Laccases of Basidiomycetes as Prospective Biocatalysts for Transformation of Antibiotics of Penicillin Series
Author(s): Ulyana A Krut*, Irina V Spichak, Irina I Oleynikova, Galina M Shaidorova, Alexandra I Radchenko and Elena V Kuzubova
Abstract
Today, one of the promising trends in modern biotechnology is the use of enzymes to obtain new compounds that have antibacterial, fungicidal and anticarcinogenic properties. As a promising biocatalyst for the synthesis and modification of various compounds, fungal laccases have recently been considered, which are a cheap and readily available enzyme for obtaining reaction components. Purpose of the study: to study the possibility of mushroom laccase use for the modification of known antibiotics to obtain their analogues with new properties. Materials and methods: The activity of laccase was determined spectrophotometrically by the rate of ABTS oxidation. The main object of the study is the laccases of the basidiomycetes of "white rot" and Lentinus strigosus 1566. The work used 2,6-dimethylphenol (Sigma-Aldrich, USA), 2,2-azino-bis (3-ethylbenzothiazoline 6-sulfonic acid) (ABTS) (Fluka, Switzerland) and glycerin (Serva, Germany). Peptone, soy flour, tetracycline hydrochloride, 6-aminopenicellanic acid, phenylglycine, potassium benzylpenicylate, pyrocatechol, hydroquinone ("Khimreaktiv", Russia). Result: According to the spectra obtained, the reaction of the antibiotic modification with laccase without the addition of hydroquinone is not carried out. Because hydroquinone is a good substrate for laccase, this compound interacts quickly with the enzyme, with the formation of an oxidation product with an absorption maximum in the range from 200 to 300 nm. An example of 6-aminopenicylic acid modification dynamics by the Lentinus strigosus 1566 laccase in the presence of hydroquinone shows the formation of new peaks in the absorption region of 250 nm, which proves the formation of new products of the antibiotic transformation. When they study the spectral characteristics of 6-aminopenicylic acid interaction with the Lentinus strigosus 1566 laccase for 120 min, no new compounds are formed, because there is no spectrum change. However, there is a significant change in the reaction spectrum of the reaction of 6-APA interaction with hydroquinone under the action of Lentinus strigosus 1566 laccase for 120 min. Conclusion: In the course of the work, the enzymatic preparation of laccase Lentinus strigosus 1566 was tested for the ability to modify the antibiotic. TLC showed the presence of three new compounds with-0.18; 0.15; 0.11; 0.07 (absent in all controls). Changes of absorption spectra in the range of 250 nm in the reaction of hydroquinone and 6-APA interaction under the action of Lentinus strigosus 1566 laccase also indicate the presence of new compounds.