绿色木霉合成纤维素酶部分培养条件的优化

Optimization of Some Culture Conditions for Biosynthesis of Cellulase by Trichoderma vivide

[目的]确定绿色木霉ZJ株产纤维素酶的最佳诱导时间和诱导物,为其实际应用提供条件。[方法]接种绿色木霉ZJ株7 d内,每天取培养物样品,采用3,5-二硝基水杨酸法检测产酶量。将绿色木霉ZJ株接种添加了不同碳源或氮源的基础培养基中,观察绿色木霉的生长情况,测定菌丝重量,检测不同培养时间的培养物中CMCase酶的产量。[结果]绿色木霉ZJ株的最适培养时间为72～96 h;绿色木霉在以单糖、双糖为碳源的培养基中均能迅速生长,CMCase酶产量在3～4 d时达到高峰,以纤维素粉的诱导效果最佳;以硫酸铵与酵母膏组成的复合氮源最适合绿色木酶ZJ菌丝的生长,产酶活力最高。[结论]接种后3～4 d收获绿色木霉ZJ株培养物可获得最大产酶量;以纤维素粉作为碳源,以硫酸铵与酵母膏组成的复合氮源为氮源,绿色木霉的产酶活力最高。

[ Objective ] To determine the best culture time and inducer for the biosynthesis of cellulase by Trichoderma vivide and thus provide the conditions for its practical application. [ Method ] Within the 7 d after the inoculation of Trichoderma vivide ZJ strain, the cultures were collected once every day, and the enzyme yield was respectively determined by 3, 5-dinitrosalicylic acid assay. The Trichoderma vivide ZJ strain was inoculated into basal medium added by different types of carbon sources or nitrogen sources, and the growth of Trichoderma viride was observed. And the mycelium weight as well as the yield of CMCase enzyme after different culture time was determined. [ Result] The optimal culture time for Trichoderma viride ZJ strain was 72 - 96 h; it grew rapidly in the medium added by monosaccharide or dtsaccharide as carbon sources, and the production of CMCase enzyme reached a peak after 3 -4 d post inoculation. Cellulose powder was the best carbon inducer. The compound nitrogen source composed of 1 g/L ammonium sulfate and 2 g/L yeast extract was the most suitable for the growth of ZJ strain and produced the highest enzyme activity. [ Conclusion] The largest enzyme yield should be obtained after 3 -4 d post the inoculation of Trichoderma viride ZJ strain. With cellulose powder as a carbon source and the complex substance composed of ammonium sulfate and yeast extract as a nitrogen source, Trichoderma viride has the highest enzyme activity.