was used to isolate the bioactive compounds from the family of benzonaphthoxanthenones, which exhibited collagenase and tyrosinase inhibitory activity.…”