摘要
红栓菌(pycnoporus cinnabarius)在发酵培养3d后出现胞外漆酶活性峰。木素类似物对红栓菌胞外漆酶活性有诱导作用,阿魏酸、香兰素,愈创木酚和DL-β-苯丙氨酸诱导24h后,发酵液中漆酶活性分别是对照的2.8、4.3、3.5和1.7倍。发酵液经(NH_4)_2S0_4沉淀,Sephadex G-150、DEAE-Sephadex A-25、Sephadex G-25柱层析纯化后,冷冻干燥。高效液相色谱(HPLC)检测为一单峰,分子量为26000,含17种氨基酸,氨基酸总量占酶组成的64.27％。等离子光谱(ICP)分析表明漆酶含有铜。该酶反应的最适温度为30℃,与邻联苯甲胺反应最适pH为4.0,Km值为385μmol/L;与丁香醛连氮反应最适pH为5.8,Km值为833μmol/L。
The activity of extracellular laccase reached a peak value three days after inoculation. It was found that the activity of laccase could be induced by ferulic acid, vallilin, guaiacol and DL-β-phenylalanine to 2.8, 4.3, 3.5, 1.7 times higher than that of the noninduced control, respectively. After( NH4)2SO4 sedimentation followed by Sephadex G-150, DEAE-Sephadex A-25 and Sephadex G-25 chromatography, we obtained purified laccase that showed a single peak on HPLC. The molecular weight of laccase detected by HPLC is 26 000. The P. cinnabarius laccase contains 17 kinds of amino acids whose total amounts accounted for 64.27% of the total molecules. ICP analysis indicates that the laccase is a copper-associated protein. The optimal reaction .temperature for P.cinnabarius laccase is 30℃ . With tolidine as substrate, the optimum pH for laccase reaction is 4.0 and Km is 385 μmol/ L, while for substrate syringaldazine, the data is 5.8 and 833μmol / L, respectively.
出处
《微生物学报》
CAS
CSCD
北大核心
1996年第5期360-366,共7页
Acta Microbiologica Sinica
关键词
红栓菌
胞外漆酶
诱导
发酵
Pycnoporus cinnabarius, Laccase, Induction
