Kannaiyan Ranjani, Mahinpey Nader, Kostenko Victoria, Martinuzzi Robert J
Department of Chemical and Petroleum Engineering, University of Calgary, Calgary, Canada.
Biotechnol Appl Biochem. 2015 Mar-Apr;62(2):173-85. doi: 10.1002/bab.1263. Epub 2014 Dec 1.
Coculturing of two white-rot fungi, Dichomitus squalens and Ceriporiopsis subvermispora, was explored for the optimization of cultivation media for simultaneous augmentation of laccase and peroxidase activities by response surface methodology (RSM). Nutrient parameters chosen from our previous studies with the monocultures of D. squalens and C. subvermispora were used to design the experiments for the cocultivation study. Glucose, arabinose, sodium nitrate, casein, copper sulfate (CuSO4 ), and manganese sulfate (MnSO4 ) were combined according to central composite design and used as the incubation medium for the cocultivation. The interaction of glucose and sodium nitrate resulted in laccase and peroxidase activities of approximately 800 U/g protein. The addition of either glucose or sodium nitrate to the medium also modifies the impact of other nutrients on the ligninolytic activity. Both enzyme activities were cross-regulated by arabinose, casein, CuSO4 , and MnSO4 as a function of concentrations. Based on RSM, the optimum nutrient levels are 1% glucose, 0.1% arabinose, 20 mM sodium nitrate, 0.27% casein, 0.31 mM CuSO4 , and 0.07 mM MnSO4 . Cocultivation resulted in the production of laccase of 1,378 U/g protein and peroxidase of 1,372 U/g protein. Lignin (16.9%) in wheat straw was degraded by the optimized enzyme mixture.
采用响应面法(RSM)研究了两种白腐真菌——黄白卧孔菌(Dichomitus squalens)和亚侧耳状拟蜡菌(Ceriporiopsis subvermispora)的共培养,以优化培养基,同时提高漆酶和过氧化物酶的活性。从我们之前对黄白卧孔菌和亚侧耳状拟蜡菌单培养的研究中选取营养参数,用于设计共培养研究的实验。根据中心复合设计将葡萄糖、阿拉伯糖、硝酸钠、酪蛋白、硫酸铜(CuSO4)和硫酸锰(MnSO4)组合,用作共培养的孵育培养基。葡萄糖和硝酸钠的相互作用产生了约800 U/g蛋白质的漆酶和过氧化物酶活性。向培养基中添加葡萄糖或硝酸钠也会改变其他营养物质对木质素分解活性的影响。阿拉伯糖、酪蛋白、CuSO4和MnSO4的浓度对两种酶的活性都有交叉调节作用。基于响应面法,最佳营养水平为1%葡萄糖、0.1%阿拉伯糖、20 mM硝酸钠、0.27%酪蛋白、0.31 mM CuSO4和待补充0.07 mM MnSO4。共培养产生了1378 U/g蛋白质的漆酶和1372 U/g蛋白质的过氧化物酶。优化后的酶混合物降解了小麦秸秆中16.9%的木质素。