Effect of stirring speed on the production of phenolic secondary metabolites and growth of Buddleja cordata cells cultured in mechanically agitated bioreactor

Abstract

Shake-flask in vitro culture of Buddleja cordata cells produces large amounts of biomass and synthetizes verbascoside (VB), linarin and hydroxycinnamic acids, bioactive phenolic secondary metabolites (PSMs). In this work, we determined the effect of stirring speed on the growth of and production of PSMs [total phenolic, phenylethanoid glycoside and flavonoid contents (PeC, PeGC and FC, respectively)] by B. cordata cells cultured in two bioreactors. Two different stirring speeds (120 and 400 rpm) were tested in two stirred-tank bioreactors: a 2 L bioreactor equipped with a ring diffuser (B2RD) and a 3 L bioreactor with a sintered diffuser (B3SD). Growth kinetics of B. cordata cells were measured in the bioreactors and shake-flask systems. The stirring speed and type of bioreactor affected phases, parameters of growth and production of PSMs. The highest production of biomass (13.62 g L −1) and PSMs [PeC of 64.63 mg gallic acid equivalents g −1 (mg GAE g −1); PeGC of 119.24 mg VB equivalents g −1 (mg VBE g −1); and FC of 5.02 mg quercetin equivalents g −1 (mg QE g −1)] occurred in B2RD at 400 rpm. These values were similar to the found in shake-flasks system. This work establishes the basis for bioprocess advances of B. cordata focused on the development of a sustainable strategy for the management of natural resources and as a source of bioactive PSMs on a large scale. Key message Buddleja cordata cells cultured in a mechanically agitated bioreactor possess an outstanding biosynthetic potential that represents a suitable biotechnological alternative for the production of bioactive phenolic secondary metabolites. Keywords Bioreactor • Buddleja cordata • Phenolic secondary metabolites • Diffuser • Stirring speed • Verbascoside Abbreviations µ Specific growth rate ANOVA Analysis of variance B2RD 2 L Bioreactor equipped with a ring diffuser B3SD 3 L Bioreactor equipped with a sintered diffuser CSCBc Cell suspension culture of B. cordata CV Cellular viability Communicated by Sergio J. Ochatt.

Photoprotective Activity of Buddleja cordata Cell Culture Methanolic Extract on UVB-irradiated 3T3-Swiss Albino Fibroblasts

“…The total phenol and total phenolic acid contents of the methanolic extract (2 mg/mL) were determined using a procedure described by Vazquez-Marquez et al [ 41 ]. Gallic acid (Sigma-Aldrich, USA) was used to build a calibration curve (0.023–0.375 mg/mL; y = 2.6609 x − 0.019, R 2 = 0.997285) for the total phenol content and the obtained results were expressed as mg gallic acid equivalents per gram of extract.…”

“…Both secondary metabolites (SMs) have been identified in several Buddleja species distributed in Mexico, namely B. americana L., B. cordata Kunth, B. parviflora Kunth, and B. scordioides Kunth) [ 39 ]. It is well documented in the literature that SM production depends on genetic, biotic, and abiotic factors as well as the physiological conditions and development of the plant [ 40 ]; thus, the concentration in vegetal material might be inconstant, becoming an advantage when vegetal material might be used as source of bioactive SMs [ 41 ]. The in vitro culture of plants represents a biotechnological tool for the production of bioactive SMs as culture conditions can be manipulated to achieve high yields of production as well as to allow mass production using bioreactors [ 41 ].…”

“…For B. cordata Kunth, several biotechnological advances have been achieved with the aim of obtaining optimal culture conditions (e.g., elicitation with methyl jasmonate and bioreactor culture, among others) for high biomass and SM productions, especially concerning verbascoside [ 27 , 41 , 42 , 43 ]. This species has a wide territorial distribution in Mexico and is used by the Mexican folk population to counteract several diseases; its medicinal properties were found to be related to their constituent SMs [ 27 ].…”

Arnica montana Cell Culture Establishment, and Assessment of Its Cytotoxic, Antibacterial, α-Amylase Inhibitor, and Antioxidant In Vitro Bioactivities

“…4.5.2. Quantification of Phenolic Compounds and Sesquiterpene Lactones TPC, TFC, and TPAC were determined according to Vazquez-Marquez et al [66]. These spectrophotometric methods are common in phytochemical analysis to determine the content of secondary metabolites based on the use of reference standards for comparison [67][68][69].…”

Shear rate as scale-up criterion of the protein production with enhanced proteolytic activity by phosphate addition in the Jacaratia mexicana cell culture

“…Buddleja cordata cells cultured in a 2-L stirred tank bioreactor enhance the biomass production (13.62 g L −1 ) and phenolic secondary metabolites (total phenolic (64.63 mg gallic acid equivalents g −1 ), phenylethanoid glycoside (119.24 mg verbascoside equivalents g −1 )) and flavonoid (5.02 mg quercetin equivalents g −1 ) at 400 rpm [11]. The highest production of biomass (13.62 g L −1 ) and PSMs [PeC of 64.63 mg gallic acid equivalents g −1 (mg GAE g −1 ); PeGC of 119.24 mg VB equivalents g −1 (mg VBE g −1 ); and FC of 5.02 mg quercetin equivalents g −1 (mg QE g −1 )] occurred in B2RD at 400 rpm [12].…”