THERMOSTABLE LACCASE III OF FUNGUS STECCHERINUM OCHRACEUM LE (BIN) 1833 D AND METHOD FOR THEIR OBTAINING

摘要

FIELD: medicine.;SUBSTANCE: invention can be used in modifying lignin-containing materials and preparing based commercially valuable compounds, wightening the paper stock and textile, treating sewage water and soils from variety of xenobiotics, polymerising phenols and some other aromatic connections, making cosmetic products for skin wigthening and hair dyeing. The method provides growing fungus Steccherinum ochraceum LE (BIN) 1833 D on a glucose-peptone medium. The prepared culture fluid is filtered from mycelium and centrifuged. The prepared supernatant is applied on a column with DE52 cellulose neutralised with 20 mM sodium-acetate buffer (buffer A). Then it is eluted with a linear gradient of 0 - 0.5 M NaCl in the buffer A. The prepared fractions with laccase activity are unitised and concentrated to desalinate and apply on the column with an ion-exchange carrier Q-Sepharose preliminary neutralised with the buffer A. Then it is eluted with a linear gradient of 0 - 0.5 M NaCl in the buffer A. The prepared two fractions with laccase activity are concentrated and desalinated. The first fraction with laccase activity prepared is concentrated and applied on column Superdex 75. It is eluted 0.1 M NaCl in the buffer A. The prepared fractions with laccase activity unitised and concentrated to prepare laccase I. The second fraction with laccase activity chromatographied on the column with Q-Sepharose is concentrated and applied on the column with the ion-exchange carrier Resource Q. It is eluted with the linear gradient 1 M NaCl in the buffer A. The prepared two fractions with laccase activity are concentrated and desalinated. The first fraction with laccase activity chromatographied on the column with Resource Q applied on the column with Superdex 75 to elute 0.1 M NaCl in the buffer A. The prepared fractions with laccase activity are unitised and concentrated to prepare laccase II. The second fraction with laccase activity chromatographied on the column with Resource Q is applied on the column with Superdex 75 and eluted 0.1 M NaCl in the buffer A. The prepared fractions with laccase activity are unitised and concentrated to produce laccase III.;EFFECT: invention improves stability of laccase.;2 cl, 4 dwg, 2 tbl, 5 ex