Production and Purification of Peroxidase from Tissue Cultures of Cauliflower (Brassica oleracea L. var. Botrytis)

The plant peroxidases are remarkable enzymes due to their widespread use in industry. Theseenzymes, which are capable of catalysing the oxidation of various organic and inorganicsubstrates, have been used in clinical diagnosis, detoxification reactions and organic synthesis.In this study, in vitro production and purification of peroxidase enzyme from cauliflower plantwas proposed. Firstly, sterile seedlings were obtained from MS/B5 nutrient medium withoutgrowth regulator from cauliflower seeds and calluses from medium containing 0.5 mg / L 2.4-D. Callus and seedlings were powdered with liquid nitrogen and then homogenized. Peroxidaseenzymes were purified from these homogenates using affinity technique. SDS-PAGEelectrophoresis was performed to determine the molecular weight of the purified enzymes andsingle bands was observed at approximately 46 kDa. In addition, KM and Vmax values of thecallus peroxidase enzyme were determined for guaiacol, pyrogallol and H2O2 substrates

Production and purification of peroxidases from callus cultures of white and red cabbage for enzymatic decolourization of reactive blue 19 and acid blue 25 dyes

The removal of textile dyes from wastewater by using plant peroxidases (PODs) offers environmentally effective solutions. Therefore, researching the availability and applicability of PODs from different sources becomes important area for the sustainable process design. In this article, PODs were produced in vitro from white and red cabbage callus cultures for evaluating their potential usage to decolourization of the Reactive Blue 19 (RB-19) and Acid Blue 25 (AB-25) textile dyes for the first time. The PODs with high activity were purified 133-fold with a specific activity of 45,083 U mg−1 and 134-fold with a specific activity of 26,785 U mg−1 from produced white and red cabbage calluses by single-step affinity chromatography. The molecular weights of purified red cabbage callus peroxidase (r-POD) and white cabbage callus peroxidase (w-POD) were found to be as 43 and 44 kDa, respectively. In the assays to determine the r-POD and w-POD dye decolourization potential, noteworthy degradation levels were recorded for RB-19 (>85) and AB-25 (>88) dyes in the reactions catalysed by w-POD. Also, r-POD exhibited maximum decolourization efficiency of % 68 for RB-19 and %73 for AB-25 dyes under the conditions at 50 °C, pH 5.5 and in the presence of 4 mM H2O2. © 2021 Informa UK Limited, trading as Taylor & Francis Group

COMPARISON OF PHYTOCHEMICALS AND ANTIOXIDANT CAPACITY OF HYPERICUM PERFORATUM; WILD PLANT PARTS AND IN VITRO SAMPLES

The aim of study is to compare the phytochemicals and free radical scavenging activities in methanol extracts of flower, leaf, stem in vitro plantlet, callus of Hypericum perforatum L. In vitro cultures of H. perforatum was established by using MS-B5 medium contained plant growth regulators such as BAP, TDZ and picloram. Total phenolics and flavonoids were analysed by spectrophotometric methods.The stem was the richest in total phenolics (228.9 mg GAE/g extract) and flavonoids (102.4 mg QE/g extract). Quinic acid, gallic acid, (+)-catechin, ferulic acid, vanillic acid, p-coumaric acid, caffeic acid, and quercetin were determinated by LC-MS/MS. Free radical scavenging activities (ABTS and DPPH) of all samples were detected as IC50 values, and was compared to standards such as trolox and ascorbic acid. As a result, the stem exhibited the stronger antioxidant activities than other samples, and vanillic acid, ferulic acid and gallic acid could be produced by in vitro culture