Enhanced Efficiency of the Removal of Cytostatic Anthracycline Drugs Using Immobilized Mycelium of Bjerkandera adusta CCBAS 930

Abstract

The aim of this study was to evaluate the bioremoval of anthracycline antibiotics (daunomycin-DNR, doxorubicin–DOX, and mitoxantrone-MTX) by immobilized mycelium of B. adusta CCBAS 930. The activity of oxidoreductases: versatile peroxidases (VP), superoxide dismutase (SOD), catalase (CAT), and glucose oxidase (GOX), and the levels of phenolic compounds (PhC) and free radicals (SOR) were determined during the biotransformation of anthracyclines by B. adusta strain CCBAS 930. Moreover, the phytotoxicity (Lepidium sativum L.), biotoxicity (MARA assay), and genotoxicity of anthracyclines were evaluated after biological treatment. After 120 h, more than 90% of anthracyclines were removed by the immobilized mycelium of B. adusta CCBAS 930. The effective biotransformation of anthracyclines was correlated with detoxification and reduced genotoxicity.

Bioremoval and Detoxification of the Anticancer Drug Mitoxantrone Using Immobilized Crude Versatile Peroxidase (icVP/Ba) Bjerkandera adusta CCBAS 930

“…Immobilized crude VP peroxidase was mixed with 50 mL of MTX solution (10 µg/mL) in 50 mM Na-malonate buffer (pH = 4.5) with 0.2 mM H 2 O 2 and 0.1 mM MnSO 4 in 50 mM Na-malonate buffer (pH = 4.5) and agitated at 130 rpm/min and 30 °C for 72 h. Decolorization was assessed by periodic measurements of absorbance (A630 nm) of the supernatants [ 39 ]. Moreover, measurements in the visible spectrum range from 300 to 700 nm were performed during MTX treatment with icVP/Ba.…”

“…The phytotoxicity assay was performed according to the producer’s protocol with assessment of root growth inhibition (RGI) and germination index (GI) [ 43 ]. The data on RGI and GI of MTX before treatment were derived from the study by Rybczyńska-Tkaczyk (2021) [ 39 ].…”

“…The Multi-species microbial assay (MARA), based on the activity of 11 living microorganisms and their ability to reduce colorless 2,3,5-triphenyltetrazoline hydrochloride to red formazan, was performed to determine biotoxicity after MTX biotransformation by icVP/Ba [ 39 ]. The data on biotoxicity of the MTX before treatment were derived from the study of Rybczyńska-Tkaczyk (2021) [ 39 ]. The results were processed using an image analysis program to facilitate the calculation of the MTC (microbial toxic concentration) value (% vol.)…”

“…In addition, bacterial alkaline phosphatase (AP) activity was used to determine bacterial cytotoxicity, i.e., the reduction factor (RF). The ratio of β-gal/AP activity was defined as the corrected induction factor (CIF = IF/RF), and it was used to indicate the degree of SOS induction for the tested samples [ 39 ]. Briefly, overnight bacterial cultures were grown in fresh LB medium (20 g tryptone/L, 10 g yeast extract/L, 20 g sodium chloride/L, pH 7.4) to an optical density of 600 nm = 0.5 and subsequently mixed ( v / v ) with the tested samples and incubated for 1.5 h at 37 °C); distilled water was used as a negative control.…”

“…The β-gal (A620 nm) and AP (A405 nm) activities were determined in a 96-well plate reader (EPOCH, Biokom). Significant genotoxic activity was defined as a corrected induction factor (CIF) equal to or greater than 1.2 [ 39 ].…”