An extracellular laccase with potent dye decolorizing ability from white rot fungus Trametes sp. LAC-01

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• 作者：Zhuo-Ren Ling^a ; Shan-Shan Wang^b ; Meng-Juan Zhu^c ; Ying-Jie Ning^a ; Shou-Nan Wang^a ; Bing Li^a ; Ai-Zhen Yang^a ; Guo-Qing Zhang^a ; ^{zhanggq1001@gmail.com" class="auth_mail" title="E-mail the corresponding author} ; Xiao-Meng Zhao^a ; ^{20027202@bac.edu.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Laccase ; Trametes sanguinea ; Dye decolorizing activity
• 刊名：International Journal of Biological Macromolecules
• 出版年：2015
• 出版时间：November 2015
• 年：2015
• 卷：81
• 期：Complete
• 页码：785-793
• 全文大小：1071 K

文摘

A novel laccase was purified from fermentation broth of white rot fungus Trametes sp. LAC-01 using an isolation procedure involving three ion-exchange chromatography steps on DEAE-cellulose, SP-Sepharose, and Q-Sepharose, and one gel-filtration step. The purified enzyme (TSL) was proved as a monomeric protein with a Mr of 59 kDa based on SDS-PAGE and FPLC. Partial amino acid sequences were obtained by LC–MS/MS sharing considerably high sequence similarity with that of other laccases. It possessed optimal pH of 2.6 and temperature of 60 °C using ABTS as the substrate. The K_m of the laccase toward ABTS was estimated to 30.28 μM at pH 2.6 and 40 °C. TSL manifested considerably high oxidizing activity toward ABTS, but was avoid of degradative activity toward benzidine, caftaric acid, etc. It was effective in the decolorization of phenolic dyes – Bromothymol Blue and Malachite Green with decolorization rate higher than 60% after 24 h of incubation. Adjunction of Cu²⁺ with the final concentration of 2.0 mmol/L significantly activated laccase production with a steady high level of 275.8-282.2 U/mL in 96-144 h. The high yield and short production period makes Trametes sp. LAC-01 and TSL potentially useful for industrial and environmental application and commercialization.