Heterologous expression and characterisation of microcystinase

Genetically Engineered Bacteria Immobilized in Alginate as an Option of Cyanotoxins Removal

“…It was indicated previously that whole cells of E. coli BL21_MlrA exhibited approximately 250 times higher MlrA activity than Sphingomonas cells [14]. Furthermore, the use of IPTG to induct the MlrA expression in BL21_MlrA was not necessary, because the cells with and without induction exhibited a similar level of activity toward MC-LR (data not presented).…”

“…The crucial step in MC detoxification by bacteria is the linearization reaction catalyzed by metalloproteinase called microcystinase (MlrA). Such a decyclization results in a significant reduction of toxicity [14]. The pathway of MC degradation was confirmed independently, with small alteration due to the different strains.…”

“…coding for MlrA was amplified by PCR using primer pairs described earlier [14]. The amplified fragments were inserted into the pTZ57R/T cloning vector and the sequence was verified.…”

“…ACM-3962 and E. coli BL21_MlrA cells was measured as described earlier using the HPLC method [14]. The rate of MCs degradation was calculated by monitoring the level of linear MC-LR.…”

“…2). For example, better biochemical characterization of these important enzymes as well as the correction of their role in MCs degradation was possible [14] - [16]. Another option is to investigate the usefulness of such recombinant strains or recombinantly expressed MlrA enzyme in biotechnological application.…”