一种丝状真菌活细胞生物量测定方法的研究

Studis on the Method for the Measurement of Fungal Biomass

研究了一种基于测定脱氢酶活(DHA)的生化方法,用以测定真菌活细胞生物量和评价菌丝活力。四唑盐(TTC)在活细胞体内经脱氢酶还原生成红色甲臜(TF)。以氢化可的松生产菌株Abasidia coerulea为模式菌,通过优化染色时间(60 min)、温度(45℃)、pH值8.0,提高了该方法的灵敏度。活细胞湿重与吸光度线形相关(r2=0.98),验证了该方法的有效性。另外,使用优化后的方法,所测脱氢酶比活力可以用来准确评价菌丝体活力。

A biochemical method, based on dehydrogenase activity（DHA） measurement, has been devel- oped for measuring viable cell biomass and assessing the viability of fungal myeelium. In the viable cell, a tetrazium salt （TTC） is reduced to a red formazan（TF） using celluar dehydrogenase. The procedure of the colorimetric assay is established and optimized by using hydrocortisone-producing strain Abasidia co- erulea as a model. The sensitivity of the method has been considerably increased by determining the optimal reaction conditions including incubation time（60min）, temperature（45℃） and pH value （8.0）. The validity of assay is proved by a correlation established between A485 and the wet weight of viable mycelium（r^2=0. 98）. Moveover, Applying the optimized procedure the specific DHA can be used for assessing the viability of fungi accurately