一株血红栓菌的分离鉴定及其产漆酶发酵条件与脱色研究

叶婷婷; 张建芬; 陈虹

doi:10.13386/j.issn1002-0306.2020.20.017

一株血红栓菌的分离鉴定及其产漆酶发酵条件与脱色研究

Isolation and Identification of a Trametes sanguinea,and Its Laccase Production Conditions and Decolorization Reasearch

摘要

摘要: 本研究从杭州西溪湿地采集到一株白腐真菌的子实体,并分离其菌丝,编号为H-1。结合形态学特征、ITS序列分析和进化树分析,菌株H-1与Trametes sanguinea的同源性最高,鉴定为血红栓菌Trametes sanguinea H-1。进一步研究在液态发酵条件下碳源、氮源、诱导剂和摇瓶装液量对T. sanguinea H-1胞外漆酶产量的影响,结果表明,以小麦淀粉为碳源,酒石酸铵为氮源,硫酸铜为诱导剂,250 mL的摇瓶装液量为100 mL,血红栓菌H-1产漆酶的酶活力最高,其酶活力达到3527.8 U/L。native-PAGE分析,其漆酶可能大小约为45 kDa的单一蛋白。血红栓菌H-1漆酶对刚果红脱色率达到65.4%,对孔雀石绿脱色率达到47.5%,对亚甲基蓝的脱色率达到31.3%。本研究可为血红栓菌H-1在染料废水脱色的应用奠定基础。

Abstract: In this study,pure culture strain(numbered as H-1)was isolated from the fruiting bodies of a rot fungus collected in the Xixi National Wetland Park. It was identified as Trametes sanguinea H-1 based on morphological,ITS sequence analysis and phylogenetic tree. Furthermore,the optimal culture conditions for extracellular laccase production were investigated in liquid fermentation,including carbon source,nitrogen source,inducer,and volume of culture media. The results showed that the highest laccase activity by T. sanguinea H-1 was attained to 3527.8 U/L when wheat starch was used as carbon source,ammonium tartrate as nitrogen source,copper sulfate as inducer and 100 mL culture media in 250 mL of shaking liquid. According to native-PAGE analysis,the laccase may be a single protein of about 45 kDa. The decolorization rate of Congo red,malachite green and methylene blue by T. sanguinea H-1 laccase was 65.4%,47.5% and 31.3%,respectively. This research established foundation to the application of T. sanguinea H-1 in decolorization of dye wastewater.