Key Laboratory of Urban Agriculture (North China) of Ministry of Agriculture, College of Biological Science and Engineering, Beijing University of Agriculture, Beijing 102206, China.
Beijing Key Laboratory for Agricultural Application and New Technique, College of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206, China.
Biochimie. 2018 May;148:46-54. doi: 10.1016/j.biochi.2018.02.015. Epub 2018 Mar 5.
A novel extracellular laccase was purified from fermentation broth of the white rot fungus Trametes sp. F1635 by a three-step protocol including two consecutive ion-exchange chromatography steps on DEAE-Sepharose and SP-Sepharose, and a final gel-filtration on Superdex 75. The purified laccase (TsL) was a monomeric protein with the molecular mass of 64.8 kDa. It demonstrated high oxidation activity of 4.00 × 10 U/mg towards ABTS. Its N-terminal amino acid sequence was AIGPVADLTIINNAV which was unique and sharing high similarity of other fungal laccases. TsL was a yellow laccase based on absorption spectrum analysis. It demonstrated an acidic pH optimum of 2.6 and temperature optimum of 50 °C towards ABTS. The K and V values towards ABTS were estimated to 18.58 μM and 1.35 μmol/min, respectively. TsL manifested effective decolorization activity towards eriochrome black T (EBT), remazol brilliant blue R (RBBR), malachite green (MG), and eriochrome black T (EBT) (over 60%). Violuric acid (VA) and acetosyringone (AS) were the optimal mediators for the laccase in dye decolorization. Results suggest that TsL demonstrates great potential for dye decolorization and water treatment.
一株白腐真菌 Trametes sp. F1635 发酵液中新型胞外漆酶的分离纯化及其酶学性质研究
采用三步分离方案，包括两次 DEAE-Sepharose 和 SP-Sepharose 离子交换层析以及最后一次 Superdex 75 凝胶过滤层析，从一株白腐真菌 Trametes sp. F1635 的发酵液中分离纯化得到了一种新型胞外漆酶（TsL）。该漆酶为单体蛋白，分子量为 64.8 kDa。TsL 对 ABTS 的氧化活性最高，为 4.00×10 U/mg。其 N 端氨基酸序列为 AIGPVADLTIINNAV，与其他真菌漆酶具有高度相似性，但序列独特。TsL 是一种黄色漆酶，根据吸收光谱分析得出。TsL 对 ABTS 的最适 pH 为 2.6，最适温度为 50°C。其对 ABTS 的 K 和 V 值分别为 18.58 μM 和 1.35 μmol/min。TsL 对媒介黑 T（EBT）、丽春红 2R（RBBR）、孔雀绿（MG）和媒介黑 T（EBT）均具有有效的脱色活性（>60%）。漆酶对染料的脱色作用以 3-羟基苯并噻唑（VA）和乙酰丁香酮（AS）为最佳。结果表明，TsL 在染料脱色和水处理方面具有巨大的应用潜力。
