The preparation, regeneration and mutagenesis of the taxol-producing fungus UV40-19 protoplasts were discussed in the experiment. Totally 42 strains displayed hygromycin resistance. Six strains were found to be positi...The preparation, regeneration and mutagenesis of the taxol-producing fungus UV40-19 protoplasts were discussed in the experiment. Totally 42 strains displayed hygromycin resistance. Six strains were found to be positive mutants when screened on plate containing 90μg/mL hygromycin. One hereditarily stable strain UN05-6 was obtained, which raised the taxol yield from （376.38 ± 8.41）μg/L to （493.12 ±11.36） μg/L. The optimal conditions for the preparation, regeneration and mutagenesis of the taxol producing fungus UV40-19 were as follows： 1 ）enzymolysis in a solution containing 3% lywallzyme, 4% snailase, 1% lysozyme and 3% cellulose at 30~C water bath, pH5.5 - 6.0 for 5h; 2） The prepared protoplasts were regenerated by using bilayer plate culturing method; 3）To mutagenize the fungus UV40-19, the protoplast suspension was treated with 0.8mg/mL NTG for 15min, followed by UV irradiation （30W, 30cm distance）for 40s under magnetic stirring. The purified products of the fungus UN05-6 fermented extracts have significant inhibitive effects on SMMC-7721 cell.展开更多
基金Supported by the National Science Foundation of China (30570025)the Fifteen Important Items of Heilongjiang (GA02C101)+3 种基金Key Scientific Program of Harbin (011421126)Research Program for Scholars Overseas by Heilongjiang Education Bureau ( 1152HZ06)Harbin Youth Science Foundation (2005AFOXJ063)Outstanding Young Scientist Foundation of Heilongjiang University
文摘The preparation, regeneration and mutagenesis of the taxol-producing fungus UV40-19 protoplasts were discussed in the experiment. Totally 42 strains displayed hygromycin resistance. Six strains were found to be positive mutants when screened on plate containing 90μg/mL hygromycin. One hereditarily stable strain UN05-6 was obtained, which raised the taxol yield from （376.38 ± 8.41）μg/L to （493.12 ±11.36） μg/L. The optimal conditions for the preparation, regeneration and mutagenesis of the taxol producing fungus UV40-19 were as follows： 1 ）enzymolysis in a solution containing 3% lywallzyme, 4% snailase, 1% lysozyme and 3% cellulose at 30~C water bath, pH5.5 - 6.0 for 5h; 2） The prepared protoplasts were regenerated by using bilayer plate culturing method; 3）To mutagenize the fungus UV40-19, the protoplast suspension was treated with 0.8mg/mL NTG for 15min, followed by UV irradiation （30W, 30cm distance）for 40s under magnetic stirring. The purified products of the fungus UN05-6 fermented extracts have significant inhibitive effects on SMMC-7721 cell.
