栓孔菌属漆酶高产菌株的初步筛选及其产酶条件的优化

Primary screening of effective Trametes strains with high laccase-productivity and optimization of conditions on laccase production

摘要:

利用显色反应对栓孔菌属(Trametes)进行了漆酶高产菌株的筛选, 并对目标菌株的产酶条件进行了优化, 在添加愈创木酚的固体培养基中, 通过显色反应初步筛选出漆酶高产菌株东方栓孔菌(Trametes orientalis) Cui 6300; 进一步通过单因子分析、正交试验和ABTS 法确定了菌株Cui 6300 的最适产酶条件: 麦芽糖15 g/L, 蛋白胨3 g/L, pH 4.8, Cu²⁺ 2.0 mmol/L, 培养温度28 °C,接种饼直径1.5 cm, 此时酶活最高可达19.923 U/mL; 同时探索了Cu²⁺浓度及添加时间对其菌丝生物量和漆酶活力的影响。研究表明, Cu²⁺最适添加浓度为2.0 mmol/L, 添加时间为接种后第3 天。

关键词:漆酶, 菌株Cui 6300, 初步筛选, 条件优化, Cu²⁺

Abstract:

Screening of effective Trametes strains with high laccase-productivity and optimization of conditions on laccase production by the diameters of colored rings by adding guaiacol to the solid medium and assaying the diameters of colored rings, we got the target strain Trametes orientalis Cui 6300. Meanwhile, the optimal conditions of strain Cui 6300 for laccase production were obtained by means of single factor analysis, orthogonal test and ABTS method: maltose 15 g/L, peptone 3 g/L, pH 4.8, Cu²⁺ 2.0 mmol/L, culture temperature 28 °C, inoculum 1.5 cm in diameter, then the highest activity was 19.923 U/mL. Besides, the influences of Cu²⁺ concentration and adding time on biomass and laccase activity were tested, and the optimal concentration and adding time were 2.0 mmol/L and on day 3 of culturing, respectively.

Key words:Laccase, Strain Cui 6300, Initial screening, Optimization, Cu²⁺