康氏木霉产酶发酵固体培养基优化研究

A study on the optimization of substrate prescription for cellulasecomplex production by solid fermentationfrom Trichoderma koningii

为利用廉价的培养基生产纤维素酶复合制剂,本实验采用培养基配方选择试验和双温度培养法对康氏木霉F244产酶特性进行了研究.在测定滤纸酶(FPA)、棉花酶、羧甲基纤维素酶(CMCase)、β-葡萄糖苷酶和果胶酶活力的基础上利用SPSS建立回归方程,全相关系数分别达到0.852,0.941,0.964,0.703,0.899,而后通过无约束规划求解找到最佳配方,并对酶活进行了预报和对比.结果表明:各酶活最大时对培养基各成分的含量要求不同;应用稻草粉质量分数20.3％,麸皮质量分数26.1％,(NH4)2SO4质量分数7.9％,水分质量分数45.7％的配方发酵时,F244的FPA、棉花酶、CMCase、β-葡萄糖苷酶、果胶酶活可望达14.1,20.1,43.9,21.6,16.8 IU/g,基本与里氏木霉Q9414在其推荐培养基上的产酶水平相当,而且该配方用料来源广泛,成本低廉,工艺简单,产品安全无毒.

In order to get the optionized substrate prescription for the production of cellulase complex of high activity from Trichoderma koningii F244, a prescription experiment is performed with two different temperature degrees, which were kept at 30℃ during the first 30h and at 23℃ for another 66h continuously. Consequently, the enzyme activities of FPA, CMCase, cotton lyase, β-glucosidase and pectinase are taken into account so as to carry out the linear regression by SPSS and for FPA, cotton lyase, CMCase, β-glucosidase and pectinase, regression models account for 85.2%, 94.1%, 96.4%, 70.3% and 89.9% of the variability, respectively. After the regression equations have been figured out by unconstraint mathematical programming, it is revealed that different ratios of rice straw, wheat bran, (NH4)2SO4 and water are required when each enzyme reaches its own maximum. A forecast of cellulase activities of different prescriptions, which correspond to the maximums of enzyme activities, and a comparison with that of Trichoderma reeseii Q9414 on its own recommended substrate clearly shows that the activities of FPA, cotton lyase, CMCase, β-glucosidase from Trichoderma koningii F244 on substrate constituted by rice straw 20.3%, wheat bran 26.1%, (NH4)2SO4 7.9%, water 45.7%, are expected to reach 14.1 IU/g, 20.1 IU/g, 43.9 IU/g, 21.6 IU/g, 16.8 IU/g respectively, almost equal to these of Q9414 on its own recommended substrate. Moreover, the materials for this prescription have an extensive source and are of low cost. The technics is simple and the production is nontoxic to livestocks.