Production of Verbascoside, Isoverbascoside and Phenolic Acids in Callus, Suspension, and Bioreactor Cultures of Verbena officinalis and Biological Properties of Biomass Extracts

Abstract

Callus, suspension and bioreactor cultures of Verbena officinalis were established, and optimized for biomass growth and production of phenylpropanoid glycosides, phenolic acids, flavonoids and iridoids. All types of cultures were maintained on/in the Murashige and Skoog (MS) media with 1 mg/L BAP and 1 mg/L NAA. The inoculum sizes were optimized in callus and suspension cultures. Moreover, the growth of the culture in two different types of bioreactors—a balloon bioreactor (BB) and a stirred-tank bioreactor (STB) was tested. In methanolic extracts from biomass of all types of in vitro cultures the presence of the same metabolites—verbascoside, isoverbascoside, and six phenolic acids: protocatechuic, chlorogenic, vanillic, caffeic, ferulic and rosmarinic acids was confirmed and quantified by the HPLC-DAD method. In the extracts from lyophilized culture media, no metabolites were found. The main metabolites in biomass extracts were verbascoside and isoverbascoside. Their maximum amounts in g/100 g DW (dry weight) in the tested types of cultures were as follow: 7.25 and 0.61 (callus), 7.06 and 0.48 (suspension), 7.69 and 0.31 (BB), 9.18 and 0.34 (STB). The amounts of phenolic acids were many times lower, max. total content reached of 26.90, 50.72, 19.88, and 36.78 mg/100 g DW, respectively. The highest content of verbascoside and also a high content of isoverbascoside obtained in STB (stirred-tank bioreactor) were 5.3 and 7.8 times higher than in extracts from overground parts of the parent plant. In the extracts from parent plant two iridoids—verbenalin and hastatoside, were also abundant. All investigated biomass extracts and the extracts from parent plant showed the antiproliferative, antioxidant and antibacterial activities. The strongest activities were documented for the cultures maintained in STB. We propose extracts from in vitro cultured biomass of vervain, especially from STB, as a rich source of bioactive metabolites with antiproliferative, antioxidant and antibacterial properties.

Antioxidant Potential and Enhancement of Bioactive Metabolite Production in In Vitro Cultures of Scutellaria lateriflora L. by Biotechnological Methods

“…The optimization of cultivation depends on the type of bioreactor used and its operation, including aeration and immersion time, which is widely reported in the literature [60,61]. This has been demonstrated by us for both Schisandra chinensis [62] and Verbena officinalis cultures [63]. It was shown that the operation conditions influenced the accumulation of flavonoids, phenolic acids, and phenylpropanoid glycosides.…”

Phenylpropanoid Glycoside and Phenolic Acid Profiles and Biological Activities of Biomass Extracts from Different Types of Verbena officinalis Microshoot Cultures and Soil-Grown Plant

“…Previously, we demonstrated the biotechnological possibilities of producing some phenylpropanoid glycosides and phenolic acids in the biomass of undifferentiating V. officinalis callus and suspension cultures that were maintained in Erlenmeyer flasks, as well as in special balloon and stirred-tank bioreactors. In these studies, we investigated the influence of different plant growth regulators (PGRs), light conditions, and inoculum size on biomass growth and metabolic profile [ 19 , 20 , 21 ]. The results showed that verbascoside was produced at an extremely high level, with a maximum amount of 9.18 g/100 g DW found in the suspension cultured in stirred-tank bioreactors [ 20 ].…”

“…In these studies, we investigated the influence of different plant growth regulators (PGRs), light conditions, and inoculum size on biomass growth and metabolic profile [ 19 , 20 , 21 ]. The results showed that verbascoside was produced at an extremely high level, with a maximum amount of 9.18 g/100 g DW found in the suspension cultured in stirred-tank bioreactors [ 20 ]. These results are plausible given the various biological properties of verbascoside, such as antioxidant, anti-inflammatory, cytoprotective, antibacterial, antiviral, and antiandrogen activities [ 22 , 23 , 24 ].…”

Bioactive constituents ofVerbena officinalisalleviate inflammation and enhance killing efficiency of natural killer cells

“…Verbena officinalis L. (V. officinalis), also known as common vervain, is a medicinal herb, widespread throughout the globe, mainly in the temperate climate zone (Kubica, Szopa et al, 2020b). In China, V. officinalis is widely distributed in the southern part of the Yellow River and has been used as traditional Chinese medicine for the treatment of rheumatism, bronchitis, depression, insomnia, anxiety, liver and gallbladder diseases (Khan, Khan et al, 2016, Kubica, Szopa et al, 2020a.…”

“…In China, V. officinalis is widely distributed in the southern part of the Yellow River and has been used as traditional Chinese medicine for the treatment of rheumatism, bronchitis, depression, insomnia, anxiety, liver and gallbladder diseases (Khan, Khan et al, 2016, Kubica, Szopa et al, 2020a. Recent reports suggest that V. officinalis has a number of scientifically proven activities, such as antioxidant, antibacterial, antifungal, anti-inflammatory, analgesic, anticonvulsant, anxiolytic, antidepressant, sedative, hypnotic, wound healing, gastro-protective, anti-cancer and insecticidal properties (Casanova, Garcia-Mina et al, 2008, Kubica et al, 2020b, Lopez-Jornet, Camacho-Alonso et al, 2014, Speroni, Cervellati et al, 2007. In addition to being used as an antimicrobial, secretolytic, expectorant and diuretic agent, V. officinalis is also widely used in food and cosmetics, especially due to its antioxidant, antibacterial, and anti-inflammatory properties (Kubica et al, 2020a).…”